Identification and functional characterisation of a novel anther-specific LTP promoter from Brassica campestris ssp chinensis
Identification and functional characterisation of a novel anther-specific LTP promoter from Brassica campestris ssp chinensis
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甘蓝型新型花药特异性LTP启动子的鉴定和功能表征
DOI：
10.1080/14620316.2016.1166992
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发表时间：
2016
影响因子：
1.9
通讯作者：
Zhang Enhui
中科院分区：
文献类型：
--
作者：
Tian Aimei;Cao Jiashu;Zhang Enhui
油菜花粉发育相关基因sltp。中国人属于脂质转移蛋白（LTP）基因家族。采用热不对称交错PCR （TAIL-PCR）分离了其sltpgene的启动子。分析了启动子（命名为Bc15）的功能元件。在该序列中还鉴定出了一些花药/花粉特异性元件，如GGTT盒和GTGA盒。为了确认启动子的活性，我们将一系列5 '截断的构建体与GUS报告基因融合，对启动子的关键调控区域进行功能分析。瞬时表达分析表明，GUS在洋葱表皮细胞−731 ~−1 bp区域具有较高的表达活性。携带- 731至- 1 bp片段的截断构建体被稳定地引入拟南芥中。组织化学分析显示，转基因植株在花药/花粉发育的不同阶段有不同的特异GUS表达。花蕾横切面进一步表明Bc15启动子特异性激活花药中报告基因的表达。这些发现阐明了花粉发育过程中LTP基因调控的分子机制，并提示Bc15启动子在工程雄性不育杂交生产中的潜在应用。
The pollen development-related genemsLTPfromBrassica campestrisssp.chinensisbelongs to the lipid transfer protein (LTP) gene family. The promoter of themsLTPgene was isolated by thermal asymmetric interlaced PCR (TAIL-PCR). The functional elements of the promoter (named Bc15) were analysed. Some anther/pollen-specific elements, such as the GGTT box and the GTGA box, were identified in the sequence. In an attempt to confirm the promoter activity, a series of 5′-truncated constructs was fused with the GUS reporter gene to functionally analyse the key regulatory regions of the promoter. Transient expression analysis showed high GUS activity in onion epidermal cells containing the region from −731 to −1 bp. The truncated construct bearing the −731 to −1 bp fragment was stably introduced intoArabidopsis. Histochemical analysis of the transgenic plants showed distinct and specific GUS expression at different stages of anther/pollen development. Transverse sections of flower buds further indicated that the Bc15 promoter activates reporter gene expression in the anther specifically. These findings elucidated the molecular mechanisms of LTP gene regulation during pollen development, and suggested a potential application of the Bc15 promoter in engineering male sterility for hybrid production.