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Initiation and development of microspore embryogenesis and plant regeneration of Brassica nigra

文献类型: 外文期刊

作者: Gu, Honghui 1 ; Sheng, Xiaoguang 1 ; Zhao, Zhenqing 1 ; Yu, Huifang 1 ; Wang, Jiansheng 1 ;

作者机构: 1.Zhejiang Acad Agr Sci, Inst Vegetables, Hangzhou 310021, Zhejiang, Peoples R China

关键词: B. nigra;Microspore culture;Microspore embryogenesis;Doubled haploid

期刊名称:IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-PLANT ( 影响因子:2.252; 五年影响因子:2.139 )

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收录情况: SCI

摘要: Brassica nigra is generally regarded as a recalcitrant species for microspore culture among Brassica crops. Conditions for reliable induction of microspore embryogenesis of B. nigra were studied in this context. Flower bud length and microspore developmental stage were correlated with further embryogenesis. The optimal bud size range was 2.0-2.5 mm for the highest proportion of totipotent, late uninucleate microspore and the highest frequency of microspore embryogenesis. Treatment of a short heat shock by incubating the microspore culture at 32 degrees C for 24 h was suitable for the microspore survival, sustained cell divisions, and further induced embryogenesis. Subsequently, the use of NLN medium with the addition of 13% sucrose and 0.1% activated charcoal (AC) provided the optimal conditions for the development of microspore-derived embryos (MDEs). The early cotyledonary (EC) stage embryos cultured on MS medium fortified with 4.6 mu M zeatin (ZT) and 0.12 mu M indole-3-acetic acid (IAA) resulted in themost efficient rates of plantlet regeneration. The ploidy levels of regenerated plants of B. nigra were determined by flow cytometry, revealing that 50.6% were diploid. The results enable the advancement of breeding programs and genetic studies in B. nigra.

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