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目的对空间诱变育成的“航茄”5号(HQ-5)进行RAPD分析,寻找特异扩增条带,试图为空间诱变育种提供分子生物学证据。方法“天水长茄”(TSCQ)和“龙果圆茄”(LGYQ)种子分别搭载“神舟”4号和“神舟”3号飞船进行空间诱变后,通过4代地面自交选育得到“天水长茄诱变第4代(TSCQ-MUT-4)”和“龙果圆茄诱变第4代(LGYQ-MUT-4)”2个突变系;利用后两者分别作为父母本进行杂交得到HQ-5。采用正交设计对其RAPD体系进行优化并进行RAPD分析。结果采用25μL扩增体系(DNA:40 ng,dNTPs:7.5 nmol,primer:10 ng和rTaq聚合酶:1.25 U)进行RAPD反应,发现“TSCQ-MUT-4”与野生型TSCQ相比产生4条差异带,LGYQ-MUT-4与野生型LGYQ相比产生1差异带;HQ-5与父母本相比均有6条差异带,有父母本共有带和特有带,并且保持航天诱变产生的变异带。结论空间诱变可能引起植物DNA水平的变异,有助于选育出新的农作物品种(系)。
OBJECTIVE: To investigate the molecular markers of space-mutation breeding using RAPD analysis of HQ-5. Methods The seeds of “TSCQ” and “LGYQ” were carried on space mutagenesis of “Shenzhou” No.4 and “Shenzhou” Self-breeding on the ground was “2 mutations (TSCQ-MUT-4)” and “LGYQ-MUT-4” Department; the use of the latter two were used as the parents of this hybrid to get HQ-5. Orthogonal design was used to optimize its RAPD system and perform RAPD analysis. Results RAPD reaction was performed using a 25 μL amplification system (DNA: 40 ng, dNTPs: 7.5 nmol, primer: 10 ng and rTaq polymerase: 1.25 U) and found to produce “TSCQ-MUT-4” 4 differential bands, 1 differential band between LGYQ-MUT-4 and wild-type LGYQ. There were 6 differential bands between HQ-5 and parents, shared with parental and endemic bands, The resulting variation band. Conclusion Spatial mutagenesis may cause variation of plant DNA level, which will help to breed new crop varieties (lines).