分室培养装置在丛枝菌根真菌研究中的应用及其发展

doi:10.3724/SP.J.1258.2014.00120

摘要/Abstract

摘要：

重点围绕玻璃珠分室培养系统、H形分室培养系统、根排斥室培养系统、供体自养植物的双分室体外培养系统、丛枝菌根(AM)真菌与普通植物根器官的双重培养系统、AM真菌与Ri T-DNA转型根的双重单胞无菌培养系统、AM真菌与Ri T-DNA转型根双重培养的改良分室单胞培养系统等7个不同的分室培养装置, 对AM真菌的培养类型及其应用进行了系统的评述。其中, 采用玻璃珠分室培养装置易于将AM真菌与培养基质分开, 能获得大量纯净的AM真菌繁殖体, 用于研究AM真菌对矿质元素和微量元素的吸收, 具有不可替代的作用。H形分室培养系统和根排斥室(RECs)培养系统均能够获得连续的、可切断的共生菌根网络(CMNs), 可用于研究植物-植物、植物-昆虫之间化感作用产生的信息交流。供体自养植物的双分室培养系统有益于研究AM真菌对宿主植物在单作和混作条件下生长效应的影响。AM真菌与植物根器官的双重培养系统为研究AM真菌的侵染过程及生理、生化特性提供了极大的方便, 同时为纯培养研究提供了重要的理论依据。AM真菌与Ri T-DNA转型根的双重单胞无菌培养体系可以获得AM真菌纯净菌体, 是研究AM真菌遗传、生理、生化等特性的理想方法。以AM真菌与Ri T-DNA转型根的双重单胞无菌培养系统为基础, 可以在菌丝生长室置换培养基、在根室中补充适量碳源, 并多次收获AM真菌繁殖体。转型根改良双重培养系统是提高AM真菌孢子接种剂产量的有效方法。综上所述, AM真菌的分室培养系统已经取得显著进展, 为开展个体、种群、群落等不同层次的菌根生态学研究提供了依据。

关键词: 丛枝菌根真菌, 宿主植物, 菌丝体网络, 分室培养装置, 孢子

Abstract:

Arbuscular mycorrhizae (AM) are an important symbiosis between vascular plants and AM fungi in terrestrial ecosystems. Many studies have focused on their species diversity, distribution, and functions in natural habitats. However, AM fungi cannot be propagated in isolation; they need to be cultured with host plants. Thus, development of the culture method has been a hotspot in the AM research. In order to facilitate the advancement of research on AM fungi, we reviewed all the culture methods for AM fungi and their applications. Seven split-compartment cultivation systems were systematically discussed, including glass bead split-compartment culture system, two-compartment H-bridge cultivation system, root exclusion compartment culture system, in vitro mycorrhizal donor plants (MDP) culture system, dual axenic culture system, dual monoaxenic culture system of AM fungi with Ri T-DNA transformed root, and the improved split-compartment monoaxenic culture system of AM fungi with Ri T-DNA transformed root. Glass bead split-compartment culture system plays an irreplaceable role in easily separating AM fungi from the medium, hence obtaining a large quantity of axenic AM fungal propagules, which can be used for studying the absorption of mineral nutrients and trace elements. The H-shaped compartment cultivation system and root exclusion compartment culture system (RECs) can be used for obtaining continuous common mycorrhizal networks (CMNs), making it possible for the study of secondary metabolites information exchange, such as the plant-plant and plant-insect allelopathy. In vitro mycorrhizal donor plants (MDP) culture system has the advantage to study the biological effects of AM fungi on monoculture of host plants or mixed cultivation with different plant species. The dual axenic culture system facilitates the study of the infection process of AM fungi and their physiological and biochemical properties, and assists with gaining theoretical understanding on pure culture of AM fungi. Dual monoaxenic culture system of AM fungi with Ri T-DNA transformed root could be used to obtain axenic mycelium of AM fungi, and for further study of its genetic, physiological and biochemical properties. Based on dual monoaxenic culture system of AM fungi with Ri T-DNA transformed root, the medium can be replaced in the hyphal compartment and carbon source could be supplemented in the mycorrhizal compartment, and thus AM fungal propagules could be harvested continuously. The improved split-compartment monoaxenic culture system of AM fungi with Ri T-DNA transformed root is an effective method to improve the production of the inoculants with AM fungi. In a word, the different split-compartment instruments have provided effective methods for mycorrhizal research in autecology, population ecology and community ecology.

Key words: arbuscular mycorrhizae fungi, host plants, network of mycelium, split-compartment instrument, spores

王强,王茜,董梅,王晓娟,张亮,金樑. 分室培养装置在丛枝菌根真菌研究中的应用及其发展. 植物生态学报, 2014, 38(11): 1250-1260. DOI: 10.3724/SP.J.1258.2014.00120

WANG Qiang,WANG Qian,DONG Mei,WANG Xiao-Juan,ZHANG Liang,JIN Liang. Application and progress of split-compartment facility in studies of arbuscular mycorrhizal fungi. Chinese Journal of Plant Ecology, 2014, 38(11): 1250-1260. DOI: 10.3724/SP.J.1258.2014.00120

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链接本文: https://www.plant-ecology.com/CN/10.3724/SP.J.1258.2014.00120

https://www.plant-ecology.com/CN/Y2014/V38/I11/1250

图/表 4

图1 常规(A)和改进(B)的玻璃珠分室培养装置。 a和d为1 mm尼龙网, b和c为30 μm尼龙网; 将装置分隔为5个分室, 其中1和5为植物生长室, 2和4为菌根生长室, 3为AM真菌菌丝生长室。在B装置的2和4菌根生长室中装满粗河砂来代替A装置中的玻璃珠。

Fig. 1 Traditional (A) and modified (B) glass bead split-compartment culture system. a and d are nylon mesh screens of 1 mm, and b and c are nylon mesh screens of 30 μm; each container was separated into five compartments which 1 and 5 are plant growing compartments, 2 and 4 are mycorrhizal growing compartments, and 3 is the AM fungal hyphae growing compartment. The 2 and 4 compartments in B filled with coarse river sands instead of glass beads in A.

图2 H形装置结构构件图(改绘自Barto et al., 2011)。 1, 30 μm尼龙网; 2, 人工化感物质注入孔; 3, 供体植物; 4, 穿孔的钢板网; 5, 30 μm尼龙网; 6, 受体植物。30 μm尼龙网和穿孔钢板网将装置分隔为两个相同的分室。

Fig. 2 The two-compartment H-bridge cultivation system (Mo- dified from Barto et al., 2011). 1, nylon mesh screens of 30 μm; 2, injection hole for artificial allelochemicals; 3, donor plants; 4, perforated steel plates; 5, nylon mesh screens of 30 μm; 6, receiver plants. The device was separated by the nylon mesh screens of 30 μm and the perforated steel plates into two identical compartments.

图3 根排斥室(RECs)培养系统示意图(改绘自Barto et al., 2011 和Babikova et al., 2013) 。 A, 根外菌丝进入REC。B, 旋转REC切断了共生菌根网络(CMNs)与RECs的联系。1, 可以产生化感物质的植物; 2, 根排斥室; 3, 根排斥室内PDMS微型管; 4, 根外菌丝; 5, 植物根系; 6, 根排斥室外PDMS微型管, 双向箭头代表旋转根排斥室。

Fig. 3 The root exclusion compartment (RECs) culture system (Modified from Barto et al., 2011 and Babikova et al., 2013). A, Extra-radical mycelium entering into REC. B, Spinning out REC to cut the communication between common mycorrhizal networks (CMNs) and RECs. 1, plants which can produce allelochemicals; 2, RECs; 3, intra-REC PDMS tubing; 4, external hyphae; 5, plant roots; 6, extra-REC PDMS tubing, and the doubled sided arrow stands for rotary RECs.

图4 供体自养植物双分室体外培养系统(改绘自Derelle et al., 2012)。 HC, 根外菌丝生长室; RC, 根系生长室。粗线代表供体自养植物的根系, 细线代表AM真菌根外菌丝。

Fig. 4 In vitro mycorrhizal donor plants culture system (Modified from Derelle et al., 2012). HC, external hyphae growing compartment; RC, root growing compartment; The thick line stands for roots of donor autotrophic plants, and the thin line stands for external hyphae.

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