Classification: Viciatetrasperma (L.) Schreb. Kingdom Plantae– Plants Subkingdom Tracheobionta – Vascular plants Superdivision Spermatophyta – Seed plants Division Magnoliophyta – Flowering plants Class Magnoliopsida– Dicotyledons Subclass RosidaeOrder FabalesFamily Fabaceae – Pea family Genus Vicia L. – vetch Specie Viciatetrasperma (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 forthedetection 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 Rh izobium 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.
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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 theso-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 peaas 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 geneactivation 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 Rhizobiumleguminosarum 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 symbioticfault.
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 Rhizobiumleguminosarum 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 plantwas eliminated by glutaraldehyde treatment.
V. tetrasperma was suggested as a suitable model plant for pea cross-inoculation group due to itsadvantages 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 theroots of both plants, a certain extent of nod gene activity was detectable in all rhizobia colonizingthe 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 rootcortex, 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 RHformation, RH curling response and rhizoplane colonization. Both mutants appeared to be blocked atthe infection thread initiation stage and in nodule initiation, consistent with the phenotype causedby 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 Rhizobiumleguminosarum 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-lacZfusion 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