中国部分地区葡萄座腔菌属及相关属的系统学研究
摘要
葡萄座腔菌属(Botryosphaeria Ces．et de Not．)真菌是一类世界广泛分布的病原真菌，可引起溃疡、枝枯、果腐、流胶等病害，严重时导致树木死亡。开展葡萄座腔菌属及其相关属系统学的研究，不仅能确定病原菌的系统发育关系，澄清以往研究中的一些混乱问题，而且对于病害流行学研究以及病害控制都有着直接或间接的参考价值。本论文着重解决以下问题：弄清引起我国树木溃疡、苹果轮纹等病害的病原种类及分类学地位；分析本属及相关属部分真菌的系统学关系和地位。
本论文对所采集的不同寄主和不同地理区域的标本以及PDA培养出的子实体进行显微结构观察，对病原进行分类和鉴定；利用ITS测序和ISSR技术对比我国的菌株和国外的菌株，对本属及相关的无性型属进行系统学研究。本论文主要得出以下结论：经形态鉴定和分子实验验证，我们所分析的引起我国树木溃疡等病害的病原菌均为葡萄座腔菌(B. dothidea)；B. dothidea的无性型是七叶树壳梭孢(Fusicoccum aesculi)，而不是长期以来所认为的聚生小穴壳菌(Dothiorella gregaria)；国内的B. dothidea菌株与国外的B. ribis菌株的系统发育分析表明二者亲缘关系较远，应该为两个不同的种，而不是同物异名；在方法学上，用单孢菌株和病斑来源的菌株DNA进行ITS测序，所得到的DNA序列完全一致，所以在ITS测序时，不必用分离的单孢菌株。
The fungi of Botryosphaeria are a kind of
worldwide pathogenic fungi that cause
canker, twig blight, fruit rots and gummosis, even result in death, on various forest trees. To research the phytogeny on the Botryosphaeria and related taxa, not only can it confirm the phylogenic relation and clarify some confusion on taxonomy before, but also it will offer valuable information, directly or indirectly, for disease epidemiology research and control the disease. The purpose of this paper is to make clear the category and taxonomic position of pathogens that cause canker and fruit rot on various tree species and analyze the phylogenic relation of Botryosphaeria and its related taxa.
In this paper, the microstructure of fruit body from PDA culture and the symptomatic materials collected from different host and different geographic regions in China were used to identify the pathogens. The ITS and ISSR technique were applied to analyze the phytogeny of this genus and related anamorphic genera by comparing the Chinese isolates to foreign isolates. The results obtained in this experiment as follows: all specimen or isolates that cause diseases such as canker on trees in China are B. dothidea through morphology observation and molecular experiment. The anamorph of B. dothidea is Fusicoccum aesculi, but not Dothiorella gregaria, which have been regarded as the anamorph of B. dothidea for a long time in China. The phylogenic analysis of B. dothidea from China and B. ribis from other countries indicates that they should be two different species, but not synonymy because they are not closely related. The nucleotide sequences of ITS derived from isolates both generated from single conidia and isol
ated from canker are completely same, so it is unnecessary to use the isolates generated from single conidia in the experiment of ITS sequencing.
本论文对所采集的不同寄主和不同地理区域的标本以及PDA培养出的子实体进行显微结构观察，对病原进行分类和鉴定；利用ITS测序和ISSR技术对比我国的菌株和国外的菌株，对本属及相关的无性型属进行系统学研究。本论文主要得出以下结论：经形态鉴定和分子实验验证，我们所分析的引起我国树木溃疡等病害的病原菌均为葡萄座腔菌(B. dothidea)；B. dothidea的无性型是七叶树壳梭孢(Fusicoccum aesculi)，而不是长期以来所认为的聚生小穴壳菌(Dothiorella gregaria)；国内的B. dothidea菌株与国外的B. ribis菌株的系统发育分析表明二者亲缘关系较远，应该为两个不同的种，而不是同物异名；在方法学上，用单孢菌株和病斑来源的菌株DNA进行ITS测序，所得到的DNA序列完全一致，所以在ITS测序时，不必用分离的单孢菌株。
The fungi of Botryosphaeria are a kind of
worldwide pathogenic fungi that cause
canker, twig blight, fruit rots and gummosis, even result in death, on various forest trees. To research the phytogeny on the Botryosphaeria and related taxa, not only can it confirm the phylogenic relation and clarify some confusion on taxonomy before, but also it will offer valuable information, directly or indirectly, for disease epidemiology research and control the disease. The purpose of this paper is to make clear the category and taxonomic position of pathogens that cause canker and fruit rot on various tree species and analyze the phylogenic relation of Botryosphaeria and its related taxa.
In this paper, the microstructure of fruit body from PDA culture and the symptomatic materials collected from different host and different geographic regions in China were used to identify the pathogens. The ITS and ISSR technique were applied to analyze the phytogeny of this genus and related anamorphic genera by comparing the Chinese isolates to foreign isolates. The results obtained in this experiment as follows: all specimen or isolates that cause diseases such as canker on trees in China are B. dothidea through morphology observation and molecular experiment. The anamorph of B. dothidea is Fusicoccum aesculi, but not Dothiorella gregaria, which have been regarded as the anamorph of B. dothidea for a long time in China. The phylogenic analysis of B. dothidea from China and B. ribis from other countries indicates that they should be two different species, but not synonymy because they are not closely related. The nucleotide sequences of ITS derived from isolates both generated from single conidia and isol
ated from canker are completely same, so it is unnecessary to use the isolates generated from single conidia in the experiment of ITS sequencing.
引文
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陈原，杨旺．北京杨溃疡病抗病性的研究．北京林业大学学报，1994，16(2)：51-57
大石亲男．栗实黑腐病的病原菌及其生态．森林防疫，1979，(2)：2-6
戴芳澜．中国真菌总汇．1979，科学出版社
杜建玲，项蔚华，沈瑞祥．诱导杨树抗溃疡病机理的研究．林业科学研究，1995，8(1)：78-81
方中达，植病研究方法．1979，农业出版社．
高克祥，刘小光等．木霉菌对杨树树皮溃疡病菌拮抗作用的研究．林业科学，2001，37(5)：82-88
何田华等译．生态与进化研究中的分子方法．科学出版社，2001，51-61
金均然，沈瑞祥．杨树林土壤微生物及优势菌对小穴壳菌抑制作用的研究．北京林业大学学报，1989，11(3)：79-84
李红叶，曹若彬．杨梅根腐病菌营养和细胞酶特性。浙江农业大学学报，1996，22(2)：196-200
李美娜，王金友，等．苹果轮纹烂果病菌的侵染时期及影响因素．中国果树，2001，5：8-12
刘红霞，杨旺．毛白杨杂种无性系对Dothiorella gregaria Sacc．抗性的研究．北京林业大学学报，1996，18(3)：55-60
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