Until now three publications have reported the development of transgenic lentil plants through protocol optimisation using the gusA gene, but there are no reports of the introduction of a gene with agronomic importance. In the present study we report the introduction of the DREB1A gene into lentil to enhance drought and salinity tolerance. Decapitated embryos were immersed in Agrobacterium suspension and then co-cultivated for 4 days. Direct organogenesis was induced from the apical meristems and cotyledonary buds. Subsequently, the explants were subjected to selection in medium containing 10 mg/L phosphinothricin for nine rounds with 2-week intervals. The putative transgenic explants were micro-grafted onto non-transformed rootstocks to establish transgenic plants. The PCR results confirmed the insertion and stable inheritance of the gene of interest and bar marker gene in the plant genome. The Southern blot analysis revealed the integration of a single copy of the transgenes. T0 plants and progeny up to T2 generations showed complete resistance to the herbicide Basta. The DREB1A gene driven by the rd29A promoter was induced in transgenic plants by salt stress from sodium chloride solution. The total RNA was extracted and cDNA synthesised. The results showed that DREB1A mRNA was accumulated and thus the DREB1A transgene was expressed in the transgenic plants, whereas no expression was detected in the non-transformed parents.

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Tilletia indica is a quarantine fungal pathogen that poses a serious biosecurity threat to wheat-exporting countries. Acquiring genetic data for the pathogenicity characters of T. indica is still a challenge for wheat breeders and geneticists. In the current study, double digest restriction-site associated-DNA genotyping by sequencing was carried out for 39 T. indica isolates collected from different locations in India. The generated libraries upon sequencing were with 3,346,759 raw reads on average, and 151 x 2 nucleotides read length. The obtained bases per read ranged from 87 Mb in Ti 25 to 1,708 Mb in Ti 39, with 505 Mb on average per read. Trait association mapping was performed using 41,473 SNPs, infection phenotyping data, population structure, and Kinship matrix, to find single nucleotide polymorphisms (SNPs) linked to virulence genes. Population structure analysis divided the T. indica population in India into three subpopulations with genetic mixing in each subpopulation. However, the division was not in accordance with the degree of virulence. Trait association mapping revealed the presence of 13 SNPs associated with virulence. Using sequences analysis tools, one gene (g4132) near a significant SNP was predicted to be an effector, and its relative expression was assessed and found upregulated upon infection.

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