Classification:

Vicia tetrasperma (L.) Schreb.

Kingdom Plantae – Plants

Subkingdom Tracheobionta – Vascular plants
Superdivision Spermatophyta – Seed plants
Division Magnoliophyta – Flowering plants
Class Magnoliopsida – Dicotyledons
Subclass Rosidae
Order Fabales
Family Fabaceae – Pea family
Genus Vicia L. – vetch
Specie Vicia tetrasperma (L.) Schreb. – lentil vetch

Economically important symbiosis of nodule bacteria with legume plants and the
associated fixation of atmospheric nitrogen are enabled, from the bacterial side, by the
so-called nodulation (nod) genes, regulated from the beginning of symbiosis by the host
stimuli. A technique was optimized for the detection of nod gene activity, as based on the
visualization of Beta-galactosidase activity transcriptionally fused to nodABC operon
using a chromogenic substrate. The technique was adapted both for the symbiosis of pea
as a standard host of Rhizobium leguminosarum bv. viciae and for a suggested laboratory
model Vicia tetrasperma. The use of the technique in early pea symbiotic mutants
carrying mutations in gene sym8 excluded disturbances in the bacterial nod gene
activation as a reason for the symbiotic fault Economically important symbiosis of nodule
bacteria with legume plants and the associated fixation of atmospheric nitrogen are
enabled, from the bacterial side, by the so-called nodulation (nod) genes, regulated from
the beginning of symbiosis by the host stimuli. A technique was optimized for
thedetection of nod gene activity, as based on the visualization of Beta-galactosidase
activity transcriptionally fused to nodABC operon using a chromogenic substrate. The
technique was adapted both for the symbiosis of pea as a standard host of Rhizobium
leguminosarum bv. viciae and for a suggested laboratory model Vicia tetrasperma. The
use of the technique in early pea symbiotic mutants carrying mutations in gene sym8
excluded disturbances in the bacterial nod gene activation as a reason for the symbiotic
fault.

Visualization of nodulation gene activity on the early stages of Rhizobium

leguminosarum bv. viciae symbiosis

Datum: 31.5.2006

Chovanec P, Novak K.

Institute of Microbiology, Academy of Sciences of the Czech Republic, 142 20 Prague, Czechia.
A technique was optimized for the in situ detection of nodulation (nod) gene activity in Rhizobium
leguminosarum bv. viciae symbiosis with compatible plant hosts Vicia tetrasperma (L.) SCHREB.
and Pisum sativum L. The transcription of nodABC-lacZ fusion was visualized as beta-galactosidase
(beta-Gal) activity after reaction with the chromogenic substrate X-Gal and subsequent light
microscopy, while the background of the indigenous beta-Gal activity of rhizobia and the host plant
was eliminated by glutaraldehyde treatment.

V. tetrasperma was suggested as a suitable model plant for pea cross-inoculation group due to its
advantages over the common model of V. hirsuta (L.) S.F. GRAY: compactness of the plant,
extremely small seeds, fast development and stable nodulation under laboratory conditions. In the
roots of both plants, a certain extent of nod gene activity was detectable in all rhizobia colonizing
the rhizoplane. In pea 1 d after inoculation (d.a.i.), the maximum was localized in the region of
emerging root hairs (RH) later (3 and 6 d.a.i.) shifting upwards from the root tip.

Nodulation genes sustained full expression even in the infection threads inside the RH and the root
cortex, independently of their association with nodule primordia. Comparison of two pea symbiotic
mutant lines, Risnod25 and Risnod27, with the wild type did not reveal any differences in the RH
formation, RH curling response and rhizoplane colonization. Both mutants appeared to be blocked at
the infection thread initiation stage and in nodule initiation, consistent with the phenotype caused
by other mutant alleles in the pea sym8 locus. Judging from the nod gene expression level and
pattern in the rhizoplane, flavonoid response upon inoculation is preserved in both pea mutants,
being independent of infection thread and nodule initiation.

Source

Visualization of nodulation gene activity on the early stages of Rhizobium

leguminosarum bv. viciae symbiosis.

A technique was optimized for the in situ detection of nodulation (nod) gene activity
in Rhizobium leguminosarum bv. viciae symbiosis with compatible plant hosts Vicia
tetrasperma (L.) SCHREB. and Pisum sativum L. The transcription of nodABC-lacZ
fusion was visualized as beta-galactosidase (beta-Gal) activity after reaction with the
chromogenic substrate X-Gal and subsequent light microscopy, while the background
of the indigenous beta-Gal activity of rhizobia and the host plant was eliminated by
glutaraldehyde treatment. V. tetrasperma was suggested as a suitable model plant
for pea cross-inoculation group due to its advantages over the common model of V.
hirsuta (L.) S.F. GRAY: compactness of the plant, extremely small seeds, fast
development and stable nodulation under laboratory conditions. In the roots of both
plants, a certain extent of nod gene activity was detectable in all rhizobia colonizing
the rhizoplane. In pea 1 d after inoculation (d.a.i.), the maximum was localized in the
region of emerging root hairs (RH) later (3 and 6 d.a.i.) shifting upwards from the
root tip. Nodulation genes sustained full expression even in the infection threads
inside the RH and the root cortex, independently of their association with nodule
primordia. Comparison of two pea symbiotic mutant lines, Risnod25 and Risnod27,
with the wild type did not reveal any differences in the RH formation, RH curling
response and rhizoplane colonization. Both mutants appeared to be blocked at the
infection thread initiation stage and in nodule initiation, consistent with the
phenotype caused by other mutant alleles in the pea sym8 locus. Judging from the
 nod gene expression level and pattern in the rhizoplane, flavonoid response upon
inoculation is preserved in both pea mutants, being independent of infection thread
and nodule initiation

Visualization of symbiotic tissue in intact root nodules of Vicia tetrasperma using GFP-
marked Rhizobium leguminosarum bv. viciae.

In rhizobial symbiosis with legume plant hosts, the symbiotic tissue in the root
nodules of indeterminate type is localized to the basal part of the nodule where the
symbiotic zones contain infected cells (IC) interspersed with uninfected cells (UC)
that are devoid of rhizobia. Although IC are easily distinguished in nodule sections
using standard histochemical techniques, their observation in intact nodules is
hampered by nodule tissue characteristics. Tagging of Rhizobium leguminosarum bv.
viciae strain 128C30 with a constitutively expressed gene for green fluorescent
protein (nonshifted mutant form cycle3) in combination with the advantages of the
tiny nodules formed by Vicia tetrasperma (L.) SCHREB . allowed for vital observation
of symbiotic tissue using fluorescence microscopy. Separation of a red-shifted
background channel and digital image stacking along z-axis enabled us to construct a
nodule image in a classical fluorescence microscopy of nodules exceeding 1 mm in
diameter. In parallel, visualization of nodule bacteria inside the symbiotic tissue by
confocal microscopy at the excitation wavelength 488 nm clearly distinguished IC/UC
pattern in the nodule virtual sections and revealed red-shifted fluorescence of
nonrhizobial origin. This signal was located on the periphery of IC and increased with
their degradation, thus suggesting accumulation of secondary metabolites,
presumably flavonoids. The simultaneous detection of bacteria and secondary
metabolites can be used for monitoring changes to intact nodule physiology in the
model legumes. The advantage of V. tetrasperma as a suggested laboratory model
for pea cross-inoculation group has been demonstrated.

Isozyme variation and phylogenetic relationships in Vicia subgenus Cracca (Fabaceae).

BACKGROUND AND AIMS: The phylogenetic relationships among 27 vetch species

belonging to the subgenus Cracca of the genus Vicia were studied in comparison with
three species of Lathyrus section Lathyrus on the basis of isozyme variation.
METHODS: Isozymes encoded by 15 putative loci of ten enzymes were resolved by
polyacrylamide gel electrophoresis and isozyme variation was analysed by using
parsimony and neighbour-joining methods. KEY RESULTS: The analyses revealed 63
parsimony-informative and 36 species-specific orthozymes. Of the latter, 23 are
monomophic and are suitable for identification of V. benghalensis, V. palaestina, V.
dumetorum, V. pisiformis, V. sylvatica, V. onobrychioides, V. cappadocica, V. cretica,
V. articulata, V. tetrasperma, V. ervilia, V. hirsuta and V. loiseleurii. Polymorphism
with heterozygous and homozygous isozyme genotypes was found for V. cracca, V.
tenuifolia, V. ochroleuca, V. villosa, V. sylvatica, V. cassubica, V. sparsiflora, V.
megalotropis, V. altissima, V. onobrychioides, V. cassia, V. cretica and L.
 heterophyllus, reflecting outcrossing in these species. By contrast, V. benghalensis,
V. palaestina, V. disperma, V. dumetorum, V. pisiformis, V. orobus, V. pauciflora, V.
tetrasperma and V. loiseleurii had only homozygous isozyme genotypes at
polymorphic loci. Isozyme-based phylogenetic trees are presented. CONCLUSIONS:
Sections Cracca, Ervum, Pedunculatae and Lenticula of traditional taxonomy are
monophyletic groups, whereas sections Oroboideae (= Vicilla) and Panduratae
appear polyphyletic and section Cassubicae is split into two species-couples linked at
a low level of support. Treatment of ervoid species in a separate subgenus Ervum is
not supported because of its polyphyly.

Larvicidal activity of leguminous seeds and grains against Aedes aegypti and Culex
pipiens pallens.

Larvicidal activity of methanol extracts of 26 leguminous seeds and 20 grains against

early 4th-stage larvae of Aedes aegypti and Culex pipiens pallens was examined. At
200 ppm of the extracts from Cassia obtusifolia, Cassia tora, and Vicia tetrasperma,
more than 90% mortality was obtained in larvae of Ae. aegypti and Cx. pipiens
pallens. Extract of C. tora gave 86.7 and 100% mortality in the larvae of Ae. aegypti
and Cx. pipiens pallens at 40 ppm but 59.2 and 78.3% mortality against larvae of
Ae. aegypti and Cx. pipiens pallens at 20 ppm, respectively. At 40 ppm, extract of C.
obtusifolia caused 51.4 and 68.5% mortality of the 4th-stage larvae of Ae. aegypti
and Cx. pipiens pallens, respectively. Larvicidal activity of extract of C obtusifolia was
significantly reduced when used at 20 ppm. Further studies of these plants as
possible agents for mosquito control are warranted.

Response of pollen germination and tube growth to cadmium with special reference to
low concentration exposure.

Cadmium is one of the most important heavy metal pollutants highly hazardous to
plants. Pollen is considered to be more sensitive to pollutants than are vegetative
parts of the plants. Five herb species were tested for responses in pollen germination
and tube growth to Cd exposure in vitro. Pollen germination of all the species was
inhibited at Cd concentrations of 2.51 microg/mL and higher, and tube growth was
inhibited at concentrations of 1.58 microg/ml and higher. Cadmium, at low
concentrations, stimulated pollen tube growth. The pollen response to Cd stress
exhibited interspecies differences. Vicia angustifolia and V. tetrasperma were
sensitive to Cd, and were inhibited in either pollen germination or tube growth by Cd
at 0.01 microg/mL. Plantago depressa was less sensitive; pollen germination and
tube growth were not inhibited until the Cd concentration reached 2.51 and 1.58
microg/mL, respectively, and its tube growth displayed two stimulatory peaks; the
one that appeared at 1.00 microg/mL showed the highest tube length in all species
tested. These results suggest that Cd, even at low concentrations, may adversely
affect plant reproduction by inhibiting pollen germination and tube growth.
 Vicia tetrasperma (four-seeded vetch) ingestion by a 3-year-old child.

Rhode Island Poison Center, Rhode Island Hospital, Providence 02903.

We report an ingestion of Vicia tetrasperma (four-seeded vetch), initially

misidentified as crown vetch (Coronilla varia). Vicia tetrasperma is not listed in
POISINDEX; little is known of its toxic effects and we found no published human case
reports. The child suffered only minor gastrointestinal effects which lasted a few
hours and had no residual effects upon 24- and 96-hour followups.

V.faba genes:\
1- gene encoding legumin
1: Z26488 Reports Links
V.faba gene encoding legumin (partial)
gi|403337|emb|Z26488.1|[403337]
Reports Links
2: Y00506
Vicia faba vicilin gene
gi|829146|emb|Y00506.1|[829146]
Reports Links
3: Z35164
V.faba (VfVCINV) mRNA for vacuolar invertase
gi|511158|emb|Z35164.1|[511158]
Reports Links
4: Z26489
V.faba gene encoding legumin
gi|403335|emb|Z26489.1|[403335]
Reports Links
5: X56240
V.faba USP gene for an unknown seed protein
gi|22042|emb|X56240.1|[22042]
Reports Links
6: X14241
Vicia faba VfLEB7 gene
gi|22024|emb|X14241.1|[22024]
Reports Links
7: X14240
Vicia faba VfLEB6 gene
gi|22020|emb|X14240.1|[22020]
Reports Links
8: X14237
Vicia faba VfLEB2 gene for legumin storage protein
gi|22013|emb|X14237.1|[22013]
Reports Links
9: X14238
Vicia faba VfLEB1 pseudogene
gi|22010|emb|X14238.1|[22010]
10: X55014 Reports Links

Vicia faba mRNA for Legumin A2 pre-pro-polypeptide

gi|22007|emb|X55014.1|[22007]
 11: X55013 Reports Links

Vicia faba mRNA for Legumin A1 pre-pro-polypeptide

gi|22006|emb|X55013.1|[22006]
Reports Links
12: AJ292221
Vicia faba sbpl gene for putative sucrose binding protein, genomic RNA
gi|12580893|emb|AJ292221.1|[12580893]
13: AJ318812 Reports Links

Vicia faba var. minor aap1 gene, promoter region

gi|15216031|emb|AJ318812.1|[15216031]
14: AJ318811 Reports Links

Vicia faba var. minor mRNA for amino acid permease AAP4 (aap4 gene)
gi|15216029|emb|AJ318811.1|[15216029]
15: AJ318810 Reports Links

Vicia faba var. minor mRNA for amino acid permease AAP3 (aap3 gene)
gi|15216027|emb|AJ318810.1|[15216027]
16: AJ318809 Reports Links

Vicia faba var. minor mRNA for amino acid permease AAP1 (aap1 gene)
gi|15216025|emb|AJ318809.1|[15216025]
17: AJ400727 Reports Links

Vicia faba partial mRNA for putative kinetochore protein (skp1-2 gene)
gi|7573586|emb|AJ400727.1|[7573586]
18: AJ400726 Reports Links

Vicia faba partial mRNA for putative kinetochore protein (skp1-1 gene)
gi|7573583|emb|AJ400726.1|[7573583]
19: X97905 Reports Links

V.faba mRNA for an RNA- or ssDNA-binding protein

gi|2104686|emb|X97905.1|[2104686]
20: X97909 Reports Links

V.faba mRNA for putative transciption factor (1399bp)

gi|2104684|emb|X97909.1|[2104684]

21: X97908 Reports Links

V.faba mRNA for putative transciption factor (2861bp)
gi|2104682|emb|X97908.1|[2104682]
22: X97907 Reports Links

V.faba mRNA for putative transciption factor (1556bp)

gi|2104680|emb|X97907.1|[2104680]
23: X97906 Reports Links

V.faba mRNA for transcription factor containing HMG-box

gi|2104678|emb|X97906.1|[2104678]
24: X97904 Reports Links

V.faba mRNA for transcription factor containing bZIP and zinc finger
gi|2104676|emb|X97904.1|[2104676]
25: X97903 Reports Links

V.faba mRNA for transcription factor containing bZIP

gi|2104674|emb|X97903.1|[2104674]
26: Z56278 Reports Links

V.faba mRNA for sucrose phosphate synthase

gi|1022364|emb|Z56278.1|[1022364]
27: Z49831 Reports Links

V.faba VFVCINV mRNA for invertase (beta-fructofuranosidase)

gi|861158|emb|Z49831.1|[861158]
28: Z35163 Reports Links

V.faba VFCWINV2 mRNA for cell wall invertase II

gi|861156|emb|Z35163.1|[861156]
29: Z35162 Reports Links

V.faba VFCWINV1 mRNA for cell wall invertase I

gi|861154|emb|Z35162.1|[861154]
30: Z48640 Reports Links

Vicia faba var. minor partial mRNA for sucrose-phosphate synthase

gi|732985|emb|Z48640.1|[732985]
31: Z36880 Reports Links

Vicia faba var. minor mRNA for alpha 1,4-glucan phosphorylase L isoform
precursor (VfPho1 gene)
gi|534971|emb|Z36880.1|[534971]
32: Z35117 Reports Links

Vicia faba var. minor mRNA for alpha 1,4-glucan phosphorylase type H, (VfPho2
gene)
gi|510931|emb|Z35117.1|[510931]
33: X76941 Reports Links

V.faba mRNA (VfAGPP) for ADP-glucose pyrophosphorylase

gi|440594|emb|X76941.1|[440594]
34: X76940 Reports Links

V.faba mRNA (VfAGPC) for ADP-glucose pyrophosphorylase

gi|440592|emb|X76940.1|[440592]
35: Z26518 Reports Links

V.faba gene encoding legumin (partial)

gi|403380|emb|Z26518.1|[403380]
36: Z26487 Reports Links

V.faba gene encoding legumin (partial)

 gi|403333|emb|Z26487.1|[403333]
37: Y00462 Reports Links

Vicia faba mRNA for vicilin storage protein

gi|22052|emb|Y00462.1|[22052]
38: X14239 Reports Links

Vicia faba VfLEB5 pseudogene

gi|22019|emb|X14239.1|[22019]
39: Z21703 Reports Links

Vicia faba mRNA for HMG box

gi|393703|emb|Z21703.1|[393703]