摘要
几丁质酶(Chitinase,CHi)是一类广泛分布在高等植物中的水解酶,作用于真菌的细胞壁,是植物抵御真菌病害的重要功能蛋白。本研究从肉桂转录组中挖掘出一个几丁质酶基因CcCHI,并对该基因进行了克隆与原核表达研究。结果表明,CcCHI基因长1206 bp,经生物信息学分析,该基因序列编码402个氨基酸,蛋白理论相对分子量为43.22 kD,等电点为9.13。经序列比对与系统进化树分析,CcCHI与沉水樟中的Ⅴ类几丁质酶亲缘关系最近。通过构建pET-32a(+)-CcCHI原核表达质粒并转化大肠杆菌中进行诱导表达后,经真菌抑菌试验发现该蛋白具有一定的抗真菌活性。本研究首次发掘了肉桂中的几丁质酶基因,并初步研究了其蛋白的抗真菌活性,为进一步研究肉桂抗真菌病害的分子机制与抗病品种选育提供理论基础。
Chitinase(CHi)is a kind of hydrolase widely distributed in higher plants with fungal cell wall hydrolysis to defense against fungal diseases.In this study,a chitinase gene CcCHI was cloned and prokaryotic expressed based on the cinnamon transcriptome.After the bioinformatics analysis,the CcCHI sequence length was 1206 bp and it encoded 402 amino acids with a theoretical molecular weight of the protein of 43.22 kD and an pI of 9.13.The sequences alignment and phylogenetic tree analysis showed that the CcCHI was close to the classⅤchitinase in submerged camphor.The p ET-32a(+)-CcCHI recombinant plasmid was constructed and transformed into the Escherichia coli,and the p ET-32a(+)-CcCHI recombinant protein was obtained via the prokaryotic expression.The antifungal activity was found by the fungal inhibition test in protein.the CcCHI gene was first cloned in this study,and we preliminarily investigate the antifungal activities of the CcCHI protein,which may provide a theoretical basis for further study on the molecular mechanisms of defensing fungal diseases and breeding disease resistant cultivars in cinnamon.
作者
闵远洋
李韵雪
吕佳
麦晋贤
廖沛然
杨全
Min Yuanyang;Li Yunxue;LüJia;Mai Jinxian;Liao Peiran;Yang Quan(Guangdong Probincial Research Center on Good Agricultural Practice&Comprehensive Agricultural Development Engineering Technology of Cantonses Medicinal Materials,Guangzhou Comprehensive Experimental Station Chinses Materia Medical Industry Technology System,Key Laboratory for Production&Development of Cantonese Medicinal Materials Under State Administration of TCM,School of Traditional Chinese,Guangdong Pharmaceutical University,Guangzhou,510006)
出处
《分子植物育种》
CAS
北大核心
2022年第7期2244-2251,共8页
Molecular Plant Breeding
基金
广东省中医药局广东省重点领域研发计划(20215007)
罗定肉桂产业发展基础研究项目(2018-028)
国家中药材产业技术体系建设专项(CARS-21)共同资助。
