植物生态学报 ›› 2009, Vol. 33 ›› Issue (3): 433-441.DOI: 10.3773/j.issn.1005-264x.2009.03.002

• 研究论文 • 上一篇    下一篇

铁皮石斛组培苗与菌根真菌共培养过程中的相互作用

金辉1,2,5(), 许忠祥3, 陈金花1,4, 韩素芬2, 葛颂1, 罗毅波1,*()   

  1. 1 中国科学院植物研究所系统与分子进化植物学国家重点实验室,北京 100093
    2 南京林业大学森林资源与环境学院,南京 210037
    3 南京出入境检验检疫局,南京 210001
    4 海南大学园艺园林学院,海南儋州 571737
    5 中国科学院研究生院,北京 100049
  • 收稿日期:2008-08-15 接受日期:2009-01-13 出版日期:2009-08-15 发布日期:2009-05-31
  • 通讯作者: 罗毅波
  • 作者简介:*E-mail: luoyb@ibcas.ac.cn
    第一联系人:

    E-mail: comefine@ibcas.ac.cn

  • 基金资助:
    国家科技支撑项目珍稀濒危鸟类和植物繁育技术与示范(2008BAC39B05);国家自然科学基金(30371190)

INTERACTION BETWEEN TISSUE-CULTURED SEEDLINGS OF DENDROBIUM OFFICINALE AND MYCORRHIZAL FUNGUS (EPULORHIZA SP.) DURING SYMBIOTIC CULTURE

JIN Hui1,2,5(), XU Zhong-Xiang3, CHEN Jin-Hua1,4, HAN Su-Fen2, GE Song1, LUO Yi-Bo1,*()   

  1. 1State Key Laboratory of Systematic and Evolutionary Botany, Chinese Academy of Sciences, Beijing 100093, China
    2College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, China
    3Entry-Exit Inspection and Quarantine Bureau, Nanjing, 210001
    4College of Horticulture and Landscape Architecture, Hainan University, Danzhou, Hainan 571737, China
    5Graduate University of Chinese Academy of Sciences, Beijing 100049, China
  • Received:2008-08-15 Accepted:2009-01-13 Online:2009-08-15 Published:2009-05-31
  • Contact: LUO Yi-Bo

摘要:

由于人为的滥采滥挖和野外生境的退化, 使得铁皮石斛 (Dendrobium officinale) 这种名贵的中药材一直处于极度濒危的状态。为了从菌根真菌的角度给人工保育铁皮石斛提供理论指导, 对铁皮石斛的组织培养苗人工接种‘GDB181’菌株 (Epulorhiza sp.) 。培养60d后, 接菌苗平均鲜重增长率比对照苗高出了84.8%。在营养元素含量方面, 接菌苗的B、Si、Fe、Cu和Mn元素含量的净增率分别为780%、533%、192%、191%和128%, 均在100%以上;其他元素含量也有不同程度的增加 (除Zn外), 结果证明两者有效地建立了共生关系。在显微和超微结构的观察中发现:真菌菌丝随机破坏铁皮石斛的根被入侵到外皮层, 并从外皮层细胞不断扩展延伸到皮层的大型细胞, 最后在大型细胞中被分解消化。在真菌侵染过程中, 被侵染的皮层细胞的细胞壁严重扭曲变形, 菌丝在皮层细胞形成菌丝结, 菌丝结常位于细胞核附近或包围细胞核。在皮层的大型细胞中, 菌丝细胞被植物的溶酶体包围, 部分或全部被消解, 出现脱壁或失去细胞质甚至成为空腔等变化, 最终形成衰败的菌丝残骸, 溶酶体也随之消失。溶酶体分布越多的部位, 菌丝细胞消解变形越严重。含有菌丝残骸的皮层细胞可被新侵染的菌丝重新定殖, 这一菌丝侵染被消化再侵染的过程在铁皮石斛生长发育过程中可不断重复发生。

关键词: 相互作用, 共生, 显微结构, 铁皮石斛, 瘤菌根菌属

Abstract:

Aims Dendrobium officinale is an invaluable Chinese medicinal herb in China that has been threatened by over-collection and habitat deterioration. Understanding the relationship between orchids and mycorrhizae is important for the conservation of orchids. Our aim is to examine the rela-tionship between seedling of D. officinale and mycorrhizae.

Methods The strain of GDB181 was identified by internal transcribed spacer (ITS) sequencing. We inoculated tissue culture seedlings of D. officinale with GDB181 (Epulorhiza sp.) and used ICP-AES to measure the mineral element content of the seedlings. We observed slices of the orchid mycorrhizae under light and electron microscopes.

Important findings Seedlings of D. officinale and the inoculation strain formed symbiosis effectively. The growth of seedlings was promoted by the mycorrhizal fungus. As compared with the control, in-oculated seedlings had 84.8% greater average rate of increase of fresh weight. The mineral contents were elevated except Zn; the contents of B, Si, Fe, Cu and Mn in inoculated seedlings were increased by780%, 533%, 192%, 191% and 128%, respectively. Fungal hyphae entered the exodermis by breaking the velamen randomly and infected other cells continuously by means of penetrating the cell wall. Lastly, the hyphae inhabited the large cells of the cortex and were digested. The cell wall of the cortical cells were distorted and deformed by infection by the hyphae, and many pelotons formed in exodermis and cortex. Pelotons were often found near or encircling the nucleus. The invaded fungal hyphae were surrounded and dissolved by lysosomes partly or completely. With changes of the hyphae from losing cell wall or cytoplasm to remaining cavum, the lysosomes disappeared gradually. More hyphae were dissolved and deformed if there were more lysosomes near them. The cortex cells containing degenerated hyphae were frequently recolonized by hyphae, and the hyphal digestion and the reinfection of cortex cells occurred repeatedly throughout all growing stage of D. officinale.

Key words: interaction, symbiosis, microstructure, Dendrobium officinale, Epulorhiza sp.