Anther and isolated microspore culture of coconut (Cocos nucifera L.)

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Dec 12, 2016 - coconut (Cocos nucifera L.) M.K. Rajesh, K.S. Muralikrishna, K.K. Sajini and Anitha Karun. ICAR - Central Plantation Crops Research Institute, ...
3rd International Symposium on Coconut Research and Development 10 - 12 December 2016, ICAR - CPCRI, Kasaragod

Abstract

S2P-02

Anther and isolated microspore culture of coconut (Cocos nucifera L.) M.K. Rajesh, K.S. Muralikrishna, K.K. Sajini and Anitha Karun ICAR - Central Plantation Crops Research Institute, Kasaragod 671124, Kerala, India ([email protected])

In vitro culturing of anthers and isolated microspores is an extremely useful technique for the production of doubled haploid in cultivated crops. In the present study, anthers in tetrad stage (-3 position) were isolated from pretreated (cold at 10 οC and heat at 40 οC for 5 and 3 days, respectively) of West Coast Tall (WCT) coconut variety and inoculated on to eight different basal medium incorporated with either 2, 4-D or picloram as auxin and BAP or TDZ as cytokinin. To isolate microspores, anthers were split into small bits and blended in 8% sucrose solution at 500-600 rpm for two minutes aseptically. The extract after filtering with 80 µm mesh was inoculated into ten different combinations of Y3 liquid media with 2, 4-D (2mg/l), picloram (100 µM), BAP (1 mg/l) and TDZ (1 mg/l). Callusing percentage in anthers was more in media supplemented with picloram alone or along with BAP. Pretreatment of anthers at heat or cold did not show significant difference for callus induction. The callus initiated from anthers did not proliferate upon sub culturing. Microspore culture did not result in callus induction or growth. However success was achieved in isolation of microspores from anthers without contamination. Further modifications are warranted, both in anther and microspore culture, for sustainable results.

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