黑松菌根共生体中真菌液泡形态构架及其活力

doi:10.3773/j.issn.1005-264x.2008.04.023

摘要/Abstract

摘要：

为探讨菌根共生体中菌根菌液泡结构及其活力变化对宿主生长的影响, 对3种外生菌根菌黄色须腹菌(Rhizopogen luteous, 简称Rl)、彩色豆马勃(Pisolithus tinctorius, 简称 Pt₂)、美味牛肝菌 (Boletus edulis, 简称Be)离体菌丝及与黑松(Pinus thunbergii)形成菌根共生体的冰冻切片进行6-羧基二乙酸荧光素(6-CFDA)染色, 观察了真菌液泡形态构架及其活力的变化。结果表明, 在Pt₂与Rl离体菌丝中既可见单个分散的大液泡, 又可见相邻液泡间由细管状连接而成的管状液泡系统, Be离体菌丝的液泡则呈细小颗粒状, 密集分布于菌丝体的各个部分; Be离体菌丝的活性强于Pt₂和Rl。上述3种菌根菌与黑松形成菌根共生体后, Pt₂与Rl对黑松的促生效果好于Be, 菌根共生体中菌套和哈氏网菌丝的活性明显强于Be, 且仍可见数个液泡连接成管状系统; 而在Be菌根中菌丝液泡系统不形成管状, 活性较低。结果暗示, 菌根共生体中真菌的管状液泡系统及其活力与对黑松的促生作用似存在一定的相关性。该研究对于进一步了解菌根的促生作用机理及菌根真菌与宿主植物的协同进化具有一定的意义。

关键词: 黑松, 外生菌根, 管状液泡, 菌丝活力, 6-羧基二乙酸荧光素

Abstract:

Aims The fungal vacuole plays a key role in nutrient transportation and storage, especially in ectomycorrhizal symbionts. Our objective was to explore the effects of fungal activity and vacuole structure changes on host plant growth to further understand the mechanism of growth-promotion of mycorrhizal fungi and coevolution between host plant and mycorrhizal fungi.

Methods Hyphae of ectomycorrhizal fungi Rhizopogen luteous (Rl), Pisolithus tinctorius (Pt₂), and Boletus edulis(Be) cultured in vitro were stained with 6-CFDA, and the morphologies of fungal vacuoles and their activities were examined. Frozen sections of the ectomycorrhizal symbiont formed by the three fungi and Pinus thunbergii seedlings were stained and observed in the same way.

Important findings Dispersive large vacuoles in hyphae of Pt₂ and Rl and many small dense vacuoles in the hyphae of Be were observed in fungi cultured in vitro, and some vacuoles of Rl and Be linked into tubular vacuole systems. The activity of Be hyphae was higher than Pt₂ and Rl, and Be grew faster than the other two. After ectomycorrhizal formation, P. thunbergii seedlings with Pt₂ or Rl grew better than those with Be, and the fungal activities vacuole morphologies had changed greatly. Hyphae in mantles and Hartig nets of ectomycorrhiza formed by Pt₂ and Rl showed higher activities than those formed by Be, which were hard to label, and tubular vacuole systems were observed sometimes in the hyphae of Hartig nets of Pt₂ and Rl symbionts. It appears that fungal activity and tubular vacuole systems are related to the growth of P. thunbergii seedlings.

Key words: Pinus thunbergii, ectomycorrhiza, tubular vacuole systems, fungal activity, 6-CFDA

郑玲, 吴小芹. 黑松菌根共生体中真菌液泡形态构架及其活力. 植物生态学报, 2008, 32(4): 932-937. DOI: 10.3773/j.issn.1005-264x.2008.04.023

ZHENG Ling, WU Xiao-Qin. MORPHOLOGY AND ACTIVITY OF ECTOMYCORRHIZAL FUNGIIN VITRO AND IN SYMBIONT WITH PINUS THUNBERGII. Chinese Journal of Plant Ecology, 2008, 32(4): 932-937. DOI: 10.3773/j.issn.1005-264x.2008.04.023

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https://www.plant-ecology.com/CN/Y2008/V32/I4/932

图/表 5

图1 3种外生菌根菌离体培养菌丝6-CFDA荧光染色后液泡的形态结构 A、B示Pt2菌丝; C、D示Rl菌丝; E、F示Be菌丝 A, B for Pt2 hyphae; C, D for Rl hyphae; E, F for Be hyphae 箭头示管状液泡系统 The arrows show the tubular vacuole systems 比例尺 Scale bars: 10 μm Pt 2、Rl、Be: 同表1 See Table 1

Fig. 1 The morphologies and structures of the three ectomycorrhizal fungi hyphae by 6-CFDA staining

表1 3种外生菌根菌离体菌丝生长速度及其活性与管状液泡丰度

Table 1 The growth rate and activity of three ectomycorrhizal fungi and abundance of tubular vacuole

菌根菌Fungi	离体菌丝生长速率 Growth rate of hyphae	荧光强度 Fluorescence intensity	菌丝活性 Hyphae activity	管状液泡分布 Distribution of tubular vacuole
菌根菌Fungi	离体菌丝生长速率 Growth rate of hyphae	荧光强度 Fluorescence intensity	菌丝活性 Hyphae activity	观察玻片数Number of total slides observed	观察视野数 Number of total visions observed	可见管状液泡视野比例 Percentage of tubular vacuole
Rl	慢 Slow	弱 Weak	++	10	50	20
Pt₂	慢 Slow	弱 Weak	++	10	50	32
Be	快 Fast	强 Strong	+++	10	50	0

图2 3种外生菌根菌对黑松苗木生长量的影响同一图表中不同小写字母表示其二者差异达到显著水平(p<0.05) Significant difference ws indicated by different letters in the same figure (p<0.05) Rl、Pt2、Be: 同表1 See Table 1

Fig. 2 The effect of different ectomycorrhizal fungi on the growth of Pinus thunbergii seedings

图3 黑松3种外生菌根冰冻切片经6-CFDA染色结果 A、B为Pt2菌根; C、D为Rl菌根; E、F为Be菌根。星号示菌套, 箭头示哈氏网 A, BC for Pt2; C, D for Rl; E, F for Be. The stars show the mantles. The arrows show the Hartig net 比例尺 Scale Bars: 10 μm Pt 2、Rl、Be: 同表1 See Table 1

Fig. 3 6-CFDA staining of frozen sections of different ectomycorrhiza of Pinus thunbergii

表2 黑松外生菌根共生体中真菌活性及管状液泡丰度

Table 2 The activity of fungi and abundance of tubular vacuole in the ectomycorrhizal symbiont of Pinus thunbergii

菌根菌 Fungi	荧光强度 Fluorescence intensity	菌丝活性 Hyphae activity	管状液泡分布 Distribution of tubular vacuole
菌根菌 Fungi	荧光强度 Fluorescence intensity	菌丝活性 Hyphae activity	观察切片总数 Number of total sections for observation	形成管状液泡的切片数 Number of sections with tubular vacuole observed	管状液泡所占比例(%) Percentage of tubular vacuole
Rl	强 Strong	+++	200	30	15
Pt₂	强 Strong	+++	200	50	25
Be	弱 Weak	+	200	0	0

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