Isolation and characterization of a novel BcMF14 gene from Brassica campestris ssp. chinensis

Isolation and characterization of a novel BcMF14 gene from Brassica campestris ssp. chinensis
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甘蓝中新型 BcMF14 基因的分离和表征。

DOI:
10.1007/s11033-010-0298-5
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发表时间:
2011-03
影响因子:
2.8
通讯作者:
李焰焰
李焰焰
中科院分区:
生物学4区
文献类型:
--
作者:
李焰焰

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从大白菜(Brassica campestris L. ssp)中分离到一个快速碱化因子(RALF)样基因(GenBank登录号EF523517),命名为BcMF14。中华白杨,syn. B. rapa ssp。利用在可育系中特异表达的cDNA- aflp差异片段快速扩增cDNA末端(RACE)。半定量逆转录聚合酶链反应(RT-PCR)发现BcMF14在可育系4期和5期花蕾中显著表达,在营养结构和不育系中不表达。详细的RT-PCR显示其在雄蕊中的强表达。成功抑制BcMF14基因表达后,正常花粉粒数量大幅减少。突变体花粉粒异常率为48.95%,萌发沟两侧花粉粒多呈皱缩状,形状不规则,萌发沟两侧花粉粒较大,呈深凹状。由于花粉管严重扭曲,花粉萌发停止。这些结果表明BcMF14基因在白菜雄性配子体发育中具有潜在的作用。
A putative RALF (rapid alkalinization factor)-like gene (GenBank accession number EF523517), named BcMF14, was isolated from Chinese cabbage (Brassica campestris L. ssp. chinensis Makino, syn. B. rapa ssp. chinensis) by rapid amplification of cDNA ends (RACE) based on a cDNA-AFLP differential fragment exclusively expressed in fertile line. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) discovered that BcMF14 was prominently expressed in stage four and five flower buds of fertile line, no expression in vegetative structures or in sterility line. Detailed RT-PCR illuminated its strong expression in stamens. Successful suppression of BcMF14 gene expression greatly reduced the normal pollen grains. The frequency of abnormal pollen grains was 48.95% in the mutant with many shriveled pollen grains with irregular shape and some larger ones with deep hollows along the germination ditch. Pollen germination was stopped because of the severely twisted pollen tubes. These results demonstrate a potential role of the BcMF14 gene in the development of male gametogenesis in Chinese cabbage.
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