Honys David, Oh Sung-Aeong, Renák David, Donders Maarten, Solcová Blanka, Johnson James Andrew, Boudová Rita, Twell David
Laboratory of Pollen Biology, Institute of Experimental Botany ASCR, Rozvojová 135, 165 02 Prague 6, Czech Republic.
BMC Plant Biol. 2006 Dec 21;6:31. doi: 10.1186/1471-2229-6-31.
The effective functional analysis of male gametophyte development requires new tools enabling the spatially and temporally controlled expression of both marker genes and modified genes of interest. In particular, promoters driving expression at earlier developmental stages including microspores are required.
Transcriptomic datasets covering four progressive stages of male gametophyte development in Arabidopsis were used to select candidate genes showing early expression profiles that were male gametophyte-specific. Promoter-GUS reporter analysis of candidate genes identified three promoters (MSP1, MSP2, and MSP3) that are active in microspores and are otherwise specific to the male gametophyte and tapetum. The MSP1 and MSP2 promoters were used to successfully complement and restore the male transmission of the gametophytic two-in-one (tio) mutant that is cytokinesis-defective at first microspore division.
We demonstrate the effective application of MSP promoters as tools that can be used to elucidate gametophytic gene functions in microspores in a male-specific manner.
对雄配子体发育进行有效的功能分析需要新的工具，以实现标记基因和感兴趣的修饰基因在空间和时间上的可控表达。特别是，需要能够驱动包括小孢子在内的早期发育阶段表达的启动子。
利用覆盖拟南芥雄配子体发育四个连续阶段的转录组数据集，筛选出显示早期表达谱且具有雄配子体特异性的候选基因。对候选基因进行启动子-GUS报告基因分析，鉴定出三个在小孢子中具有活性、对雄配子体和绒毡层具有特异性的启动子（MSP1、MSP2和MSP3）。MSP1和MSP2启动子成功地互补并恢复了配子体二合一（tio）突变体的雄配子传递，该突变体在第一次小孢子分裂时存在胞质分裂缺陷。
我们证明了MSP启动子作为工具的有效应用，可用于以雄性特异性方式阐明小孢子中配子体基因的功能。
