Functional characterization of E3 ubiquity ligase Bra015092 in pollen development of Brassica campestris ssp. Chinensis
Functional characterization of E3 ubiquity ligase Bra015092 in pollen development of Brassica campestris ssp. Chinensis
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E3 泛在连接酶 Bra015092 在油菜花粉发育中的功能表征。
DOI：
10.1111/ppl.13808
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发表时间：
2022
影响因子：
6.4
通讯作者：
Zhuoyue Cui
中科院分区：
文献类型：
--
作者：
Aimei Tian;Hui Yu;Zhuoyue Cui
花粉发育在种子型植物的有性生殖中起着重要作用。蛋白质的泛素化是蛋白质翻译后修饰的重要环节。E3泛素连接酶是蛋白质泛素化途径中识别底物的关键蛋白。以大白菜（Brassica campestrisL.）ssp.Chinensis）作为试验材料。结果表明，该基因为Bra 015092，大小为642 bp。利用半定量（RT-PCR）和定量真实的-时间PCR（qRT-PCR）技术分析了Bra 015092在大白菜两用系中的表达。结果表明，Bra 015092在花序中表达量较高。亚细胞定位分析表明，Bra 015092与GFP融合表达蛋白广泛存在于烟草表皮细胞中，将Bra 015092转化“优青49”甘蓝，获得Bra 015092 OE过表达转基因植株。对Bra 015092 OE植株的形态学观察表明，BcMF 29 OE植株的花粉畸形、失活，营养核和生殖核发育异常。离体萌发试验表明，Bra 015092 OE植株中约有24.5%的花粉不能萌发。半薄切片结果表明，Bra 015092 OE植株的花粉发育在双核花粉粒阶段出现异常。透射电镜观察发现，Bra 015092 OE植株的花粉粒从双核期逐渐退化到三核期，花粉内壁发育异常，表明Bra 015092在花粉发育过程中起主要作用。
Pollen development plays an important role in the sexual reproduction of seed‐type plants. Ubiquitination of proteins is an essential link in the post‐translational modification of proteins. E3 ubiquity ligase is a key protein that recognizes substrates in the protein ubiquitination pathway. The hybrid line “Bcajh97‐01A/B” of Chinese cabbage (Brassica campestrisL. ssp.Chinensis) was used as test material. The geneBra015092,with a size of 642 bp, was amplified. Semi‐quantitative (RT‐PCR) and quantitative real‐time PCR (qRT‐PCR) techniques were utilized to analyze the expression ofBra015092in the dual‐purpose line of Chinese cabbage. It was found thatBra015092had a higher expression level in inflorescence. Subcellular localization analysis showed thatBra015092and GFP fusion expression protein widely exist in tobacco epidermal cells.Bra015092was transformed into “Youqing49” cabbage to obtainBra015092OEoverexpressing transgenic lines. The morphological observation ofBra015092OEplants showed that the pollen ofBcMF29OEplants became deformed and inactive, and the vegetative and reproductive nuclei were abnormally developed. The in vitro germination experiments showed that about 24.5% of the pollen inBra015092OEplants could not germinate. The results of the semi‐thin section showed that the pollen development ofBra015092OEplants was abnormal at the stage of binuclear pollen grains. Transmission electron microscopy revealed that the pollen grains ofBra015092OEplants gradually degraded from the binuclear to the trinucleate pollen grain stage, and the pollen inner wall was abnormally developed, indicating thatBra015092plays a major role in the process of pollen development.