实验室条件下五唇兰菌根真菌专一性研究

doi:10.3773/j.issn.1005-264x.2010.12.009

摘要/Abstract

摘要：

利用从高原温带兰科植物菌根中获得的22个菌根真菌菌株, 对五唇兰(Doritis pulcherrima)进行了室内种子萌发、原球茎分化和组培苗回接试验, 从交叉回接的角度对附生兰科植物与菌根真菌的生理专一性进行了探讨。经过20周的共生培养, 只有编号为Cf1和Mm1的两个菌株使种子表现出种胚明显膨大的萌发迹象; 9个菌株能够促使原球茎较好地分化发育出根叶; 11个菌株处理苗的平均鲜重增长率高于对照组(156.25%), 其中Mm1的效果达到极显著水平(p = 0.01)。通过根切片显微观察, 在原球茎分化根和回接效果良好的处理苗的根皮层组织发现典型的菌丝团结构, 表明菌根体系已成功建立。温带地生兰菌根真菌对五唇兰种子萌发、原球茎发育和幼苗生长等3个重要生长阶段影响的试验显示, 五唇兰的种子和菌根真菌的共生萌发效果不佳, 而原球茎及幼株更容易与之建立良好的共生关系。同时, 也没有发现同一个真菌菌株能够对五唇兰的种子、原球茎和幼苗均产生促进作用。研究结果表明, 五唇兰的菌根真菌专一性因生理生长阶段的不同而存在差异。

关键词: 附生兰, 菌根真菌, 专一性, 室内种子萌发, 共生培养, 生存策略

Abstract:

Aims The degree of specificity of mycorrhizal association in tropical orchids has been controversial for many years. Our objective is to address this issue.
Methods Twenty-two endophyte fungal strains isolated from the roots of wild temperate terrestrial orchids were inoculated with the seeds, protocorms and seedlings of tropical epiphytic orchid species Doritis pulcherrima in flasks. Transverse sections of roots were observed by microscope.
Important findings After 20-weeks cultivation, only the fungal strains Cf1 and Mm1 caused swelling of the embryo, and nine caused differentiation of the protocorms into leaves or roots. The other fungal strains were pathogenic or had no function in seed germination and protocorm differentiation. In the seedling-fungi symbiotic treatment, the survival rate of the seedlings reached 100% and 11 of 22 fungal strains significantly enhanced the seedling growth. The fungal strain Mm1 had the most significant effect on increasing fresh mass of seedling. The existence of pelotons in the cortex of the roots of the protocorms and seedlings indicated successful infection. The effects of 22 endophyte fungal strains on seed germination, protocorm development and seedling growth of D. pulcherrima showed that the endophyte fungi did not enhance seed germination but were associated with the protocorms and seedlings; However, no one fungus was associated with all three key stages simultaneously. Our study suggested that the degree of specificity of tropical epiphyte orchids is influenced by the development stages of the plant.

Key words: epiphytic orchid, mycorrhizal, specificity, symbiotic germination in vitro, symbiotic cultivation, survival strategy

侯天文, 金辉, 刘红霞, 罗毅波. 实验室条件下五唇兰菌根真菌专一性研究. 植物生态学报, 2010, 34(12): 1433-1438. DOI: 10.3773/j.issn.1005-264x.2010.12.009

HOU Tian-Wen, JIN Hui, LIU Hong-Xia, LUO Yi-Bo. Mycorrhizal specificity of Doritis pulcherrima in in-vitro research. Chinese Journal of Plant Ecology, 2010, 34(12): 1433-1438. DOI: 10.3773/j.issn.1005-264x.2010.12.009

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https://www.plant-ecology.com/CN/Y2010/V34/I12/1433

图/表 3

参考文献 23

[1]	Arditti J, Ernst R, Yam TW, Glabe C (1990). The contributions of orchid mycorrhizal fungi to seed germination: a speculative review. Lindleyana, 5, 249-255.
[2]	Bayman P, Gonzalez EJ, Fumero JJ, Tremblay RL (2002). Are fungi necessary? How fungicides affect growth and survival of the orchid Lepanthes rupestris in the field. Journal of Ecology, 90, 1002-1008.
[3]	Bonnardeaux Y, Brundrett M, Batty A (2007). Diversity of mycorrhizal fungi of terrestrial orchids: compatibility webs, brief encounters, lasting relationships and alien invasions. Mycological Research, 3, 51-61.
[4]	Chen JH (陈金花), Hu MJ (胡美娇), Song XQ (宋希强), He MG (何明高), Luo YB (罗毅波) (2010). Microscopic observation on mycorrhiza of Doritis pulcherrima. Mycosystema (菌物学报), 29, 26-30. (in Chinese with English abstract)
[5]	Chen XC (陈心启), Tsi ZH (吉占和) (1998). The Chinese Orchids (中国兰花全书). China Forestry Publishing House, Beijing. (in Chinese)
[6]	Dearnaley JDW (2007). Further advances in orchid mycorrhizal research. Mycorrhiza, 17, 475-486. DOI URL PMID
[7]	Dijk E, Willems JH, van Andel J (1997). Nutrient responses as a key factor to the ecology of orchid species. Acta Botanica Neerlandica, 46, 339-363.
[8]	Fan L (范黎), Guo SX (郭顺星) (1998). Research development of orchid mycorrhizal fungi. Microbiology (微生物学通报), 24, 227-230. (in Chinese with English abstract)
[9]	He MG (何明高), Song SQ (宋松泉), Song XQ (宋希强), Wang RX (王瑞霞) (2009). A Kind of Testing Method of Orchid Seeds Vitality (一种兰花种子活力的测试方法). China Patent No.: 200810118434.4. 2009. 4. 8
[10]	Hou TW (侯天文) (2010). Mycorrhizal Fungi Diversity of the Dominant Orchids in the Huanglong Valley, Sichuan (四川黄龙沟优势兰科植物菌根真菌多样性). Master Dissertation, Beijing Forestry University, Beijing. (in Chinese with English abstract)
[11]	Ke HL (柯海丽), Song XQ (宋希强), Tan ZQ (谭志琼), Liu HX (刘红霞), Luo YB (罗毅波) (2007). Endophytic fungi diversity in root of Doritis pulcherrima (Orchidaceae). Biodiversity Science (生物多样性), 15, 456-462. (in Chinese with English abstract)
[12]	Ke HL (柯海丽), Song XQ (宋希强), Luo YB (罗毅波), Zhu GP (朱国鹏), Ling XB (凌绪柏) (2008). Seedling cultivation of Doritis pulcherrima Lindl. with mycorrhizal fungi. Acta Horticulturae Sinica (园艺学报), 35, 571-576. (in Chinese with English abstract)
[13]	Otero JT, Ackerman JD, Bayman P (2002). Diversity and host specificity of endophytic Rhizoctonia-like fungi from tropical orchids. American Journal of Botany, 89, 1852-1858. DOI URL PMID
[14]	Otero JT, Ackerman JD, Bayman P (2004). Differences in mycorrhizal preferences between two tropical orchids. Molecular Ecology, 13, 2393-2404. DOI URL PMID
[15]	Otero JT, Flanagan NS, Herre EA, Ackerman JD, Bayman P (2007). Widespread mycorrhizal specificity correlates to mycorrhizal function in the neotropical, epiphytic orchid Ionopsis utricularioides(Orchidaceae). American Journal of Botany, 94, 1944-1950. DOI URL PMID
[16]	Rasmussen HN (2002). Recent development in the study of OM. Plant and Soil, 244, 149-163.
[17]	Rasmussen HN, Whigham DF (1998). The underground phase: a special challenge in studies of terrestrial orchid populations. Botanical Journal of the Linnean Society, 126, 49-64.
[18]	Suárez JP, Weiß M, Abele A, Garnica S, Oberwinkler F, Kottke I (2006). Diverse tulasnelloid fungi form mycorrhizas with epiphytic orchids in an Andean cloud forest. Mycological Research, 110, 1257-1270. DOI URL PMID
[19]	Tan LY, Aung T, Yam TW (2004). Studies on the growth rate of tropical orchid seeds in symbiotic cultures. http://staff.science.nus.edu.sg/~scilooe/srp_2003/sci_paper/botanic/research_paper/tan_liyang.pdf. Cited July 10, 2009.
[20]	Taylor DL, Bruns TD (1999). Population, habitat and genetic correlates of mycorrhizal specialization in the ‘cheating’ orchids Corallorhiza maculate and C. mertensiana. Molecular Ecology, 8, 1719-1732. URL PMID
[21]	Tremblay RL, Ackerman JD, Zimmerman JK, Calvo RN (2004). Variation in sexual reproduction in orchids and its evolutionary consequences: a spasmodic journey to diversification. Biological Journal of the Linnean Society, 84, 1-54.
[22]	Warcup JH (1981). The mycorrhizal relationships of Australian orchids. New Phytologist, 87, 371-381.
[23]	Zettler LW, Burkhead JC, Marshall JA (1999). Use of a mycorrhizal fungus from Epidendrum conopseum to germinate seed of Encyclia tampensis in vitro. Lindleyana, 14, 102-105.

阶段 Stage	描述 Description
0	未萌发 No germination
1	种胚膨大 Swelling of the embryo
2	基毛出现 Development of radical hairs
3	叶原基出现 Development of a leaf primodium
4	第一片叶子出现 Development of the first leaf
5	第二片叶子出现 Development of the second leaf
6	根萌发 Development of roots

阶段 Stage	描述 Description
0	未萌发 No germination
1	种胚膨大 Swelling of the embryo
2	基毛出现 Development of radical hairs
3	叶原基出现 Development of a leaf primodium
4	第一片叶子出现 Development of the first leaf
5	第二片叶子出现 Development of the second leaf
6	根萌发 Development of roots

处理 Treatment	鉴定信息 Identified information	种子萌发 Seeds germination	原球茎分化 Protocorm differentiation	处理苗平均鲜重增长率 Average increasement of fresh mass (%)
Control	无真菌处理 No fungus treatment	×	无明显分化 No obvious differentiation	156.25
Cf1	Cryptosporiopsis radicicola	√	死亡 Dead	0
Cf2	未知真菌 Unknown fungus	×	同对照 As control treatment	86.10
Ct1	康宁木霉 Trichoderma koningii	×	死亡 Dead	265.89
Ct2	冬虫夏草 Cordyceps sinensis	×	同对照 As control treatment	185.98
Ct3	胶膜菌 Tulasnella sp.	×	死亡 Dead	272.22
Gd1	绿色肉座菌 Hypocrea virens	×	死亡 Dead	177.08
Mm1	瘤菌根菌 Epulorhiza sp.	√	被污染 Polluted	619.84^**
Mm2	烧地环锈伞 Pholiota carbonaria	×	同对照 As control treatment	135.10
Mm3	帚状弯孢聚壳 Eutypella scoparia	×	死亡 Dead	51.87
Mm4	丛赤壳科真菌 Nectriaceae sp.	×	分化旺盛 Well differentiated	110.42
Na1	杜鹃菌根菌 Ericoid mycorrhizal fungus	×	分化旺盛 Well differentiated	112.70
Na2	柱捕单顶孢菌 Monacrosporium cionopagum	×	分化旺盛 Well differentiated	79.80
Na3	未知担子菌 Unknown basidiomycete	×	分化旺盛 Well differentiated	0
Pc1	毛霉属真菌 Mortierella sp.	×	死亡 Dead	302.67
Pd1	胶膜菌 Tulasnellaceae isolate	×	分化旺盛 Well differentiated	285.00
Pd2	镰刀菌 Fusarium tricinctum	×	分化旺盛 Well differentiated	65.50
Pm1	炭角菌 Xylaria arbuscula	×	死亡 Dead	0
Ts1	丝核菌 Rhizoctonia sp.	×	分化旺盛 Well differentiated	201.80
Ts2	柔膜菌目外生菌根真菌 Ectomycorrhizal fungi of Helotiales	×	分化旺盛 Well differentiated	161.10
Ts3	粪盘菌 Ascobolus crenulatus	×	同对照 As control treatment	266.67
Ts4	Creosphaeria sassafras	×	分化旺盛 Well differentiated	347.04
Ts5	奥氏蜜环菌 Armillaria ostoyae	×	死亡 Dead	145.90

处理 Treatment	鉴定信息 Identified information	种子萌发 Seeds germination	原球茎分化 Protocorm differentiation	处理苗平均鲜重增长率 Average increasement of fresh mass (%)
Control	无真菌处理 No fungus treatment	×	无明显分化 No obvious differentiation	156.25
Cf1	Cryptosporiopsis radicicola	√	死亡 Dead	0
Cf2	未知真菌 Unknown fungus	×	同对照 As control treatment	86.10
Ct1	康宁木霉 Trichoderma koningii	×	死亡 Dead	265.89
Ct2	冬虫夏草 Cordyceps sinensis	×	同对照 As control treatment	185.98
Ct3	胶膜菌 Tulasnella sp.	×	死亡 Dead	272.22
Gd1	绿色肉座菌 Hypocrea virens	×	死亡 Dead	177.08
Mm1	瘤菌根菌 Epulorhiza sp.	√	被污染 Polluted	619.84^**
Mm2	烧地环锈伞 Pholiota carbonaria	×	同对照 As control treatment	135.10
Mm3	帚状弯孢聚壳 Eutypella scoparia	×	死亡 Dead	51.87
Mm4	丛赤壳科真菌 Nectriaceae sp.	×	分化旺盛 Well differentiated	110.42
Na1	杜鹃菌根菌 Ericoid mycorrhizal fungus	×	分化旺盛 Well differentiated	112.70
Na2	柱捕单顶孢菌 Monacrosporium cionopagum	×	分化旺盛 Well differentiated	79.80
Na3	未知担子菌 Unknown basidiomycete	×	分化旺盛 Well differentiated	0
Pc1	毛霉属真菌 Mortierella sp.	×	死亡 Dead	302.67
Pd1	胶膜菌 Tulasnellaceae isolate	×	分化旺盛 Well differentiated	285.00
Pd2	镰刀菌 Fusarium tricinctum	×	分化旺盛 Well differentiated	65.50
Pm1	炭角菌 Xylaria arbuscula	×	死亡 Dead	0
Ts1	丝核菌 Rhizoctonia sp.	×	分化旺盛 Well differentiated	201.80
Ts2	柔膜菌目外生菌根真菌 Ectomycorrhizal fungi of Helotiales	×	分化旺盛 Well differentiated	161.10
Ts3	粪盘菌 Ascobolus crenulatus	×	同对照 As control treatment	266.67
Ts4	Creosphaeria sassafras	×	分化旺盛 Well differentiated	347.04
Ts5	奥氏蜜环菌 Armillaria ostoyae	×	死亡 Dead	145.90