摘要
由枝状枝孢霉(Cladosporium cladosporioides)侵染引起的杧果露水斑病是近年杧果上新发生的病害,主要为害果实。明确该菌能否穿透果皮侵入果肉,可为防治该病害选择合适的药剂提供依据。本研究将绿色荧光蛋白基因gfp和潮霉素抗性基因hygB表达盒插入了经定点突变后Cl.cladosporioides的ITS序列中,成功构建了pITGH重组载体,将其用PEG介导方法转化杧果露水斑病菌原生质体,获得了在菌丝、分生孢子、萌发的分生孢子中稳定发出绿色荧光的GFP标记菌株,该标记菌株与野生菌的菌落形态、颜色及致病力无明显差异,生长速度明显慢于野生菌。用该标记菌株观察侵染过程,发现杧果露水斑病菌可以在杧果果皮上侵染和扩展,但不能扩展至果肉中。该结果为选择农药类型提供了依据,也为进一步研究该病菌在活体杧果上的生物学、预测病害和通过观察定殖情况确定杧果品种的抗病性等奠定了良好的材料基础。
Mango sooty blotch disease, caused by the fungal pathogen Cladosporium cladosporioides, is a major disease of mango that only infects mango fruit. To provide a basis for selecting fungicides to control this disease, it is necessary to confirm whether Cl. cladosporioides could penetrate skin into flesh. The green fluorescent protein（GFP）-expression vector pITGH was constructed by inserting a gfp-hygB cassette into rDNA-ITS of Cl. cladosporioides using PCR-based site-directed mutagenesis, then was used to transform the Cl. cladosporioides isolate by a PEG-mediated method. GFP-marked Cl. cladosporioides with stable green fluorescence in hyphae, germinating conidia was acquired. There were no difference between GFP-marked Cl. cladosporioides and wild type in morphology, color, and virulence, but GFP-marked Cl. cladosporioides was slower than the wild type strain in growth rate. The infection process of the GFP-marked strains indicated that Cl. cladosporioides only infected and developed on mango skin, but could not extend to mango flesh. The results provided a basis for selecting fungicide types, and would be also useful for further study on the biology in vivo, predicting the mango sooty blotch disease, and evaluating resistance of mango varieties to Cl. cladosporioides by means of the colonization of GFP-marked Cl. cladosporioides on mango.
作者
刘晓妹
杨永利
张贺
李鸿鹏
吴秋玉
钟昌开
蒲金基
LIU Xiaomei;YANG Yongli;ZHANG He;LI Hongpeng;WU Qiuyu;ZHONG Changkai;PU Jinji(Institute of Tropical Agriculture and Forestry,Hainan University / Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests,Ministry of Education,Haikou,Hainan 570228,China;Environment and Plant Protection Institute,Chi-nese Academy of Tropical Agricultural Sciences / Key Laboratory of Integrated Pest Management of Tropical Crops,Ministry of Agriculture,Haikou,Hainan 571101,China)
出处
《热带作物学报》
CSCD
北大核心
2018年第11期2235-2240,共6页
Chinese Journal of Tropical Crops
基金
海南省自然科学基金(No.317031)
海南省重大科技专项(No.ZDKJ2017003)
海南省高等学校教育教学改革研究项目(No.Hnjg2016ZD-2
No.hdjy1606)
关键词
杧果
露水斑病
ITS定点突变
GFP标记
侵染部位
mango
sooty blotch
site-directed mutagenesis of rDNA-ITS
GFP marker
infection site
