摘要
在高山地区和许多地区的低温季节里,植物的冷害是一个普遍的农业灾害。冷胁迫导致种子发芽率和活力下降,对种苗和植株的生长以及授粉都造成了不利影响,这将导致生物产量的降低。PEG引发能够提高种子的冷忍耐力,但是PEG引发的机制仍然未能得到明确的解答。为了对大豆种子冷害和PEG引发获得更好的理解,我们在生理生化研究的基础上应用蛋白质组学方法进行分析。
生理生化研究结果表明,冷处理导致种子发芽率下降到5%,活力指数,脱氢酶活性大幅度降低,电导率明显上升,而PEG引发的种子在冷处理后却能够使种子的各项指标恢复到正常水平,且使发芽率提高了90%。
通过适于大豆种子蛋白质组学的研究方法,我们获得了以下研究结果:
(1)通过双向凝胶电泳,我们从对照(CK)、冷处理(CT)、PEG处理(PT)、PEG处理后冷处理(PC)的大豆种子下胚轴中分别得到706、585、670、658个蛋白质点。
(2)25个差异蛋白和9个感兴趣的高丰度蛋白共34个蛋白质点得到质谱鉴定,这些蛋白主要涉及细胞的生长与分裂、疾病与防御、蛋白质定位与贮藏。
(3)发现核苷二磷酸激酶、Os12g0160400转录蛋白被冷胁迫诱导且只和冷胁迫相关,不受渗透胁迫的影响。
(4)热激蛋白90、微管蛋白A、35 kDa种子成熟蛋白、钼蝶呤合成酶、大豆伴豆球蛋白A1ab1b前体、大豆胰蛋白酶抑制剂都能被PEG引发所诱导,与渗透调控紧密相关,它们参与了种子的抗冷。
(5)种子成熟蛋白PM25与冷害和PEG引发都相关,可能是一种新的胁迫蛋白。
对于大豆种子冷害与渗透调控的研究结果进行分析,我们发现冷胁迫导致蛋白质点数量的下降,而PEG引发能够提高大约70个蛋白点,发现两个冷胁迫蛋白核苷二磷酸激酶、Os12g0160400转录蛋白,以及一种新的胁迫蛋白种子成熟蛋白PM25。这些研究结果对种子的冷害提供了一种认识的新视角,也为渗透调控提高种子抗冷提供了证据和可能的机制,对种子的防护具有重要作用。
Chilling injury is a prevalent agricultural disaster in alp zones and low temperature seasons of many areas. Chilling stress can reduce seed germination, seed vigor, seedling growth, plant growth and pollination, which will result in yield-decrease. PEG priming could induce chilling tolerance in seeds. The mechanism of PEG priming is yet to be resolved. To gain better understanding of chilling injury and PEG priming in soybean (Glycine max) seeds, we conducted a proteomic analysis based on the study of the physiological and biochemical levels.
Chilling treatment resulted in decreased germination rate, vigor index, dehydrogenase activity, and increased relative electrical conductivity, which were recuperated by PEG priming at subsequently chilling treatment. Germination rate improved 90%.
The results on the proteomic aspects were obtained through the method fitted for soybean seed proteome.
(1) Approximately 706, 585, 670, 658 protein spots from control (CK), chilling treatment (CT), PEG treatment (PT), and chilling treatment after PEG priming (PC) were separated, respectively.
(2) Twenty-five distinct and nine high abundant proteins were identified. These proteins mainly involved in cell growth/division, disease/defense, protein destination and storage.
(3) We found that nucleoside diphosphate kinase and Os12g0160400 could have more expression at chilling, and have no influence by osmoconditioning.
(4) A number of proteins like heat-shock protein 90, tubulin A, 35 kDa seed maturation protein, molybdopterin synthase, soybean proglycinin A1ab1b homotrimer, soybean trypsin inhibitor, induced by PEG priming, further resisted chilling stress.
(5) The seed maturation protein PM25 was induced both chilling and PEG priming suggested that the protein related with these two factors, which may be a novel stress protein.
To summary, we found that chilling stress lead to decline quantity of protein spots, but enhanced approximately 70 protein spots by PEG priming. And nucleoside diphosphate kinase and Os12g0160400 were chilling stress protein, as well as seed maturation protein PM25 is a novel stress protein.
The results provided not only new insights into chilling injury but also the evidence and mechanism of osmoconditioning in preventing chilling injury in seeds.
引文
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