小麦耐水分胁迫突变体生理及RAPD特性分析

检验小麦耐水分胁迫突变体对渗透胁迫环境的耐受能力并对其生理生化及分子特性进行研究。测定突变体愈伤组织在甘露醇,NaCl和聚乙二醇(PEG-6000)模拟的渗透胁迫下的相对生长量及其在20%甘露醇胁迫下游离脯氨酸含量、可溶性糖含量、可溶性蛋白含量等生理生化指标,并进一步检测了突变体再生苗的K /Na 含量及可溶性蛋白质组成和基因组DNA RAPD多态性等生化和分子特征。突变细胞系可以在对照不能生长的20%甘露醇、1.5%NaCl和20%PEG-6000胁迫条件下,分别表现出14.5%,12.8%,41.8%的相对生长量;在20%甘露醇胁迫条件下突变细胞系游离脯氨酸积累量为对照的80%,可溶性糖积累量为对照的1.2倍,可溶性蛋白含量为对照的1.3倍。在相同浓度的甘露醇模拟的渗透胁迫环境中突变体再生植株比对照植株相能维持较高的K /Na 比值。与对照相比,耐水分胁迫突变体再生植株可溶性蛋白SDS-PAGE发生显著变化:6条新可溶性蛋白谱带出现在突变体再生植株中,同时对照系中的1条可溶性蛋白谱带在突变株中缺失。突变体植株与对照株RAPD带型呈现一定的多态性。所研究的小麦耐水分胁迫突变体是一个具有较强渗透胁迫耐受能力,可用于进一步育种工作的良好中间材料。

Physiology and RAPD Identification of a Water-stress Resistance Mutant of Wheat

To test the tolerance-resistant ability of a water-stress resistance mutant of wheat and study it's physiology and molecular Identifications.The relative growth rate of the mutant calluses were tested under the stress of mannitol,NaCl and PEG-6000.The accumulation of free praline,soluble sugars and soluble proteins in the mutant calluses also were tested under the stress of 20% mannitol.Sds-PAGE and RAPD were carried out to study the physiology and molecular identifications of the regenerated mutant plants.The result showed that the mutant calluses could retain the relative growth rate at 14.5%,12.8% and 41.8%,when they were planted on the media contain 20% mannitol,1.5% NaCl or 20% PEG-6000,while the control calluses could not grow under each of these stress circumstances.The accumulation of free praline in the mutant calluses is about 80% of that of the control under the stress of 20% mannitol.On the contrary,the accumulation of soluble sugars and soluble proteins in the mutant calluses is about 1.2 and 1.3 times of that of the control.Growing in a osmotic-stressed environment triggered by same mannitol concentration,the regenerated mutant plants could maintain higher K /Na than that of control.6 new soluble proteins appeared and 1 protein of the control disappeared in the SDS-PAGE map of the mutant.Mutant variations were found at DNA level through the RAPD analyzing.The results demonstrated these mutants could be used as new water-stress resistance seeding resources.