BcMF13, a new reproductive organ-specific gene from Brassica rapa. ssp chinensis, affects pollen development
BcMF13, a new reproductive organ-specific gene from Brassica rapa. ssp chinensis, affects pollen development
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DOI：
10.1007/s11033-007-9072-8
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发表时间：
2008-06-01
影响因子：
2.8
通讯作者：
Xiang, Xun
中科院分区：
文献类型：
--
作者：
Li, Yanyan;Cao, Jiashu;Xiang, Xun
在大白菜(Brassica campestris L.SSP.)野生型花蕾中积累了一个转录衍生片段(GenBank登录号为DN237920.1)。[医]中华草本植物白菜型油菜B.Rapa Ssp.Chinensis)，并对其进行了进一步的研究。通过快速扩增cDNA端，克隆了该片段的全长DNA和cDNA。该基因BcMF13编码一个由73个氨基酸组成的蛋白质，并被一个106个碱基的内含子打断(GenBank登录号EF158459)。Southern杂交分析表明，BcMF13基因可能是大白菜基因组中的单拷贝基因。序列BLAST分析表明，BcMF13是一个新基因。在EST数据库中，这些序列与BcMF13的同源性均为96-98%，均来自油菜的花蕾、小孢子、花药，证明了BcMF13同源基因与油菜雄配子发生密切相关。RT-PCR发现，该基因仅在可育系的第四、五期花蕾中表达，在雄蕊中强烈表达。通过RNA反义策略成功地抑制了BcMF13基因的表达，大大减少了正常的花粉粒数，表明BcMF13基因在油菜属植物的花粉发育中是必需的。
A transcript-derived fragment (GenBank accession number DN237920.1) accumulated in the wild-type flower buds of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) was isolated and further investigated. The full length DNA and cDNA of the fragment were cloned by rapid amplification of cDNA ends. The gene, BcMF13, encodes a protein of 73 amino acids and is interrupted by an intron of 106 bp (GenBank accession number EF158459). Southern blot analysis revealed that BcMF13 could be a single-copy gene in the Chinese cabbage genome. Sequence blast analysis showed that BcMF13 was a new gene. In EST database, those sequences share 96 - 98% identity with BcMF13 cDNA all came from flower buds, microspores, anthers of Brassica, which proved BcMF13 homologs closely related to the development of male gametogenesis in Brassica. RT-PCR discovered that it is exclusively expressed in stage four and five flower buds of fertile line, strongly expressed in stamens. Successful suppression of BcMF13 gene expression by RNA antisense strategy greatly reduced the normal pollen grains, suggesting that BcMF13 was essential in pollen development in Brassica.