Comparative Transcriptome Profile of the Cytoplasmic Male Sterile and Fertile Floral Buds of Radish (Raphanus sativus L.).
Comparative Transcriptome Profile of the Cytoplasmic Male Sterile and Fertile Floral Buds of Radish (Raphanus sativus L.).
复制标题
萝卜细胞质雄性不育和可育花芽的转录组谱比较（Raphanus sativus L.）
DOI：
10.3390/ijms17010042
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发表时间：
2016-01-06
影响因子：
5.6
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Mei S;Liu T;Wang Z
萝卜细胞质雄性不育（CMS）在萝卜属和芸苔属植物育种中有着广泛的应用。然而，雄性不育的详细调控网络仍有待确定。我们以前的工作表明，在细胞质雄性不育萝卜的异常出现后不久，四分体阶段时，小孢子畸形和绒毡层细胞生长异常大。组织学分析表明，萝卜花蕾长约1.5 mm时，花药处于四分体时期。此外，高通量RNA测序技术被用来表征长度约为1.5毫米的萝卜芽的转录组从两个CMS诱导orf138基因和相应的近等基因保持系。总共对67，140个单基因进行了功能注释。这些基因的功能术语在55个基因本体（GO）组和323个京都基因和基因组百科全书（KEGG）途径中显著丰富。该转录组检测到萝卜叶绿体基因组中编码的总共79个蛋白质基因中的72个的转录本。相比之下，萝卜线粒体基因组包含34个蛋白质基因，但从转录组中只检测到16个蛋白质转录本。CMS和近等基因保持系之间的转录组比较显示了539个差异表达基因（DEG），表明使用具有不同核背景的两组CMS/保持系进行比较转录组分析的假阳性率明显降低。127个转录本在不育系中的表达量增加，412个转录本的表达量减少。除了CMS诱导orf138的转录水平没有变化，确定从线粒体和叶绿体基因组。对与CMS相关的MYB和bHLH转录因子、五肽重复序列（PPR）蛋白、热休克转录因子（HSFs）和热休克蛋白（HSPs）等DEG进行了讨论。该转录组数据集和比较分析将为进一步了解萝卜花药发育、细胞质雄性不育机制和分子育种提供重要的资源。
Radish cytoplasmic male sterility (CMS) has been widely used for breeding in Raphanus and Brassica genera. However, the detailed regulation network of the male sterility remains to be determined. Our previous work has shown that the abnormalities in a CMS radish appeared shortly after the tetrad stage when microspores were malformed and the tapetal cells grew abnormally large. In this work, histological analysis shows that anthers are at the tetrad stage when the radish buds are about 1.5 mm in length. Furthermore, a high throughput RNA sequencing technology was employed to characterize the transcriptome of radish buds with length about 1.5 mm from two CMS lines possessing the CMS-inducing orf138 gene and corresponding near-isogenic maintainer lines. A total of 67,140 unigenes were functionally annotated. Functional terms for these genes are significantly enriched in 55 Gene Ontology (GO) groups and 323 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The transcriptome detected transcripts for 72 out of a total of 79 protein genes encoded in the chloroplast genome from radish. In contrast, the radish mitochondrial genome contains 34 protein genes, but only 16 protein transcripts were detected from the transcriptome. The transcriptome comparison between CMS and near-isogenic maintainer lines revealed 539 differentially expressed genes (DEGs), indicating that the false positive rate for comparative transcriptome profiling was clearly decreased using two groups of CMS/maintainer lines with different nuclear background. The level of 127 transcripts was increased and 412 transcripts were decreased in the CMS lines. No change in levels of transcripts except CMS-inducing orf138 was identified from the mitochondrial and chloroplast genomes. Some DEGs which would be associated with the CMS, encoding MYB and bHLH transcription factors, pentatricopeptide repeat (PPR) proteins, heat shock transcription factors (HSFs) and heat shock proteins (HSPs), are discussed. The transcriptome dataset and comparative analysis will provide an important resource for further understanding anther development, the CMS mechanism and to improve molecular breeding in radish.