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Optimising protocol for successful development of haploids in marigold (Tagetes spp.) through in vitro androgenesis

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Title Optimising protocol for successful development of haploids in marigold (Tagetes spp.) through in vitro androgenesis
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Creator K. Ravindra Kumar · Kanwar Pal Singh · Reeta Bhatia · D. V. S. Raju · Sapna Panwar
 
Subject African marigold · French marigold · Anther culture · Haploid · Flow cytometry · Cytology · Chloroplast
 
Description Not Available
For the frst time, we have developed a protocol for rapid and successful induction of haploids in Tagetes spp. using in vitro androgenesis techniques. Various factors afecting the in vitro androgenesis response in Tagetes spp. were optimised. At frst, a correlation was established between foret size/bud size and stage of microspore development in diferent genotypes of Tagetes erecta and Tagetes patula. The foret size of 3.5–4.0 mm (French marigold) and 3.0–3.5 mm (African marigold) contained the highest percent of microspores at early-uninucleate to early-binucleate stage. The frequency of shoot formation from anthers ranged from 0 to 78.6% in T. patula cv. Pusa Arpita and 0 to 63.9% in T. erecta cv. Pusa Basanti Gainda. The French marigold cultivars exhibited higher androgenic response over the African marigold cultivars. The efects of basal media, plant growth regulator combinations and sucrose concentration for direct diferentiation of shoot buds and
haploid induction were studied. Marigold anthers cultured on newly formulated EMS1 basal media (enriched with coconut water, AgNO3, PVP etc.) exhibited a very high androgenic response over the other commercially available media. We have also optimised the cold temperature pre-treatments and culture condition for high frequency androgenesis in marigold. Ten days cold pre-treated anthers along with 20 days dark incubation of cultures proved highly benefcial for haploid induction. Among a total of 424 anther-regenerants, 56 plants were randomly selected for fow cytometry and cytological analysis. The ploidy analysis revealed 14.3% of anther regenerants as dihaploids, 66.1% as tetraploid (similar to donor mother plant) and 19.6% as polyploid. The determination of ploidy level by counting the number of chloroplast in stomatal guard cells of marigold was also established for rapid screening of haploids. The identifed haploids were successfully diploidised and are
being utilised in the hybrid breeding programme at our institute. The developed protocol will facilitate doubled haploids based breeding in Tagetes spp.
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Date 2020-04-15T08:57:28Z
2020-04-15T08:57:28Z
2019-03-30
 
Type Research Paper
 
Identifier Not Available
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http://krishi.icar.gov.in/jspui/handle/123456789/34798
 
Language English
 
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Publisher Not Available