矮化核桃枝枯病病原菌的形态与分子鉴定
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摘要
为了明确河南省洛阳地区核桃枝条一种新病害的病原种类,对采自田间的典型症状枝条进行了常规组织分离,对单孢菌株进行了形态鉴定、rDNA ITS序列分析及致病性测定。结果表明,病原菌在PDA培养基上菌落初为白色,随着色素的积累,由中心向边缘变为灰绿色,再变成墨绿色,最终全部变成黑色;气生菌丝棉絮毛状,菌落边缘不整齐,近圆形或不规则形。分生孢子器散生或聚生,呈黑色小粒状,分生孢子器球形或近球形,深褐色,具孔口;分生孢子梗短缺,无色,杆状,末端产生分生孢子;分生孢子梭形、纺锤形,无色,无隔,基部钝圆,顶部稍尖,单孢(24.5～29.0)μm×(4.8～7.5)μm。离体枝条接种结果表明,该菌能侵染核桃枝条引起与田间相同的症状。据此将分离物鉴定为葡萄座腔菌Botryosphaeria dothidea。
In order to clarify the pathogen of a new disease of walnut branch in Luoyang, Henan province, routine tissue separation of typical symptomatic branches from the field was carried out, and the morphological identification, rDNA-ITS sequence analysis and pathogenicity determination were performed for single spore strains. The results showed that the colony of the pathogen was initially white in the PDA medium, and with the accumulation of pigment, it was gray-green from the center to the edge; then it turned atrovirens, and eventually it turned black; the aerial hyphae were floccular and hairy, with irregular colony edge and nearly round or irregular shape. Pycnidia were scattered or aggregated, with small black granules, spherical or nearly spherical, dark brown, with orifice; conidiophores were short, colorless, rod-shaped, and at the termina were conidia; conidium was fusiform, colorless, without septum, with obtuse-rounded base, slightly pointed apex, and single spores(24.5-29.0)μm×(4.8-7.5)μm. The results of inoculation in excised branches showed that the pathogen could infect the walnut branches and cause the same symptoms as those in the field. Accordingly, the isolate was identified as Botryosphaeria dothidea.
In order to clarify the pathogen of a new disease of walnut branch in Luoyang, Henan province, routine tissue separation of typical symptomatic branches from the field was carried out, and the morphological identification, rDNA-ITS sequence analysis and pathogenicity determination were performed for single spore strains. The results showed that the colony of the pathogen was initially white in the PDA medium, and with the accumulation of pigment, it was gray-green from the center to the edge; then it turned atrovirens, and eventually it turned black; the aerial hyphae were floccular and hairy, with irregular colony edge and nearly round or irregular shape. Pycnidia were scattered or aggregated, with small black granules, spherical or nearly spherical, dark brown, with orifice; conidiophores were short, colorless, rod-shaped, and at the termina were conidia; conidium was fusiform, colorless, without septum, with obtuse-rounded base, slightly pointed apex, and single spores(24.5-29.0)μm×(4.8-7.5)μm. The results of inoculation in excised branches showed that the pathogen could infect the walnut branches and cause the same symptoms as those in the field. Accordingly, the isolate was identified as Botryosphaeria dothidea.
引文
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[15] 余仲东,赵官成,淡静雅,等.葡萄座腔菌属ITS-nrDNA的分子系统学分析[J].菌物学报,2010,29(2):285-293.
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[21] EI-GOORANI M A.Dieback of grapevine by Botryodiplodia theobromae Pat.In Egypt [J].Phytopathologia Mediterranea,1972,11:210-211.
[22] 陈海燕,田呈明,梁军,等.引起树木溃疡病病原菌Botryosphaeria属及相关无性态分类研究[J].西北林学院学报,2006(6):145-150.
[2] 鲍剑松.核桃主要病害及综合防治措施[J].乡村科技,2017(8):38-39.
[3] 杨克强,程三虎,牛亚胜,等.若干个核桃品种(系)对白粉病的抗性[J].果树科学,1998,15(2):154-157.
[4] 刘世骐.核桃溃疡病的研究[J].安徽农学院学报,1986(2):1-6.
[5] 郭开发,王刚,吴彩兰,等.新疆南疆核桃树腐烂病菌鉴定[J].新疆农业科学,2016(3):496-501.
[6] 韩敏.南疆核桃叶斑病病原与防治技术研究[D].乌鲁木齐:新疆农业大学,2014.
[7] 王璇,马良进,吕全,等.山核桃干腐病病原菌的鉴定[J].浙江农林大学学报,2014,31(2):238-245.
[8] 尹万瑞,朱天辉.核桃枝枯病病原菌生物学特性及药剂防治[J].东北林业大学学报,2016(7):98-101.
[9] 吴跃开,余金勇,朱秀娥.贵州核桃病原菌的初步鉴定[J].贵州林业科技,2016,44(4):1-6.
[10] 赵嘉平,梁军,吕全,等.葡萄座腔菌属(Botryosphaeria)系统分类评述[J].林业科学研究,2007,20(3):415-422.
[11] 方中达.植病研究方法(第三版)[M].北京:中国农业出版社,1998.
[12] 赵丹,康业斌.牡丹溃疡病—葡萄座腔菌引起的新病害[J].植物病理学报,2012(5):528-531.
[13] 赵丹.牡丹根部茎部真菌病害及病原鉴定[D].洛阳:河南科技大学,2012.
[14] 徐成楠,周宗山,迟福梅,等.越橘葡萄座腔菌枝枯病的病原菌鉴定[J].园艺学报,2013(2):231-236.
[15] 余仲东,赵官成,淡静雅,等.葡萄座腔菌属ITS-nrDNA的分子系统学分析[J].菌物学报,2010,29(2):285-293.
[16] 程燕林,梁军,吕全,等.葡萄座腔菌科研究进展—鉴定,系统发育学和分子生态学[J].生态学报,2011(11):3197-3207.
[17] 李诚,蒋军喜,赵尚高,等.猕猴桃灰霉病病原菌鉴定及室内药剂筛选[J].植物保护,2014,40(3):48-52.
[18] WISSEMANN V.Molecular evidence for allopolyploid origin of the Rosa canina complex (Rosaceae,Rosoidae)[J].Journal of Applied Botany 2002,76 (6):176-178.
[19] CHOI I Y.First report of bark dieback on blueberry caused by Botryosphaeria dothidea in Korea [J].Plant Disease,2011,95(2):227.
[20] TANG W,DING Z,ZHOU Z Q,etc.Phylogenetic and pathogenic analyses show that the causal agent of apple ring rot in China is Botryosphaeria dothidea[J].Plant Disease,2012,96:486-496.
[21] EI-GOORANI M A.Dieback of grapevine by Botryodiplodia theobromae Pat.In Egypt [J].Phytopathologia Mediterranea,1972,11:210-211.
[22] 陈海燕,田呈明,梁军,等.引起树木溃疡病病原菌Botryosphaeria属及相关无性态分类研究[J].西北林学院学报,2006(6):145-150.