A low molecular weight proteome comparison of fertile and male sterile 8 anthers of Zea mays.
A low molecular weight proteome comparison of fertile and male sterile 8 anthers of Zea mays.
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玉米可育和雄性不育 8 个花药的低分子量蛋白质组比较。
DOI：
10.1111/j.1467-7652.2012.00721.x
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发表时间：
2012
影响因子：
13.8
通讯作者：
Walbot,Virginia
中科院分区：
文献类型：
--
作者：
Wang,Dongxue;Adams,ChristopherM;Fernandes,JohnF;Egger,RachelL;Walbot,Virginia
在玉米花药发育过程中，体细胞室细胞分化以支持花粉母细胞的减数分裂。减数分裂是花药生长过程中的一个重要事件，对于植物育性至关重要，因为花粉含有单倍体精子。玉米雄性不育突变体的一个子集表现出减数分裂失败，包括ms8（雄性不育8），其中性母细胞停滞为二分体，并且小室体细胞表现出多种缺陷。在生物三次重复和技术三次重复中分析了系统蛋白质组图谱，将 8 个花药与减数分裂前和减数分裂阶段的可育同胞样本进行比较；通过 1-D PAGE 分离 3.5 至 20 kDa 的蛋白质，用 Lys-C 裂解，然后使用 LTQ Orbitrap Velos MS 范例进行测序。 359 个蛋白质被鉴定为具有两个或多个指定肽，其中每个肽都被计数至少两次或多次（肽错误发现率 (FDR) 为 0.4%，蛋白质 FDR 为 0.2%）； 2761 个蛋白质被鉴定为具有一种或多种指定肽（0.4% 肽 FDR 和 7.6% 蛋白质 FDR）。阶段特异性蛋白质表达为早期花药发育提供了候选阶段标记，与不育花药相比，在可育花药中特异性表达的蛋白质提供了有关减数分裂调节的重要线索。这项研究检测到的蛋白质中有 49% 对于独立的完整花药蛋白质组来说是新的，并且许多被自动玉米基因组注释遗漏的小蛋白质也得到了验证；这些结果表明关注低分子量蛋白质的价值。讨论了独特表达蛋白质的作用以及低分子量蛋白质的质谱分析方法。
During maize anther development, somatic locular cells differentiate to support meiosis in the pollen mother cells. Meiosis is an important event during anther growth and is essential for plant fertility as pollen contains the haploid sperm. A subset of maize male sterile mutants exhibit meiotic failure, includingms8(male sterile 8) in which meiocytes arrest as dyads and the locular somatic cells exhibit multiple defects. Systematic proteomic profiles were analysed in biological triplicates plus technical triplicates comparingms8anthers with fertile sibling samples at both the premeiotic and meiotic stages; proteins from 3.5 to 20 kDa were fractionated by 1‐D PAGE, cleaved with Lys‐C and then sequenced using a LTQ Orbitrap Velos MS paradigm. Three hundred and 59proteins were identified with two or more assigned peptides in which each of those peptides were counted at least two or more times (0.4% peptide false discovery rate (FDR) and 0.2% protein FDR); 2761 proteins were identified with one or more assigned peptides (0.4% peptide FDR and 7.6% protein FDR). Stage‐specific protein expression provides candidate stage markers for early anther development, and proteins specifically expressed in fertile compared to sterile anthers provide important clues about the regulation of meiosis. 49% of the proteins detected by this study are new to an independent whole anther proteome, and many small proteins missed by automated maize genome annotation were validated; these outcomes indicate the value of focusing on low molecular weight proteins. The roles of distinctive expressed proteins and methods for mass spectrometry of low molecular weight proteins are discussed.