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为提高小麦的耐盐性,以农杆菌介导的生长点转化法将TaCHP基因导入小麦品种‘济南17’和‘济麦22’中。经潮霉素涂抹和PCR检测,T0代转基因植株的阳性率分别为6.7%和5.8%;T1代阳性率为11.1%和19.0%;T2代阳性率为4.3%(JN17,1个株系)和9.4%(JM22,5个株系)。转基因株系现已种植到T5代,遗传分析表明该基因已在T5代转基因株系中稳定表达。T5代阳性株系显示了明显耐盐性,已进行盐碱地的安全性中间试验。抗逆生理指标分析显示,NaCl胁迫下,与亲本小麦对照相比,转基因株系的脯氨酸含量升高,MDA含量降低,POD和CAT酶活都有所增加。表明TaCHP基因通过渗透调节和维持氧化还原稳态提高了小麦对盐胁迫的耐受性。
In order to improve the salt tolerance of wheat, Agrobacterium tumefaciens-mediated growth point transformation was used to transfer the TaCHP gene into wheat varieties ’Jinan 17’ and ’Jimai 22’. The positive rates of T0 transgenic plants were 6.7% and 5.8% respectively by hygromycin smear and PCR; the positive rates of T1 were 11.1% and 19.0%; the positive rates of T2 were 4.3% (JN17 and 1 strain) And 9.4% (JM22, 5 lines). Transgenic lines have been planted to T5 generation, genetic analysis showed that the gene has been stably expressed in T5 transgenic lines. T5 generation positive lines show significant salt tolerance and have been tested for safety in saline-alkali lands. The analysis of physiological indexes showed that proline content, MDA content and POD and CAT activity of transgenic lines were increased under NaCl stress compared with the parental wheat control. The results showed that TaCHP increased salt tolerance of wheat through osmotic adjustment and maintain of redox homeostasis.