Rhizobium meliloti purine auxotrophs arenod + but defective in nitrogen fixation

Several purine auxotrophs were isolated inRhizobium meliloti and characterized for their nutritional requirements. They were found to produce small, irregular nodules lacking any detectable nitrogenase activity onMedicago sativa. The symbiotic aberration manifests itself only in the late developmental stage, for, (i) these purine auxotrophs infect theMedicago sativa root hairs by forming normal infection threads, and (ii) the mutants are recovered from the root nodules induced by them. External supplementation of the plant growth substrate with purines or their biosynthetic intermediates fails to restore symbiosis. This, and the failure of complementation of these auxotrophs with the known symbiotic genes, demonstrates that these mutants perhaps define a new set of genes influencing the symbiotic process inRhizobium meliloti.     

Fine Mapping of Aroma QTLs in Basmati Rice (Oryza sativa L) on Chromosomes 3, 4 and 8

Aroma is one of the most important quality traits of basmati rice (Oryza sativa L) that leads to high consumer acceptance. Earlier three significant QTLs for aroma, namely aro3-1, aro4-1 and aro8-1, have been mapped on rice chromosomes 3, 4 and 8, respectively, using a population of recombinant inbred lines (RILs) derived from a cross between Pusa 1121 (a basmati quality variety) and Pusa 1342 (a non-aromatic variety). For fine mapping of these QTLs, 184 F₆ RILs were grown in the Kharif season of 2005 at New Delhi and Karnal, India. A total of 115 new SSR markers covering the three QTL intervals were designed and screened for parental polymorphism. Of these, 26 markers were polymorphic between parents, eight for the interval aro3-1, eight for the interval aro4-1 and ten for the interval aro8-1, thus enriching the density of SSR markers in these QTL intervals. Revised genetic maps were constructed by adding 23 of these new markers to the earlier map, by giving physical order of the markers in the pseudomolecules a preference. In the revised maps, the interval for QTL aro4-1 could not be improved further but QTL aro3-1 was narrowed down to an interval of 390 kbp from the earlier reported interval of 8.6 Mbp and similarly the QTL aro8-1 was narrowed down to a physical interval of 430 kbp. The numbers of candidate genes in the aro3-1 and aro8-1 intervals have now been reduced to 51 and 66, respectively. The badh2 gene on chromosome 8 was not associated with the aroma QTL on this chromosome.     

Fine mapping of grain length QTLs on chromosomes 1 and 7 in Basmati rice (Oryza sativa L.)

Grain dimensions (length, breadth and length/breadth ratio) are important quality attributes of Basmati rice for its high consumer acceptance. Earlier we identified two significant quantitative trait loci (QTL) intervals on chromosomes 1 and 7 for grain dimensions in Basmati rice using a population of recombinant inbred lines (RILs) from cross between Basmati variety Pusa 1121 and a short grain non-aromatic variety Pusa 1342. For fine mapping of these QTLs, 184 F₆ RILs were grown and phenotyped in the normal rice growing season at two different locations. Forty-nine new SSR markers targeting these QTL intervals were tested and nine were found polymorphic between the parents. Using revised genetic maps adding new markers, the grain length QTL qGRL1.1 on chromosome 1 was narrowed down to 108?kbp from the earlier reported 6,133?kbp. There were total 13 predicted gene models in this interval which includes the probable candidate gene for the exceptionally high grain length of Basmati variety Pusa 1121. Similarly, two tandem QTL intervals qGRL7.1 and qGRL7.2 on chromosome 7 were merged into a single one narrowed down to 2,390?kbp from the earlier reported length of 5,269?kbp. This region of chromosome 7 also has co-localized QTLs for grain breadth and length to breadth ratio. SSR markers tightly linked to the QTL at a map distance of ??0.2?cM were developed for the qGRL1.1 and qGRL7.1 loci that could be used for marker-assisted breeding. Validation of earlier published markers for the grain length gene GS3 on chromosome 3 showed no difference between Pusa 1121 and Pusa 1342, highlighting the significance of qGRL1.1 and qGRL7.1 for the extra grain length of Basmati variety Pusa 1121.     

Endophytic bacterial flora in the stem tissue of a tropical maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) genotype: isolation,identification and enumeration

The genotypic diversity of indigenous bacterial endophytes within stem of tropical maize (Zea mays L.) was determined in field and greenhouse experiments. Strains were isolated from stem tissues of a tropical maize cultivar (PEHM-1) by trituration and surface disinfestation and their population dynamics was determined. Endophytes were found in most of the growing season at populations ranging from 1.36–6.12 × 10⁵ colony-forming units per gram fresh weight (c.f.u./gm fw) of stem. Analysis of these bacterial endophytes using Gas Chromatography—Fatty Acid Methyl Ester (GC-FAME) led to the identification of Bacillus pumilus, B. subtilis, Pseudomonas aeruginosa and P. fluorescens as the relatively more predominant group of bacterial species residing in maize stem. When the maize seedlings grown in a greenhouse were inoculated with these four isolates individually, their population densities decreased (1.6–3.1 × 10⁵ c.f.u./gm fw of stem) as compared to the field-grown maize (1.8–3.8 × 10⁵ c.f.u./gm fw of stem). The highest persistence, however, was recovered in the case of B. subtilis with a population density of 3.1 × 10⁵ c.f.u./gm fw of stem tissue on 28 days after emergence (DAE). This is the first report on population dynamics of bacterial endophytes from tropical maize and the results establish that symptomless populations of bacteria exist in the maize stem.     

Pleiotropic effects of purine auxotrophy inRhizobium meliloti on cell surface molecules

Rhizobial purine auxotrophs have earlier been shown to be defective in symbiosis, though the exact reason for this failure is not clear. Using various dyes that specifically bind different cell surface molecules, we show that there are multiple changes in the cell surface molecules associated with different purine auxotrophs. Affected molecules in different purine auxotrophs that were tested include (i) acidic exopolysaccharides, (ii) cellulose fibrils, and (iii) beta (1–3) glucans. Our results show that the symbiotic deficiency of purine auxotrophs is likely to be a result of these associated changes on the cell surface     

Phenotypic and molecular characterization of indigenous rhizobia nodulating chickpea in India

In a combined approach of phenotypic and genotypic characterization, 28 indigenous rhizobial isolates obtained from different chickpea growing regions in peninsular and northern India were analyzed for diversity. The field isolates were compared to two reference strains TAL620 and UPM-Ca142 representing M. ciceri and M. mediterraneum respectively. Phenotypic markers such as resistance to antibiotics, tolerance to salinity, temperature, pH, phosphate solubilization ability, growth rate and also symbiotic efficiency showed considerable diversity among rhizobial isolates. Their phenotypic patterns showed adaptations of rhizobial isolates to abiotic stresses such as heat and salinity. Two salt tolerant strains (1.5% NaCl by T1 and T4) with relatively high symbiotic efficiency and two P-solubilising strains (66.7 and 71 microg/ml by T2 and T5) were identified as potential bioinoculants. Molecular profiling by 16S ribosomal DNA Restriction Fragment Length Polymorphism (RFLP) revealed three clusters at 67% similarity level. Further, the isolates were differentiated at intraspecific level by 16S rRNA gene phylogeny. Results assigned all the chickpea rhizobial field isolates to belong to three different species of Mesorhizobium genus. 46% of the isolates grouped with Mesorhizobium loti and the rest were identified as M. ciceri and M. mediterraneum, the two species which have been formerly described as specific chickpea symbionts. This is the first report on characterization of chickpea nodulating rhizobia covering soils of both northern and peninsular India. The collection of isolates, diverse in terms of species and symbiotic effectiveness holds a vast pool of genetic material which can be effectively used to yield superior inoculant strains.     

Characterization of intrinsic variability of Mesorhizobium ciceri isolates of cultivated fields

A large number of putative rhizobial isolates were obtained from the root nodules of various chickpea cultivars growing in agricultural research fields. Of these, thirty were selected and characterized for traits, such as, generation time, intrinsic azide resistance and several symbiotic characters.     

Use of nitrogen-fixing bacteria as biofertiliser for non-legumes: prospects and challenges

The potential of nitrogen-fixing (NF) bacteria to form a symbiotic relationship with leguminous plants and fix atmospheric nitrogen has been exploited in the field to meet the nitrogen requirement of the latter. This phenomenon provides an alternative to the use of the nitrogenous fertiliser whose excessive and imbalanced use over the decades has contributed to green house emission (N(2)O) and underground water leaching. Recently, it was observed that non-leguminous plants like rice, sugarcane, wheat and maize form an extended niche for various species of NF bacteria. These bacteria thrive within the plant, successfully colonizing roots, stems and leaves. During the association, the invading bacteria benefit the acquired host with a marked increase in plant growth, vigor and yield. With increasing population, the demand of non-leguminous plant products is growing. In this regard, the richness of NF flora within non-leguminous plants and extent of their interaction with the host definitely shows a ray of hope in developing an ecofriendly alternative to the nitrogenous fertilisers. In this review, we have discussed the association of NF bacteria with various non-leguminous plants emphasizing on their potential to promote host plant growth and yield. In addition, plant growth-promoting traits observed in these NF bacteria and their mode of interaction with the host plant have been described briefly.     

Mutation in the lysA gene impairs the symbiotic properties of Mesorhizobium ciceri

A Tn5-induced mutant of Mesorhizobium ciceri, TL28, requiring the amino acid lysine for growth on minimal medium was isolated and characterized. The Tn5 insertion in the mutant strain TL28 was located on a 6.8-kb EcoRI fragment of the chromosomal DNA. Complementation analysis with cloned DNA indicated that 1.269 kb of DNA of the 6.8-kb EcoRI fragment restored the wild-type phenotype of the lysine-requiring mutant. This region was further characterized by DNA sequence analysis and was shown to contain a coding sequence homologous to lysA gene of different bacteria. The lys ^? mutant TL28 was unable to elicit development of effective nodules on the roots of Cicer arietinum L. There was no detectable level of lysine in the root exudates of chickpea. However, addition of lysine to the plant growth medium restored the ability of the mutant to produce effective nodules with nitrogen fixation ability on the roots of C. arietinum.     

Natural occurrence of Bacillus thuringiensis in leguminous phylloplanes in the New Delhi region of India

Bacillus thuringiensis (Bt) isolates were present on the phylloplanes of chickpea (Cicer arietinum), pigeon pea (Cajanus cajan), pea (Pisum sativum) and mung bean (Vigna radiata). Bt index (ratio of the number of Bt colonies to the total number of spore-forming colonies per g of leaves) differed significantly among these plants, with the highest (0.20) in the chickpea phylloplane, followed by pigeon pea (0.17). Bt population of the chickpea phylloplane varied with plant age, being maximal in 45-day-old plants. Diversity was observed among Bt isolates for growth (up to 10-fold difference), antibiotic resistance, PCR product profile and toxicity to Helicoverpa armigera. Two isolates with high activity towards H. armigera were found. This revised version was published online in July 2006 with corrections to the Cover Date.