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Clinical Studies
Abstracts are presented below for clinical
studies on Black Gram.
Plant Phytonutrient Profile
1: Genome. 2008 Aug;51(8):628/37.
Construction of a genetic linkage map of black gram, Vigna mungo (L.) Hepper,
based on molecular markers and comparative studies.
Gupta SK, Souframanien J, Gopalakrishna T.
Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre,
Trombay, Mumbai 400 085, India. gupta_sk@hotmail.com
A genetic linkage map of black gram, Vigna mungo (L.) Hepper, was constructed
with 428 molecular markers using an F9 recombinant inbred population of 104
individuals. The population was derived from an inter/subspecific cross between a
black gram cultivar, TU94/2, and a wild genotype, V. mungo var. silvestris. The
linkage analysis at a LOD score of 5.0 distributed all 428 markers (254 AFLP, 47
SSR, 86 RAPD, and 41 ISSR) into 11 linkage groups. The map spanned a total
distance of 865.1 cM with an average marker density of 2 cM. The largest linkage
group spanned 115 cM and the smallest linkage group was of 44.9 cM. The number of
markers per linkage group ranged from 11 to 86 and the average distance between
markers varied from 1.1 to 5.6 cM. Comparison of the map with other published
azuki bean and black gram maps showed high colinearity of markers, with some
inversions. The current map is the most saturated map for black gram to date and
will provide a useful tool for identification of QTLs and for marker/assisted
selection of agronomically important characters in black gram.
Publication Types:
Comparative Study
PMID: 18650952 [PubMed / indexed for MEDLINE]
2: J Gen Appl Microbiol. 2008 Apr;54(2):93/9.
Characterization of a symbiotically effective Rhizobium resistant to arsenic:
Isolated from the root nodules of Vigna mungo (L.) Hepper grown in an
arsenic/contaminated field.
Mandal SM, Pati BR, Das AK, Ghosh AK.
Department of Biotechnology, Indian Institute of Technology, Kharagpur, India.
Bacteria were isolated from the root nodules of Vigna mungo (L.) Hepper, grown in
an arsenic/contaminated field and the strain was selected by its nodulation
ability as well as better arsenic tolerant capacity compared to others. The
selected strain was identified as Rhizobium by 16S rDNA sequencing and designated
as VMA301. Phylogenetic analysis of the gene sequences showed its close
relatedness with Sinorhizobium fredii. LC(50) value of arsenate for the bacteria
as determined by flow cytometry was found to be 2.8 mM and arsenic uptake was
measured by atomic absorption spectrometry as 0.048 mg g(/1) biomass. The high
amount of arsenic was toxic to the cell, which changed the morphology of the
bacteria to an elongated shape. Presence of a transcriptional regulatory gene
(ArsR) of the ars genetic system was confirmed by amplification and sequencing.
The symbiotic property of the isolate was also confirmed by amplification and
sequencing of the NodC gene. These results indicate that the isolated Rhizobium
bacteria may exert dual roles in the environment, arsenic bioremediation from the
soil as well as increase of soil fertility through nitrogen fixation.
PMID: 18497483 [PubMed / indexed for MEDLINE]
3: Ann Bot (Lond). 2007 Nov;100(5):903/24. Epub 2007 May 10.
Contrasting patterns in crop domestication and domestication rates: recent
archaeobotanical insights from the Old World.
Fuller DQ.
Institute of Archaeology, University College London, 31/34 Gordon Square, London
WC1H 0PY, UK. d.fuller@ucl.ac.uk
BACKGROUND: Archaeobotany, the study of plant remains from sites of ancient human
activity, provides data for studying the initial evolution of domesticated
plants. An important background to this is defining the domestication syndrome,
those traits by which domesticated plants differ from wild relatives. These
traits include features that have been selected under the conditions of
cultivation. From archaeological remains the easiest traits to study are seed
size and in cereal crops the loss of natural seed dispersal. SCOPE: The rate at
which these features evolved and the ordering in which they evolved can now be
documented for a few crops of Asia and Africa. This paper explores this in
einkorn wheat (Triticum monococcum) and barley (Hordeum vulgare) from the Near
East, rice (Oryza sativa) from China, mung (Vigna radiata) and urd (Vigna mungo)
beans from India, and pearl millet (Pennisetum glaucum) from west Africa. Brief
reference is made to similar data on lentils (Lens culinaris), peas (Pisum
sativum), soybean (Glycine max) and adzuki bean (Vigna angularis). Available
quantitative data from archaeological finds are compiled to explore changes with
domestication. The disjunction in cereals between seed size increase and
dispersal is explored, and rates at which these features evolved are estimated
from archaeobotanical data. Contrasts between crops, especially between cereals
and pulses, are examined. CONCLUSIONS: These data suggest that in domesticated
grasses, changes in grain size and shape evolved prior to non/shattering ears or
panicles. Initial grain size increases may have evolved during the first
centuries of cultivation, within perhaps 500/1000 years. Non/shattering
infructescences were much slower, becoming fixed about 1000/2000 years later.
This suggests a need to reconsider the role of sickle harvesting in
domestication. Pulses, by contrast, do not show evidence for seed size increase
in relation to the earliest cultivation, and seed size increase may be delayed by
2000/4000 years. This implies that conditions that were sufficient to select for
larger seed size in Poaceae were not sufficient in Fabaceae. It is proposed that
animal/drawn ploughs (or ards) provided the selection pressure for larger seeds
in legumes. This implies different thresholds of selective pressure, for example
in relation to differing seed ontogenetics and underlying genetic architecture in
these families. Pearl millet (Pennisetum glaucum) may show some similarities to
the pulses in terms of a lag/time before truly larger/grained forms evolved.
Publication Types:
Historical Article
Research Support, Non/U.S. Gov't
Review
PMID: 17495986 [PubMed / indexed for MEDLINE]
4: Biotechnol Lett. 2007 Aug;29(8):1271/5. Epub 2007 May 9.
Production and composition of extracellular polysaccharide synthesized by a
Rhizobium isolate of Vigna mungo (L.) Hepper.
Mandal SM, Ray B, Dey S, Pati BR.
Department of Microbiology, Vidyasagar University, Midnapore, WB, 721102, India.
An extracellular polysaccharide (EPS) was produced by a Rhizobium sp. isolated
from the root nodules of Vigna mungo (L.) Hepper. Maximum EPS production (346 mg
l(/1)) was when the yeast extract basal medium was supplemented with mannitol
(1%), biotin (1.5 mg l(/1)) and asparagine (0.3%). Ribose (53%) and mannose (47%)
were the principle monomers of the EPS. Chemical, chromatographic and
spectroscopic analysis showed that this polymer, which has Man(4)Rib(1) as an
oligomeric subunit, has an apparent molecular mass of 750 kDa.
PMID: 17487547 [PubMed / indexed for MEDLINE]
5: J Theor Biol. 2007 Jun 7;246(3):564/73. Epub 2007 Jan 25.
New motifs within the NB/ARC domain of R proteins: probable mechanisms of
integration of geminiviral signatures within the host species of Fabaceae family
and implications in conferring disease resistance.
Pal A, Chakrabarti A, Basak J.
Plant Molecular and Cellular Genetics, Bose Institute, P 1/12 CIT Scheme VIIM,
Kolkata 700054, India. amita@bic.boseinst.ernet.in
The Gemini viruses are a group of plant infectious agents, of which mungbean
yellow mosaic India virus (MYMIV) belongs to the bipartite subgroup of Gemini
virus and causes serious yield penalty in the leguminous group of plants. In this
investigation we have isolated two resistant gene homologues (RGHs; AY301990,
AY301991) from two MYMIV/resistant lines of Vigna mungo and V. radiata that have
high homology with a MYMIV/resistant linked marker, VMYR1 (AY 297425). These
three resistance factors also have similarity with 221 reported R gene/RGH
sequences in the NB/ARC domain of the family Fabaceae. NB/ARC domain is an
ancient, highly conserved domain of a class of plant disease resistance
genes/proteins. Out of 221 in silico translated protein sequences, multialignment
of 188 sequences without large insertion or truncation, unlike that of the rest
33, illustrated presence of both TIR and non/TIR subfamilies of NB/ARC domain. A
critical analysis of these sequences revealed eight new conserved motifs, in
addition to the reported conserved motifs within the NB/ARC domains, which are
hitherto not reported. Further analysis of these eight motifs with the aid of
PRINTS and PROSITE databases revealed signatures of geminiviral coat protein
(GVCP) within the favoured allele, R gene or RGHs. GVCP signatures are absent
within the NB/ARC domain of three species of Medicago, which are non/host to
Gemini virus. These observations tempted us to predict probable mechanism of
integration of GVCP within the plant R gene/RGHs and their implications in
conferring geminiviral disease resistance to the host plants. Our conjecture is
that these signatures were integrated during plant pathogen interaction and are
being maintained within this conserved domain through active selection of the
favoured allele. We comprehensively addressed the biological significance of GVCP
signatures, which probably provides additional defense against Gemini viruses
through degradation of homologous transcript of the virus.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 17320114 [PubMed / indexed for MEDLINE]
6: Theor Appl Genet. 2006 Nov;113(7):1261/9. Epub 2006 Aug 24.
Development of a black gram [Vigna mungo (L.) Hepper] linkage map and its
comparison with an azuki bean [Vigna angularis (Willd.) Ohwi and Ohashi] linkage
map.
Chaitieng B, Kaga A, Tomooka N, Isemura T, Kuroda Y, Vaughan DA.
Department of Horticulture, Faculty of Agriculture, Ubonratchathani University,
Ubonratchathani, Thailand.
The Asian Vigna group of grain legumes consists of six domesticated species,
among them black gram is widely grown in South Asia and to a lesser extent in
Southeast Asia. We report the first genetic linkage map of black gram [Vigna
mungo (L.) Hepper], constructed using a BC(1)F(1) population consisting of 180
individuals. The BC(1)F(1) population was analyzed in 61 SSR primer pairs, 56
RFLP probes, 27 AFLP loci and 1 morphological marker. About 148 marker loci could
be assigned to the 11 linkage groups, which correspond to the haploid chromosome
number of black gram. The linkage groups cover a total of 783 cM of the black
gram genome. The number of markers per linkage group ranges from 6 to 23. The
average distance between adjacent markers varied from 3.5 to 9.3 cM. The results
of comparative genome mapping between black gram and azuki bean show that the
linkage order of markers is highly conserved. However, inversions, insertions,
deletions/duplications and a translocation were detected between the black gram
and azuki bean linkage maps. The marker order on parts of linkage groups 1, 2 and
5 is reversed between the two species. One region on black gram linkage group 10
appears to correspond to part of azuki bean linkage group 1. The present study
suggests that the azuki bean SSR markers can be widely used for Asian Vigna
species and the black gram genetic linkage map will assist in improvement of this
crop.
Publication Types:
Comparative Study
PMID: 16932883 [PubMed / indexed for MEDLINE]
7: J Biomol Struct Dyn. 2006 Oct;24(2):123/30.
Theoretical model of the three/dimensional structure of a disease resistance gene
homolog encoding resistance protein in Vigna mungo.
Basak J, Bahadur RP.
Plant Molecular and Cellular Genetics Section, Bose Institute, P/1/12, CIT Scheme
VII M, Kolkata 700054. India. jolly.basak@ese.u/psud.fr
Plant disease resistance (R) genes, the key players of innate immunity system in
plants encode 'R' proteins. 'R' protein recognizes product of avirulance gene
from the pathogen and activate downstream signaling responses leading to disease
resistance. No three dimensional (3D) structural information of any 'R' proteins
is available as yet. We have reported a 'R' gene homolog, the 'VMYR1', encoding
'R' protein in Vigna mungo. Here, we describe the homology modeling of the
'VMYR1' protein. The model was created by using the 3D structure of an
ATP/binding cassette transporter protein from Vibrio cholerae as a template. The
strategy for homology modeling was based on the high structural conservation in
the superfamily of P/loop containing nucleoside triphosphate hydrolase in which
target and template proteins belong. This is the first report of theoretical
model structure of any 'R' proteins.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16928135 [PubMed / indexed for MEDLINE]
8: Carbohydr Res. 2006 Sep 4;341(12):2156/60. Epub 2006 May 23.
Isolation and structural analysis of ajugose from Vigna mungo L.
Kotiguda G, Peterbauer T, Mulimani VH.
Department of Biochemistry, Gulbarga University, Gulbarga 585106, India.
The hexasaccharide ajugose,
alpha/D/galactopyranosyl/(1//>6)/alpha/D/galactopyranosyl/(1//>6)/O/alpha/D/galac
topyranosyl/(1//>6)/alpha/D/galactopyranosyl/(1//>6)/alpha/D/glucopyranosyl/(1<//
>2)/beta/D/fructofuranoside, generally uncommon in legumes, was detected in the
seeds of Vigna mungo L. by TLC and paper chromatography. Ajugose was then
isolated by silica gel chromatography and its structure was established by acid
and enzymatic hydrolysis, fast atom bombardment mass spectrometry and both one/
and two/dimensional 1H and 13C NMR techniques.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 16716272 [PubMed / indexed for MEDLINE]
9: J Environ Sci Eng. 2004 Apr;46(2):151/8.
Growth, nodulation and dry matter yield of blackgram cultivars under nickel
stress.
Vijayarengan P.
Division of Environmental Science, Department of Botany, Annamalai University,
Annamalainagar.
Four cultivars of blackgram (Vigna mungo (L.) Hepper) were grown in soil amended
with nickel (0, 50, 100, 150 and 200 mg/kg) and analysed on the 45th day. Nickel
at all levels tested reduced the length of root and shoot, number of nodules,
area of leaves and dry matter yield of root and shoot of blackgram cultivars. The
reduction was lower in AB/1903 and ADT/3 than in ADT/4 and ADT/5. Cultivars
AB/1903 and ADT/3 were less sensitive than ADT/4 and ADT/5. The accumulation of
nickel was higher in roots than in shoots. The increase of nickel content in
roots of blackgram did not differ with cultivars. However, the accumulation of
nickel in shoots of blackgram differed with cultivars. The sensitive cultivar
ADT/4 and ADT/5 accumulate more nickel in their shoots than the other cultivars,
AB/ 1903 and ADT/3.
PMID: 16649606 [PubMed / indexed for MEDLINE]
10: Indian J Exp Biol. 2006 Mar;44(3):250/3.
Host range nodulation and adaptation in frenchbean rhizobia.
Dhar B, Mishra A, Singh MK.
Laboratory of Biological Nitrogen Fixation, Department of Genetics and Plant
Breeding, Institute of Agricultural Sciences, B.H.U., Varanasi 221 005, India.
The host range nodulation efficiency of four genetically marked frenchbean
rhizobial strains (HURR/3, Raj/2, Raj/5 and Raj/6) was studied with five legume
hosts namely, frenchbean (Phageolus vulgaris L.), pigeonpea [Cajanus cajan (L.)
Millsp.], mungbean [Vigna radiata (L.) Wilezek.], urdbean [Vigna mungo (L.)
Hepper.] and soybean [Glycine max (L.) Merril.]. Except soybean and pigeonpea,
all other legume hosts were nodulated by two or more frenchbean rhizobial strains
tested. Rhizobia were isolated from nodules produced by strains, HURR/3 and
Raj/5, on main (frenchbean) and different (mungbean and urdbean) hosts. There was
marked improvement in host range nodulation and nitrogen fixation efficiency of
rhizobial strains, HURR/3 and Raj/5. after their isolation from chance nodules on
different hosts. This is clearly evident from the ability of such isolates to
form nodules on pigeonpea besides mungbean and urdbean, and higher nodulation in
all the above three different hosts. The phage/susceptibility pattern and
intrinsic antibiotic resistance (used as markers) of the two strains did not
change after their passage through different hosts. The results indicate that
frenchbean rhizobia had undergone some modification in symbiotic behaviour to
adapt to wide host range during their passage through different (alternate?)
hosts.
PMID: 16538866 [PubMed / indexed for MEDLINE]
11: Plant Foods Hum Nutr. 2005 Dec;60(4):173/80.
Oligosaccharins of black gram (Vigna mungo L.) as affected by processing methods.
Girigowda K, Prashanth SJ, Mulimani VH.
Department of Biochemistry, Gulbarga University, Gulbarga, 585106, India.
The oligosaccharide content was determined in 12 different cultivars of black
gram. The effect of various treatments such as soaking, cooking, and enzyme
treatment on the raffinose family oligosaccharides of dry seeds and flour was
studied. Ajugose, a higher oligosaccharide (DP 6) found in trace quantities in
seeds, was shown in black gram by HPLC. The percent reduction of raffinose,
stachyose, verbascose, and ajugose after soaking for 16 hr was 41.66%, 47.61%,
28.48%, and 26.82%, respectively in Local/I variety and 43.75%, 20.58%, 23.60%,
and 15.88%, respectively in Local/II variety. Cooking for 60 min resulted in
decrease of 100% for raffinose, 76.19% for stachyose, 36.39% for verbascose, and
60.97% for ajugose in Local/I variety and 100% for raffinose, 55.88% for
stachyose, 48.52% for verbascose, and 56.07% for ajugose in Local/II variety.
Thin layer chromatographic analysis of 3 hr enzyme/treated samples revealed
almost complete hydrolysis of raffinose family of oligosaccharides. Among the
different methods employed, enzyme treatment was found to be the most effective
for removing alpha/galactosides in black gram.
PMID: 16395628 [PubMed / indexed for MEDLINE]
12: Bull Environ Contam Toxicol. 2005 Jun;74(6):1126/33.
Effect of copper and lead on photosynthesis and plant pigments in black gram
[Vigna mungo (L.) Hepper].
Bibi M, Hussain M.
Botany Department, University of Agriculture, Faisalabad, Pakistan.
PMID: 16158851 [PubMed / indexed for MEDLINE]
13: Plant Cell Rep. 2005 Jun;24(3):164/71. Epub 2005 Apr 7.
Transformation of a recalcitrant grain legume, Vigna mungo L. Hepper, using
Agrobacterium tumefaciens/mediated gene transfer to shoot apical meristem
cultures.
Saini R, Jaiwal PK.
Department of Biosciences, M.D. University, Rohtak, India.
The efficiency of Vigna mungo L. Hepper transformation was significantly
increased from an average of 1% to 6.5% by using shoot apices excised from
embryonic axes precultured on 10 microM benzyl/6/aminopurine (BAP) for 3 days and
wounded prior to inoculation in Agrobacterium tumefaciens strain EHA105 carrying
the binary vector pCAMBIA2301, which contains a neomycin phosphotransferase gene
(nptII) and a beta/glucuronidase (GUS) gene (gusA) interrupted by an intron. The
transformed green shoots that were selected and rooted on medium containing
kanamycin, and which tested positive for nptII gene by polymerase chain reaction,
were established in soil to collect seeds. GUS activity was detected in whole
T(0) shoots and T(1) seedlings. All T(0) plants were morphologically normal,
fertile and the majority of them transmitted transgenes in a 3:1 ratio to their
progenies. Southern analysis of T(1) plants showed integration of nptII into the
plant genome.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 15815929 [PubMed / indexed for MEDLINE]
14: Plant Foods Hum Nutr. 2004 Summer;59(3):123/8.
The in vivo synthesis and accumulation of lectin in developing seeds of black
gram (Vigna mungo L. Hepper).
Suseelan KN, Mitra R, Bhatia CR, Gopalakrishna T.
Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre,
Trombay. Mumbai/400 085, India. kns@magnum.barc.ernet.in
Black gram (Vigna mungo L. Hepper) seed contains two D/galactose/specific lectin
species, BGL/I and BGL/II, identified on the basis of elution from ion exchange
column and immunochemical cross/reactivity. BGL/I consisted of two monomeric
lectins, BGL/I/1 and BGL/1/2, of relative molecular weights 94 and 89 kDa,
respectively. BGL/II is another monomeric lectin with a molecular weight of 83
kDa. The in vivo synthesis studies using pulse/chase experiment showed that
BGL/II lectin was synthesized as early as 14 days after flowering (DAF). The
94/kDa BGL/I/1 lectin was synthesized around 17 DAF. There was no cotranslational
or posttranslational modification of the lectin proteins. The amount of lectin in
developing seeds was determined by radial immunodiffusion assay technique. The
maximum amount of lectin per seed was found at 28 DAF.
PMID: 15678718 [PubMed / indexed for MEDLINE]
15: Plant Biol (Stuttg). 2005 Jan;7(1):41/8.
Specificity patterns indicate that auxin exporters and receptors are the same
proteins.
Hössel D, Schmeiser C, Hertel R.
Institut Biologie III, Albert/Ludwig/Universität, Schänzlestrasse 1, 79104
Freiburg, Germany.
A study of transport and action of synthetic auxin analogues can help to identify
transporters and receptors of this plant hormone. Both aspects//transportability
and action on growth//were tested with 2/naphthoxyacetic acid (2/NOA) and
compared across several plant species. 2/NOA stimulates elongation effectively at
low concentrations in petioles of the gymnosperm Ginkgo biloba L., in hypocotyls
or internodes of the dicot legumes, mung bean (Vigna mungo L.) and pea (Pisum
sativum L.), in cotyledons of onion (Allium cepa L.) and in leaf bases of chive
(Allium schoenoprasum L.), the latter two of the monocot order Asparagales. In
contrast, elongation of coleoptile segments of maize (Zea mays L.) is poorly
responsive to 2/NOA. Significant auxin/like transport of 2/NOA was observed in
segments of mung bean hypocotyls, pea internodes, and chive leaf bases, but not
in segments of the grass coleoptiles. Thus, for the two assays, elongation and
polar transportability, the same difference in ligand specificity was observed
between the grass and all other species assayed. This finding supports the
hypothesis that a common protein mediates auxin efflux as well as auxin action on
elongation.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 15666213 [PubMed / indexed for MEDLINE]
16: J Biosci. 2004 Sep;29(3):297/308.
Infectivity analysis of two variable DNA B components of Mungbean yellow mosaic
virus/Vigna in Vigna mungo and Vigna radiata.
Balaji V, Vanitharani R, Karthikeyan AS, Anbalagan S, Veluthambi K.
Centre for Plant Molecular Biology, School of Biotechnology, Madurai Kamaraj
University, 625 021, India.
Mungbean yellow mosaic virus/Vigna (MYMV/Vig), a Begomovirus that causes yellow
mosaic disease, was cloned from field/infected blackgram (Vigna mungo). One DNA A
clone (KA30) and five different DNA B clones (KA21, KA22, KA27, KA28 and KA34)
were obtained. The sequence identity in the 150/nt common region (CR) between DNA
A and DNA B was highest (95%) for KA22 DNA B and lowest (85.6%) for KA27 DNA B.
The Rep/binding domain had three complete 11/nt (5'/TGTATCGGTGT/3') iterons in
KA22 DNA B (and KA21, KA28 and KA34), while the first iteron in KA27 DNA B
(5'/ATCGGTGT/3') had a 3/nt deletion. KA27 DNA B, which exhibited 93.9% CR
sequence identity to the mungbean/infecting MYMV, also shared the 3/nt deletion
in the first iteron besides having an 18/nt insertion between the third iteron
and the conserved nonanucleotide. MYMV was found to be closely related to KA27
DNA B in amino acid sequence identity of BV1 (94.1%) and BC1 (97.6%) proteins and
in the organization of nuclear localization signal (NLS), nuclear export signal
(NES) and phosphorylation sites. Agroinoculation of blackgram (V. mungo) and
mungbean (V. radiata) with partial dimers of KA27 and KA22 DNA Bs along with DNA
A caused distinctly different symptoms. KA22 DNA B caused more intense yellow
mosaic symptoms with high viral DNA titre in blackgram. In contrast, KA27 DNA B
caused more intense yellow mosaic symptoms with high viral DNA titre in mungbean.
Thus, DNA B of MYMVVig is an important determinant of host/range between V. mungo
and V. radiata.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 15381851 [PubMed / indexed for MEDLINE]
17: Theor Appl Genet. 2004 Nov;109(8):1687/93. Epub 2004 Sep 11.
A comparative analysis of genetic diversity in blackgram genotypes using RAPD and
ISSR markers.
Souframanien J, Gopalakrishna T.
Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre,
Trombay, Mumbai, 400085, India. souf@scientist.com
Random amplified polymorphic DNA (RAPD) and inter/simple sequence repeat (ISSR)
markers were used to study the DNA polymorphism in elite blackgram genotypes. A
total of 25 random and 16 ISSR primers were used. Amplification of genomic DNA of
the 18 genotypes, using RAPD analysis, yielded 104 fragments that could be
scored, of which 44 were polymorphic, with an average of 1.8 polymorphic
fragments per primer. Number of amplified fragments with random primers ranged
from two (OPA/13) to nine (OPK/4) and varied in size from 200 bp to 2,500 bp.
Percentage polymorphism ranged from 16.6% (OPK/7) to a maximum of 66.6% (OPE/5,
OPH/2, and OPK/8), with an average of 42.7%. The 16 ISSR primers used in the
study produced 101 bands across 18 genotypes, of which 55 were polymorphic. The
number of amplified bands varied from two (ISSR 858) to ten (ISSR 810), with a
size range of 200/2,200 bp. The average numbers of bands per primer and
polymorphic bands per primer were 6.3 and 3.4, respectively. Percentage
polymorphism ranged from 25% (ISSR 885) to 100% (ISSR 858), with an average
percentage polymorphism of 57.5% across all the genotypes. The 3'/anchored
primers based on poly(GA) and poly(AG) motifs produced high average polymorphisms
of 54.98% and 58.32%, respectively. ISSR markers were more efficient than the
RAPD assay, as they detected 57.4% polymorphic DNA markers in Vigna mungo as
compared to 42.7% for RAPD markers. The Mantel test between the two Jaccard's
similarity matrices gave r=0.32, showing low correlation between RAPD/ and
ISSR/based similarities. Clustering of genotypes within groups was not similar
when RAPD and ISSR derived dendrogram were compared, whereas the pattern of
clustering of the genotypes remained more or less the same in ISSR and combined
data of RAPD and ISSR.
Publication Types:
Comparative Study
PMID: 15368042 [PubMed / indexed for MEDLINE]
18: Arch Virol. 2004 Aug;149(8):1643/52. Epub 2004 Apr 5.
Analysis of an isolate of Mungbean yellow mosaic virus (MYMV) with a highly
variable DNA B component.
Karthikeyan AS, Vanitharani R, Balaji V, Anuradha S, Thillaichidambaram P,
Shivaprasad PV, Parameswari C, Balamani V, Saminathan M, Veluthambi K.
Centre for Plant Molecular Biology and Department of Plant Biotechnology, School
of Biotechnology, Madurai Kamaraj University, Madurai, India.
One DNA A (KA30) and five different DNA B components (KA21, KA22, KA27, KA28 and
KA34) of a geminivirus, Mungbean yellow mosaic virus/Vigna (MYMV/Vig) were cloned
from a pooled sample of field/infected Vigna mungo plants from Vamban, South
India. MYMV/Vig DNA A (KA30) and one of the DNA B components (KA27) exhibited 97%
and 95% sequence identities, respectively, to those of MYMV reported from
Thailand. However, the DNA B components KA21, KA22, KA28 and KA34 exhibited only
71 to 72% sequence identity to MYMV DNA B. Co/existence of multiple DNA B
components in field/infected V. mungo was proved by Southern and PCR analyses.
Each of the five DNA B components was infective together with the DNA A upon
agroinoculation. Agroinoculation with mixed cultures of Agrobacterium with
partial dimers of DNA A and all five DNA Bs proved that all five DNA B components
can co/infect a single V. mungo plant.
Publication Types:
Comparative Study
Research Support, Non/U.S. Gov't
PMID: 15290387 [PubMed / indexed for MEDLINE]
19: J Plant Physiol. 2004 May;161(5):563/71.
Correlation of endogenous free polyamine levels with root nodule senescence in
different genotypes in Vigna mungo L.
Lahiri K, Chattopadhyay S, Ghosh B.
Department of Botany, Krishnanagar Government College, Krishnanagar 741 101,
Nadia, West Bengal, India. kajarilahiri@yahoo.com
Endogenous free polyamines, nitrogenase (EC 1.1.8.6.1, acetylene reduction), and
leghaemoglobin (pyridine/hemochrome assay) levels were compared among five
genotypes of developing Vigna root nodules grown under field conditions.
Nitrogenase activity and leghaemoglobin level attained a peak at the flowering
stage and gradually declined thereafter. Individual and total polyamine also
followed the same pattern. Ranking on the basis of legume yield and other
morphometric attributes was PDU/2 > UH/28 > UH/82 > T/9 > Sardhomash. Except
spermine, the levels of putrescine, spermidine, and total polyamine showed
significant differences (p<0.05) amongst the genotypes, particularly from
flowering to mid/pod development stage. Genotype, development stage, and their
interaction between the two had significant (p<0.01) effects on individual as
well as total polyamines. Moreover, significant high linear correlations were
found between total free polyamine and putrescine with conventional nodule
senescence marker like nitrogenase (R2 = 0.94 and R2 = 0.92, respectively).
Putrescine had an overall positive correlation with high legume yield. The
results strongly suggest a relationship between polyamine and nodule senescence.
Endogenous free polyamine and putrescine may be considered as genotypic markers
for nodule senescence in field grown V. mungo. It is suggested that the flowering
stage is more suitable for selection.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 15202713 [PubMed / indexed for MEDLINE]
20: J Exp Bot. 2004 Apr;55(398):825/35. Epub 2004 Feb 27.
Molecular cloning and characterization of a cDNA encoding asparaginyl
endopeptidase from sweet potato (Ipomoea batatas (L.) Lam) senescent leaves.
Chen HJ, Hou WC, Liu JS, Yang CY, Huang DJ, Lin YH.
Department of Horticulture, Chinese Culture University, 111 Taipei, Taiwan.
Asparaginyl endopeptidase is a cysteine endopeptidase that has strict substrate
specificity toward the carboxyl side of asparagine residues, and is possibly
involved in the post/translational processing of proproteins. In this report one
full/length cDNA, SPAE, was isolated from senescent leaves of sweet potato
(Ipomoea batatas (L.) Lam). SPAE contained 1479 nucleotides (492 amino acids) in
the open reading frame, and exhibited high amino acid sequence homologies (c.
61/68%) with asparaginyl endopeptidases of Vicia sativa, Phaseolus vulgaris,
Canavalia ensiformis, and Vigna mungo. SPAE probably encoded a putative precursor
protein. Via cleavage of the N/ and C/termini, it produced a mature protein
containing 325 amino acids (from the 51st to the 375th amino acid residues), the
conserved catalytic residues (the 173rd His and 215th Cys amino acid residues),
and the putative N/glycosylation site (the 332nd Asn amino acid residue).
Semi/quantitative RT/PCR and western blot hybridization showed that SPAE gene
expression was enhanced significantly in natural senescent leaves and in dark/
and ethephon/induced senescent leaves, but was much less in mature green leaves,
stems, and roots. Phylogenic analysis showed that SPAE displayed close
association with vacuolar processing enzymes (legumains/asparaginyl
endopeptidases), which function via cleavage for proprotein maturation in the
protein bodies during seed maturation and germination. In conclusion, sweet
potato SPAE is probably a functional, senescence/associated gene and its mRNA and
protein levels were significantly enhanced in natural and induced senescent
leaves. The possible role and function of SPAE associated with bulk protein
degradation and mobilization during leaf senescence were also discussed.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 14990624 [PubMed / indexed for MEDLINE]
21: Plant Physiol. 2003 Aug;132(4):1892/900.
C/terminal KDEL sequence of a KDEL/tailed cysteine proteinase
(sulfhydryl/endopeptidase) is involved in formation of KDEL vesicle and in
efficient vacuolar transport of sulfhydryl/endopeptidase.
Okamoto T, Shimada T, Hara/Nishimura I, Nishimura M, Minamikawa T.
Department of Biological Sciences, Tokyo Metropolitan University, Hachioji,
Tokyo, 192/0397 Japan. okamoto/takashi@c.metro/u.ac.jp
Sulfhydryl/endopeptidase (SH/EP) is a papain/type vacuolar proteinase expressed
in cotyledons of germinated Vigna mungo seeds, and the enzyme possesses a
C/terminal propeptide containing KDEL tail, an endoplasmic reticulum retention
signal for soluble proteins. SH/EP is transported to vacuoles via a KDEL vesicle
(KV) through a Golgi complex/independent route. To see the function of the KDEL
sequence of SH/EP, wild/type SH/EP and its KDEL deletion mutant (SH/EPDeltaKDEL)
were heterologously expressed in Arabidopsis and in cultured tobacco Bright
Yellow 2 cells, and their intracellular transport pathways and localizations were
analyzed. A combination of the results from analyses for transformed Arabidopsis
and tobacco (Nicotiana tabacum) cells indicated that wild/type SH/EP is packed
into KV/like vesicles through the KDEL sequence and is transported to vacuoles in
the cells of transformants. In contrast, KV was not formed/induced in the cells
expressing SH/EPDeltaKDEL, and the mutant protein was mainly secreted. Therefore,
the C/terminal KDEL sequence of the KDEL/tailed cysteine proteinase is thought to
be involved in the formation of KV, and in the efficient vacuolar transport of
the proteins through KV.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 12913146 [PubMed / indexed for MEDLINE]
22: Plant Cell Rep. 2003 Jun;21(9):851/9. Epub 2003 Mar 22.
Erratum in:
Plant Cell Rep. 2003 Sep;22(2):166. Jaiwal S [corrected to Sonia].
Stable genetic transformation of Vigna mungo L. Hepper via Agrobacterium
tumefaciens.
Saini R, Sonia, Jaiwal PK.
Department of Biosciences, M.D. University, 124001, Rohtak, India.
Vigna mungo is one of the large/seeded grain legumes that has not yet been
transformed. We report here for the first time the production of morphologically
normal and fertile transgenic plants from cotyledonary/node explants inoculated
with Agrobacterium tumefaciens carrying binary vector pCAMBIA2301, the latter of
which contains a neomycin phosphotransferase ( nptII) gene and a
beta/glucuronidase (GUS) gene ( uidA) interrupted with an intron. The transformed
green shoots, selected and rooted on medium containing kanamycin, tested positive
for nptII and uidA genes by polymerase chain reaction (PCR) analysis. These
shoots were established in soil and grown to maturity to collect the seeds.
Mechanical wounding of the explants prior to inoculation with Agrobacterium, time
lag in regeneration due to removal of the cotyledons from explants and a second
round of selection at the rooting stage were found to be critical for
transformation. Analysis of T(0) plants showed the expression and integration of
uidA into the plant genome. GUS activity in leaves, roots, flowers, anthers and
pollen grains was detected by histochemical assay. PCR analysis of T(1) progeny
revealed a Mendelian transgene inheritance pattern. The transformation frequency
was 1%, and 6/8 weeks were required for the generation of transgenics.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 12789502 [PubMed / indexed for MEDLINE]
23: Theor Appl Genet. 2002 Dec;105(8):1215/1219. Epub 2002 Oct 19.
Assessment of peroxidase isozyme marker/based model for cross identifications in
hybrids (F(1)) of urdbean [ Vigna mungo (L.) Hepper].
Upadhyay R, Shukla A, Gaur K.
Department of Genetics and Plant Breeding, G.B. Pant University of Agriculture
and Technology, Pantnagar/263145, India, deanag@gbpuat.eruet.in
Four hybrids (4 F(1)s) were chosen out of crosses in the urdbean [ Vigna mungo
(L.) Hepper, 2n = 22] having contrasting morphological characters. Zymograms for
isozyme peroxidase were drawn from the patterns obtained from parents and their
respective F(1) hybrids on the basis of relative similarities to parental bands.
The selfed or crossed nature of hybrid pods was determined from the zymograms and
their analysis. The number of bands and their intensities gave an idea about the
extent of crossing in F(1) populations. Genetic identity (I) values were
indicative of their selfed nature. Dendrograms were constructed on the basis of
genetic identity values to display the relative similarities between the
populations. Analysis was based on individual pods to confirm their hybrid or
selfed nature. Possible use of this technique for identification of F(1) pods and
elimination of selfed pods might be implemented to shorten the breeding
operations during crossing.
PMID: 12582901 [PubMed / as supplied by publisher]
24: Planta. 2002 Dec;216(2):293/301. Epub 2002 Aug 24.
Species differences in ligand specificity of auxin/controlled elongation and
auxin transport: comparing Zea and Vigna.
Zhao H, Hertel R, Ishikawa H, Evans ML.
Collaborators: Evans ML.
Department of Plant Biology, Ohio State University, Columbus OH 43210, USA.
The plant hormone auxin affects cell elongation in both roots and shoots. In
roots, the predominant action of auxin is to inhibit cell elongation while in
shoots auxin, at normal physiological levels, stimulates elongation. The question
of whether the primary receptor for auxin is the same in roots and shoots has not
been resolved. In addition to its action on cell elongation in roots and shoots,
auxin is transported in a polar fashion in both organs. Although auxin transport
is well characterized in both roots and shoots, there is relatively little
information on the connection, if any, between auxin transport and its action on
elongation. In particular, it is not clear whether the protein mediating polar
auxin movement is separate from the protein mediating auxin action on cell
elongation or whether these two processes might be mediated by one and the same
receptor. We examined the identity of the auxin growth receptor in roots and
shoots by comparing the response of roots and shoots of the grass Zea mays L. and
the legume Vigna mungo L. to indole/3/acetic acid, 2/naphthoxyacetic acid,
4,6/dichloroindoleacetic acid, and 4,7/dichloroindoleacetic acid. We also studied
whether or not a single protein might mediate both auxin transport and auxin
action by comparing the polar transport of indole/3/acetic acid and
2/naphthoxyacetic acid through segments from Vigna hypocotyls and maize
coleoptiles. For all of the assays performed (root elongation, shoot elongation,
and polar transport) the action and transport of the auxin derivatives was much
greater in the dicots than in the grass species. The preservation of ligand
specificity between roots and shoots and the parallels in ligand specificity
between auxin transport and auxin action on growth are consistent with the
hypothesis that the auxin receptor is the same in roots and shoots and that this
protein may mediate auxin efflux as well as auxin action in both organ types.
Publication Types:
Comparative Study
Research Support, U.S. Gov't, Non/P.H.S.
PMID: 12447543 [PubMed / indexed for MEDLINE]
25: Planta. 2002 Oct;215(6):980/8. Epub 2002 Sep 5.
Basipetal propagation of gravity/induced surface pH changes along primary roots
of Lepidium sativum L.
Monshausen GB, Sievers A.
Botanisches Institut I, Universität Bonn, Venusbergweg 22, 53115 Bonn, Germany.
gabriele.monshausen@bio/geo.uni/karlsruhe.de
While there is ample evidence for a role of auxin in root gravitropism, the
seeming rapidity of gravi/induced changes in electrical parameters has so far
been an argument against auxin being a primary signal in gravitropic signal
transmission. To address this problem, we re/investigated the effect of
gravistimulation on membrane voltages of Lepidium sativum L. and Vigna mungo L.
root cells. In our hands, gravistimulation did not induce changes in membrane
voltage in cells of the root cap statenchyma, root meristem or apical elongation
zone that can be correlated with the orientation of the cells relative to the
gravity vector. While these results challenge a model of rapid electrically based
signal transmission, there is evidence for a slower signal propagation along
gravistimulated L. sativum roots. Using multiple proton/selective microelectrodes
to simultaneously measure surface pH on opposite root flanks at different
distances from the root tip, we observed gravi/induced asymmetric pH changes at
the surface of all investigated root zones. Upon gravistimulation, the surface pH
decreased on the physically upper root flank and increased on the lower flank.
The pH asymmetry appeared first [2.1+//0.4 min (mean +// SD) after tilting] at
the root cap and then / with incrementing lag times / at the meristem (after
2.5+//0.3 min at 300 micro m from root tip; after 3.7+//0.4 min at 700 micro m)
and apical elongation zone (4.8+//0.5 min at 1,000 micro m), suggesting a
basipetal progression of differential surface acidification at a rate of 250/350
micro m min(/1), consistent with reported auxin transport rates.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 12355158 [PubMed / indexed for MEDLINE]
26: Bull Environ Contam Toxicol. 2001 Nov;67(5):729/32.
Lindane and fenvalerate residues in blackgram (Vigna mungo).
Parihar NS, Gupta A.
Department of Entomology, Agricultural Research Station, Durgapura, Jaipur,
India.
PMID: 11911643 [PubMed / indexed for MEDLINE]
27: Indian J Exp Biol. 2001 Sep;39(9):916/20.
In vitro high frequency regeneration of plantlets of Vigna mungo and their ex
vitro growth.
Agnihotri S, Singh RR, Chaturvedi HC.
Tissue Culture Laboratory, National Botanical Research Institute, Lucknow, India.
Of the five explants of V. mungo var. T9 used, the excised shoot tips gave best
response with regard to offshoot formation followed by the embryonal axis
explants. While a treatment comprising 0.5 mgL(/1) BAP, 0.5 mgL(/1) 2iP and 0.1
mgL(/1) NAA induced differentiation of an average 10 offshoots in shoot tip
explants, only 3 offshoots were formed in the explants of embryonal axis in a
treatment containing 0.5 mgL(/1) BAP and 0.1 mgL(/1) NAA, found optimum for them.
Multiple shoots differentiated when explants with earlier regenerated and growing
offshoots were first cultured in a treatment containing 0.1 mgL(/1) BAP, 0.25
mgL(/1) IAA and 5 mgL(/1) CCC and then subcultured in the same treatment but
having only 1 mgL(/1) CCC. The isolated shoots rooted in 0.5 mgL(/1) IAA resulted
in the formation of complete plantlets of an average height of 15 cm in 20 days.
The in vitro/regenerated plants grew normally under field conditions and came to
flowering as well.
PMID: 11831376 [PubMed / indexed for MEDLINE]
28: Protoplasma. 2001;218(3/4):144/53.
Degradation of ribulose/bisphosphate carboxylase by vacuolar enzymes of senescing
French bean leaves: immunocytochemical and ultrastructural observations.
Minamikawa T, Toyooka K, Okamoto T, Hara/Nishimura I, Nishimura M.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/ohsawa
1/1, Hachioji, Tokyo 192/0397, Japan.
The possible involvement of vacuolar cysteine proteinases in degradation of
ribulose/bisphosphate carboxylase (Rubisco) in senescing French bean leaves was
studied by ultrastructural and immunocytochemical analyses with antibodies raised
against the large subunit (LSU) of Rubisco and SH/EP, a cysteine proteinase from
Vigna mungo that is immunologically identical to one of the major proteinases of
French bean plants. Primary leaves of 10/day/old plants were detached and placed
at 25 degrees C in darkness for 0, 4, and 8 days to allow their senescence to
proceed. The leaves at each senescence stage were subjected to the conventional
electron microscopic and immunocytochemical studies. The results indicated that
the chloroplasts of senescing French bean leaves were separated from the
cytoplasm of the cell periphery and taken into the central vacuole and that the
Rubisco LSU in the chloroplasts was degraded by vacuolar enzymes such as an
SH/EP/related cysteine proteinase that developed in senescing leaves. The present
results together with the results of previous biochemical studies using vacuolar
lysates support the view that Rubisco is degraded through the association of
chloroplasts with the central vacuole during the senescence of leaves that were
detached and placed in darkness.
PMID: 11770431 [PubMed / indexed for MEDLINE]
29: Plant Cell Physiol. 2001 Nov;42(11):1290/3.
Involvement of gibberellins in expression of a cysteine proteinase (SH/EP) in
cotyledons of Vigna mungo seedlings.
Taneyama M, Okamoto T, Yamane H, Minamikawa T.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/ohsawa,
Hachioji, Tokyo, 192/0397 Japan.
The expression of a papain/type proteinase, designated SH/EP, in cotyledons of
Vigna mungo seedlings has been shown to require some factors in the embryonic
axes. Gibberellin A1 (GA(1)) and GA(20) were identified by GC/MS in embryonic
axes of V. mungo seedlings. The level of accumulation of SH/EP in cotyledons of
V. mungo seedlings was greatly reduced by treatment of the seeds with
uniconazole/P, an inhibitor for GA biosynthesis. The reduced level of
accumulation of SH/EP in cotyledons by uniconazole/P was recovered by exogenous
application of GA(1) and GA(20) to the seedlings.
PMID: 11726715 [PubMed / indexed for MEDLINE]
30: Plant Cell Environ. 2000 Nov;23(11):1275/80.
Two distinct regions of response drive differential growth in Vigna root
electrotropism.
Wolverton C, Mullen JL, Ishikawa H, Evans ML.
Collaborators: Wolverton CS, Evans ML.
Department of Plant Biology, The Ohio State University, Columbus, OH 43210, USA.
Although exogenous electric fields have been reported to influence the
orientation of plant root growth, reports of the ultimate direction of
differential growth have been contradictory. Using a high/resolution image
analysis approach, the kinetics of electrotropic curvature in Vigna mungo L.
roots were investigated. It was found that curvature occurred in the same root
toward both the anode and cathode. However, these two responses occurred in two
different regions of the root, the central elongation zone (CEZ) and distal
elongation zone (DEZ), respectively. These oppositely directed responses could be
reproduced individually by a localized electric field application to the region
of response. This indicates that both are true responses to the electric field,
rather than one being a secondary response to an induced gravitropic stimulation.
The individual responses differed in the type of differential growth giving rise
to curvature. In the CEZ, curvature was driven by inhibition of elongation,
whereas curvature in the DEZ was primarily due to stimulation of elongation. This
stimulation of elongation is consistent with the growth response of the DEZ to
other environmental stimuli.
Publication Types:
Research Support, U.S. Gov't, Non/P.H.S.
PMID: 11706855 [PubMed / indexed for MEDLINE]
31: Plant Cell Physiol. 2001 Oct;42(10):1062/70.
Isolation of a putative receptor for KDEL/tailed cysteine proteinase (SH/EP) from
cotyledons of Vigna mungo seedlings.
Tsuru/Furuno A, Okamoto T, Minamikawa T.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/osawa
1/1, Hachioji, Tokyo, 192/0397 Japan.
SH/EP is the major papain/type proteinase expressed in cotyledons of germinated
Vigna mungo seeds. The proteinase possesses a KDEL sequence at the C/terminus
although the mature form of SH/EP is localized in vacuoles. It has also been
shown that the proform of SH/EP is accumulated at the edge or middle region of
the endoplasmic reticulum, and the accumulated proSH/EP is directly transported
to vacuoles via the KDEL/tailed cysteine proteinase/accumulating vesicle, KV. In
this study, to address the transport machinery of proSH/EP through KV, putative
receptor for proSH/EP was isolated from membrane proteins of cotyledons of V.
mungo seedlings using a proSH/EP/immobilized column. The deduced amino acid
sequence from cDNA to the protein revealed that the putative receptor for
proSH/EP is a member of vacuolar sorting receptor, VSR, that is known to be
localized in the Golgi/complex and/or clathrin coated vesicle. We carried out
subcellular fractionation of cotyledon cells and subsequently conducted
SDS/PAGE/immunoblotting and immunocytochemistry with anti/V. mungo VSR (VmVSR) or
SH/EP antibody. The results showed that VmVSR is co/localized in the fraction of
the gradient in which KV existed.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 11673621 [PubMed / indexed for MEDLINE]
32: J Cell Biol. 2001 Sep 3;154(5):973/82. Epub 2001 Aug 27.
Cotyledon cells of Vigna mungo seedlings use at least two distinct autophagic
machineries for degradation of starch granules and cellular components.
Toyooka K, Okamoto T, Minamikawa T.
Department of Biological Sciences, Tokyo Metropolitan University, Tokyo, 192/0397
Japan.
alpha/Amylase is expressed in cotyledons of germinated Vigna mungo seeds and is
responsible for the degradation of starch that is stored in the starch granule
(SG). Immunocytochemical analysis of the cotyledon cells with anti/alpha/amylase
antibody showed that alpha/amylase is transported to protein storage vacuole
(PSV) and lytic vacuole (LV), which is converted from PSV by hydrolysis of
storage proteins. To observe the insertion/degradation processes of SG into/in
the inside of vacuoles, ultrastructural analyses of the cotyledon cells were
conducted. The results revealed that SG is inserted into LV through autophagic
function of LV and subsequently degraded by vacuolar alpha/amylase. The autophagy
for SG was structurally similar to micropexophagy detected in yeast cells. In
addition to the autophagic process for SG, autophagosome/mediated autophagy for
cytoplasm and mitochondria was detected in the cotyledon cells. When the embryo
axes were removed from seeds and the detached cotyledons were incubated, the
autophagosome/mediated autophagy was observed, but the autophagic process for the
degradation of SG was not detected, suggesting that these two autophagic
processes were mediated by different cellular mechanisms. The two distinct
autophagic processes were thought to be involved in the breakdown of SG and cell
components in the cells of germinated cotyledon.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 11524437 [PubMed / indexed for MEDLINE]
33: Plant Foods Hum Nutr. 2001;56(3):265/73.
Tannin contents and protein digestibility of black grams (Vigna mungo) after
soaking and cooking.
Zia/Ur/rehman, Shah WH.
Biotechnology and Food Research Centre, PCSIR Laboratories Complex, Lahore,
Pakistan.
The objective of this research was to ascertain the effects of soaking black
grams (Cultivar AARI/5732) in different salt solutions at different temperatures
and different time periods, and different methods of cooking on the tannin
content and protein digestibility. Tannin content of black grams was reduced to
various extents by soaking at 30 degrees and 100 degrees C for different time
periods. However, soaking at 100 degrees C increased the rate of extraction and
reduced the extraction time of tannins. Soaking black grams in water at 100
degrees C reduced tannins by 22.14% in 45 minutes whereas about 2.5 times more
tannin was reduced after soaking in sodium bicarbonate solution with or without
sodium chloride. Maximum improvement in protein digestibility was also observed
after soaking black grams in sodium bicarbonate solution. Tannin contents were
further reduced along with improvement in protein digestibility as a result of
cooking.
PMID: 11442226 [PubMed / indexed for MEDLINE]
34: Plant Cell Physiol. 2001 Jun;42(6):635/41.
A TGACGT motif in the 5'/upstream region of alpha/amylase gene from Vigna mungo
is a cis/element for expression in cotyledons of germinated seeds.
Yamauchi D.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/ohsawa
1/1, Hachioji, Tokyo, 192/0397 Japan. yamauchi/daisuke@c.metro/u.ac.jp
Alpha/amylase is expressed at high levels in cotyledons of germinated seeds of
Vigna mungo. The mRNA for alpha/amylase appeared in cotyledons of the seeds at 1
d after imbibition started (DAI). Two TGACGT motifs at /445 and at /125 in the
promoter region of the gene interacted with nuclear proteins from cotyledons of
dry seeds and the activities were detected until 3 DAI. A transient assay with
particle bombardment showed that the downstream region from /135 in the promoter
was required for high level expression in the cotyledons and the activity was
reduced by mutation of the TGACGT motif at /125. The activities to bind the
TGACGT motifs were detected in the axes of the seeds at 1 DAI but disappeared at
4 DAI, although the mRNA for alpha/amylase in the axes appeared at 4 DAI and
increased in level by 6 DAI. A transient assay experiment showed that a positive
regulatory element for the expression in the axes was located in the region from
/630 to /453. These results indicated that the TGACGT motif at /125 was required
for high level expression of the gene in the cotyledons of the germinated seeds.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 11427683 [PubMed / indexed for MEDLINE]
35: J Biol Chem. 2001 Jan 5;276(1):742/51.
Identification of a membrane/associated cysteine protease with possible dual
roles in the endoplasmic reticulum and protein storage vacuole.
Okamoto T, Toyooka K, Minamikawa T.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/osawa
1/1, Hachioji, Tokyo, 192/0397 Japan. okamoto/takashsi@c.metro/u.ac.jp
SH/EP is a vacuolar cysteine proteinase from germinated seeds of Vigna mungo. The
enzyme has a C/terminal propeptide of 1 kDa that contains an endoplasmic
reticulum (ER) retention signal, KDEL. The KDEL/tail has been suggested to
function to store SH/EP as a transient zymogen in the lumen of the ER, and the
C/terminal propeptide was thought to be removed within the ER or immediately
after exit from the ER. In the present study, a protease that may be involved in
the post/translational processing of the C/terminal propeptide of SH/EP was
isolated from the microsomes of cotyledons of V. muno seedlings. cDNA sequence
for the protease indicated that the enzyme is a member of the papain superfamily.
Immunocytochemistry and subcellular fractionation of cotyledon cells suggested
that the protease was localized in both the ER and protein storage vacuoles as
enzymatically active mature form. In addition, protein fractionations of the
cotyledonary microsome and Sf9 cells expressing the recombinant protease
indicated that the enzyme associates with the microsomal membrane on the luminal
side. The protease was named membrane/associated cysteine protease, MCP. The
possibility that a papain/type enzyme, MCP, exists as mature enzyme in both ER
and protein storage vacuoles will be discussed.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 11022031 [PubMed / indexed for MEDLINE]
36: Int J Food Sci Nutr. 2000 May;51(3):169/74.
Effect of cooking and supplementation with different kinds of meats on the
nutritional value of mash (Vigna mungo).
Bhatty N, Gilani AH, Nagra SA.
Department of Rural Home Economics, University of Agriculture, Faisalabad,
Pakistan.
The study was conducted to determine the nutritional value of mash (Vigna mungo)
in raw and cooked forms and as effected by supplementation with different kind of
meat, i.e. poultry, mutton and beef at 10, 15 and 20% levels. Nutritional
assessment of all mash/containing diets (without or with supplementation) was
made by chemical analysis as well as through rat assay. Mash contained 23.83%
protein. Cooking resulted in minor changes in nutrients. Mash had 1.79% lysine
which was reduced by 35% on cooking. All other amino acids also showed losses
during cooking. Protein efficiency ratio (PER) of diet containing raw mash was
1.9% and cooking improved it to 2.8%. True digestibility (TD) also showed a
significant improvement. Supplementation of mash with different kinds of meat did
not improve the PER significantly over unsupplemented diet containing cooked mash
only. TD, however, was improved from 74.89% in cooked to 75.58/87.06% in
supplemented diets. Similarly net protein utilization (NPU), as a result of meat
supplementation, improved from 43.54% in cooked to 42.88%/51.96%. Higher PER, TD
and NPU values were observed in diets containing mash supplemented with 20% level
of different meats.
PMID: 10945112 [PubMed / indexed for MEDLINE]
37: J Cell Biol. 2000 Feb 7;148(3):453/64.
Mass transport of proform of a KDEL/tailed cysteine proteinase (SH/EP) to protein
storage vacuoles by endoplasmic reticulum/derived vesicle is involved in protein
mobilization in germinating seeds.
Toyooka K, Okamoto T, Minamikawa T.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/osawa
1/1, Hachioji, Tokyo, 192/0397 Japan.
A vacuolar cysteine proteinase, designated SH/EP, is expressed in the cotyledon
of germinated Vigna mungo seeds and is responsible for the degradation of storage
proteins. SH/EP is a characteristic vacuolar proteinase possessing a
COOH/terminal endoplasmic reticulum (ER) retention sequence, KDEL. In this work,
immunocytochemical analysis of the cotyledon cells of germinated V. mungo seeds
was performed using seven kinds of antibodies to identify the intracellular
transport pathway of SH/EP from ER to protein storage vacuoles. A proform of
SH/EP synthesized in ER accumulated at the edge or middle region of ER where the
transport vesicle was formed. The vesicle containing a large amount of proSH/EP,
termed KV, budded off from ER, bypassed the Golgi complex, and was sorted to
protein storage vacuoles. This massive transport of SH/EP via KV was thought to
mediate dynamic protein mobilization in the cotyledon cells of germinated seeds.
We discuss the possibilities that the KDEL sequence of KDEL/tailed vacuolar
cysteine proteinases function as an accumulation signal at ER, and that the mass
transport of the proteinases by ER/derived KV/like vesicle is involved in the
protein mobilization of plants.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 10662772 [PubMed / indexed for MEDLINE]
38: Eur J Biochem. 1999 Aug;264(1):223/32.
Erratum in:
Eur J Biochem 1999 Dec;266(2):697. Mimamikawa T [corrected to Minamikawa T].
Asparaginyl endopeptidase (VmPE/1) and autocatalytic processing synergistically
activate the vacuolar cysteine proteinase (SH/EP).
Okamoto T, Yuki A, Mitsuhashi N, Minamikawa T.
Department of Biological Sciences, Tokyo Metopolitan University, Hachioji, Japan.
okamoto/takashi@c.metro/u.ac.jp
A vacuolar cysteine proteinase, designated SH/EP, is synthesized in cotyledons of
germinated Vigna mungo seeds and is responsible for degradation of the seed
proteins accumulated in protein bodies (protein storage vacuoles). SH/EP belongs
to the papain proteinase family and has a large N/terminal prosegment consisting
of 104 amino acid residues and a C/terminal prosegment of 10 amino acid residues.
It has been suggested that an asparaginyl endopeptidase, V. mungo processing
enzyme 1 (VmPE/1), is involved in the N/terminal post/translational processing of
SH/EP. The recombinant proform of SH/EP (rSH/EP) was produced in Escherichia coli
cells, purified to homogeneity and refolded by stepwise dialysis. 31P/NMR
analysis of intact germinated cotyledons revealed that the vacuolar pH of
cotyledonary cells changes from 6.04 to 5.47 during seed germination and early
seedling growth. rSH/EP was converted in vitro to the mature form through
autocatalytic processing at a pH mimicking the vacuolar pH at the mid and late
stages of seed germination, but not at the pH of the early stage. VmPE/1
accelerated the rate of processing of rSH/EP in vitro at the pH equivalent to the
vacuolar pH at the early and mid stages of germination. In addition, the cleavage
sites of the in vitro processed intermediates and the mature form of SH/EP were
identical to those of SH/EP purified from germinated cotyledons of V. mungo. We
propose that the asparaginyl endopeptidase (VmPE/1)/mediated processing mainly
functions in the activation of proSH/EP at the early stage of seed germination,
and both VmPE/1/mediated and autocatalytic processings function synergistically
in the activation of proSH/EP in cotyledons at the mid and late stages.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 10447692 [PubMed / indexed for MEDLINE]
39: J Mass Spectrom. 1999 Aug;34(8):804/12.
Analysis of saponins from black bean by electrospray ionization and fast atom
bombardment tandem mass spectrometry.
Lee MR, Chen CM, Hwang BH, Hsu LM.
Department of Chemistry, National Chung/Hsing University, Taichung, Taiwan.
mrlee@mail.nchu.edu.tw
Saponins from black bean (Vigna mungo L. Hepper) were analyzed using positive and
negative ion fast atom bombardment mass spectrometry (FAB/MS) and liquid
chromatography/mass spectrometry. Methanol was used to extract the saponins from
defatted black bean, which was partially purified by extraction with n/butanol,
and the extract was dialyzed with 3000 M(r) cut/off tubing. The dialyzate was
analyzed using mass spectrometry. According to FAB/MS/MS, mixtures from black
bean contain soyasaponin I as the predominant saponin. In addition, MS/MS
analysis was performed in which the structures of saponins of black bean
cotyledon were determined to be soyasaponin I, soyasaponin II, soyasaponin V,
3/O/[alpha/L/rhamnopyranosyl/(1 //> 2)/beta/D/galactopyranosyl/(1 //>
2)/beta/D/glucuronopyranosyl]complogenin (saponin A) and
3/O/[alpha/L/rhamnopyranosyl/(1 //> 2)/beta/D/glucopyranosyl/(1 //>
2)/beta/D/glucopyranosyl]oleanolic acid (saponin B). For the black bean shell and
the root of black bean sprout, analysis confirmed the saponins of soyasaponin I,
soyasaponin II, soyasaponin V, saponin A, saponin B, acetylsoyasaponin A(4) and
soyasaponin beta(g). Moreover, all the studied saponins were found in the stem
and leaves of the black bean sprouts, except soyasaponin beta(g) and
acetylsoyasaponin A(4), respectively. Copyright 1999 John Wiley & Sons, Ltd.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 10423561 [PubMed / indexed for MEDLINE]
40: J Biol Chem. 1999 Apr 16;274(16):11390/8.
Erratum in:
J Biol Chem 1999 Aug 27;274(35):25188. Okomoto T [corrected to Okamoto T].
Posttranslational removal of the carboxyl/terminal KDEL of the cysteine protease
SH/EP occurs prior to maturation of the enzyme.
Okamoto T, Minamikawa T, Edward G, Vakharia V, Herman E.
Department of Biological Sciences, Tokyo Metropolitan University, Minami/osawa,
Hachioji, Tokyo, 192/0397 Japan. okamoto/takashi@c.metro/u.ac.jp
SH/EP is a cysteine protease from germinating mung bean (Vigna mungo) that
possesses a carboxyl/terminal endoplasmic reticulum (ER) retention sequence,
KDEL. In order to examine the function of the ER retention sequence, we expressed
a full/length cDNA of SH/EP and a minus/KDEL control in insect Sf/9 cells using
the baculovirus system. Our observations on the synthesis, processing, and
trafficking of SH/EP in Sf/9 cells suggest that the KDEL ER/retention sequence is
posttranslationally removed either while the protein is still in the ER or
immediately after its exit from the ER, resulting in the accumulation of proSH/EP
minus its KDEL signal. It is this intermediate form that appears to progress
through the endomembrane system and is subsequently processed to form mature
active SH/EP. The removal of an ER retention may regulate protein delivery to a
functional site and present an alternative role for ER retention sequences in
addition to their well established role in maintaining the protein composition of
the ER lumen.
Publication Types:
Research Support, Non/U.S. Gov't
Research Support, U.S. Gov't, Non/P.H.S.
PMID: 10196232 [PubMed / indexed for MEDLINE]
41: Bull Environ Contam Toxicol. 1999 Apr;62(4):502/7.
Metal binding properties of ferritin in Vigna mungo (L.) Hepper (black gram):
possible role in heavy metal detoxification.
Rama Kumar T, Prasad MN.
Department of Plant Sciences, University of Hyderabad, Hyderabad 500046, India.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 10094736 [PubMed / indexed for MEDLINE]
42: Plant Mol Biol. 1999 Jan;39(1):63/73.
Molecular cloning and characterization of Vigna mungo processing enzyme 1
(VmPE/1), an asparaginyl endopeptidase possibly involved in post/translational
processing of a vacuolar cysteine endopeptidase (SH/EP).
Okamoto T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Hachioji, Japan.
Asparaginyl endopeptidase is a cysteine endopeptidase that has strict substrate
specificity toward the carboxy side of asparagine residues. Vigna mungo
processing enzyme 1, termed VmPE/1, occurs in the cotyledons of germinated seeds
of V. mungo, and is possibly involved in the post/translational processing of a
vacuolar cysteine endopeptidase, designated SH/EP, which degrades seed storage
protein. VmPE/1 also showed a substrate specificity to asparagine residues, and
its enzymatic activity was inhibited by NEM but not E/64. In addition, purified
VmPE/1 had a potential to process the recombinant SH/EP precursor to its
intermediate in vitro. cDNA clones for VmPE/1 and its homologue, named VmPE/1A,
were identified and sequenced, and their expressions in the cotyledons of V.
mungo seedlings and other organs were investigated. VmPE/1 mRNA and SH/EP mRNA
were expressed in germinated seeds at the same stage of germination although the
enzymatic activity of VmPE/1 rose prior to that of SH/EP. The level of VmPE/1A
mRNA continued increasing as germination proceeded. In roots, stems and leaves of
fully grown plants, and in hypocotyls, VmPE/1 and VmPE/1A were little expressed.
We discuss possible functions of VmPE/1 and VmPE/1A in the cotyledons of
germinated seeds.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 10080709 [PubMed / indexed for MEDLINE]
43: Plant Cell Physiol. 1998 Nov;39(11):1145/55.
Isolation and characterization of a cDNA clone of UDP/galactose: flavonoid
3/O/galactosyltransferase (UF3GaT) expressed in Vigna mungo seedlings.
Mato M, Ozeki Y, Itoh Y, Higeta D, Yoshitama K, Teramoto S, Aida R, Ishikura N,
Shibata M.
Department of Floriculture, National Research Institute of Vegetables, Ornamental
Plants and Tea, Mie, Japan.
Four cDNA clones were isolated from Vigna mungo seedlings by the screening with
cDNA encoding UDP/glucose:flavonoid 3/O/glucosyltransferase (UF3GT) of
Antirrhinum majus as a probe; the product of the gene corresponding to one cDNA
was more highly expressed in the first simple leaves than in stems. Nucleotide
sequence analysis revealed 1,691 bp (including 326 bp non/reading) containing an
open reading frame of 455 amino acids. The deduced amino acid sequence showed 42%
and 23% identity with those of A. majus UDP/glucose:flavonoid
3/O/glucosyltransferase (UF3GT) and Petunia hybrida UDP/rhamnose:anthocyanidin
3/O/glucoside rhamnosyltransferase (RT), respectively. One region of the cDNA
(amino acids 325 to 387) showed similarity to ceramide UDP/galactosyltransferases
of mice, rats and humans. A crude extract from Escherichia coli, in which the
protein was expressed from the cDNA, showed high UF3GaT activity but low UF3GT
activity, and was similar in K(m), optimal pH and substrate specificity to UF3GaT
from V. mungo. We conclude that we have obtained UDP/galactose:flavonoid
3/O/galactosyltransferase (UF3GaT) cDNA from V. mungo.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 9891414 [PubMed / indexed for MEDLINE]
44: Planta. 1998 Oct;206(3):466/75.
A cysteine endopeptidase with a C/terminal KDEL motif isolated from castor bean
endosperm is a marker enzyme for the ricinosome, a putative lytic compartment.
Schmid M, Simpson D, Kalousek F, Gietl C.
Lehrstuhl für Botanik, Technische Universität München, Germany.
A papain/type cysteine endopeptidase with a molecular mass of 35 kDa for the
mature enzyme, was purified from germinating castor bean (Ricinus communis L.)
endosperm by virtue of its capacity to process the glyoxysomal malate
dehydrogenase precursor protein to the mature subunit in vitro (C. Gietl et al.,
1997, Plant Physiol 113: 863/871). The cDNA clones from endosperm of germinating
seedlings and from developing seeds were isolated and sequence analysis revealed
that a very similar or identical peptidase is synthesised in both tissues.
Sequencing established a presequence for co/translational targeting into the
endoplasmic reticulum, an N/terminal propeptide and a C/terminal KDEL motif for
the castor bean cysteine endopeptidase precursor. The 45/kDa pro/enzyme stably
present in isolated organelles was enzymatically active. Immunocytochemistry with
antibodies raised against the purified cysteine endopeptidase revealed highly
specific labelling of ricinosomes, organelles which co/purify with glyoxysomes
from germinating Ricinus endosperm. The cysteine endopeptidase from castor bean
endosperm, which represents a senescing tissue, is homologous to cysteine
endopeptidases from other senescing tissues such as the cotyledons of germinating
mung bean (Vigna mungo) and vetch (Vicia sativa), the seed pods of maturing
French bean (Phaseolus vulgaris) and the flowers of daylily (Hemerocallis sp.).
Publication Types:
Research Support, Non/U.S. Gov't
Research Support, U.S. Gov't, P.H.S.
PMID: 9763713 [PubMed / indexed for MEDLINE]
45: Plant Mol Biol. 1998 Mar;36(4):565/71.
Analysis of the expression of two thiolprotease genes from daylily (Hemerocallis
spp.) during flower senescence.
Guerrero C, de la Calle M, Reid MS, Valpuesta V.
Departamento de Bioquímica y Biología Molecular, Universidad de Málaga, Spain.
A cDNA clone encoding a daylily (Hemerocallis spp.) thiolprotease (SEN11), whose
expression is strongly upregulated in flower tepal senescence, has been isolated.
The amino acid sequence, deduced from the nucleotide sequence, showed highest
similarity to plant thiolproteases of Vigna mungo, Phaseolus vulgaris and
Hemerocallis (SEN102), and contains a putative ER retention signal that has been
described in Vigna mungo. SEN102 and SEN11 transcripts were not detectable in
flower buds at the opening stage, but two peaks of transcripts were seen after 9
h and 19 h, in both petals and sepals, when wilting symptoms were apparent. The
pattern of protease activity migrating on a 26.3 kDa protein was similar to the
SEN102 and SEN11 transcript profiles. These two genes were also expressed in
stamens and leaves, but their transcripts were undetectable in carpels and
rhizomes. The expression of SEN102 was lower in the senescent leaf than in the
green leaf. The pattern of expression of these genes suggests their involvement
in the protein hydrolysis occurring in tepals at the late senescence stage,
whereas in leaves they could be involved in the constitutive protein turnover
machinery. Exogenous gibberellic acid application to cut flowers increased
transcripts of both genes.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 9484451 [PubMed / indexed for MEDLINE]
46: Eur J Biochem. 1997 Sep 1;248(2):304/12.
Proteinase A, a storage/globulin/degrading endopeptidase of vetch (Vicia sativa
L.) seeds, is not involved in early steps of storage/protein mobilization.
Becker C, Senyuk VI, Shutov AD, Nong VH, Fischer J, Horstmann C, Müntz K.
Institut für Pflanzengenetik und Kulturpflanzenforschung, Gatersleben, Germany.
Proteinase A is a papain/like cysteine endopeptidase of vetch (Vicia sativa L.)
which was assumed to initiate storage/globulin breakdown just after the onset of
seed germination. This enzyme was purified from cotyledons of vetch seedlings. On
gelatin/containg SDS gels, active proteinase A migrated with an apparent
molecular mass of 21 kDa, whereas after heat denaturation its molecular size on
SDS/PAGE was 29 kDa. Although proteinase A is capable of hydrolyzing storage
globulins in vitro it could not be localized in the protein/body fraction of
cotyledons from germinating seeds. cDNA clones encoding proteinase A precursor
have been obtained by PCR. The precursor is composed of an N/terminal signal
sequence followed by a propeptide, the region encoding mature proteinase A, and a
C/terminal KDEL sequence. Mature proteinase A with a derived molecular mass of
25,244 Da does not have the KDEL sequence. The derived amino acid sequence of the
proteinase A precursor is 78.2% identical to sulfhydryl/endopeptidase (SH/EP), a
cysteine endopeptidase from germinating Vigna mungo seedlings. Northern blot
analysis indicated that proteinase A mRNA appears de novo in cotyledons of
1/day/germinated vetch seeds, where its amount increases up to day 6. No
proteinase A mRNA was detected in other vetch organs, not even in the embryo
axis, which contains stored globulins. By means of antibodies raised against the
purified and against recombinantly produced proteinase A, the 29/kDa bands of
mature proteinase A were detected in cotyledon extracts of 6/day/germinated seeds
when globulin degradation has already far proceeded. The reported data do not
agree with the proposed triggering role of proteinase A in storage/globulin
breakdown during germination.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 9346282 [PubMed / indexed for MEDLINE]
47: Biochim Biophys Acta. 1997 Aug 29;1336(2):323/30.
Theophylline metabolism in higher plants.
Ito E, Crozier A, Ashihara H.
Department of Biology, Ochanomizu University, Faculty of Science, Tokyo, Japan.
Metabolism of [8/(14)C]theophylline was investigated in leaf segments from
Camellia sinensis (tea), Camellia irrawadiensis, Ilex paraguariensis (maté) and
Avena sativa, root segments of Vigna mungo seedlings and cell suspension cultures
of Catharanthus roseus. There was extensive uptake and metabolism of
[8/(14)C]theophylline by leaves of tea and Camellia irrawadiensis and, to a
lesser extent, maté. These purine alkaloid/containing species converted
[8/(14)C]theophylline into 3/methylxanthine, xanthine, the ureides allantoin and
allantoic acid, and CO2. With the other test systems, which were from species
that do not produce purine alkaloids, there were low levels of
[8/(14)C]theophylline uptake which were accompanied by incorporation of
relatively small amounts of label into 3/methylxanthine, xanthine and CO2. None
of the higher plants converted [8/(14)C]theophylline to either 1/methyluric acid
or 1,3/dimethyluric acid, which are the main catabolites of theophylline in
mammals. The data indicate that the main route of theophylline degradation in
higher plants involves a theophylline //> 3/methylxanthine //> xanthine //> uric
acid //> allantoin //> allantoic acid //> //> CO2 + NH3 pathway. In tea and mate,
large amounts of [8/(14)C]theophylline were also converted to theobromine and
caffeine via a theophylline //> 3/methylxanthine //> theobromine //> caffeine
salvage pathway. The diversity of theophylline metabolism in higher plants and
mammals is discussed.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 9305805 [PubMed / indexed for MEDLINE]
48: Plant Physiol. 1997 Mar;113(3):863/71.
A cysteine endopeptidase isolated from castor bean endosperm microbodies
processes the glyoxysomal malate dehydrogenase precursor protein.
Gietl C, Wimmer B, Adamec J, Kalousek F.
Institute of Botany, Technical University of Munich, Germany.
gietl@botanik.biologie.tumuenchen.de
A plant cysteine endopeptidase with a molecular mass of 35 kD was purified from
microbodies of germinating castor bean (Ricinus communis) endosperm by virtue of
its capacity to specifically process the glyoxysomal malate dehydrogenase
precursor protein to the mature subunit in vitro. Processing of the glyoxysomal
malate dehydrogenase precursor occurs sequentially in three steps, the first
intermediate resulting from cleavage after arginine/13 within the presequence and
the second from cleavage after arginine/33. The endopeptidase is unable to remove
the presequences of prethiolases from rape (Brassica napus) glyoxysomes and rat
peroxisomes at the expected cleavage site. Protein sequence analysis of
N/terminal and internal peptides revealed high identity to the mature papain/type
cysteine endopeptidases from cotyledons of germinating mung bean (Vigna mungo)
and French bean (Phaseolus vulgaris) seeds. These endopeptidases are synthesized
with an extended pre//prosequence at the N terminus and have been considered to
be processed in the endoplasmic reticulum and targeted to protein/storing
vacuoles.
Publication Types:
Research Support, Non/U.S. Gov't
Research Support, U.S. Gov't, P.H.S.
PMID: 9085576 [PubMed / indexed for MEDLINE]
49: Eur J Biochem. 1996 Jun 1;238(2):317/24.
Successive amino/terminal proteolysis of the large subunit of ribulose
1,5/biphosphate carboxylase/oxygenase by vacuolar enzymes from French bean
leaves.
Yoshida T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
Mainly using the protein immunoblot technique, we observed the decrease in
amounts of the large subunit (LSU) and the small subunit (SSU) of ribulose
1,5/biphosphate carboxylase/oxygenase (Rubisco) in detached primary leaves of
French bean plants during senescence under the light or in darkness, but detected
no significant degradation products of these subunits. Treatment of the detached
leaves with 0.6% (mass/vol.) dimethyl sulfoxide, 0.05% (mass/vol.) Tween 80
considerably promoted the senescence, as estimated by the reduction in content of
the soluble protein, and also in the amounts of LSU and SSU, but no degradation
product of either subunit was found. When extracts prepared from the primary
leaves were incubated at pH 5.4 or pH 7.4, the amount of LSU of 53 kDa decreased
and concurrently 50/kDa and 42/kDa polypeptides were formed. Since the results
suggested that Rubisco may be degraded by vacuolar enzymes, we incubated Rubisco
with vacuolar lysates prepared from the senescing primary leaves and found that
the LSU, but not SSU, was degraded to a 41/kDa polypeptide through three
intermediates of 50 kDa, 48 kDa and 42 kDa. Determination of amino/terminal amino
acid sequences of these fragments indicated that each of the proteolysis steps
occurred by removal of a small amino/terminal peptide. Experiments with various
inhibitors of proteases as well as with a purified Vigna mungo vacuolar protease,
termed SH/EP [Mitsuhashi, W. & Minamikawa, T. (1989) Plant Physiol. 89, 274/279]
suggested the involvement of two types of proteases in these steps: a cysteine
protease that is the same type of enzyme as SH/EP catalyzes the steps from the
LSU to the 48/kDa polypeptide through the 50/kDa polypeptide, and a serine
protease catalyzes the steps from the 48/kDa polypeptide to the 41/kDa
polypeptide through the 42/kDa polypeptide.
PMID: 8681940 [PubMed / indexed for MEDLINE]
50: Plant Mol Biol. 1996 May;31(2):239/54.
A major cysteine proteinase, EPB, in germinating barley seeds: structure of two
intronless genes and regulation of expression.
Mikkonen A, Porali I, Cercos M, Ho TH.
Dept. of Biology, Washington University, St. Louis, MO 63130, USA.
The barley cysteine proteinase B (EPB) is the main protease responsible for the
degradation of endosperm storage proteins providing nitrogenous nutrients to
support the growth of young seedlings. The expression of this enzyme is induced
in the germinating seeds by the phytohormone, gibberellin, and suppressed by
another phytohormone, abscisic acid. In situ hybridization experiments indicate
that EPB is expressed in the scutellar epithelium within 24 h of seed
germination, but the aleurone tissue surrounding the starchy endosperm eventually
becomes the main tissue expressing this enzyme. The EPB gene family of barley
consists of two very similar genes, EPB1 and EPB2, both of which have been mapped
to chromosome 3. The sequences of EPB1 and EPB2 match with the two previously
published cDNA clones indicating that both genes are expressed. Interestingly,
neither of these genes contain any introns, a rare phenomenon in which all
members of a small gene family are active intronless genes. Sequence comparison
indicates that the barley EPB family can be classified as cathepsin L/like
endopeptidases and is most closely related to two legume cysteine proteinases
(Phaseolus vulgaris EP/C1 and Vigna mungo SHEP) which are also involved in seed
storage protein degradation. The promoters of EPB1 and EPB2 have been linked to
the coding sequence of a reporter gene, GUS, encoding beta/glucuronidase, and
introduced into barley aleurone cells using the particle bombardment method.
Transient expression studies indicate that EPB promoters are sufficient to confer
the hormonal regulation of these genes.
Publication Types:
Comparative Study
Research Support, Non/U.S. Gov't
Research Support, U.S. Gov't, Non/P.H.S.
PMID: 8756590 [PubMed / indexed for MEDLINE]
51: Plant Mol Biol. 1996 Jan;30(2):321/9.
Promoter regions of cysteine endopeptidase genes from legumes confer
germination/specific expression in transgenic tobacco seeds.
Yamauchi D, Terasaki Y, Okamoto T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
Cysteine endopeptidases, SH/EP from Vigna mungo and EP/C1 from Phaseolus
vulgaris, act to degrade seed storage protein during seed germination. Using
transgenic tobacco plants, expression of SH/EP and promoter activity of the EP/C1
gene were analyzed in transgenic tobacco plants. The promoters of the two genes
in tobacco seeds showed germination/specific activation, although
post/translational processing of SH/EP and regulatory regions of promoter of the
gene for EP/C1 were found to differ between leguminous seeds and transgenic
tobacco seeds.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 8616255 [PubMed / indexed for MEDLINE]
52: Eur J Biochem. 1995 Jul 15;231(2):300/5.
Purification of a processing enzyme (VmPE/1) that is involved in
post/translational processing of a plant cysteine endopeptidase (SH/EP).
Okamoto T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
A cysteine endopeptidase, designated SH/EP, occurs in the cotyledons of
germinated seeds of Vigna mungo and acts to degrade the seed storage protein in
protein storage vacuoles. SH/EP is synthesized on membrane/bound ribosomes as an
inactive 45/kDa precursor, which is cotranslationally processed to a 43/kDa
intermediate through cleavage of the signal sequence; the 43/kDa intermediate of
SH/EP is further processed to the 33/kDa mature enzyme via 39/kDa and 36/kDa
intermediates [Mitsuhashi, W. & Minamikawa, T. (1989) Plant Physiol. 89,
274/279]. The present in vitro processing experiments indicated that at least two
processing enzymes, designated VmPE/1 and VmPE/2 (V. mungo processing enzymes 1
and 2), were involved in post/translational processing of SH/EP in cotyledons of
V. mungo seedlings. VmPE/1 was purified from the cotyledons as a protease that
was involved in the processing of the 43/kDa intermediate to the 36/kDa
intermediate. The enzyme has a molecular mass of 33 kDa as estimated by
SDS/polyacrylamide gel electrophoresis, and showed high similarity to the
jackbean asparaginyl endopeptidase in terms of the primary structure and
substrate specificity. We discuss the function of VmPE/1 in the processing of
SH/EP and related proteases in the cotyledons of germinated seeds.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 7635141 [PubMed / indexed for MEDLINE]
53: FEBS Lett. 1994 Aug 29;351(1):31/4.
Erratum in:
FEBS Lett 1994 Dec 12;356(1):152.
Posttranslational processing of a carboxy/terminal propeptide containing a KDEL
sequence of plant vacuolar cysteine endopeptidase (SH/EP)
Okamoto T, Nakayama H, Seta K, Isobe T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
A plant cysteine endopeptidase, designated SH/EP, is a major protease occurring
in cotyledons of Vigna mungo seedlings, and acts to degrade seed globulin stored
in protein bodies. Here we show that the 43 kDa intermediate of SH/EP formed in
the endoplasmic reticulum is transported to protein bodies and processed to the
33 kDa mature form during transport or thereafter, and that the COOH/terminal
propeptide of 10 amino acid residues containing a KDEL sequence, which is known
as a retention signal for the endoplasmic reticulum lumen, is processed to form
the mature SH/EP.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 8076688 [PubMed / indexed for MEDLINE]
54: Plant Cell Physiol. 1994 Jun;35(4):705/11.
Structure and expression of alpha/amylase gene from Vigna mungo.
Yamauchi D, Takeuchi H, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
A single copy of the alpha/amylase gene, composed of three introns and four
exons, was found in Vigna mungo. Examination of levels of alpha/amylase and its
mRNA in detached cotyledons indicated that attachment of the embryonic axis is
not required for expression of the gene in cotyledons of germinating seeds.
Publication Types:
Comparative Study
Research Support, Non/U.S. Gov't
PMID: 8075901 [PubMed / indexed for MEDLINE]
55: Plant Physiol. 1993 Dec;103(4):1459.
Nucleotide sequence of the alpha/amylase gene from Vigna mungo.
Takeuchi H, Yamauchi D, Wada S, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
PMID: 8290640 [PubMed / indexed for MEDLINE]
56: Biochim Biophys Acta. 1993 Feb 13;1156(2):123/7.
Pyrophosphate: fructose/6/phosphate 1/phosphotransferase and biosynthetic
capacity during differentiation of hypocotyls of Vigna seedlings.
Ashihara H, Sato F.
Department of Biology, Faculty of Science, Ochanomizu University, Tokyo, Japan.
The relationship between the activity of pyrophosphate:fructose/6/phosphate
1/phosphotransferase (PFP) and the capacity for biosynthesis of macromolecules
was examined in segments from different parts of hypocotyls of etiolated
seedlings of Vigna mungo and V. radiata. The relative ratio of the maximum
activity of PFP to that of ATP/dependent phosphofructokinase (PFK) (PFP/PFK
ratio) was high in young tissues and decreased with differentiation and ageing of
the tissues. The highest level of fructose/2,6/bisphosphate was observed in the
youngest part of hypocotyls of V. mungo. The level was markedly decreased with
ageing of tissues. The levels of PPi and ATP were also higher in younger parts
than in older parts of the hypocotyls, but the ratio of the level of PPi to that
of ATP was almost constant in all parts of the hypocotyl. A good correlation was
found between the PFP/PFK ratio and the biosynthetic capacity, as estimated from
the rate of incorporation of [U/14C]sucrose into ethanol/insoluble
macromolecules.
Publication Types:
Comparative Study
Research Support, Non/U.S. Gov't
PMID: 8381302 [PubMed / indexed for MEDLINE]
57: Plant Physiol. 1993 Feb;101(2):421/428.
Expression of an Endopeptidase (EP/C1) in Phaseolus vulgaris Plants.
Tanaka T, Minamikawa T, Yamauchi D, Ogushi Y.
Department of Biology, Tokyo Metropolitan University, Minami/ohsawa, Hachioji,
Tokyo 192/03, Japan.
Endopeptidase activity increases continually in pods of maturing fruits of French
bean (Phaseolus vulgaris L. cv Goldstar) plants and is thought to participate in
the protein mobilization in pods during the development of seeds (M. Endo, T.
Minamikawa, D. Yamauchi, W. Mitsuhashi [1987] J Exp Bot 38: 1988/1995). In the
present studies, one of the major endopeptidase forms, designated EP/C1, was
purified as a 34/kD polypeptide from pods of maturing French bean fruits. EP/C1
was found to be immunologically distinguished from other forms in extracts from
pods, but homologous to SH/EP, the major cysteine endopeptidase expressed in
cotyledons of germinating Vigna mungo seeds (W. Mitsuhashi, T. Minamikawa [1989]
Plant Physiol 89: 274/279). The level of endopeptidase that reacted with the
antiserum to EP/C1 increased in pods as the fruit maturation proceeded. EP/C1 was
also immunologically detected in stems of French bean plants bearing fruits of
later maturation stages. Protein immunoblotting showed that a 34/kD polypeptide
corresponding to EP/C1 in molecular mass occurred in extracts from 7/ to 9/d
cotyledons of germinating French bean seeds. In addition, two other polypeptides
with slightly higher molecular masses were observed in extracts from 3/ to 5/d
cotyledons. We suggest that these two polypeptides are intermediates involved in
posttranslational processing of EP/C1. RNA blot hybridization with EP/C1 cDNA as
a probe showed that EP/C1 mRNA occurred in pods of fruits at later maturing
stages and also in cotyledons of 3/ to 7/d germinating seeds.
PMID: 12231697 [PubMed / as supplied by publisher]
58: Indian J Biochem Biophys. 1991 Oct/Dec;28(5/6):439/43.
A monomeric protein with hemagglutinating activity from seeds of Vigna mungo
(Phaseolus mungo).
Singh SS, Rao SL.
Department of Biochemistry, Osmania University, Hyderabad, India.
Black gram (Vigna mungo) seeds are shown to contain a lectin with certain unusual
features. The lectin agglutinates only trypsinized red cells, and its sugar
specificity is complex as none of the common sugars, oligosaccharides or complex
polysaccharides exhibit any affinity for the lectin. The purified lectin has a
molecular weight of 58 kDa and is a monomer. Unlike other plant lectins,
antibodies to the P. mungo lectin do not exhibit any immunological cross
reactivity. The clot forming ability of the lectin is unusual in that the clot
once formed is rapidly disaggregated indicated that it induces, as yet undefined,
certain membrane alterations.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 1812079 [PubMed / indexed for MEDLINE]
59: Environ Pollut. 1991;69(2/3):131/49.
Sensitivity of three leguminous crops to O3 as influenced by different stages of
growth and development.
Kasana MS.
Department of Biological Sciences, University of Lancaster, Bailrigg, Lancaster,
LA1 4YQ, UK.
The sensitivity of Cicer arietinum, Vigna mungo and Trigonella foenum/graecum to
O(3) has been assessed at different stages of growth and development. Plants of
different ages (0, 1, 2, 4 and 6 weeks old) were fumigated with 0 and 120 nl
litre(/1) O(3), from 09.30 h to 16.30 h each day for four weeks, in hemispherical
chambers located out/of/doors. Seed germination was not affected by O(3) in any
of the species, but there were responses (differing between species) on the
cotyledons. True leaves were fairly resistant when young but later they became
more sensitive. Premature senescence and earlier abscission of leaves (in C.
arietinum and T. foenum/graecum) and flowers and abortive fruit drop (in C.
arietinum) were also observed. Of the five growth stages examined, 2/ and
4/week/old plants seemed to be most sensitive except for Trigonella where
sensitivity decreased with increasing age of the plants. The partitioning and
distribution of dry matter among different plant parts was also significantly
disturbed and root, leaf and stem were adversely affected in a decreasing order.
However, the percentage reductions in dry weight per plants for Cicer and Vigna
increased with age up to four weeks, then declined abruptly. Growth reductions at
the 0/ and 6/week/old stages differed only slightly and were very small in
magnitude. It may, therefore, be suggested that the plants of these legumes in
early stages of exponential growth are more vulnerable to O(3) damage and that
the developmental or physiological age is an important factor in O(3)
sensitivity.
PMID: 15092157 [PubMed]
60: Plant Foods Hum Nutr. 1990 Oct;40(4):243/7.
Temporal variation in protein content and yield of Vigna mungo (L.) Hepper
leaves.
Pandey VN, Srivastava AK.
Botany Department, University of Gorakhpur, India.
Temporal variation in total protein and soluble protein contents and protein
yield of Vigna mungo leaves at intervals of every three hours during day and
night was studied. The study was done with the view to ascertain the hour of
harvesting the leaves for maximum yield of leaf protein concentrate. Observations
reveal that the total protein and soluble protein contents in the leaves are
minimum during 3.00 to 6.00 hrs, which steadily rise with time to reach the
maximum values during 12.00 to 15.00 hrs, after which the same shows a steady
decrease with time.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 2247432 [PubMed / indexed for MEDLINE]
61: Nucleic Acids Res. 1990 Jul 25;18(14):4250.
Nucleotide sequence of cDNA for alpha/amylase from cotyledons of germinating
Vigna mungo seeds.
Yamauchi D, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
PMID: 2377468 [PubMed / indexed for MEDLINE]
62: Plant Cell Physiol. 1990 Jun;31(4):457/62.
Gravity/induced changes in intracellular potentials in elongating cortical cells
of mung bean roots.
Ishikawa H, Evans ML.
Collaborators: Evans ML.
Department of Botany, The Ohio State University, Columbus 43210.
Gravity/induced changes in intracellular potentials in primary roots of 2/day/old
mung bean (Vigna mungo L. cv. black matpe) seedlings were investigated using
glass microelectrodes held by 3/dimensional hydraulic micro/drives. The
electrodes were inserted into outer cortical cells within the elongation zone.
Intracellular potentials, angle of root orientation with respect to gravity, and
position within the root of the impaled cortical cell were measured
simultaneously. Gravistimulation caused intracellular potential changes in
cortical cells of the elongation zone. When the roots were oriented vertically,
the intracellular potentials of the outer cortical cells (2 mm behind the root
apex) were approximately / 115 mV. When the roots were placed horizontally
cortical cells on the upper side hyperpolarized to / 154 mV within 30 s while
cortical cells on the lower side depolarized to about / 62 mV. This electrical
asymmetry did not occur in cells of the maturation zone. Because attempts to
insert the electrode into cells of the root cap were unsuccessful, these cells
were not measured. The hyperpolarization of cortical cells on the upper side was
greatly reduced upon application of N,N'/dicyclohexylcarbodiimide (DCCD), an
inhibitor of respiratory energy coupling. When stimulated roots were returned to
the vertical, the degree of hyperpolarization of cortical cells on the previous
upper side decreased within 30 s and approached that of cortical cells in
non/stimulated roots. This cycle of hyperpolarization/loss of hyperpolarization
was repeatable at least ten times by alternately turning the root from the
vertical to the horizontal and back again. The very short (<30 s) lag period of
these electrical changes indicates that they may result from stimulus/perception
and transduction within the elongation zone rather than from transmission of a
signal from the root cap.
Publication Types:
Research Support, Non/U.S. Gov't
Research Support, U.S. Gov't, Non/P.H.S.
PMID: 11537168 [PubMed / indexed for MEDLINE]
63: Nucleic Acids Res. 1990 Apr 11;18(7):1892.
Nucleotide sequence of the gene for the Vigna mungo sulfhydryl/endopeptidase
(SH/EP).
Akasofu H, Yamauchi D, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Setagaya/ku, Japan.
PMID: 2336365 [PubMed / indexed for MEDLINE]
64: Biochem Int. 1989 Oct;19(4):745/53.
Properties of a cyclic 3'5'/nucleotide phosphodiesterase from Vigna mungo.
Lee CH, Abidin UZ.
Dept. of Biochemistry and Microbiology, Univ. of Agriculture Malaysia, Selangor.
Cyclic AMP phosphodiesterase (PDE) partially purified from roots of Vigna mungo
exhibited optimum activity at pH 5.5 to 6.0 and maximum enzyme activity at 50
degrees C. Levels of PDE activity in roots remained relatively constant from the
first to the eleventh day after germination; on the twelfth day there was a 400%
increase in PDE activity. The enzyme was stable for at least 48 hours at 28
degrees C, retaining 92% of its original activity. Plant growth hormones
including gibberellic acid, indoleacetic acid and kinetin at 1.0 and 10.0 microM
concentrations did not have any significant effect on enzyme activity.
Nucleotides tested including cyclic 2'3' AMP, cyclic 2'3' GMP completely
abolished enzyme activity at 1.0mM while cyclic 3'5' GMP, cyclic 3'5' GMP,
2'deoxy 5' ATP, 2'deoxy 5'GTP and 5'ADP were also inhibitory to the enzyme. The
enzyme was stimulated by Mg2+, Fe2+ and NH4+ while Cu2+ and Fe3+ were inhibitory.
Theophylline, caffeine, phosphate, pyrophosphate and EDTA were inhibitory to the
enzyme.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 2559728 [PubMed / indexed for MEDLINE]
65: Plant Foods Hum Nutr. 1989 Sep;39(3):257/66.
Antinutrients in amphidiploids (black gram x Mung bean): varietal differences and
effect of domestic processing and cooking.
Kataria A, Chauhan BM, Punia D.
Department of Foods and Nutrition, Haryana Agricultural University, Hisar, India.
Phytic acid, saponin and polyphenol contents in grains of various varieties of
black gram (Vigna mungo) Mung bean (Vigna radiata L.) amphidiploids ranged from
697 to 750, 2746 to 2972 and 702 to 783 mg/100 g, respectively. Domestic
processing and cooking methods including soaking, ordinary and pressure cooking
of soaked and unsoaked seeds, and sprouting significantly lowered phytic acid,
saponin and polyphenol contents of the amphidiploid seeds. Soaking for 18 h
removed 31 to 37% of the phytic acid; the extent of removal was higher with long
periods of soaking. Saponins and polyphenols were relatively less affected. Loss
of the antinutrients was greater when soaked instead of unsoaked seeds were
cooked. Pressure cooking had a greater effect than ordinary cooking. Antinutrient
concentrations declined following sprouting; the longer the period of germination
the greater was the reduction.
Publication Types:
In Vitro
PMID: 2608635 [PubMed / indexed for MEDLINE]
66: Nucleic Acids Res. 1989 Aug 25;17(16):6733.
Nucleotide sequence of cDNA for sulfhydryl/endopeptidase (SH/EP) from cotyledons
of germinating Vigna mungo seeds.
Akasofu H, Yamauchi D, Mitsuhashi W, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Japan.
PMID: 2780300 [PubMed / indexed for MEDLINE]
67: Plant Foods Hum Nutr. 1989 Jun;39(2):149/54.
Protein digestibility (in vitro) of chickpea and blackgram seeds as affected by
domestic processing and cooking.
Jood S, Chauhan BM, Kapoor AC.
Department of Foods & Nutrition, Haryana Agricultural University, Hisar, India.
Protein digestibility (in vitro) of grains of chickpea (Cicer arietinum) and
blackgram (Vigna mungo) cultivars varied from 48 to 53% and 52 to 58%,
respectively. Soaking, cooking (both of unsoaked and soaked seeds), autoclaving
and sprouting improved significantly the protein digestibility of all the
cultivars of chickpea and blackgram. Autoclaving was found to be most effective
followed by cooking and sprouting; cooking of sprouts had only marginal effect.
Protein digestibility was higher when soaked instead of unsoaked grains were
cooked.
PMID: 2762243 [PubMed / indexed for MEDLINE]
68: J Biochem. 1989 Feb;105(2):265/70.
Detection and characterization of p/coumaric acid hydroxylase in mung bean, Vigna
mungo, seedlings.
Kojima M, Takeuchi W.
Institute for Biochemical Regulation, Faculty of Agriculture, Nagoya University,
Aichi.
A new p/coumaric acid (4/hydroxycinnamic acid) hydroxylase was detected in mung
bean seedlings treated with tentoxin, a fungal toxin, in which polyphenol oxidase
that hydroxylates a wide variety of monophenols in vitro was completely
eliminated. The enzyme required molecular oxygen and showed a pH optimum of 5.0.
The enzyme acted only on p/coumaric acid (Km, 3.0 X 10(/5) M), while its
specificity for the electron donor was rather broad. The Km value for NADPH (1.5
X 10(/4) M) was much lower than that for L/ascorbic acid (1.0 X 10(/2) M),
although the Vmax value was almost the same with both electron donors. The enzyme
was potently inhibited by beta/mercaptoethanol (Ki, 3.5 X 10(/6) M) and
diethyldithiocarbamate (Ki, 2.3 X 10(/4) M), but was insensitive to
p/chloromercuribenzoate. The enzyme was localized in the cell organelles which
sedimented between mitochondria and endplasmic reticulum on sucrose density
gradient centrifugation. The enzyme activity in the seedling was changed in
response to induction by light in a manner suggesting its involvement in
biosynthesis of phenolic compounds in mung bean seedlings.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 2498300 [PubMed / indexed for MEDLINE]
69: Plant Physiol. 1989 Jan;89(1):274/279.
Synthesis and Posttranslational Activation of Sulfhydryl/Endopeptidase in
Cotyledons of Germinating Vigna mungo Seeds.
Mitsuhashi W, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Setagaya/ku, Tokyo 158,
Japan.
A sulfhydryl/endopeptidase was purified as a 33 kilodalton (kD) mass polypeptide
from cotyledons of Vigna mungo seedlings. Immunoblot analysis with antiserum made
against the purified enzyme showed that the sulfhydryl/endopeptidase was
synthesized only in the cotyledons during germination and that the amount of the
enzyme increased until 4 days after imbibition and decreased thereafter. Next, an
RNA fraction was prepared from cotyledons of 3 day old seedlings and translated
in a wheat germ system. The synthesis of a 45 kD polypeptide was shown by the
analysis of its translation products by immunoprecipitation with the antiserum to
the endopeptidase and gel electrophoresis. When the RNA fraction was translated
in the presence of canine microsomal membranes, a smaller polypeptide, having a
43 kD molecular mass, was detected as the translation product. When
membrane/bound polysomes, but not free polysomes, prepared from cotyledons were
used for translation in the wheat germ system, both the 43 and 45 kD polypeptides
were synthesized. By incubation of a crude enzyme extract from cotyledons at 5
+// 1 degrees C at neutral pH, the 43 kD polypeptide was sequentially cleaved to
the 33 kD polypeptide via 39 and 36 kD intermediate polypeptides. The
endopeptidase was activated simultaneously with the processing. Two/dimensional
polyacrylamide gel electrophoresis showed that the 33 kD polypeptide was the
fully activated form of the enzyme, whereas little or no activity was detected in
other forms. From the present results, we postulate that the
sulfhydryl/endopeptidase is first synthesized as the 45 kD precursor with a 2 kD
signal peptide being cleaved, and that the 43 kD polypeptide is further cleaved
to give the 33kD mature enzyme.
PMID: 16666526 [PubMed / as supplied by publisher]
70: Plant Foods Hum Nutr. 1988;38(1):75/81.
Proximate composition and antinutritional factors in rice bean (Vigna umbellata).
Malhotra S, Malik D, Dhindsa KS.
Department of Chemistry and Biochemistry, Haryana Agricultural University, Hisar,
India.
Thirteen promising strains of Rice bean (Vigna umbellata) were analysed for their
proximate compositions and antinutritional factors. Protein content in these
varieties ranged from 17.50 to 23.10 per cent, ash from 3.06 to 4.48 per cent,
ether extract from 2.4 to 3.9 per cent and crude fibre from 1.70 to 4.25 per
cent. Trypsin inhibitor activity ranged from 112.63 to 163.98 units/g and
polyphenols ranged from 0.58 to 1.19 per cent. Phytohemagglutinating activity was
present in all the strains, except one, RB/32. Oligosaccharides, viz., raffinose,
stachyose and verbascose, ranged from 0.32 to 0.91, 0.95 to 1.98 and 1.40 to 2.58
per cent, respectively. Attempts have been made to compare the results with a
standard variety each of cowpea (Vigna unguiculata), moong (Vigna radiata) and
mash (Vigna mungo).
PMID: 3231596 [PubMed / indexed for MEDLINE]
71: Plant Foods Hum Nutr. 1988;38(2):163/73.
Characterization of seed storage proteins of urdbean (Vigna mungo).
Mahajan R, Malhotra SP, Singh R.
Department of Chemistry and Biochemistry, Haryana Agricultural University, Hisar,
India.
Dehulled and defatted flour of urdbean (Vigna mungo), Var T/9, contained 25%
protein with maximum contribution by globulins (63%). Albumins and glutelins
contributed 12% and 21% respectively, whereas prolamins were present only in
traces (1%). Globulins were further fractionated into legumin and vicilin type
proteins which were present in the ratio of 4:1. All the protein fractions were
heterogenous in nature as revealed by high performance liquid chromatography.
SDS/polyacrylamide gel electrophoresis revealed the total protein sample to
contain 21 different components with molecular weights ranging from 8.92 to
117.49 kd. Albumins, globulins, prolamins and glutelins resolved into 4, 8, 6 and
13 different sized components of molecular weights ranging from 10.23 to 25.53,
10.84 to 112.72, 10.33 to 51.52 and 8.91 to 112.72 kd, respectively. Amino acid
analysis of all fractions revealed that glutamic acid was present in maximum
concentration followed by aspartic acid and lysine. Just like other pulse
proteins, the urdbean proteins were also deficient in sulphur containing amino
acids.
PMID: 3200802 [PubMed / indexed for MEDLINE]
72: Plant Foods Hum Nutr. 1988;38(2):115/20.
Gamma/rays and EMS induced pentaphyllous mutant in black gram (Vigna mungo).
Singh RK, Raghuvanshi SS, Prakash D.
Botany Department, Lucknow University, India.
Pentaphyllous mutants in black gram were isolated in M2 generation of a
segregating family, irradiated at 20 kR. The genetic nature of mutants was tested
by hybridizing with controls, and chi/square tests applied to the F2 population,
proved it to be a monogenic recessive. The pentaphyllous mutant had a greater
number of pods and leaves per plant and larger and more root nodules. It also
showed improved nutritional value with increased seed protein percentage and no
increase in TIA (trypsin inhibitor activity).
PMID: 3200796 [PubMed / indexed for MEDLINE]
73: Plant Physiol. 1986 Mar;80(3):628/634.
Separation and Characterization of Two Endopeptidases from Cotyledons of
Germinating Vigna mungo Seeds.
Mitsuhashi W, Koshiba T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Setagaya/ku, Tokyo 158,
Japan.
Two major endopeptidases were present in cotyledons of germinating Vigna mungo
seeds, as detected by the zymogram after polyacrylamide gel electrophoresis. They
were not detectable in cotyledons of dry seeds, but their intensities on the
zymogram increased during germination. During incubation of detached cotyledons,
however, the activities showed only a slight increase for 5 days. These two
endopeptidases could be separated by Sephacryl S/200 column chromatography. One
of them was found to be a serine/endopeptidase as judged by
phenylmethylsulfonylfluoride and diisopropyl fluorophosphate inhibition. The
other was a sulfhydryl/endopeptidase because of its dependency on
2/mercaptoethanol and inhibition by leupeptin, chymostatin, and antipain.
Analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis indicatd
that the two endopeptidases digested the Vigna mungo seed globulin subunits at
different rates. The serine enzyme digested the 56 kilodalton subunit at first,
but the sulfhydryl enzyme digested the 54 kilodalton peptide more efficiently
than the 56 kilodalton peptide. The pattern of digestion of globulin by the
combination of the serine/ and sulfhydryl/endopeptidases was similar to that
using crude enzyme extracts.
PMID: 16664675 [PubMed / as supplied by publisher]
74: Plant Physiol. 1985 Dec;79(4):939/942.
Biosynthesis of alpha/Amylase in Vigna mungo Cotyledon.
Tomura H, Koshiba T.
Department of Biology, Tokyo Metropolitan University, Setagaya/ku, Tokyo 158,
Japan.
In vitro translation of RNA extracted from Vigna mungo cotyledons showed that
alpha/amylase is synthesized as a polypeptide with a molecular mass of 45,000,
while cotyledons contain a form of alpha/amylase with a molecular mass of 43,000.
To find out whether the 45,000 molecular mass polypeptide is a precursor to the
43,000 found in vivo, the cell free translation systems were supplemented with
canine microsomal membrane; when mRNA was translated in the wheat germ system
supplemented with canine microsomes, the 45,000 molecular mass form was not
processed to a smaller form but the precursor form was partly processed in the
membrane/supplemented reticulocyte lysate system. When V. mungo RNA was
translated in Xenopus oocyte system, only the smaller form (molecular mass
43,000) was detected. Involvement of contranslational glycosylation in the
maturating process of the alpha/amylase was ruled out because there was no effect
of tunicamycin, and the polypeptide was resistant to endo/beta/H or endo/beta/D
digestion. We interpret these results to mean that the 45,000 molecular mass form
is a precursor with a signal peptide or transit sequence, and that the 43,000
molecular mass is the mature form of the protein.
PMID: 16664549 [PubMed / as supplied by publisher]
75: Plant Physiol. 1985 Dec;79(4):935/938.
Imunohistochemical Localization of alpha/Amylase in Cotyledons of Vigna mungo
Seedlings.
Tomura H, Koshiba T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Setagaya/ku, Tokyo 158,
Japan.
We studied the localization of alpha/amylase with indirect fluorescence
microscopy in transversely sectioned cotyledons of Vigna mungo seedlings. Tissue
sections were fixed in periodate/lysine/paraformaldehyde and treated with
anti/alpha/amylase immunoglobulin G followed by fluorescein isothiocyanate
labeled goat anti/rabbit immunoglobulin G. alpha/Amylase appeared in the cells
farthest from vascular bundles on the second day of growth and appeared gradually
closer to the vascular bundles as growth progressed. The pattern of alpha/amylase
appearance was similar in detached cotyledons, indicating that attachment of the
embryonic axis has no effect on this pattern. However, in attached cotyledons,
alpha/amylase disappeared from the regions where starch grains had been digested,
but in detached cotyledons there was no disappearance of alpha/amylase, and
digestion was slower than in intact cotyledons.
PMID: 16664548 [PubMed / as supplied by publisher]
76: J Bacteriol. 1985 May;162(2):784/9.
Legume agglutinins that bind to Rhizobium meliloti.
Seegers R, LaRue TA.
A protein found in seeds and roots of alfalfa (Medicago sativa) was implicated in
the specificity of the infection process, based on its binding to the symbiont
Rhizobium meliloti. We found an agglutinin with similar properties in seeds and
roots of sweet clover (Melilotis alba). The sweet clover differed from alfalfa in
nodulation by a mutant strain of R. meliloti, but the agglutinins were
indistinguishable by sodium dodecyl sulfate/polyacrylamide gel electrophoresis,
Rhizobium agglutination, and cross/reactivity to antibodies. Similar agglutinins
binding R. meliloti were found in seeds of legumes from different
cross/inoculation groups, including soybean (Glycine max), cowpea (Vigna
unguiculata), pea (Pisum sativum L), and mung bean (Vigna mungo). The agglutinins
from these legumes were recognized by antibodies raised against the agglutinins
of alfalfa and sweet clover. Seeds of corn (Zea mays) and tomato (Lycopersicon
esculentum) contained a protein similar to the legume agglutinin, but it did not
react with the antibodies. We conclude that the alfalfa agglutinin is
representative of a common legume protein and that there is no evidence for its
role in specificity or nodule initiation.
PMID: 3988714 [PubMed / indexed for MEDLINE]
77: Appl Environ Microbiol. 1984 Jul;48(1):232/233.
Occurrence of a Lysogenic Streptomyces sp. on the Nodule Surface of Black Gram
(Vigna mungo (L.) Hepper).
Rangarajan M, Ravindran AD, Hariharan K.
Centre of Advanced Studies in Agricultural Microbiology, Tamil Nadu Agricultural
University, Coimbatore 641 003, Tamil Nadu, India.
A lysogenic Streptomyces sp., strain NS.A4, which was isolated from the nodule
surface of black gram (Vigna mungo (L.) Hepper), was found to inhibit rhizobia of
fast/and slow/growing strains of cowpeas and soybeans. It exhibited plaques when
there was a change in cultural conditions. Repeated culturing of the organism in
nutrient agar and broth confirmed the infection of Streptomyces sp. strain NS.A4
by an actinophage. Addition of the culture filtrate of Streptomyces sp. strain
NS.A4 to shaken broth cultures of three other Streptomyces spp. resulted in phage
infection.
PMID: 16346593 [PubMed / as supplied by publisher]
78: Res Exp Med (Berl). 1984;184(2):85/90.
Influence of black gram (Vigna mungo) trypsin inhibitory fraction on the hepatic
protein catabolism in male albino mice.
Kamalakannan V, Sathyamoorthy AV, Motlag DB.
The effect of black gram and black gram trypsin inhibitor on the protein
catabolism of male albino mice has been investigated. Group 1 was given
autoclaved black gram (control), Group II raw black gram and Group III the
autoclaved black gram incorporated with 1% black gram trypsin inhibitor. Blood as
well as urinary urea and creatine were found to be elevated in Groups II and III.
Increased levels of arginase, ornithine transcarbamylase and transaminases were
noted in Groups II and III. The results suggested an enhanced catabolism of
proteins evoked by the native black gram trypsin inhibitor.
Publication Types:
Research Support, Non/U.S. Gov't
PMID: 6473902 [PubMed / indexed for MEDLINE]
79: Plant Physiol. 1983 Dec;73(4):869/873.
A Pathway for Photosynthetic Carbon Flow to Mannitol in Celery Leaves : Activity
and Localization of Key Enzymes.
Rumpho ME, Edwards GE, Loescher WH.
Department of Botany, Washington State University, Pullman, Washington
99164/4230.
In the polyol producing plant, celery (Apium graveolens L.), mannitol is a major
photosynthetic product and a form in which carbohydrate is translocated.
Measurements of whole leaf extracts of celery indicated substantial activity of
the following enzymes: mannose/6/P reductase, mannose/6/P isomerase, mannitol/1/P
phosphatase, and nonreversible glyceraldehyde/3/P dehydrogenase. The activities
of these enzymes were either undetectable or very low in the nonpolyol producing
plants, Secale cereale L. (rye) and Vigna mungo (L.) Hepper (black
gram).Mesophyll protoplasts were enzymically isolated from celery leaves, broken
with a Yeda press and the intracellular localization of the above enzymes for
mannitol synthesis studied following differential and/or sucrose density gradient
centrifugation of the protoplast extract. These data suggested the enzymes
involved in mannitol synthesis are exclusively localized in the cytoplasm.
Ninety/five to 100% of the activity of these enzymes, along with the cytoplasmic
marker enzyme phosphoenolpyruvate carboxylase, was found in the cytosolic
fraction.We propose the pathway of photosynthetic carbon flow from triose/P to
mannitol in celery occurs via fructose/6/P, mannose/6/P, and mannitol/1/P; these
final reactions being catalyzed by the cytoplasmic enzymes, mannose/6/P
isomerase, NADPH/dependent mannose/6/P reductase, and mannitol/1/P phosphatase,
respectively. The requirement for NADPH may be met via the cytoplasmically
located NADP/linked nonreversible glyceraldehyde/3/P dehydrogenase.
PMID: 16663332 [PubMed / as supplied by publisher]
80: Plant Physiol. 1983 Sep;73(1):82/86.
Appearance and Disappearance of Cyanide/Resistant Respiration in Vigna mungo
Cotyledons during and following Germination of the Axis.
Morohashi Y, Matsushima H.
Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu,
Tokyo 183, Japan.
Mitochondrial preparations isolated from black gram (Vigna mungo L.) cotyledons
exhibited cyanide/resistant respiration which was of mitochondrial origin. The
appearance and the disappearance of this alternative respiration took place
during and following imbibition. During the first 6 hours of imbibition, the
respiration was completely inhibited by cyanide, but after this time the
alternative respiration markedly developed, reaching a maximal cyanide/resistance
12 to 16 hours after the start of imbibition. Subsequently, the alternative
respiration gradually disappeared. The actions of cycloheximide and
chloramphenicol indicated that the appearance was dependent on cytoplasmic
protein synthesis and that the disappearance depended on both cytoplasmic and
mitochondrial protein synthesis. The alternative pathway contributed to state 4
respiration, but not to state 3 respiration, in mitochondria from 1/day/old
cotyledons. On day 3, it contributed to neither state 3 nor state 4.
PMID: 16663192 [PubMed / as supplied by publisher]
81: Plant Physiol. 1983 Jan;71(1):173/176.
In Vivo Synthesis and Turnover of alpha/Amylase in Attached and Detached
Cotyledons of Vigna mungo Seeds.
Koshiba T, Minamikawa T.
Department of Biology, Tokyo Metropolitan University, Setagaya/ku, Tokyo 158,
Japan.
alpha/Amylase activity increased in attached cotyledons of germinated Vigna mungo
seeds until the 5th day after imbibition and decreased thereafter, whereas in
detached and incubated cotyledons the activity continuously increased and, at the
6th day, reached the value more than three times that of the maximum activity of
attached cotyledons. Zymograms of the activities and Ouchterlony double
immunodiffusion test on the activities of attached and detached cotyledons showed
that the increase of activity in detached cotyledons was due to the identical
enzyme as in attached tissues. alpha/Amylase contents, determined by single
radial immunodiffusion method, changed in parallel with enzyme activity in both
attached and detached cotyledons, which also suggested the de novo synthesis of
alpha/amylase in V. mungo cotyledons.The rate of incorporation of the label from
[(3)H]leucine into alpha/amylase and the ratios of dpm in alpha/amylase/dpm in
trichloroacetic acid/insoluble fraction did not show significant difference
between attached and detached cotyledons. The results indicated that in attached
cotyledons fluctuation of alpha/amylase activity was regulated by both synthesis
and degradation of the enzyme, whereas in detached cotyledons alpha/amylase was
synthesized and accumulated, because of low degrading activity during incubation.
PMID: 16662780 [PubMed / as supplied by publisher]
82: J Sci Food Agric. 1981 Dec;32(12):1172/6.
Studies on black gram (Vigna mungo) trypsin inhibitor.
Kamalakannan V, Sathyamoorthy AV, Motlag DB.
Publication Types:
In Vitro
Research Support, Non/U.S. Gov't
PMID: 7321531 [PubMed / indexed for MEDLINE]
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