Molecular Cloning and Characterization of a Novel Polygalacturonase Gene, BcMF24, Involved in Pollen Development of Brassica campestris ssp. chinensis
Molecular Cloning and Characterization of a Novel Polygalacturonase Gene, BcMF24, Involved in Pollen Development of Brassica campestris ssp. chinensis
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DOI：
10.1007/s11105-013-0666-0
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发表时间：
2014-09
影响因子：
2.1
通讯作者：
Youjian Yu;M. Lv;Ying Liang;X. Xiong;Jiashu Cao
中科院分区：
文献类型：
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作者：
Youjian Yu;M. Lv;Ying Liang;X. Xiong;Jiashu Cao
本研究从芸苔属油菜（Brassica campestris L.）中国菜心（Brassica rapassp.chinensis）。序列分析表明，该基因含有PG蛋白的三个经典保守结构域（I、II和IV）。系统发育分析表明，该菌与其他几种植物的PG聚为PG家族的E支。定量逆转录-聚合酶链反应（qRT-PCR）和原位杂交结果表明，该基因B. campestris Male Fertility 24（BcMF 24）在小孢子、双核期部分降解的绒毡层和成熟花粉中特异表达。对BcMF 245 ′上游1，489 bp序列的生物信息学分析表明，该基因含有多个典型的顺式作用元件和花粉特异性元件。瞬时表达分析表明，BcMF 24启动子可以驱动洋葱表皮细胞中的绿色荧光蛋白（GFP）表达，转基因拟南芥的β-葡萄糖醛酸酶（GUS）检测表明，BcMF 24启动子可以特异性地驱动双核期和成熟花粉期花蕾花药中的基因表达。据我们所知，目前的研究是第一次证明PG基因，它属于E支，参与花粉发育。
In the present study, a polygalacturonase (PG) gene was isolated fromBrassica campestrisL. ssp.chinensisMakino (syn.Brassica rapassp.chinensis). Sequence analysis indicated that it contained three classical conserved domains (I, II, and IV) of PG proteins. Phylogenetic analysis demonstrated that it was clustered into the clade E of PG family with several other PGs from different plant species. The results obtained from quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and in situ hybridization showed that the gene,B. campestris Male Fertility 24(BcMF24), was specifically expressed in the microspores, in the partly degraded tapetum at binucleate stage and in the pollen grains at mature pollen stage. Bioinformatics analysis of a 1,489-bp sequence in theBcMF245′-upstream region illustrated that the said gene contained several typicalcis-acting elements and pollen-specific elements. Transient expression analysis revealed thatBcMF24can drive the green fluorescent protein (GFP) expression in onion epidermal cells.β-glucuronidase (GUS) assay of theBcMF24promoter–GUS constructs in transgenicArabidopsis thalianashowed that it could specifically drive gene expression in the anther of flower buds at binucleate and mature pollen stages. To the best of our knowledge, the current study is the first demonstration of a PG gene, which belongs to clade E that is involved in pollen development.