一株铁皮石斛内生真菌的鉴定及对株型的影响

doi:10.19303/j.issn.1008-0384.2019.07.013

福建农业学报 ›› 2019, Vol. 34 ›› Issue (7): 837-841.doi: 10.19303/j.issn.1008-0384.2019.07.013

一株铁皮石斛内生真菌的鉴定及对株型的影响

王伟英, 邱珊莲, 邹晖, 戴艺民, 林江波

福建省农业科学院亚热带农业研究所, 福建漳州 363005

收稿日期:2018-12-17 修回日期:2019-03-16 出版日期:2019-07-20 发布日期:2019-10-14
通讯作者: 林江波(1976-),男,硕士,副研究员,主要从事农业生物技术研究(E-mail:linjiangbo1976@163.com) E-mail:linjiangbo1976@163.com
作者简介:王伟英(1980-),女,硕士,助理研究员,主要从事农业生物技术研究(E-mail:weiyingwang178@126.com)
基金资助:
福建省农业科学院科技创新项目（PC2018-13）；福建省科技计划项目——省属公益类科研院所基本科研专项（2016R1012-5）

Identification of an Endophytic Fungus and Its Morphological Effect on Dendrobium officinale

WANG Wei-ying, QIU Shan-lian, ZOU Hui, DAI Yi-min, LIN Jiang-bo

Subtropical Agriculture Research Institute, Fujian Academy of Agricultural Sciences, Zhangzhou, Fujian 363005, China

Received:2018-12-17 Revised:2019-03-16 Online:2019-07-20 Published:2019-10-14

摘要/Abstract

摘要： [目的] 对连城冠豸山上生长较好的野生铁皮石斛进行内生真菌的分离，以获得具有促生效果的菌株，为进一步研究瘤菌根菌属与铁皮石斛共生关系奠定基础。[方法] 将连城冠豸山野生铁皮石斛根进行消毒后在PDA培养基上，对长出的内生真菌分离纯化，将一株纯化菌株标记为T11，并通过形态特征与5.8SrDNA-ITS分子生物学方法进行鉴定，并将T11菌株接入液体培养基震荡培养7 d，成菌液后浇灌无菌石斛盆栽苗进行共培养6个月，观察石斛株型的变化。[结果] PDA平板上培养的T11菌株的菌落乳白色，形成同心环，菌丝细胞为双核，有隔，含有椭圆形念珠状细胞，即从形态特征初步鉴定为瘤菌根菌属Epulorhiza sp.，为无性态的胶膜菌Tulasnella calospora。以提取T11菌株DNA为模板进行5.8SrDNA-ITS序列扩增，扩增产物片段长度为630 bp，通过序列比对和系统进化树分析，T11与大花蕙兰植物菌根真菌胶膜菌聚类在同一分支，亲缘关系最为相近，其同源性达到99%。接菌后的石斛苗长势较好，其株高、叶片数和叶宽都显著增加。[结论] 从形态特征和分子生物学鉴定得出T11菌株为兰科菌根真菌担子菌的瘤菌根菌属，瘤菌根菌能促进铁皮石斛苗的生长。

关键词: 铁皮石斛, 内生真菌, 5.8SrDNA-ITS

Abstract: [Objective] An endophytic fungus from its symbiotic Dendrobium officinale plant was identified and studied for its effect on the morphology of the host.[Method] The fungus was isolated from the disinfected roots of a wild D. officinale plant grown on top of Mt. Guanzhai in Liancheng county. It was further purified on a PDA medium and labeled as T11 to be identified by morphological and 5.8 SrDNA-ITS molecular biological methods. After a 7 d-culture in a shaking flask, the inoculated liquid was added to the aseptically potted seedlings of D. officinale. Growth of the seedlings was observed for 6 months.[Result] The milky white T11 colonies on the PDA plate had concentric rings on them. The T11 mycelial cells were binucleate, septate, and elliptical in shape. It was preliminarily identified as an Epulorhiza sp., an asexual plastic film fungus of Tulasnella calospora. Subsequently, the 5.8SrDNA-ITS sequence amplified from the DNA extracted from T11 as template showed the PCR fragment to be 630bp in length. The sequence alignment and phylogenetic tree clustered T11 in the same branch as the T. calospora of Cymbidium, with a 99% genetic homology. With the T11 inoculation, the D. officinale seedlings grew well with significantly increased plant height, leaf count, and leaf width.[Conclusion] The morphological and molecular biological characteristics of T11 indicated it to be an Epulorhiza sp., the Basidiomycetes of orchidaceae mycorrhizal fungi, which is known to enhance the growth of D. officinale.

Key words: Dendrobium officinale, endophytic fungus, 5.8 SrDNA-ITS

中图分类号:

R282

王伟英, 邱珊莲, 邹晖, 戴艺民, 林江波. 一株铁皮石斛内生真菌的鉴定及对株型的影响[J]. 福建农业学报, 2019, 34(7): 837-841.

WANG Wei-ying, QIU Shan-lian, ZOU Hui, DAI Yi-min, LIN Jiang-bo. Identification of an Endophytic Fungus and Its Morphological Effect on Dendrobium officinale[J]. Fujian Journal of Agricultural Sciences, 2019, 34(7): 837-841.

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一株铁皮石斛内生真菌的鉴定及对株型的影响

Identification of an Endophytic Fungus and Its Morphological Effect on Dendrobium officinale

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