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1Proteomic analysis of seed germination under salt stress in soybeans显示文摘Soybean (Glycine max (L.) Merrill) is a salt-sensitive crop,and its production is severely affected by saline soils.Therefore,the response of soybean seeds to salt stress during germination was investigated at both physiological and proteomic levels.The salt-tolerant cultivar Lee68 and salt-sensitive cultivar N2899 were exposed to 100 mmol/L NaCl until radicle protrusion from the seed coat.In both cultivars,the final germination percentage was not affected by salt,but the mean germination times of Lee68 and N2899 were delayed by 0.3 and 1.0 d,respectively,compared with controls.In response to salt stress,the abscisic acid content increased,and gibberellic acid (GA1+3) and isopentenyladenosine decreased.Indole-3-acetic acid increased in Lee68,but remained unchanged in N2899.The proteins extracted from germinated seeds were separated using two-dimensional gel electrophoresis (2-DE),followed by Coomassie brilliant blue G-250 staining.About 350 protein spots from 2-DE gels of pH range 3 to 10 and 650 spots from gels of pH range 4 to 7 were reproducibly resolved,of which 18 protein spots showed changes in abundance as a result of salt stress in both cultivars.After matrix-assisted laser desorption ionization-time of flight-mass spectroscopy (MALDI-TOF-MS) analysis of the differentially expressed proteins,the peptide mass fingerprint was searched against the soybean UniGene database and nine proteins were successfully identified.Ferritin and 20S proteasome subunit β-6 were up-regulated in both cultivars.Glyceraldehyde 3-phosphate dehydrogenase,glutathione S-transferase (GST) 9,GST 10,and seed maturation protein PM36 were down-regulated in Lee68 by salt,but still remained at a certain level.However,these proteins were present in lower levels in control N2899 and were up-regulated under salt stress.The results indicate that these proteins might have important roles in defense mechanisms against salt stress during soybean seed germination.Xiao-yan XU Rui FAN Rui ZHENG Chun-mei LI De-yue YU 2011Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)2011,12,7:10
2水稻丙酮酸脱羧酶基因OsPDC3功能的初步研究显示文摘植物花粉中存在着活跃的有氧发酵过程。丙酮酸脱羧酶(PDC)作为发酵途径中的关键酶,参与花粉中的能量和物质代谢,在花粉萌发过程中起重要作用。通过反向遗传学的方法对水稻丙酮酸脱羧酶基因OsPDC3的功能进行了初步分析。OsPDC3是一个单外显子基因,编码蛋白与另外4个水稻PDC蛋白间的序列一致性达到77%~82%。表达模式分析显示OsPDC3在花粉中特异表达,GUS组织染色进一步证实其启动子具有花粉特异表达活性。超量表达OsPDC3会使转基因植株叶片中的PDC酶活性上升,表明OsPDC3在体内具有活性功能,能够参与花粉内的生理活动。反义抑制下调了OsPDC3在花粉中的表达水平,但对花粉的离体萌发率没有产生影响,推测这是由于PDC基因间的冗余造成的。陈波 韩斌 2011中国水稻科学2011,25,6:2
3运用BioID技术筛选水稻GS3互作蛋白显示文摘水稻异源三聚体G蛋白系统中的非典型γ亚基GS3,是一个控制籽粒大小的主效数量效应基因座,在调节籽粒大小中发挥负调控因子的功能。BioID(proximity-dependent biotin identification)为邻近蛋白标记技术,其工作原理是生物素连接酶能使其周围的蛋白带上生物素,同时生物素又能和链霉亲和素紧密结合,所以能够利用链霉亲和素偶联的磁珠富集目标蛋白。该技术具有灵敏、高效和周期短等特点,为筛选互作蛋白提供了新方法。为了解析GS3的蛋白调控网络,该研究以水稻原生质体为材料,采用BioID技术对GS3在水稻中的互作蛋白进行了筛选。Western-blot结果表明:融合蛋白Bir AG-GS3在原生质体中成功表达并生物素化GS3邻近蛋白。使用链霉亲和素磁珠富集生物素化后的蛋白,并进行蛋白质谱测序,获得了与GS3邻近的可能存在直接或间接互作的蛋白。将获得的蛋白进行功能富集与注释,并构建蛋白-蛋白互作网络。对部分蛋白进行了BiFC验证,发现GS3可能与ICL、PPDK、RPN7和RH15发生相互作用,涉及能量代谢的调节、种子淀粉物质的储存、泛素-蛋白酶体系统以及凋亡途径等生物过程。杜荣裕 周泽娇 冯嘉琳 刘赛 阳辉勇 李洪清 2018广西植物2018,38,6:1
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