火龙果赤斑病病原菌的分离与鉴定
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摘要
通过对广东省湛江南亚热带植物园的火龙果病害调查,发现1种新的病害。其症状表现为病斑红色或褐色,圆形或近圆形,少数不规则形,稍凹陷,外缘有黄色晕圈,严重时可呈现褐色坏死斑。从田间感病火龙果茎的病健交界处用PDA培养基分离,获得了菌株BJ28,并对其进行了致病性测定、形态学观察和分子序列分析。结果表明,经柯赫氏法则验证菌株BJ28为致病病原菌。采用引物ITS1和ITS4对致病菌的ITS1-5.8S-ITS2基因片段进行扩增结果显示,该致病菌的ITS1-5.8S-ITS2序列与HQ608063(Nigrospora sphaerica)同源性最高,达到100%。通过培养形状、形态学观察和ITS1-5.8S-ITS2序列分析,将火龙果赤斑病病原菌鉴定为球黑孢菌[Nigrospora sphaerica(Sacc.)E.W.Mason]。
A new disease on pitaya was observed in South Asia Tropical Plant Park in Zhanjiang of Guangdong province in China. The typical symptoms were round reddish-brown necrotic lesions surrounded by yellow haloes, and as the disease progressed, smaller lesions coalesced into larger irregularly shaped areas that became gray to dark brown in color.BJ28 strain was isolated from boundary of the spot on rotted stems of field pitaya plants with PDA. Pathogenicity test,morphology observation and molecular sequence analysis were conducted. The results showed that the strain BJ28 was the pathogen by Koch′s Postulates. Based on ITS1-5.8 S-ITS2 sequences analysis with primers of ITS1 and ITS4, sequences of the pathogen showed the highest homology with Nigrospora sphaerica(Gen Bank Accession No. HQ608063), and the value reached 100%. And the pathogen was identified as Nigrospora sphaerica(Sacc.) E.W. Mason.
A new disease on pitaya was observed in South Asia Tropical Plant Park in Zhanjiang of Guangdong province in China. The typical symptoms were round reddish-brown necrotic lesions surrounded by yellow haloes, and as the disease progressed, smaller lesions coalesced into larger irregularly shaped areas that became gray to dark brown in color.BJ28 strain was isolated from boundary of the spot on rotted stems of field pitaya plants with PDA. Pathogenicity test,morphology observation and molecular sequence analysis were conducted. The results showed that the strain BJ28 was the pathogen by Koch′s Postulates. Based on ITS1-5.8 S-ITS2 sequences analysis with primers of ITS1 and ITS4, sequences of the pathogen showed the highest homology with Nigrospora sphaerica(Gen Bank Accession No. HQ608063), and the value reached 100%. And the pathogen was identified as Nigrospora sphaerica(Sacc.) E.W. Mason.
引文
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[3]Lan G B,He Z F,Xi P G,et al.First report of brown spot disease cau sed by Neoscytalidium dimidiatum on Hylocereus undatus in Guangdong,Chinese Mainland[J].Plant Disease,2012,96(11):702.
[4]易润华,甘罗军,晏冬华,等.火龙果溃疡病病原菌鉴定及生物学特性[J].植物保护学报,2013,40(2):102-108.
[5]Liou M R,Huang C L,Liou R F.First report of Cactus virus X on Hylocereus undatus(Cactaceae)in Taiwan[J].Plant Disease,2001,85(2):229.
[6]梁秋玲,韦健,李孝云,等.火龙果茎腐病病原鉴定及室内药剂毒力测定[J].中国南方果树,2011,40(1):9-12.
[7]崔志婧,王奕文,于岳,等.上海市进口火龙果软腐病病害分析[J].微生物学通报,2011,38(10):1499-1506.
[8]陈圆,严婉荣,赵志祥,等.海南省火龙果炭疽菌病病原鉴定及有效药剂筛选[J].基因组学与应用生物学,2017,36(2):638-643.
[9]林武镇,廖富荣,陈细红,等.火龙果细菌性软腐病菌的分离与鉴定[J].植物病理学报,2015,45(2):220-224.
[10]朱迎迎,李敏,高兆银,等.火龙果炭疽病病原菌的鉴定及生物学特性研究[J].南方农业学报,2016,47(1):59-66.
[11]方中达.植病研究方法[M].第3版.北京:中国农业出版社,1998.
[12]Zhang L X,Song J H,Tan G J,et al.First report of leaf blight caused by Nigrospora sphaerica on curcuma in China[J].Plant Disease,2011,95(9):1190.
[13]Domsch K H,Gams W,Anderson T H.Compendium of Soil Fungi[M].London:Academic Press,1980:515-516.
[14]Zhao H,Liu H Y,Yang X.S,et al.First report of Nigrospora leaf blight on sesame caused by Nigrospora sphaerica in China[J].Plant Disease,2014,98(6):842.
[15]Dutta J,Gupta S,Thakur D,et al.First report of Nigrospora leaf blight on tea caused by Nigrospora sphaerica in India[J].Plant Disease,2015,99(3):417.
[16]Li YG,Huang M H,Sun L P,et al.Occurrence of leaf spot of calabash caused by Nigrospora sphaerica in Georgia[J].Plant Disease,2016,100(7):1506.
[17]赵世学,符碧,海戴俊,等.10种杀细菌剂对火龙果细菌性茎腐病菌的毒力测定[J].中国南方果树,2016,45(2):84-86.
[18]郑伟,王彬,马骁,等.6种杀菌剂防治火龙果茎斑病的田间药效[J].江苏农业科学,2015,43(2):140-141.
[19]李敏,胡美姣,高兆银,等.杀菌剂对火龙果革节孢属病菌的室内毒力测定[J].热带农业科学,2012,32(4):72-75.