CAJ | 학술논문

采用基因枪转化法用带有β-萄糖醛酸酶(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 blot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.
채용기인창전화법용대유β-도당철산매(GUS)보고기인적질립pGUS6L20전화소맥조수균,건립료우화적전화체계;재전화당대적조수균중득도료GUS기인순시표체적균주;련속계대전접배양후,재전화후제1대(T1)화제2대(T2)조수균중검측도료GUS기인은정표체적균주;이용GUS보고기인화독성표기상결합적방법,경과3대사선감정,재소맥조수균중국감별기주우피Ⅱ호화항인655상,각획득1개은정적독성돌변체,경PCR급PCR-southern blot증실외원편단이정합도독성돌변균주적기인조중,해연구표명기인창전화법시획득조수균독성돌변체적유효방법.
To obtain virulent mutant stains, the optimized transformation system was established and plasmid pGUS6L20 containing the GUS gene was transformed into urediniospores of Puccinia striiformis f. sp. tritici via particle bombardment . The transient expression of GUS gene was identified in the transformed uredinios-pores. The GUS-carrying spores were cultured and increased consecutively. GUS gene was expressed in the uredia and urediniospores of T1 and T2 generation of the stripe rust fungus. Two insertion mutants of increased virulence were identified on wheat cultivars Jubilejina Ⅱ and Kangyin 655, respectively. The presence of GUS gene was detected by polymerase chain reaction (PCR) and Southern blot in these two virulent mutants. The results showed that the particle bombardment was an effective way to obtain the virulent strains of P. striifor-mis.