三种镰刀菌引起的板栗内腐病病原菌鉴定

为明确板栗内腐病的病原，采用组织分离法分离纯化BL-5、BL-7和BL-9共3株病原菌，通过柯赫氏法则、形态学特征及分子生物学技术对病原菌进行鉴定。结果显示，BL-5在PDA培养基上初期菌丝白色，菌落中心部位有淡红色素，后期菌落逐渐变成深红色，菌落中心可见黄色菌丝；BL-7在PDA培养基上菌落呈白色、绒毛状，分生孢子长椭圆形或梨形、颜色透明、具隔膜；BL-9在PDA培养基上菌丝呈棉絮状，初期菌落呈粉红色，随着菌落的生长，后期菌落颜色逐渐变为浅黄色或褐色，分生孢子中间粗、两头较细长，似镰刀形，弯曲或直立，具有厚垣孢子。BL-5、BL-7、BL-9的ITS序列分别与Fusarium graminearum KJ847741、Fusarium proliferatum MW686898、Fusarium equiseti KU984711的序列同源性均达99%。因此，结合形态学特征和分子生物学特征，判断引起板栗内腐病的3种镰刀菌分别为禾谷镰刀菌（Fusarium graminearum）、层出镰孢菌（Fusarium proliferatum）、木贼镰刀菌（Fusarium equiseti），为板栗内腐病的有效防治提供了依据。

Identification of Pathogens of Three Kinds of Fusarium Causing Seed Rot of Chestnut

To identify the pathogen of the chestnut endosepsis， 3 pathogens including BL-5， BL-7 and BL-9 were isolated and purified by tissue isolation method， and were identified by Koch’?s postulate， morphological characteristics and molecular biological techniques. The results showed that， in the initial stage， the hypha of BL-5 on PDA medium was white with light red pigment in the center of the colony， and the colony gradually turned dark red with yellow hypha observed in the center of the colony in the later stage. The colony of BL-7 on PDA medium was white and villous， and the conidia were oval or pear shaped with transparent color and septate. In addition， the hypha of BL-9 was cotton flocculent on PDA medium， the colony was pink in the initial stage. With the growth of the BL-9 colony， the color of the colony gradually changed to light yellow or brown， and the conidia was slender at both ends and thick in the middle， sickle like， curved or erect shape with chlamydospores. Furthermore， the ITS sequences of BL-5， BL-7 and BL-9 had 99% homology with Fusarium graminearum KJ847741， Fusarium proliferatum MW686898 and Fusarium equiseti KU984711， respectively. So， combining with morphological characteristics and molecular biology technology， BL-5， BL-7 and BL-9 were belonged to Fusarium graminearum， Fusarium proliferatum and Fusarium equiseti， respectively， which provided basis for the development of the effective pathogen control technology system.