CROP IMPROVEMENT Quarterly Report March - June 2013 Genetic Resources Management M.

.acuminata from Baramurah hills and Behula and Batheeshwar from Nagicherra, Tripura were collected and added to the in vitro genebank of NRCB, Trichy.. Thirty two batches of tissue cultured banana cv. Grand Naine from M/s. Jain Irrigation Systems Ltd., Jalgaon, Maharashtra have been tested for their fidelity using SSR and ISSR markers under DBT ATL and issued test reports for quality certification purpose. 650 plants of Udhayam have been taken for hardening. 80 Grand Naine and 38 Rasthali virus free immature male flower buds were supplied to partner institutes (NABI and BARC), while 50 buds were initiated at NRCB for the development of ECS. Agricultural Biotechnology 2D-IEF condition was standardized for calli derived protein samples. Seventy five plants mutated Rasthali were challenge inoculated with spores of Fusarium oxysporum fsp. cubense for in vivo screening purpose. Digital Gene Expression studies were performed for Sigatoka, Drought and Nematode transcripts and were annotated with Banana Genome sequences. Results of the gene expression studies indicated cytochrome oxidase, lipoxygenase, metallothionein, falvin containing monoxygenase were up-regulated only in Sigatoka (M.eumusae) resistant cultivar, Manoranjitham. Auxin responsive factor, serine glyoxylate, retrotransposon ty1-copia were found to be highly up-regulated in resistant cultivar while a slight upregulation was observed in susceptible cultivar, Grand Naine. Brown hopper induced resistant protein were upregulated only in susceptible cultivar. Ethylene responsive factor, senescence-associated protein, aleurain-like protease and zinc finger were found to be highly upregulated in susceptible cultivar and slight up-regulation in resistant cultivar also. Gene expression studies were also performed for different developmental stages of banana, and the following genes which were upregulated in various stages - Shoot tip culture Short root (SHR), Macropropagation WUSCHEL (WUS), Leafy cotyledon I& II (LEC I& II), Enhanced Shoot Regeneration I (ESR I), Auxin Response Factor (ARF), Embryogenic cell suspension - Short root (SHR), WUSCHEL (WUS), Leafy Cotyledon I (LEC I), Cup shaped cotyledon (CUC), Direct Regeneration - WUSCHEL (WUS), Enhanced Shoot Regeneration I (ESR I). Out of 84813 unigenes, 3013 defense related genes were identified from the whole transcriptome sequences of Banana. Un-annotated 24873 transcripts were further annotated using TBLASTX in BLAST2Go program. 4780 SSRs were identified and annotated from 82413 unigenes of Banana transcriptome. 523 primers were designed for these SSRs. 30 SSR primers which are related to plant defense mechanism were synthesized and PCR conditions were standardized for 17 primers.

Varietal Improvement Hybrid embryos of following combinations have been regenerated with hormonal priming of seeds (GA3 for 48 hours). Saba (ABB) X Pisang Lilin (AA) Saba (ABB) X Pisang Jajee (AA) Bangrier (ABB) X Pisang Lilin (AA) Udhayam (ABB) X Pisang Lilin (AA) A histological study of pollen tube penetration into the ovary through the stylar region was carried out between seed fertile and sterile cultivars and seed set was successfully observed in the cv. Alpon (AAB). Open pollinated seeds of Sommarani Monthan and M.acuminata ssp. microcarpa and the following four different combinations of hybrid seeds were germinated under in vitro. Progeny ( Matti based cross) X cv.Rose Bangrier (ABB) X Pisang Lilin (AA) Udhayam (ABB) X Pisang Lilin (AA) Kapur (ABB) X Pisang Jajee (AA) Malformed embryos from floating seeds of 12 hybrids, and cultivars were imitated on embryo initiation medium to study the regeneration efficiency. Although most of them failed to germinate, those germinated (Matti X cv. Rose, Kothia X Calcutta-4, Bankela X Pisang Lilin, open pollinated seeds of cvs. Saba, Bhat Manohar and of M. velutina) resulted in callus mediated production of multiple shoots. No of shoots ranged from 5-10 from a single embryo.

(S.UMA) Principal Scientist