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Identification of differentially accumulating pistil proteins associated with self-incompatibility of non-heading Chinese cabbage

机译:与非抽穗大白菜自交不亲和性相关的雌蕊差异积累蛋白的鉴定

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摘要

Non-heading Chinese cabbage (Brassica campestris L. ssp. chinensis Makino), an important vegetable crop in China, exhibits a typical sporophytic self-incompatibility (SI) system. To better understand the mechanism of SI response and identify potential candidate proteins involved in the SI system of this vegetable crop, the proteomic approach was taken to identify differential accumulating pistil proteins. Pistils were collected at 0 h and 2 h after self-pollination at anthesis in self-incompatible and compatible lines of non-heading Chinese cabbage, and total proteins were extracted and separated by two-dimensional gel electrophoresis (2-DE). A total of 25 protein spots that displayed differential abundance were identified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry (MALDI–TOF/TOF MS) and peptide mass fingerprinting (PMF). Among them, 22 protein spots were confidently established. The mRNA levels of the corresponding genes were detected by quantitative RT-PCR. The 22 identified protein spots are involved in energy metabolism (four), protein biosynthesis (three), photosynthesis (six), stress response and defence (five), and protein degradation (four). Among these potential candidate proteins, UDP-sugar pyrophosphorylase could be involved in sucrose degradation to influence pollen germination and growth. Glutathione S–transferases could be involved in pollen maturation, and affect pollen fertility. Senescence-associated cysteine protease, which is related to programmed cell death, could be mainly related to self pollen recognition of non-heading Chinese cabbage. The study will contribute to further investigations of molecular mechanism of sporophytic SI in Brassicaceae.
机译:无头大白菜(Brassica campestris L. ssp。chinensis Makino)是中国重要的蔬菜作物,具有典型的孢子自交不亲和性(SI)系统。为了更好地理解SI应答的机制并确定该蔬菜作物SI系统中涉及的潜在候选蛋白,采用了蛋白质组学方法来鉴定差异积累的雌蕊蛋白。自花授粉后0小时和2小时,在无花粉大白菜的自交不亲和和自交系中收集雌蕊,并通过二维凝胶电泳(2-DE)提取和分离总蛋白。通过基质辅助激光解吸/电离飞行时间质谱(MALDI–TOF / TOF MS)和肽质量指纹图谱(PMF)鉴定出总共25个显示出不同丰度的蛋白质斑点。其中,有22个蛋白质斑点被确定地建立。通过定量RT-PCR检测相应基因的mRNA水平。鉴定出的22个蛋白质斑点涉及能量代谢(四个),蛋白质生物合成(三个),光合作用(六个),应激反应和防御(五个)和蛋白质降解(四个)。在这些潜在的候选蛋白中,UDP糖焦磷酸化酶可能与蔗糖降解有关,从而影响花粉的萌发和生长。谷胱甘肽S-转移酶可能参与花粉的成熟,并影响花粉的繁殖力。与程序性细胞死亡有关的衰老相关的半胱氨酸蛋白酶可能主要与无标题大白菜的自花粉识别有关。这项研究将有助于进一步研究十字花科孢子状SI的分子机制。

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