Factors Affecting Microspore Embryogenesis and Early Ploidy Level Determination of Regenerated Plants in Brassica napus

doi:10.7668/hbnxb.2011.05.020

Abstract

Abstract: The breeding materials of Brassica napus grown in different ecological regions and years were used to determine their microspore embryogenesis capacity.The microspores of 11 genotypes grown in field condition of Wuhan,Hubei Province,were isolated at different flowering stages and cultured.The aim of this study was to investigate the effects of genotype,ecological condition and sampling time on microspore embryogenesis,as well as the colchicine treatment on chromosome doubling of regenerated seedlings.Ploidy level of regenerated plants was determined according to DNA content measured by flow cytometric analysis and confirmed by field observation.The results showed that the microspore embryogenesis was significantly different among the genotypes and in different ecological conditions.The plants sown in the summer in Xining,Qinghai Province,had significantly higher microspore embryogenesis than those sown in the spring in Wuhan,Hubei Province.The microspores isolated at early flowering stage(1-3 days)gave rise to significantly higher embryogenesis than those isolated at full flowering stage(12-15 days after flowering).The treatment of 50 mg/L colchicine directly on isolated microspores for 24 h could significantly increase the number of diploids in regenerated plants.The spontaneous chromosome doubling rate of regenerated plants ranged from 8.62% to 20.88% for different genotypes,whereas the colchicine treatment could raise the chromosome doubling rate to 60%-77.59%.The ploidy level of regenerated plants determined by flow cytometry was consistent with the result observed in the field,so the flow cytometry provided a useful means for early dihaploid discrimination of microspore-derived plants.

Key words: Oilseed rape, Microspore culture, DNA content, Ploidy identification

CLC Number:

S565.4

MI Zhe, LI Yun-chang, MEI De-sheng, LI Ying-de, XU Yu-song, CHEN Yu-feng, HU Qiong. Factors Affecting Microspore Embryogenesis and Early Ploidy Level Determination of Regenerated Plants in Brassica napus[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(5): 97-102. doi: 10.7668/hbnxb.2011.05.020.

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