滇西槽舌兰内生真菌多样性及其共生真菌对叠鞘石斛种子萌发的效应

doi:10.13304/j.nykjdb.2023.0078

中国农业科技导报 ›› 2023, Vol. 25 ›› Issue (10): 91-98.DOI: 10.13304/j.nykjdb.2023.0078

滇西槽舌兰内生真菌多样性及其共生真菌对叠鞘石斛种子萌发的效应

李爱花¹(), 王涛¹, 王苗苗¹, 邓军育¹, 刘佳¹, 亚吉东², 张毓¹()

^1.国家植物园北园，植物迁地保护国家林业与草原局重点实验室，北京市花卉园艺工程技术研究中心，北京 100093
^2.中国科学院昆明植物研究所中国西南野生生物种质资源库，昆明 650201

收稿日期:2023-02-08 接受日期:2023-04-12 出版日期:2023-10-15 发布日期:2023-10-27
通讯作者: 张毓
作者简介:李爱花 Email：liaihua@chnbg.cn；
基金资助:
北京市植物园园管课题项目(BZ202103);北京市公园管理中心课题项目(ZX2022009);国家林草局机构运行保障专项(XM-JZ03)

Diversity of Endophytic Fungi in Holcoglossum rupestre and Effects of Symbiotic Fungi on Seed Germination of Dendrobium denneanum

Aihua LI¹(), Tao WANG¹, Miaomiao WANG¹, Junyu DENG¹, Jia LIU¹, Jidong YA², Yu ZHANG¹()

^1.Key Laboratory of National Foresty and Grassland Adminstration on Plant Ex situ Conservation，Beijing Floriculture Engineering Technology Research Center，North Garden of China National Botanical Garden，Beijing 100093，China
^2.Germplasm Bank of Wild Species in Southwest China，Kunming Institute of Botany，Chinese Academy of Sciences，Kunming 650201，China

Received:2023-02-08 Accepted:2023-04-12 Online:2023-10-15 Published:2023-10-27
Contact: Yu ZHANG

摘要/Abstract

摘要：

兰科植物种子萌发甚至整个生长阶段依赖于共生真菌。滇西槽舌兰（Holcoglossum rupestre）是我国特有极度濒危物种，为了更好地开展其种质资源保护，采集云南香格里拉、楚雄、昆明盘龙、昆明呈贡的滇西槽舌兰植株，基于ITS rDNA序列对其根部的内生真菌进行分离、鉴定和多样性分析，并研究这些内生真菌对叠鞘石斛（Dendrobium denneanum）种子萌发的影响。结果表明，滇西槽舌兰4个分布地植株的根内共分离、鉴定出68株内生真菌，分别属于粪壳菌纲（Sordriomycetes）、伞菌纲（Agaricomycetes）和座囊菌纲（Dothideomycetes）3个纲18个属。其中，镰刀菌属（Fusarium）、间座壳属（Diaporthe）、胶膜菌属（Tulasnella）、角担菌属（Ceratobasidium）的相对丰度分别为27.94%、10.29%、8.82%、8.82%，为主要优势内生真菌。胶膜菌属和角担菌属为滇西槽舌兰首次分离到的兰科共生菌，且胶膜菌属的P2和P47菌株可以显著促进叠鞘石斛种子萌发进程及萌发一致性。

关键词: 滇西槽舌兰, 内生真菌, 多样性, 胶膜菌属, 共生萌发

Abstract:

The germination and growth of orchid seeds depend on symbiotic fungi. Holcoglossum rupestre is an endemic， narrowed distributed and critical endangered species of Orchidaceae. Aiming to better ex situ conserve H. rupestre， the plants were collected from Shangrila， Chuxiong， Kunming Panlong and Kunming Chenggong in Yunnan. The endophytic fungi in its roots were isolated and identified based on ITS rDNA sequences， and their effect on seed germination was studied. The results showed that 68 culturable fungal endophytes were isolated and identified from roots of H. rupestre from 4 localities of Yunnan province， which belonged to 18 genera of 3 classes， i.e.， Sordriomycetes， Agaricomycetes and Dothideomycetes. Among all strains， Fusarium， Diaportha， Tulasnella and Ceratobasidium were the dominant fungal endophytes with 27.94%， 10.29%， 8.82% and8.82% of relative abundance. Some orchids mycorrhizal fungi as well as Tulasnella and Ceratobasidium were firstly found in H. rupestre roots. And Tulasnella strains P2 and P47 could significantly promote symbiotic germination of Dendrobium denneanum seeds by advancing germination process and improving germination consistency.

Key words: Holcoglossum rupestre, endophytic fungi, diversity, Tulasnella, symbiotic germination

中图分类号:

S182

李爱花, 王涛, 王苗苗, 邓军育, 刘佳, 亚吉东, 张毓. 滇西槽舌兰内生真菌多样性及其共生真菌对叠鞘石斛种子萌发的效应[J]. 中国农业科技导报, 2023, 25(10): 91-98.

Aihua LI, Tao WANG, Miaomiao WANG, Junyu DENG, Jia LIU, Jidong YA, Yu ZHANG. Diversity of Endophytic Fungi in Holcoglossum rupestre and Effects of Symbiotic Fungi on Seed Germination of Dendrobium denneanum[J]. Journal of Agricultural Science and Technology, 2023, 25(10): 91-98.

图/表 5

表1 4个分布地/栽培地滇西槽舌兰根部内生真菌分离结果

Table 1 Isolation results of fungal isolates from roots of Holcoglossum rupestre in 4 localities

地点Locality	样品编号Sample No.	分离率IF/%	菌落数Number of fungi
香格里拉Shangrila	D	36.67	22
楚雄Chuxiong	C	25.45	12
呈贡Chenggong	K	16.00	8
盘龙Panlong	P	20.80	52

表2 不同地区滇西槽舌兰根部内生真菌的种类及数量

Table 2 Fungal taxon and amount in roots of Holcoglossum rupestre from different regions

属 Genus	香格里拉Shangrila	楚雄 Chuxiong	呈贡 Chenggong	盘龙 Panlong	合计 Total
镰孢属Fusarium	3	7	5	4	19
间座壳属Diaporthe	2	0	0	5	7
角担菌属Ceratobasidium	6	0	0	0	6
胶膜菌属Tulasnella	0	0	0	6	6
交链孢属Alternaria	1	2	2	1	6
Biscogniauxia	1	3	0	0	4
拟盘多毛孢属Pestalotiopsis	0	0	0	4	4
木霉属Trichoderma	1	0	2	0	3
新拟盘多毛孢属Neopestalotiopsis	0	0	0	2	2
炭角菌属Xylaria	0	0	0	2	2
新壳梭孢属Neofusicoccum	1	0	0	1	2
刺盘孢属Colletotrichum	1	0	0	0	1
生赤壳属Clonostachys	1	0	0	0	1
旋孢腔菌属Cochliobolus	1	0	0	0	1
炭垫菌属Nemania	0	0	0	1	1
毛壳菌属Chaetomium	0	0	0	1	1
拟茎点霉属Phomopsis	0	0	0	1	1
Obolarina	0	0	0	1	1
总计Total	18	12	9	29	68
Shannon多样性指数 Shannon’s diversity index	2.93	1.38	1.44	3.23	—

图1 不同地区滇西槽舌兰根部内生真菌的相对丰度

Fig. 1 Relative abundance of cultured endomycetes in root of Holcolglossum repestre from different regions

图2 滇西槽舌兰根部内生真菌的系统聚类树注：内圈不同颜色表示科水平；外圈不同颜色表示目水平。

Fig. 2 Phylogenetic tree of cultured endomycetes in root of Holcolglossum rupestreNote： Different colors of inner circular indicate the family level； different colors of outer circular indicate the order level.

图3 胶膜菌菌株P2与叠鞘石斛种子的共生萌发A：菌株P2；B：P2菌丝；C：阴性对照种子；D：P2共生萌发原球茎；E：非共生萌发原球茎；F：不同阶段种子萌发统计

Fig. 3 Myco-germination of Dendrobium denneanum with mycorrhiza fungus P2A： Fungus strain P2； B： Euhypha of P2； C： Nagative control； D： Symbiotic germinating protocorms with P2； E： Non-symbiotic germinating protocorms； F： Germination percentage of different stages

参考文献 28

1	RODRIGUEZ R J, WHITE JR J F, ARNOLD A E, et al.. Fungal endophytes:diversity and functional roles [J]. New Phytol., 2009, 182(2):314-330.
2	NAIK B S. Functional roles of fungal endophytes in host fitness during stress conditions [J]. Symbiosis, 2019, 79:99-115.
3	BARBERIS L, MICHALET S, PIOLA F, et al.. Root fungal endophytes: identity, phylogeny and roles in plant tolerance to metal stress [J]. Fung. Biol., 2021, 125(4):326-345.
4	MCCORMICK M K, WHIGHAM D F, O'NEILL J P, et al.. Abundance and distribution of Corallorhiza odontorhiza reflect variations in climate and ectomycorrhizae [J]. Ecol. Monographs, 2009, 79(4):619-635.
5	SMITH S E, READ D J. Mycorrhizal symbiosis [M]. 3rd ed. London: Academic Press, 2008:1-175.
6	WATERMAN R J, BIDARTONDO M I, STOFBERG J, et al.. The effects of above- and belowground mutualisms on Orchid speciation and coexistence RID A-6126-2008 [J]. Am. Nat., 2011, 177(2):54-68.
7	SWARTS N D, SINCLAIR E A, FRANCIS A, et al.. Ecological specialization in mycorrhizal symbiosis leads to rarity in an endangered orchid [J]. Mol. Ecol., 2010, 19(15):3226-3242.
8	金效华,覃海宁.槽舌兰属的濒危状况调查[C]//中国植物学会七十周年年会论文摘要汇编.成都:高等教育出版社,2003:32-33.
9	TAN X M, CHEN X M, WANG C L, et al.. Isolation and identification of endophytic fungi in roots of nine Holcoglossum plants (Orchidaceae) collected from Yunnan, Guangxi, and Hainan Provinces of China [J]. Curr. Microbiol., 2012, 64(2):140-147.
10	魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979:1-804.
11	CHEN J, HU K X, HOU X Q, et al.. Endophytic fungi assemblages from 10 Dendrobium medicinal plants (Orchidaceae) [J]. World J. Microbiol. Biotechnol., 2011, 27:1009-1016.
12	MÁRQUEZ M S S, BILLS G F, ZABALGOGEAZCOA I. Diversity and structure of the fungal endophytic assemblages from two sympatric coastal grasses [J]. Fungal Diver., 2008, 33(11):87-100.
13	KUMAR S, TAMURA K, NEI M. MEGA3: Integrated software for molecular evolutionary genetics analysis and sequence alignment [J]. Briefings Bioinform., 2004, 5(2):150-163.
14	YUAN Z L, CHEN Y C, YANG Y. Diverse non-mycorrhizal fungal endophytes inhabiting an epiphytic, medicinal orchid (Dendrobium nobile):estimation and characterization [J]. World J. Microbiol. Biotechnol., 2009, 25(2):295-303.
15	STEWART S L, ZETTLER L W. Symbiotic germination of three semi-aquatic rein orchids ( Habenaria repens, H. quinquiseta, H. macroceratitis) from Florida [J]. Aquat. Bot., 2002:72(1):25-35.
16	STONE J K, BACON C W, WHITE J F. An overview of endophytic microbes [C]// BACON C W, WHITE J F. Microbial endophytes. New York: Marcel Dekker, 2000:3-29.
17	王艳,郭良栋,程虎印,等.不同生境重楼内生真菌及土壤真菌多样性比较[J].微生物学通报, 2020, 47(9):2867-2876.
	WANG Y, GUO L D, CHENG H Y, et al.. Comparison of endophytic and soil fungi of Paris polyphylla diversity from different habitat [J]. Microbiol. China, 2020, 47(9):2867-2876.
18	ERCOLE E, ADAMO M, RODDA M, et al.. Temporal variation in mycorrhizal diversity and carbon and nitrogen stable isotope abundance in the wintergreen meadow orchid Anacamptis morio [J]. New Phytol., 2015, 205(11):1308-1313.
19	EDWARDS J, JOHNSON C, SANTOS-MEDELLÍN C, et al.. Structure, variation, and assembly of the root-associated microbiomes of rice [J]. Proc. Natl. Acad. Sci. USA, 2011, 8:911-920.
20	侯天文,金辉,刘红霞,等.四川黄龙沟优势兰科植物菌根真菌多样性及其季节变化[J].生态学报, 2010, 30(13):3424-3432.
	HOU T W, JIN H, LIU H X, et al.. The variations of mycorrhizal fungi fiversity among different growing periods of the dominant orchids from two habitats in the Huanglong valley, Sichuan [J]. Acta Ecol. Sin., 2010, 30(13):3424-3432.
21	JACQUEMYN H, HONNAY O, CAMMUE BPA, et al.. Low specificity and nested subset structure characterize mycorrhizal associations in five closely related species of the genus Orchis [J]. Mol. Ecol., 2010, 19(18):4086-4095.
22	TAN X M, WANG C L, CHEN X M, et al.. In vitro seed germination and seedling growth of an endangered epiphytic orchid, Dendrobium officinale, endemic to China using mycorrhizal fungi (Tulasnella sp.) [J]. Sci. Hortic., 2014, 165:62-68.
23	RAMMITSU K, YUKAWA T, YAMASHITA Y, et al.. The mycorrhizal community of the epiphytic orchid Thrixspermum japonicum is strongly biased toward a single Ceratobasidiaceae fungus, despite a wide range of fungal partners [J]. Am. J. Bot., 2020, 107(12):1654-1662.
24	RAMMITSU K, KAJITA T, IMAI R, et al.. Strong primer bias for Tulasnellaceae fungi in metabarcoding:specific primers improve the characterization of the mycorrhizal communities of epiphytic orchids [J]. Mycoscience, 2021, 62:356-363.
25	SALMAN R, PRENDERGAST G, ROBERTS P. Germination of Dactylorhiza fuchsii seeds using fungi from non-orchid sources [C]// KINDLMANN P, WILLEMS J H, WHIGHAM D F. Trends and fluctuations and Underlying Mechanisms in Terrestrial Orchid Populations. Leiden: Backhuys, 2002.
26	YANG Q Y, XU L J, XIA W, et al.. Mycorrhizal compatibility and germination-promoting activity of Tulasnella species in two species of orchid (Cymbidium mannii and Epidendrum radicans [J/OL]. Horticulturae, 2021, 7(11):472 [2023-03-28]. .
27	HUANC H, ZI X M, LIN H, et al.. Host-specificity of symbiotic mycorrhizal fungi for enhancing seed germination, protocorm formation and seedling development of over-collected medicinal orchid, Dendrobium devonianum [J]. J. Microbiol., 2018, 56(1):42-48.
28	HADLEY G. Non-specificity of symbotic infection in orchid mycorrhiza [J]. New Phytol., 2010, 69(4):1015-1023.

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滇西槽舌兰内生真菌多样性及其共生真菌对叠鞘石斛种子萌发的效应

Diversity of Endophytic Fungi in Holcoglossum rupestre and Effects of Symbiotic Fungi on Seed Germination of Dendrobium denneanum

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