ICAR-Indian Institute of Pulses Research (IIPR)

Collaborating Centre Principal Investigator (CC-PI)

Dr. Alok Das

Senior Scientist

Principal Investigators involved in the project

Dr. Alok Das

Senior Scientist

Neetu Singh Kushwah

Senior Scientist

Dr. Meenal Rathore

Principal Scientist & Head

Dr. Shanmugavadivel P S

Senior Scientist

Dr. Aravinda K Konda

Senior Scientist

Co-Principal Investigators (Co-PIs)

Chickpea

Dr. Avinash Srivastava

Principal Scientist

Dr. SenthilKumar M

Principal Scientist

Dr. Shanmugavadivel

Senior Scientist

Dr. Neetu S Kushwaha

Senior Scientist

Grasspea

Dr. Meenal Rathore

Principal Scientist & Head

Dr. Vaibhav Kumar

Scientist

Dr. Archana Singh

Principal Scientist

Mr. Sudhir K Jha

Scientist

Pigeonpea

Dr. Shanmugavadivel

Senior Scientist

Dr. Antara Das

Scientist

Dr. Abhishek Bohra

Senior Scientist

Dr. Kuldeep Kumar

Scientist

Blackgram

Meenal Rathore

Principal Scientist & Head

Dr Kuldeep Kumar

Scientist

Dr Aravinda K Konda

Senior Scientist

Dr Debjyoti Sengupta

Senior Scientist

Lentil

Dr Jitendra Kumar

Principal Scientist

Dr Neetu Singh Kushwaha

Senior Scientist

Dr Antara Das

Scientist

Dr Shanmugavadivel

Senior Scientist

Target Traits in Pulses & Legumes

Research Methods & Expected Outcomes

Chickpea Component

• Allele mining across chickpea genotypes and wild species indicated a largely conserved coding region, with a unique single-base substitution observed in one wild species, linking it with seed number per pod trait.
• RT-PCR showed constitutive expression across developmental stages in selected cultivars, with up-regulation after flowering. Prime editing guide RNAs were designed for the conserved regulatory domain.
• Bioinformatics analysis of a seed-related locus identified gene structure; multiple oligo sets were designed for amplification in contrasting genotypes. Guide RNAs for targeted deletion were also developed.

Grasspea Component

• Transformation was attempted without BAP and kanamycin; 499 explants were transformed using CRISPR/Cas constructs, resulting in 122 putative transformants. T1 seeds were harvested from 95 T0 plants.
• Alternative transformation strategies are being tested to improve establishment rate using newly developed vectors.
• Sequence analysis of a candidate gene in Mahateora variety revealed a 195 bp deletion in one intron and multiple SNPs in the coding region compared to the reference genome.

Pigeonpea Component

• Contrasting parents (2, 4, and 6 seeds/pod) were sown for spatiotemporal sampling. Transcriptome libraries were prepared from embryo sac (0 DAA), seed/pod wall (5–20 DAA), and reproductive leaf tissues.
• No significant sequence variation was detected among contrasting parents, including IPA15-6 and Asha.
• Minimum inhibitory concentration for Basta was identified in IPA15-6 for leaf paint assay and in vitro bioassay to explore alternate selection pressure in transformation.

Blackgram Component

• Candidate genes related to cytokinin regulation were identified from the genome assembly of IPU0243; two showed multiple binding domains. Four SNPs were detected in one candidate gene when comparing IPU13-01 with IPU0243.
• In vitro regeneration was optimized via direct organogenesis using dicotyledonary node (DCN) explants; shoot induction was achieved from decapitated embryonic axis explants (IPU13-01).
• Agrobacterium-mediated in-planta transformation was attempted in IPU13-01 and IPU10-26 using a CRISPR/Cas9-based plasmid.

Lentil Component

• Genome-wide identification of candidate genes associated with cytokinin regulation in lentil showed a dispersed distribution. Expression analysis via transcriptome and proteomics is in progress.
• In vitro regeneration was achieved via direct organogenesis using cotyledonary explants with embryonic axis. Best results were obtained on MS+B5+CaCl₂ media with TDZ (4 µM), 2iP (10 µM), KIN (2 µM) for shoot induction, and IAA (2 mg/l) for rooting.
• Phylogenetic analysis showed lentil candidate genes clustering with Arabidopsis counterparts, though no close homologs were detected for some reference genes.