摘要
利用农杆菌(Agrobacterium tumefaciens)介导的转化方法,建立香蕉枯萎病菌1号生理小种(Fusariumoxysporum f.sp cubense race 1)遗传转化体系,并对影响转化效率的因子进行优化,明确了高效转化的条件为:农杆菌OD600为0.15,AS诱导浓度为150μmol/L,诱导时间为7 h,Focr1-N2孢子浓度为1×106个/mL,潮霉素B筛选的浓度为150μg/mL,诱导培养基pH值为5.5,共培养温度为25℃,共培养时间为48 h为最佳条件。构建了包含1 200个突变体的突变体库,并对1 200个突变体进行了致病性测定,初步筛选出致病性丧失突变体20个,致病性显著减弱突变体18个,致病性严重增强突变体53个。
Agrobacterium（Agrobacterium tumefaciens） mediated transformation method was used to establish the genetic transformation system of Fusarium oxysporum f.sp cubense race1,and the factors affecting the transformation efficiency were optimized.Optimized high efficiency conversion conditions were as： OD600 for agrobacterium 0.15,AS（Acetosyringone） inducing concentration 150 μmol/L,inducing time： 7 hours,Focr1 N2 spore concentration 1×106/mL,hygromycin B 150 μg/mL,inducing medium pH5.5,cultivation temperature 25 ℃ and cultivation time 48 hours.A mutant library containing 1 200 mutants was established.There were 20 mutants that had lost their pathogenicity,18 with significantly reduced pathogenicity,53 with a marked increase of pathogenicity.
出处
《热带作物学报》
CSCD
2010年第10期1790-1796,共7页
Chinese Journal of Tropical Crops
基金
国家自然科学基金项目(No.30860160)
科技部国际科技合作项目(No.2007DFA31460)
国家公益性行业(农业)科研专项课题(No.20093049-2)资助
关键词
香蕉枯萎病菌1号生理小种
突变体
遗传转化
Fusarium oxysporum f. sp. cubense （FOC） racel
Mutant
Genetic transformation
