转化药用野生稻TAC克隆的水稻芽期耐盐性分析

Analysis of Salinity-Tolerance at Germination Stage in Transgenic Rice Transformed an Oryza officinalis TAC Clone

为了解转药用野生稻TAC(Transformation-competent artificial chromosome)基因克隆的水稻株系芽期耐盐性,本研究分别以TAC载体插入片段两端的抗性标记Hpt和SacB检测T_(4)代16个转化株系,并利用0.3%和0.6%的NaCl溶液处理转化TAC克隆的阳性株系及其受体的种子,研究盐胁迫对转基因株系种子萌发相关性状的影响。结果表明,Hpt和SacB均从16个转基因株系中扩增出目的片段。在盐胁迫下,16个转基因株系和受体亲本的发芽势、发芽率、芽长、主根长和根数等性状都显著下降,其中根数和主根长的下降程度最大,减少达72.0%和69.6%。相对发芽势、相对发芽率、相对芽长、相对主根长和相对根数等指标,以‘华粳籼74’(HJX74)为受体的7个转基因株系与其受体亲本相比,全部或大部分株系无显著差异;而以‘日本晴’为受体的9个转基因株系与其受体亲本相比,部分株系与受体相比差异显著,其中TR6-1-6-1、TR19-3-5-6、NAC3-2-4-2、NAC17-1-6-3、STK2-1-2-2和STK33-5-3-1等株系在0.3%或0.6%盐浓度胁迫下的相对发芽率(相对盐害率)显著低于受体‘日本晴’。结合相对盐害率和5个性状的隶属函数值评价表明,TR19-3-5-6和NAC3-2-4-2的耐盐性显著优于其受体亲本。

In order to evaluate the seed germination salt tolerance in transgenic rice transformed an Oryza officinalis TAC(transformation-competent artificial chromosome)clone,sixteen transgenic rice lines of the T_(4) generation were verified by the Hpt and SacB markers flanking the TAC.In this study,and the seeds of positive lines with their recipients were treated by 0.3%and 0.6%NaCl solutions respectively for investigating the characteristics of seed germination under salt stress.The results showed that 16 transgenic lines were confirmed by the Hpt and SacB.The germination rate,shoot length,main root length,and root number of the 16 transgenic lines and recipient parents decreased significantly under salt stress.Especially,the root number and main root length recorded the highest reduction up to 72.0%and 69.6%respectively.All or most of the 7 transgenic lines with’Huajingxian74’(HJX74)as the recipient showed no significant difference in terms of relative germination energy,relative germination rate,relative shoot length,relative main root length and relative root number compared with the recipient.Interestingly,some of 9 transgenic lines with’Nipponbare’as the recipient were significantly different from the recipient,the rel ative germination rate of TR6-1-6-1,TR19-3-5-6,NAC3-2-4-2,NAC17-1-6-3,STK2-1-2-2 and STK33-5-3-1 were significantly lower when treated with 0.3%or 0.6%NaCl.Combining the relative salt damage rate and the evaluation of the membership function value of 5 traits,TR19-3-5-6 and NAC3-2-4-2 were significantly more salt tolerant than their recipient parents.