橡胶树健康和患病叶片的叶际微生物差异分析

Comparative Analysis of the Phyllosphere Microbiota on Healthy and Diseased Rubber Tree Leaves

摘要

橡胶树是重要的战略物资，也是海南最重要的植被类型，橡胶树健康对橡胶产业健康发展有重要意义。叶际微生物能调控叶片功能，保障植物健康，并维持多种生态系统功能。本研究利用高通量测序技术，分析健康和患病橡胶树叶际微生物群落组成、多样性及网络结构，系统比较两类叶片微生物群落的差异，为海南橡胶产业的健康发展提供参考。结果显示：在物种组成方面，橡胶树健康叶片的叶际细菌以放线菌纲和芽孢杆菌纲（如短小杆菌属）为主，真菌则以座囊菌纲和囊担菌纲（如枝孢菌属和尤氏菌属）为主。橡胶树患病叶片的细菌优势类群为γ-变形菌纲和α-变形菌纲（如泛菌属和微小杆菌属），其真菌则主要由锤舌菌纲（如白粉菌属）构成。主坐标分析（基于Bray-Curtis距离指数）和置换多因素方差分析（PERMANOVA）均表明，橡胶树健康和患病叶片的真菌群落相似性低，物种组成结构存在差异；在多样性方面，橡胶树健康叶片叶际微生物的Shannon多样性指数和Chao多样性指数均显著高于患病叶片；在相关性方面，橡胶树健康叶片叶际细菌的模块化结构明显，网络结构更稳定。橡胶树患病叶片的叶际真菌模块化结构更明显。橡胶树健康叶片的叶际细菌群落以确定性过程为主导，而患病叶片的叶际细菌则主要受随机过程影响。与之不同，无论是健康还是患病叶片，其叶际真菌群落的构建均以随机过程为主导。综上所述，橡胶树健康和患病叶片叶际微生物群落在组成和多样性上存在显著差异，橡胶树可能招募有益微生物来抵御病原真菌。

Abstract

The rubber tree ( Hevea brasiliensis) is a significant strategic material and the most prominent vegetation type in Hainan. Therefore, its health status is vital for the sustainable development of the local rubber industry. The phyllosphere microbiota plays a key role in regulating leaf function, promoting plant health, and sustaining multiple ecosystem services. This study employs high-throughput sequencing to analyze the composition, diversity, and network structures of the phyllosphere microbial communities between healthy and diseased rubber tree leaves. It systematically elucidates the differences between these communities to provide insights for the healthy development of Hainan's rubber industry. The results reveal distinct taxonomic profiles: healthy leaves are predominantly colonized by bacterial classes such as Actinobacteria and Bacilli (e.g., Curtobacterium), and fungal classes including Dothideomycetes and Agaricomycetes (e.g., Cladosporium and Uwebraunia). In contrast, diseased leaves are primarily dominated by the bacterial classes of Gammaproteobacteria and Alphaproteobacteria (e.g., Pantoeaand Exiguobacterium), and the fungi class of Leotiomycetes (e.g., Oidium) . Both principal coordinate analysis (based on Bray–Curtis distance) and permutational multivariate analysis of variance (PERMANOVA) confirm that the fungal communities on healthy and diseased rubber tree leaves are low in similarity, with differences in species composition structure; in terms of diversity, the Shannon and Chao diversity indices of the microbial community on healthy rubber tree leaves are significantly higher than those on diseased leaves; in terms of correlation, the bacterial community on healthy rubber tree leaves exhibits a modular structure with a more stable network structure. The fungal community on diseased rubber tree leaves has a more pronounced modular structure. Furthermore, the bacterial community on healthy rubber tree leaves is dominated by deterministic processes, whereas the bacterial community on diseased leaves is mainly influenced by stochastic processes. In contrast, fungal community assembly is mainly driven by stochastic processes in both healthy and diseased leaves. In summary, significant differences exist in the composition and diversity of phyllosphere microbiota between healthy and diseased rubber tree leaves, suggesting that rubber trees may recruit beneficial microorganisms to counteract pathogenic fungi.

关键词

橡胶树 / 健康 / 患病 / 叶际微生物

Key words

Rubber plantations / healthy / diseased / phyllosphere communities

参考文献

[1] Xu J, Grumbine R E, Beckschafer P. Landscape transformation through the use of ecological and socioeconomic indicators in Xishuangbanna, Southwest China, Mekong Region[J]. Ecological Indicators, 2014, 36: 749-756.

[2] Lan G, Wu Z, Chen B, et al. Species diversity in a naturally managed rubber plantation in Hainan Island, South China[J]. Tropical Conservation Science, 2017, 10: 1940082917712427.

[3] Zhai D L, Cannon C H, Ferry Slik J W, et al. Rubber and pulp plantations represent a double threat to Hainan’s natural tropical forests[J]. Journal of Environmental Management, 2012, 96: 64-73.

[4] 孙向前, 黎晟沅. 云南天然橡胶产业高质量发展策略与思考[J]. 橡胶工业, 2023, 70(06): 474-479.

[5] 曾卫宝. 特重病年情况下如何做好橡胶"两病"防治[J]. 中国热带农业, 2013(04): 38-40.

[6] Wei Y, Lan G, Wu Z, et al. Phyllosphere fungal communities of rubber trees exhibited biogeographical patterns, but not bacteria[J]. Environmental Microbiology, 2022, 24(08): 3777-3790.

[7] Shoemaker W R, Locey K J, Lennon J T. A macroecological theory of microbial biodiversity[J]. Nature Ecology and Evolution, 2017, 1(05): 0107.

[8] Morris C E. Phyllosphere[M]//New Jersey: John Wiley and Sons, Ltd, 2002.

[9] Vacher C, Cordier T, Vallance J. Phyllosphere fungal communities differentiate more thoroughly than bacterial communities along an elevation gradient[J]. Microbial Ecology, 2016, 72(01): 1-3.

[10] Jia T, Wang R, Fan X, et al. A comparative study of fungal community structure, diversity and richness between the soil and the phyllosphere of native grass species in a copper tailings dam in Shanxi Province, China[J]. Applied Sciences, 2018, 8(08): 1297.

[11] Shakir S, Zaidi S S E A, De Vries F T, et al. Plant genetic networks shaping phyllosphere microbial community[J]. Trends in Genetics, 2021, 37(04): 306-316.

[12] Sivakumar N, Sathishkumar R, Selvakumar G, et al. Phyllospheric microbiomes: diversity, ecological significance, and biotechnological applications[M]// Plant microbiomes for sustainable agriculture. Cham: Springer International Publishing, 2020: 113.

[13] Innerebner G, Knief C, Vorholt J A. Protection of arabidopsis thaliana against leaf-pathogenic pseudomonas syringae by sphingomonas strains in a controlled model system[J]. Applied and Environmental Microbiology, 2011, 77(10): 3202-3210.

[14] Reed S C, Townsend A R, Cleveland C C, et al. Microbial community shifts influence patterns in tropical forest nitrogen fixation[J]. Oecologia, 2010, 164(02): 521-531.

[15] Chen T, Nomura K, Wang X, et al. A plant genetic network for preventing dysbiosis in the phyllosphere[J]. Nature, 2020, 580(7805): 653-657.

[16] 肖 莹 . 海南省橡胶树白粉菌群体遗传结构研究[D]. 武汉: 华中农业大学, 2024.

[17] Wu H, Pan Y, Di R, et al. Molecular identification of the powdery mildew fungus infecting rubber trees in China[J]. Forest Pathology, 2019, 49(05): e12519.

[18] 余卓桐, 王绍春. 橡胶树白粉病流行过程和流行结构分析[J]. 热带作物学报, 1988(01): 83-89.

[19] 吴 华. 中国区橡胶树白粉菌分类进化与遗传多样性研究[D]. 海口: 海南大学, 2020.

[20] 张 哲, 邓琳梅, 鲁绍凤, 等. 马铃薯不同抗性品种叶际微生物群落结构及多样性分析[J]. 江西农业学报, 2025, 37(07): 60-73.

[21] 黄 宇, 汪汉成, 陈乾丽, 等. 感染白粉病烟株叶片真菌的群落结构与多样性[J]. 贵州农业科学, 2020, 48(05): 54-58.

[22] Tian X, Xu D, Shi T, et al. Plant resistance and leaf chemical characteristic jointly shape phyllosphere bacterial community[J]. World Journal of Microbiology and Biotechnology, 2020, 36(9): 139.

[23] 马 瑞, 王贞霖, 芮 凯. 槟榔黄化病叶际微生物群落结构与多样性[J]. 微生物学报, 2025, 65(08): 3600-3614.

[24] 庞 通, 李玉武, 兰国玉, 等. 基于宏基因组技术的橡胶树根际细菌多样性及驱动因素[J]. 西南农业学报, 2024, 37(03) : 604-611.

[25] 许心诺, 全 飞, 魏亚情, 等. 海南和西双版纳植胶区橡胶树根际真菌群落多样性分析[J]. 南方农业学报, 2022, 53(09): 2392-2402.

[26] Ji K, Wei Y, Lan G. Geographic location affects the bacterial community composition and diversity more than species identity for tropical tree species[J]. Plants, 2024, 13(11): 1565.

[27] Sampson T R, Debelius J W, Thron T, et al. Gut microbiota regulate motor deficits and neuroinflammation in a model of parkinson's disease[J]. Cell, 2016, 167(06): 1469-1480.

[28] Walters W, Hyde E R, Berg-Lyons D, et al. Improved bacterial 16S rRNA gene (V4 and V4-5) and fungal internal transcribed spacer marker gene primers for microbial community surveys[J]. Msystems, 2016, 1(01): e00009-15.

[29] 王 欣, 兰国玉, 舒常庆, 等. 热带雨林树种陆均松和青梅根际真菌群落多样性分析[J]. 西南农业学报, 2024, 37(07): 1563-1571.

[30] Adams R I, Miletto M, Taylor J W, et al. Dispersal in microbes: Fungi in indoor air are dominated by outdoor air and show dispersal limitation at short distances[J]. The ISME Journal, 2013, 7(07): 1262-1273.

[31] De Vries F T, Griffiths R I, Knight C G, et al. Soil bacterial networks are less stable under drought than fungal networks[J]. Nature Communications, 2018, 9(01): 3033.

[32] Ning D, Deng Y, Tiedje J M, et al. A general framework for quantitatively assessing ecological stochasticity[J]. Proceedings of the National Academy of Sciences, 2019, 116(34): 201904623.

[33] Röttjers L, Faust K. From hairballs to hypotheses-biological insights from microbial networks[J]. FEMS Microbiology Reviews, 2018, 42(06): 761-780.

[34] Berry D, Widder S. Deciphering microbial interactions and detecting keystone species with co-occurrence networks[J]. Frontiers in Microbiology, 2014, 5: 219.

[35] Hernandez D J, David A S, Menges E S, et al. Environmental stress destabilizes microbial networks[J]. The ISME Journal, 2021, 15(06): 1722-1734.

[36] Coyte K Z, Schluter J, Foster K R. The ecology of the microbiome: networks, competition, and stability[J]. Science, 2015, 350(6261): 663-666.

[37] 赵 珊. 锤舌菌纲真菌综合分类学研究及其抑菌活性物质的分离纯化[D]. 保定: 河北大学, 2025.

[38] Hongsanan S, Hyde K D, Phookamsak R, et al. Refined families of Dothideomycetes: orders and families incertae sedis in Dothideomycetes[J]. Fungal Diversity, 2020, 105(01): 17-318.

[39] 蒋淑华, 魏江春, 魏鑫丽. 座囊菌纲地衣型真菌系统学研究进展[J]. 菌物研究, 2022, 20(04): 285-291.

[40] Batista T M, Hilario H O, De Brito G A M, et al. Whole-genome sequencing of the endemic Antarctic fungus Antarctomyces pellizariae reveals an ice-binding protein, a scarce set of secondary metabolites gene clusters and provides insights on Thelebolales phylogeny[J]. Genomics, 2020, 112(05): 2915-2921.

[41] Mitra D, Mondal R, Khoshr u B, et al. Actinobacteria-enhanced plant growth, nutrient acquisition, and crop protection: Advances in soil, plant, and microbial multifactorial interactions[J]. Pedosphere, 2022, 32(01): 149-170.

[42] 施玉萍, 刘一贤, 李岚岚, 等. 橡胶树白粉病拮抗放线菌的筛选及田间防效评价[J]. 植物保护, 2024, 50(04): 347-355.

[43] Tedersoo L, Bahram M, Põlme S, et al. Global diversity and geography of soil fungi[J]. Science, 2014, 346(6213): 1256688.

[44] 白穆冉, 于 婷, 陈 培, 等. 芽胞杆菌抑菌物质作用研究进展 [J]. 中国抗生素杂志 , 2024, 49(05): 481-489.

[45] Sarris A, Ntouros M, Aguilar M G L, et al. Whole genome sequencing and root colonization studies reveal novel insights in the biocontrol potential and growth promotion by Bacillus subtilis MBI 600 on cucumber[J]. Frontiers in Microbiology, 2021, 11: 600393.

[46] Radovanović N, Milutinović M, Mihajlovski K, et al. Biocontrol and plant stimulating potential of novel strain bacillus sp. PPM3 isolated from marine sediment[J]. Microbial Pathogenesis, 2018, 120: 71-78.

[47] Singh P, Xie J, Qi Y, et al. A thermotolerant marine bacillus amyloliquefaciens S185 producing iturin A5 for antifungal activity against fusarium oxysporum f. sp. cubense[J]. Marine Drugs, 2021, 19(09): 516.

[48] 黄有航. 橡胶树白粉病生防菌研究[D]. 海口: 海南大学, 2012.

[49] 樊兰艳. 橡胶树内生枯草芽孢杆菌(bacillus subtilis) Czk1 抗菌活性物质的初步研究[D]. 海口: 海南大学, 2016.

[50] 贺春萍, 樊兰艳, 吴 贺, 等. 枯草芽孢杆菌Czk1 脂肽物质对橡胶树炭疽病和白粉病的抑制效果研究[J]. 中国农业科技导报, 2023, 25(06): 126-134.

[51] Sun W, Liu Q, Chen H, et al. Rice phyllospheric Pantoea spp. suppress blast and bacterial blight diseases[J]. Environmental Microbiome, 2025, 20(01): 137.

[52] 刘宇星, 邵秋雨, 葛 伟, 等. 健康与患病刺梨植株可培养叶际真菌菌群差异比较[J]. 菌物学报, 2021, 40(10): 2620-2640.

[53] 周慧娜, 张 涛, 徐自恒, 等. 山桐子叶际微生物群落多样性、结构及功能预测分析[J]. 经济林研究, 2022, 40(04): 163-172.

[54] Wang W, Portal-Gonzalez N, Wang X, et al. Metabolome-driven microbiome assembly determining the health of ginger crop (Zingiber officinale L. Roscoe) against rhizome rot [J]. Microbiome, 2024, 12(01): 167.

[55] 岳雪华, 范深厚, 杜雪柯, 等. 巨杉健康和感病叶片表面微生物组成和多样性比较[J]. 福建农林大学学报(自然科学版), 2020, 49(01): 10-17.

[56] Zhou Z X, Jiang H, Yang C, et al. Microbial community on healthy and diseased leaves of an invasive plant eupatorium adenophorum in southwest China[J]. The Journal of Microbiology, 2010, 48(02): 139-145.

[57] 杨尚东, 任奎瑜, 谭宏伟. 甘蔗宿根矮化病感病与非感病植株养分含量、根系生长及内生细菌群落的差异[J]. 植物营养与肥料学报, 2020, 26(09): 1591-1599.

[58] Qin C, Tao J, Liu T, et al. Responses of phyllosphere microbiota and plant health to application of two different biocontrol agents[J]. AMB Express, 2019, 9(01): 42.

[59] Ahmed W, Dai Z, Liu Q, et al. Microbial cross-talk: dissecting the core microbiota associated with flue-cured tobacco (Nicotiana tabacum) plants under healthy and diseased state[J]. Frontiers in Microbiology, 2022, 13: 845310.

[60] Yuan T, Qazi I H, Li J, et al. Analysis of changes in bacterial diversity in healthy and bacterial wilt mulberry samples using metagenomic sequencing and culture-dependent approaches[J]. Frontiers in Plant Science, 2023, 14: 1206691.

[61] Chen B, Guo W, Yang H, et al. Photosynthetic properties and biochemical metabolism of Cucurbita moschata genotypes following infection with powdery mildew[J]. Journal of Plant Pathology, 2020, 102(04): 1-7.

[62] Jiao S, Chu H, Zhang B, et al. Linking soil fungi to bacterial community assembly in arid ecosystems[J]. iMeta, 2022, 1(01): e2.

[63] Tao J, Meng D, Qin C, et al. Integrated network analysis reveals the importance of microbial interactions for maize growth[J]. Applied Microbiology and Biotechnology, 2018, 102(08): 3805.

[64] Zhong Y, Sorensen P O, Zhu G, et al. Differential microbial assembly processes and co-occurrence networks in the soil-root continuum along an environmental gradient[J]. iMeta, 2022, 1(02): e18.

[65] Qi G, Ma G, Chen S, et al. Microbial network and soil properties are changed in bacterial wilt-susceptible soil[J]. Applied and Environmental Microbiology, 2019, 85(13): e00162-19.

[66] Ma H, Pineda A, Hannula S E, et al. Steering root microbiomes of a commercial horticultural crop with plant-soil feedbacks[J]. Applied Soil Ecology, 2020, 150: 103468.

[67] Bai B, Liu W, Qiu X, et al. The root microbiome: Community assembly and its contributions to plant fitness[J]. Journal of Integrative Plant Biology, 2022, 64(02): 230-243.

[68] 綦 洋, 李凯旋, 赵紫玥, 等. 黄萎病对棉花根内细菌组成和群落构建的影响[J]. 中国生态农业学报(中英文), 2024, 32(07): 1183-1192.

[69] 綦 洋. 大丽轮枝菌侵染对棉花根系细菌组成及群落构建的影响[D]. 合肥: 安徽农业大学, 2024.

基金资助

国家自然科学基金项目(32271603)

海南省重点研发项目(ZDYF2024SHFZ096)

现代农业产业技术体系建设专项(CARS-33-ZP3)

中央级公益性科研院所基本科研业务费专项(1630022024013)