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采用基因枪转化法用带有β-葡糖醛酸酶(GUS)报告基因的质粒pGUS6L20转化小麦条锈菌,建立了优化的转化体系;在转化当代的条锈菌中得到了GUS基因瞬时表达的菌株;连续继代转接培养后,在转化后第1代(T1)和第2代(T2)条锈菌中检测到了GUS基因稳定表达的菌株;利用GUS报告基因和毒性标记相结合的方法,经过3代筛选鉴定,在小麦条锈菌中国鉴别寄主尤皮Ⅱ号和抗引655上,各获得1个稳定的毒性突变体,经PCR及PCR-southern b lot证实外源片段已整合到毒性突变菌株的基因组中。该研究表明基因枪转化法是获得条锈菌毒性突变体的有效方法。
The plasmid pGUS6L20 with β-glucuronidase (GUS) reporter gene was used to transform wheat stripe rust by using gene gun method, and an optimized transformation system was established. The transient expression of GUS gene was obtained in the transformation of contemporary stripe rust Strain GUS gene was stably expressed in the first passage (T1) and second generation (T2) of the transformed bacterium after continuous subculture; the combination of the GUS reporter gene and the toxicity marker Method. After three generations of screening, a stable virulent mutant was obtained from the two Chinese virulence testatives, Yupi II and Yikou 655, respectively. The PCR products were confirmed by PCR and Southern blot analysis Into the genome of the virulent mutant strain. This study shows that particle gun transformation is an effective method to obtain stripe rust mutant.