CAJ | 학술논문

以携带潮霉素磷酸转移酶基因的pBIG3C为转化载体，根癌农杆菌EHA105为转化介体，对樱桃干腐病菌LXS230101分生孢子进行转化。结果表明，樱桃干腐病菌的最优转化体系为：α型分生孢子浓度为106个/mL，培养基中添加200μmol/mL乙酰丁香酮（AS），共培养温度和时间分别为25℃和72 h，其转化效率为1×106个α型分生孢子产生686个转化子。随机选取转化子进行PCR和Southern blot 鉴定，发现T-DNA已整合进樱桃干腐病菌基因组中；转化子在不含潮霉素的PDA培养基中连续培养5代后，仍表现出对潮霉素的抗性，表明外源基因能在樱桃干腐病菌中稳定遗传。
이휴대조매소린산전이매기인적pBIG3C위전화재체，근암농간균EHA105위전화개체，대앵도간부병균LXS230101분생포자진행전화。결과표명，앵도간부병균적최우전화체계위：α형분생포자농도위106개/mL，배양기중첨가200μmol/mL을선정향동（AS），공배양온도화시간분별위25℃화72 h，기전화효솔위1×106개α형분생포자산생686개전화자。수궤선취전화자진행PCR화Southern blot 감정，발현T-DNA이정합진앵도간부병균기인조중；전화자재불함조매소적PDA배양기중련속배양5대후，잉표현출대조매소적항성，표명외원기인능재앵도간부병균중은정유전。
We developed an Agrobacterium-mediated transformation system for P.perniciosa by using the α-conidia of strain LXS230101 as transformation recipients ,A.tumefacien strain EHA105 carring plasmid pBIG3C har-boring the hygromycin B phosphotransferase gene ( hph) .Successful transformation of P.perniciosa was performord and the highest effiency reached on 686 transformants per 1 ×106 spores.The optimal transformation conditions were that 1 ×106 spores per milliliter of P.perniciosa α-conidia suspension were co-cultured with Agrobacterium cells at 25 ℃for 72 h,in the presence of Co-culture medium containing acetosyringone ( AS) at 200 μmol/mL.The trans-formants were verified by PCR amplification and by Southern blot analysis with the hph primers and probe ,respec-tively.The results showed that all the detected transformants could be amplified the target bands and the T -DNA was inserted into the genome of P.perniciosa.In addition,the transformants were stable when grown on PDA medium without hygromycin for five times .