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新型隐性核基因雄性不育不结球白菜(芸薹属白菜亚种) WS24-3A 系的转录组比较分析和细胞学观察。

Comparative transcript profiling and cytological observation of the newly bred recessive genic male sterility non-heading Chinese cabbage (Brassica rapa ssp. chinensis) line WS24-3A.

机构信息

Wuhan Vegetable Research Institute, Wuhan Academy of Agriculture Science and Technology, Wuhan, 430345, China.

Biotechnology and Genetic Resources Institute, Yunnan Academy of Agricultural Sciences, Kunming, People's Republic of China.

出版信息

Genes Genomics. 2019 Dec;41(12):1475-1492. doi: 10.1007/s13258-019-00867-x. Epub 2019 Oct 1.

DOI:10.1007/s13258-019-00867-x
PMID:31576519
Abstract

BACKGROUND

WS24-3A is a newly bred non-heading Chinese cabbage genic male-sterile line, in which sterility is controlled by a recessive gene, designated as Bra2ms. WS24-3A has been used for hybrid breeding.

OBJECTIVE

To reveal the underlying molecular mechanisms responsible for the sterility of WS24-3A.

METHODS

Cytological observation of the process of sterile/fertile anther development was performed to determine the tissue and stage in which sterility occurs. Phenotyping and transcriptomic analyses were performed to identify differentially expressed genes (DEGs) between sterile and fertile flower buds at different stages.

RESULTS

Cytological analysis revealed no tetrads at stage 7 or at later stages of anther development, and the degradation of callose was delayed. Abnormal meiocytes were surrounded by sustaining callose that degenerated gradually in WS24-3A. Comparative transcript profiling identified 3282 DEGs during three anther developmental stages, namely, pre-meiotic anther, meiotic anther, and anthers with single-celled pollen stage. The difference in DEG percentage between up-regulated and down-regulated at meiotic anther stage was obviously larger than at the other two stages; further, most DEGs are important for male meiosis, callose synthesis and dissolution, and tapetum development. Ten DEGs were found to be involved in anther and pollen development, which were analyzed by quantitative PCR.

CONCLUSION

Bra2ms affected gene expression in meiocytes and associated with callose synthesis, degradation and tapetum development. Our results provide clues to elucidate the molecular mechanism of genic male sterility in non-heading Chinese cabbage.

摘要

背景

WS24-3A 是一种新培育的不结球白菜隐性核雄性不育系,其不育性由一个隐性基因 Bra2ms 控制。WS24-3A 已被用于杂交育种。

目的

揭示 WS24-3A 不育的潜在分子机制。

方法

通过细胞学观察不育/可育花药发育过程,确定发生不育的组织和阶段。表型分析和转录组分析用于鉴定不同发育阶段不育和可育花蕾之间的差异表达基因(DEGs)。

结果

细胞学分析显示,7 期或后期花药发育过程中没有四分体,且胼胝质的降解延迟。在 WS24-3A 中,异常减数分裂小孢子被持续的胼胝质包围,这些胼胝质逐渐退化。比较转录谱分析在三个花药发育阶段(花粉母细胞前期、减数分裂期和单核花粉期)共鉴定出 3282 个 DEGs。减数分裂期上调和下调基因的 DEG 百分比差异明显大于其他两个阶段;此外,大多数 DEGs 对减数分裂、胼胝质合成和溶解以及绒毡层发育至关重要。发现 10 个 DEG 与花药和花粉发育有关,并用定量 PCR 进行了分析。

结论

Bra2ms 影响减数分裂细胞中的基因表达,并与胼胝质合成、降解和绒毡层发育有关。我们的研究结果为阐明不结球白菜基因雄性不育的分子机制提供了线索。

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