关键词: 白菜, BcMF12, BcA9启动子, 反义RNA, 基因表达, GUS活性

Abstract: 【Objective】The study analyzed the expression of BcMF12 gene regulated by BcA9 promoter in the transgenic Chinese cabbage, and confirmed the effect of antisense BcMF12 gene on the pollen development.【Method】An conserved BcMF12 gene fragment was amplified from the cDNA of flower buds in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino), and was fused to the anther specific BcA9 promoter. The plant antisense expression vector was constructed and then introduced into Chinese cabbage via Agrobacterium-mediated transformation. The transgenic plants were screened by antibiotics and molecular analysis.【Result】The analysis of GUS activity revealed that the expression level of antisense BcMF12-GUS fusion gene regulated by the BcA9 promoter was strongest in the anther, while very weak or no activity could be detected in other organs of transgenic plants. It showed that antisense BcMF12-GUS fusion genes could be tissue-specifically expressed in transgenic Chinese cabbage when driven by the BcA9 promoter. Northern blot suggested that the expression of BcMF12 gene in pollen was down-regulated significantly. The pollen germinated rate of transgenic plants contained the antisense BcMF12 gene decreased as compared with that of the control plants.【Conclusion】The expression of the gene BcMF12 related to the pollen development was inhibited by the antisense BcMF12 driven by the anther-specific BcA9 promoter, which affected the pollen development in Chinese cabbage.

Key words: Brassica campestris ssp. chinensis, BcMF12, BcA9 promoter, antisense RNA, gene expression, GUS activity