Comparative transcript profiling of fertile and sterile flower buds from multiple-allele-inherited male sterility in Chinese cabbage (Brassica campestris L. ssp pekinensis)
Comparative transcript profiling of fertile and sterile flower buds from multiple-allele-inherited male sterility in Chinese cabbage (Brassica campestris L. ssp pekinensis)
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大白菜（Brassica Campestris L. ssp pekinensis）多等位基因遗传雄性不育的可育和不育花芽的比较转录谱分析
DOI：
10.1007/s00438-017-1324-2
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发表时间：
2017
影响因子：
3.1
通讯作者：
Feng Hui
中科院分区：
文献类型：
--
作者：
Zhou Xue;Liu Zhiyong;Ji Ruiqin;Feng Hui
研究了大白菜多等位基因遗传雄性不育的原因。北京亚种（sp.pekinensis）的花粉育性差异表达基因（DEGs）。在这项工作中，我们在显微镜下验证了不育的阶段，然后使用Illumina HiSeq 2000平台测序对从可育和不育芽中分离的mRNA进行转录组分析。约2.29亿个高质量双端读段中的约80%被唯一映射到参考基因组。在不育芽中，699个基因表达显著上调，4096个基因表达显著下调。其中，花粉细胞壁相关基因28个，转录因子基因54个，植物发育相关基因45个，花药和花粉相关基因20个，特异表达转录本212个，定位于连锁群A07的DEG 417个。其中BrAMS、BrMS 1、BrbHLH 089、BrbHLH 091、BrAtMYB 103和BrANAC 025等6个转录因子基因被鉴定为不育相关基因。BrABP 1调控的生长素弱信号可能是导致花粉不育的关键因素之一。此外，在这项工作中鉴定了几个显著富集的GO术语，如“细胞壁组织或生物发生”（GO：0071554）、“膜固有”（GO：0031224）、“膜整合”（GO：0016021）、“水解酶活性，作用于酯键”（GO：0016788）和一个显著富集的途径“淀粉和蔗糖代谢”（ath 00500）。qRT-PCR、PCR和原位杂交实验验证了我们的RNA-seq转录组分析的准确性和可靠性。本研究为阐明大白菜雄性不育的分子机理奠定了基础，为进一步研究大白菜AB 01的多等位基因遗传雄性不育性提供了有价值的信息。
We studied the underlying causes of multiple-allele-inherited male sterility in Chinese cabbage (Brassica campestrisL. ssp.pekinensis) by identifying differentially expressed genes (DEGs) related to pollen sterility between fertile and sterile flower buds. In this work, we verified the stages of sterility microscopically and then performed transcriptome analysis of mRNA isolated from fertile and sterile buds using Illumina HiSeq 2000 platform sequencing. Approximately 80% of ~229 million high-quality paired-end reads were uniquely mapped to the reference genome. In sterile buds, 699 genes were significantly up-regulated and 4096 genes were down-regulated. Among the DEGs, 28 pollen cell wall-related genes, 54 transcription factor genes, 45 phytohormone-related genes, 20 anther and pollen-related genes, 212 specifically expressed transcripts, and 417 DEGs located on linkage group A07 were identified. Six transcription factor genesBrAMS,BrMS1,BrbHLH089,BrbHLH091,BrAtMYB103, andBrANAC025were identified as putative sterility-related genes. The weak auxin signal that is regulated byBrABP1may be one of the key factors causing pollen sterility observed here. Moreover, several significantly enriched GO terms such as “cell wall organization or biogenesis” (GO:0071554), “intrinsic to membrane” (GO:0031224), “integral to membrane” (GO:0016021), “hydrolase activity, acting on ester bonds” (GO:0016788), and one significantly enriched pathway “starch and sucrose metabolism” (ath00500) were identified in this work. qRT-PCR, PCR, and in situ hybridization experiments validated our RNA-seq transcriptome analysis as accurate and reliable. This study will lay the foundation for elucidating the molecular mechanism(s) that underly sterility and provide valuable information for studying multiple-allele-inherited male sterility in the Chinese cabbage line ‘AB01’.