In our endeavor to improve upon nitrogen fixation efficiency of a soil diazotroph that would be unaffected by synthetic nitrogenous fertilizers, we have deleted a part of the negative regulatory gene and constitutively expressed the positive regulatory gene in the chromosome of CBD15, a strain isolated from the local field soil. No antibiotic resistance gene or other foreign gene was there in the chromosome of the engineered strain. Wheat seeds inoculated with this engineered strain, which we have named HKD15, were tested for three years in pots and one year in the field. Yield of wheat was enhanced by ∼60% due to inoculation of seeds by HKD15 in the absence of any urea application. Ammonium only marginally affected acetylene reduction by the engineered strain. When urea was also applied, the same wheat yield could be sustained by using seeds inoculated with HKD15 and using ∼85kg less urea (∼40kg less nitrogen) than the usual ∼257 kg urea (∼120 kg nitrogen) per hectare. Wheat plants arising from the seeds inoculated with the engineered strain, exhibited far superior overall performance, had much higher dry weight and nitrogen content and assimilated molecular N much better. A nitrogen balance experiment also revealed much higher total nitrogen content. IAA production by the wild type and the engineered strain was about the same. Inoculation of the wheat seeds with HKD15 did not adversely affect the microbial population in the field rhizosphere soil.IMPORTANCE Application of synthetic nitrogenous fertilizers is a standard agricultural practice to augment crop yield. Plants, however, utilize only a fraction of the applied fertilizers, while the unutilized fertilizers cause grave environmental problems. Wild type soil diazotrophic microrganisms cannot replace synthetic nitrogenous fertilizers, as these reduce atmospheric nitrogen very inefficiently and almost none at all in presence of added nitrogenous fertilizers. If the nitrogen fixing ability of soil diazotrophs could be improved upon and sustained even in the presence of synthetic nitrogenous fertilizers, then a mixture of the bacterial and reduced quantity of chemical nitrogenous fertilizers could be employed to obtain the same grain yield, but at much reduced environmental cost. The engineered that we have reported here has considerably enhanced nitrogen fixation and excretion abilities and can replace ∼85 kg of urea per hectare, but sustain same wheat yield, if the seeds are inoculated with it before sowing.

Wheat is one of the most important cereals used worldwide in the form of a range of products. Crop landraces have been an immense source of diversity for the breeders. In the present study, 517 Indian wheat landraces have been observed for the difference in bread making quality by assessing allelic behaviour of high molecular weight-glutenin subunits (HMW-GS). A total of 33 Glu-1 alleles (3 at Glu-A1, 15 at Glu-B1 and 15 at Glu-D1) were detected in wheat landraces. Allelic frequency of HMW-GS allelic band pattern null, 17 + 18, 2 + 12 (24.75%) was found to be the highest. Allelic frequency of HMW-GS allele null (68.27%) at Glu-A1, 17 + 18 (49.14%) at Glu-B1, and 2 + 12 (72.81%) at Glu-D1 was found to be the highest Five Novel alleles were identified at Glu-D1 locus, 12*, 12.1, 12.1*, 12.2 and 12.3. As Glu-D1 has highest quality contribution as compared to Glu-A1 and Glu-B1, reporting novel alleles at Glu-D1 represents that genetic variability available for selection is increased and it will provide tools for breeders to further improve dough properties and bread making quality.

The use of glutamine has been shown to increase the frequency of organogenesis and regeneration in the in vitro culture of several plants. The effect of glutamine on hormone-induced multiple shoot formation in desi and kabuli genotypes of chickpea (C-235 and PUSA-1053) were evaluated. Embryo axes with or without attached cotyledons were cultured in thidiazuron (TDZ) or 6-benzylaminopurine (BAP)-containing medium, respectively, with various concentrations of glutamine. Glutamine improved and prolonged the multiple shoot regeneration ability of the embryo axis. Chickpea embryo axis with attached cotyledon and cultured in TDZ-containing medium showed improved and prolonged shoot regeneration with 5 mM glutamine, while embryo axis without cotyledon and cultured in BAP-containing medium showed prolonged regeneration ability in 10 mM glutamine. Glutamine, however, did not serve as a substitute for cotyledon. Desi genotype (C-235) showed better response for multiple shoot formation as compared to the kabuli genotype (PUSA-1053). Glutamine at a concentration of 5 mM also improved root formation in excised in vitro shoots.

Tomato (Lycopersicon esculentum) is important widely grown vegetable in India and its productivity is affected by bacterial wilt disease infection caused by Ralstonia solanacearum. To prevent this disease infection a study was conducted to isolate and screen effective plant growth promoting rhizobacteria (PGPR) antagonistic to R. solanacearum. A total 297 antagonistic bacteria were isolated through dual culture inoculation technique, out of which forty-two antagonistic bacteria were found positive for phlD gene by PCR amplification using two primer sets Phl2a:Phl2b and B2BF:BPR4. The genetic diversity of phlD (+) bacteria was studied by amplified 16S rDNA restriction analysis and demonstrated eleven groups at 65% similarity level. Out of these 42 phlD (+) antagonistic isolates, twenty exhibited significantly fair plant growth promoting activities like phosphate solubilization (0.92-5.33%), 25 produced indole acetic acid (1.63-7.78 μg ml(-1)) and few strains show production of antifungal metabolites (HCN and siderophore). The screening of PGPR (phlD (+)) for suppression of bacterial wilt disease in glass house conditions was showed ten isolated phlD (+) bacteria were able to suppress infection of bacterial wilt disease in tomato plant (var. Arka vikas) in the presence R. solanacearum. The PGPR (phlD (+)) isolates s188, s215 and s288 was observed to be effective plant growth promoter as it shows highest dry weight per plant (3.86, 3.85 and 3.69 g plant(-1) respectively). The complete absence of wilt disease symptoms in tomato crop plants was observed by these treatments compared to negative control. Therefore inoculation of tomato plant with phlD (+) isolate s188 and other similar biocontrol agents may prove to be a positive strategy for checking wilt disease and thus improving plant vigor.