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BcMF26a and BcMF26b Are Duplicated Polygalacturonase Genes with Divergent Expression Patterns and Functions in Pollen Development and Pollen Tube Formation in Brassica campestris

机译:BcMF26a和BcMF26b是重复的聚半乳糖醛酸酶基因在油菜花粉发育和花粉管形成中具有不同的表达模式和功能

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摘要

Polygalacturonase (PG) is one of the cell wall hydrolytic enzymes involving in pectin degradation. A comparison of two highly conserved duplicated PG genes, namely, Brassica campestris Male Fertility 26a (BcMF26a) and BcMF26b, revealed the different features of their expression patterns and functions. We found that these two genes were orthologous genes of At4g33440, and they originated from a chromosomal segmental duplication. Although structurally similar, their regulatory and intron sequences largely diverged. QRT-PCR analysis showed that the expression level of BcMF26b was higher than that of BcMF26a in almost all the tested organs and tissues in Brassica campestris. Promoter activity analysis showed that, at reproductive development stages, BcMF26b promoter was active in tapetum, pollen grains, and pistils, whereas BcMF26a promoter was only active in pistils. In the subcellular localization experiment, BcMF26a and BcMF26b proteins could be localized to the cell wall. When the two genes were co-inhibited, pollen intine was formed abnormally and pollen tubes could not grow or stretch. Moreover, the knockout mutants of At4g33440 delayed the growth of pollen tubes. Therefore, BcMF26a/b can participate in the construction of pollen wall by modulating intine information and BcMF26b may play a major role in co-inhibiting transformed plants.
机译:聚半乳糖醛酸酶(PG)是涉及果胶降解的细胞壁水解酶之一。比较两个高度保守的重复PG基因,即油菜雄性育性26a(BcMF26a)和BcMF26b,揭示了它们表达模式和功能的不同特征。我们发现这两个基因是At4g33440的直系同源基因,它们起源于染色体节段重复。尽管在结构上相似,但是它们的调控序列和内含子序列差异很大。 QRT-PCR分析表明,在甘蓝型油菜的几乎所有被测器官和组织中,BcMF26b的表达水平均高于BcMF26a。启动子活性分析表明,在生殖发育阶段,BcMF26b启动子在绒毡层,花粉粒和雌蕊中有活性,而BcMF26a启动子仅在雌蕊中有活性。在亚细胞定位实验中,BcMF26a和BcMF26b蛋白可以定位在细胞壁上。当这两个基因被共同抑制时,花粉短链形成异常,花粉管无法生长或伸展。此外,At4g33440的敲除突变体延迟了花粉管的生长。因此,BcMF26a / b可以通过调节常规信息参与花粉壁的构建,而BcMF26b可能在共抑制转化植物中起主要作用。

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