摘要
【目的】鉴定西番莲顶枯病病原菌种类,明确其分类地位,为西番莲顶枯病的科学防控及西番莲产业的持续快速发展提供理论依据。【方法】采用植物组织分离法对首次在广西南宁市发现为害西番莲的新病害——西番莲顶枯病进行病原菌分离,测定其致病性,观察其形态特征,分析其核糖体转录间隔区(ITS)、钙调蛋白(CAL)和交配型基因(ApMat)的基因序列,鉴定引起西番莲顶枯病的病原菌。【结果】从具有典型西番莲顶枯病症状植株中分离到同一形态真菌,命名为TK2A2CLA6菌株,其孢子与自然发病西番莲枝条上切片观察到的病原菌孢子形态一致,进行顶芽接种致病性测定均能发病,且发病症状与自然发病症状一致,再分离得到的病原菌与原接种病原菌相同。TK2A2CLA6菌株菌落圆形或近圆形,正面产生发达且较致密的灰白色气生菌丝;菌落背面初期为浅黄色,后期中央浅墨色、周围浅黄色,在PDA培养基上培养7 d后偶有橙色孢子堆产生;菌丝平均生长速率为18.89 mm/d;分生孢子无色,近圆柱状钝圆,另一端钝圆或略细,一般具1~2个油球,大小为(13.03~17.75)μm×(3.75~5.25)μm。对TK2A2CLA6菌株的ITS V1~V4区、CAL和ApMat基因序列进行PCR扩增,将基因序列在NCBI中进行BLAST比对分析,发现其与暹罗炭疽菌(Colletotrichum siamense)多个序列的同源性达100%。以ITS、CAL和ApMat基因序列进行多基因系统发育进化树分析,TK2A2CLA6菌株与C.siamense LF177和C.siamense JH-Coll001以100%的自展支持率聚为独立的1支,确定引起西番莲顶枯病的致病菌为暹罗炭疽菌。【结论】使用形态学方法结合多基因系统进化树分析,明确危害广西南宁西番莲顶枯病的病原菌为暹罗炭疽菌。
【Objective】The present experiment aimed to identify the pathogen species of top blight of Passiflora edulis and clarify its taxonomic status, so as to provide a theoretical basis for the scientific prevention and control of top blight of P. edulis and the sustainable and rapid development of the P. edulis industry.【Method】The pathogen of P. edulis top blight, a new disease found in Nanning for the first time, was isolated by plant tissue isolation method, its pathogenicity was determined, and its morphological characteristics were observed, and the genetic sequence of its ribosomal transcribed spacer(ITS),calmodulin(CAL) and mating type gene(ApMat) were analyzed. To identify the pathogen causing top blight of P. edulis.【Result】The same form of fungi was isolated from the plants with typical symptoms of top blight of P.caerulea,named TK2 A2 CLA6 strain, and its spore morphology was consistent with the pathogen spore morphology observed from the slices of natural diseased branches of P.caerulea,which could be infected by the pathogenicity test of terminal bud inoculation, and the disease symptoms were consistent with the natural disease symptoms. The pathogenic bacteria obtained by re-separation were the same as the original inoculated pathogenic bacteria. The colony of TK2 A2 CLA6 was round or nearly round, and developed and dense gray-white aerial hyphae were produced on the front of the colony, while the back of the colony was light yellow at the early stage. At the later stage, the center of the fungus was light black and the periphery was light yellow, and orange spore piles were occasionally produced after 7 days of culture on PDA medium. 18.89 mm/d;Colorless conidia was nearly cylindrical and obtuse, the other end was obtuse or slightly thin, generally with 1-2 oil globules, the size was(13.03-17.75)μm×(3.75-5.25)μm. The genetic sequence of TK2 A2 CLA6 strain was amplified by PCR,and the genetic sequence was analyzed by BLAST in NCBI. The results showed that the homology between TK2 A2 CLA6 st
作者
黄艳花
崔忠吉
欧善生
黄远光
蒙成
李杨秀
HUANG Yan-hua;CUI Zhong-ji;OU Shan-sheng;HUANG Yuan-guang;MENG Cheng;LI Yang-xiu(Guangxi Agricultural Vocational and Technical University,Nanning 530007,China;Daoshui Town Agricultural Technology Extension Station,Changzhou District,Wuzhou City,Guangxi,Wuzhou,Guangxi 543006,China)
出处
《西南农业学报》
CSCD
北大核心
2022年第8期1826-1832,共7页
Southwest China Journal of Agricultural Sciences
基金
国家现代农业产业技术体系广西创新团队建设项目(nycytxgxcxtd-17-03)
广西农业科技自筹经费项目(Z202006)。
