科微学术

微生物学通报

丛枝菌根真菌繁殖体的高效扩繁
作者:
基金项目:

山东省重大科技创新工程项目(2021CXGC010801);国家农业产业技术体系项目(CARS-10-B11);青岛市科技惠民项目(21-1-4-ny-13-nsh)


Efficient propagation of arbuscular mycorrhizal fungal propagules
Author:
  • 摘要
  • | |
  • 访问统计
  • |
  • 参考文献 [40]
  • |
  • 相似文献 [20]
  • | | |
  • 文章评论
    摘要:

    【背景】丛枝菌根(arbuscular mycorrhiza, AM)真菌能够和大多数植物形成互利共生体系,以促进植物生长、提高抗逆能力,在生产中具有重要作用,但AM真菌的繁殖技术限制了其应用。【目的】构建AM真菌的高效繁殖体系。【方法】于温室盆栽条件下,将根内根孢囊霉(Rhizophagus intraradice)接种于由3种寄主植物高粱(Sorghum bicolor)、玉米(Zea mays)、红三叶草(Trifolium repens)与5种培养基质(沸石、河砂、草炭、珍珠岩和蛭石)构建的4种繁殖体系中进行培养。研究不同繁殖体系对根内根孢囊霉侵染程度、产孢量的影响;然后利用高粱接种扩繁的菌剂进行AM真菌侵染能力的测定以验证其扩繁效果;最后基于筛选出的最优扩繁条件探讨对其他种类AM真菌摩西斗管囊霉(Funneliformis mosseae)、幼套近明球囊霉(Clariodeoglous etunicatum)、地表多样孢囊霉(Diversispora versiformis)和脆无梗囊霉(Acaulospora delicate)的扩繁效果。【结果】基质为河砂+蛭石+草炭(体积比为1:4:1),寄主植物为玉米+红三叶草处理的根内根孢囊霉产孢量最高,达到 1 912个/g-基质且接种高粱后表现出较好的侵染潜力,同时显著提高了高粱的地上部生物量。此外,利用该组合扩繁其他种类AM真菌,发现孢子数均得到了显著的提高,扩繁后摩西斗管囊霉、幼套近明球囊霉、地表多样孢囊霉和脆无梗囊霉的孢子数较扩繁前分别增加了6.24、2.92、35.18和4.18倍。【结论】以玉米+红三叶草为寄主植物,以河砂+蛭石+草炭(1:4:1)为基质,即通气孔隙为3.57%、持水孔隙为48.19%、容重为1.03 g/cm3、电导率为152.5 μS/cm、pH值为5.61、速效磷为5.6 mg/kg、碱解氮为80 mg/kg、速效钾为449.8 mg/kg、有机质为56.11 g/kg的条件有助于根内根孢囊霉达到最佳的扩繁效果,同时对其他AM真菌扩繁效果也相对较好,能够达到高效扩繁AM真菌繁殖体的目的。

    Abstract:

    [Background] Arbuscular mycorrhizal (AM) fungi can form mutualistic symbiosis with most plants to promote the growth and enhance the stress resistance of plants, playing an important role in production. However, the immature propagation technology limits the application of AM fungi. [Objective] To establish an efficient propagation system of AM fungi. [Methods] A pot experiment was conducted. Specifically, Rhizophagus intraradice was inoculated into four propagation systems constructed with three host plants (Sorghum bicolor, Zea mays, and Trifolium repens) and five substrates (zeolite, river sand, peat, perlite, and vermiculite) for culture. The effects of different propagation systems on the infection and sporulation of R. intraradice were studied. Then, we measured the infection ability of AM fungi by inoculating S. bicolor with propagation inoculum to verify the propagation effect. Finally, the optimal propagation conditions were employed to explore the propagation of other species of AM fungi: Funneliformis mosseae, Clariodeoglous etunicatum, Diversispora versiformis, and Acaulospora delicate. [Results] The propagation system with river sand+vermiculite+peat (1:4:1) as the substrate and Z. mays+T. repens, R. intraradice as the host plants had the highest spore production of 1 912 spores/g-dry substrate. Moreover, the AM fungi propagated with this system showed better infection potential and remarkably increased the aboveground biomass of S. bicolor. In addition, the propagation with this system increased the sporulation of F. mosseae, C. etunicatum, D. versiformis, and A. delicate by 6.24, 2.92, 35.18, and 4.18 times, respectively, compared with that before propagation. [Conclusion] The propagation system with Z. mays+T. repens as the host plants and river sand+vermiculite+peat (1:4:1) as the substrate had the optimum results. This system had the aeration porosity of 3.57%, the water-holding porosity of 48.19%, the bulk density of 1.03 g/cm3, the electrical conductivity of 152.5 μS/cm, pH 5.61, the available phosphorus of 5.6 mg/kg, the available nitrogen of 80 mg/kg, the available potassium of 449.8 mg/kg, and the organic matter of 56.11 g/kg, which are conducive to the efficient propagation R. intraradice and other AM fungi.

    参考文献
    [1] WANG HX, HAO ZP, ZHANG X, XIE W, CHEN BD. Arbuscular mycorrhizal fungi induced plant resistance against Fusarium wilt in jasmonate biosynthesis defective mutant and wild type of tomato[J]. Journal of Fungi:Basel, Switzerland, 2022, 8(5):422.
    [2] SCHUBRET R, WERNER S, CIRKA H, RÖDEL P, TANDRON MOYA Y, MOCK HP, HUTTER I, KUNZE G, HAUSE B. Effects of arbuscular mycorrhization on fruit quality in industrialized tomato production[J]. International Journal of Molecular Sciences, 2020, 21(19):7029.
    [3] 郭开发, 杨宇彤, 陈慧, 刘红彦, 谢丹, 周琦, 金晨钟. 适合G. mosseae扩繁的最佳培养基质和最适宿主植物[J]. 湖南农业科学, 2020(1):51-53. Guo KF, YANG YT, CHEN H, LIU HY, XIE D, ZHOU Q, JIN CZ. The best medium and host plant for G.mosseae propagation[J]. Hunan Agricultural Sciences, 2020(1):51-53(in Chinese).
    [4] JONGEN M, ALBADRAN B, BEYSCHLAG W, UNGER S. Can arbuscular mycorrhizal fungi mitigate drought stress in annual pasture legumes?[J]. Plant and Soil, 2022, 472(1/2):295-310.
    [5] HANSCH F, JASPAR H, von SIVERS L, BITTERLICH M, FRANKEN P, KÜHN C. Brassinosteroids and sucrose transport in mycorrhizal tomato plants[J]. Plant Signaling & Behavior, 2020, 15(2):1714292.
    [6] BASU S, RABARA RC, NEGI S. AMF:the future prospect for sustainable agriculture[J]. Physiological and Molecular Plant Pathology, 2018, 102:36-45.
    [7] 郝志鹏, 谢伟, 陈保冬. 丛枝菌根真菌在农业中的应用:研究进展与挑战[J]. 科技导报, 2022, 40(3):87-98. HAO ZP, XIE W, CHEN BD. Application of arbuscular mycorrhizal fungi in agriculture:research progress and challenges[J]. Science & Technology Review, 2022, 40(3):87-98(in Chinese).
    [8] SELVAKUMAR G, KRISHNAMOORTHY R, KIM K, SA TM. Propagation technique of arbuscular mycorrhizal fungi isolated from coastal reclamation land[J]. European Journal of Soil Biology, 2016, 74:39-44.
    [9] FASUSI OA, AMOO AE, BABALOLA OO. Propagation and characterization of viable arbuscular mycorrhizal fungal spores within maize plant (Zea mays L.)[J]. Journal of the Science of Food and Agriculture, 2021, 101(14):5834-5841.
    [10] IJDO M, CRANENBROUCK S, DECLERCK S. Methods for large-scale production of AM fungi:past, present, and future[J]. Mycorrhiza, 2011, 21(1):1-16.
    [11] 李媛媛, 王晓娟, 豆存艳, 林双双, 罗巧玉, 崔慧君, 孙莉, 金樑. 四种宿主植物及其不同栽培密度对AM真菌扩繁的影响[J]. 草业学报, 2013, 22(5):128-135. LI YY, WANG XJ, DOU CY, LIN SS, LUI QY, CUI HJ, SUN L, JIN L. Effects of four host plants and different cultivation densities on the propagation of arbuscular mycorrhizal fungi[J]. Acta Prataculturae Sinica, 2013, 22(5):128-135(in Chinese).
    [12] SCHLEMPER TR, STÜRMER SL. On farm production of arbuscular mycorrhizal fungi inoculum using lignocellulosic agrowastes[J]. Mycorrhiza, 2014, 24(8):571-580.
    [13] SMITH SE, READ DJ. Mycorrhizal Symbiosis[M]. NewYork:Academicpress, 2010:185-187.
    [14] 王亚军, 安巍, 罗青, 马萍, 石志刚, 赵建华. 丛枝菌根真菌菌剂扩繁及菌根化枸杞育苗技术研究[J]. 北方园艺, 2014(5):139-143. WANG YJ, AN W, LUO Q, MA P, SHI ZG, ZHAO JH. Research on enlargment reproducing of AMF microbial inoculum and growing seedling technology of wolfberry with mycorrhization[J]. Northern Horticulture, 2014(5):139-143(in Chinese).
    [15] HARIKUMAR VS. A new method of propagation of arbuscular mycorrhizal fungi in field cropped sesame (Sesamum indicum L.)[J]. Symbiosis, 2017, 73(1):71-74.
    [16] 周霞, 崔明, 秦永胜, 于萌, 黄建国, 周金星. 扩繁条件对3种丛枝菌根真菌(AMF)的影响[J]. 中国农学通报, 2012, 28(12):83-87. ZHOU X, CUI M, QIN YS, YU M, HUANG JG, ZHOU JX. The effects of the propagation condition on the three kinds of arbuscular mycorrhizal fungi[J]. Chinese Agricultural Science Bulletin, 2012, 28(12):83-87(in Chinese).
    [17] LI YD, NAN ZB, DUAN TY. Rhizophagus intraradices promotes alfalfa (Medicago sativa) defense against pea aphids (Acyrthosiphon pisum) revealed by RNA-Seq analysis[J]. Mycorrhiza, 2019, 29:623-635.
    [18] MATHIMARAN N, RUH R, VULLIOUD P, FROSSARD E, JANSA J. Glomus intraradices dominates arbuscular mycorrhizal communities in a heavy textured agricultural soil[J]. Mycorrhiza, 2005, 16(1):61-66.
    [19] 王幼珊, 张淑彬, 张美庆. 中国丛枝菌根真菌资源与种质资源[M]. 北京:中国农业出版社, 2012. WANG YS, ZHANG SB, ZHANG MQ. Fungi Resources and Germplasm Resources of Arbuscular Mycorrhizal in China[M]. Beijing:Chinese Agriculture Press, 2012(in Chinese).
    [20] PHILLIPS JM, HAYMAN DS. Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection[J]. Transactions of the British Mycological Society, 1970, 55(1):158-161.
    [21] 刘润进, 陈应龙. 菌根学[M]. 北京:科学出版社, 2007. LIU RJ, CHEN YL. Mycorrhizology[M]. Beijing:Science Press, 2007(in Chinese).
    [22] 周浓, 丁博, 冯源, 戚文华, 张华, 郭冬琴, 向军. 接种不同AM真菌对滇重楼菌根侵染率和入药品质的影响[J]. 中国中药杂志, 2015, 40(16):3158-3167. ZHOU N, DING B, FENG Y, QI WH, ZHANG H, GUO DQ, XIANG J. Effects of mycorrhizal colonization and medicine quality of Paris polyphylla var. yunnanensis inoculated by different foreign AM fungi specie[J]. China Journal of Chinese Materia Medica, 2015, 40(16):3158-3167(in Chinese).
    [23] 陈宁, 王幼珊, 蒋家珍, 杨延杰, 林多, 仇宏伟, 张美庆. 培养基质对丛枝菌根(AM)真菌生长发育的影响[J]. 农业工程学报, 2007, 23(9):205-207. CHEN N, WANG YS, JIANG JZ, YANG YJ, LIN D, QIU HW, ZhANG MQ. Effects of culture substrates on development of arbuscular mycorrhizal fungi[J]. Transactions of the Chinese Society of Agricultural Engineering, 2007, 23(9):205-207(in Chinese).
    [24] 孙向伟, 王晓娟, 陈牧, 豆存艳, 高飞翔, 金樑. 生态环境因子对AM真菌孢子形成与分布的作用机制[J]. 草业学报, 2011, 20(1):214-221. SUN XW, WANG XJ, CHEN M, DOU CY, GAO FX, JIN L. Effects of eco-environmental factors on the production and distribution of arbuscular mycorrhizal fungal spores[J]. Acta Prataculturae Sinica, 2011, 20(1):214-221(in Chinese).
    [25] TANAKA S, HASHIMOTO K, KOBAYASHI Y, YANO K, MAEDA T, KAMEOKA H, EZAWA T, SAITO K, AKIYAMA K, KAWAGUCHI M. Asymbiotic mass production of the arbuscular mycorrhizal fungus Rhizophagus clarus[J]. Communcations Biology, 2022, 5:43.
    [26] 毕银丽, 孙欢, 郭楠, 胡晶晶, 龚云丽, 裘浪. 不同基质和菌种组合对丛枝菌根真菌扩繁效果的影响[J]. 应用与环境生物学报, 2017, 23(4):616-621. BI YL, SUN H, GUO N, HU JJ, GONG YL, QIU L. Propagate-effects of different substrates and strain combinations on arbuscular mycorrhizal fungi[J]. Chinese Journal of Applied and Environmental Biology, 2017, 23(4):616-621(in Chinese).
    [27] 左龙亚, 郭冬琴, 周浓, 丁博, 张丹, 王雪, 阮神清. 8属22种AM真菌增殖技术的初步研究[J]. 资源开发与市场, 2014, 30(11):1281-1283. ZUO LY, GUO DQ, ZHOU N, DING B, ZHANG D, WANG X, RUAN SQ. Preliminary study of propagation technology of twenty-two species of arbuscular mycorrhizal fungi belonging to eight genera[J]. Resource Development & Market, 2014, 30(11):1281-1283(in Chinese).
    [28] WU QS, HE JD, SRIVASTAVA AK, ZHANG F, ZOU YN. Development of propagation technique of indigenous amf and their inoculation response in citrus[J]. Indian Journal of Agricultural Sciences, 2019, 89(7):1190-1194.
    [29] 马继芳. 丛枝菌根(AM)真菌高效繁殖条件及应用的研究[D]. 金华:浙江师范大学硕士学位论文, 2011. MA JF. High efficient propagation and application of arbuscular mycorrhiza (AM) fungi[D]. Jinhua:Master's Thesis of Zhejiang Normal University, 2011(in Chinese).
    [30] 郭辉娟, 李改丽. 扩繁条件对柠条土著AM真菌生长发育的影响[J]. 黑龙江农业科学, 2012(12):44-47. GUO HJ, LI GL. Effects of propagation condition on growth and development of arbuscular mycorrhizal fungi in the rhizosphere of Caragana korshinskii[J]. Heilongjiang Agricultural Sciences, 2012(12):44-47(in Chinese).
    [31] DOUDS DD JR, NAGAHASHI G, HEPPERLY PR. Production of inoculum of indigenous AM fungi and options for diluents of compost for on farm production of AM fungi[J]. Bioresource Technology, 2010, 101:2326-2330.
    [32] RODRÍGUEZ-ECHEVERRÍA S, FREITAS H. Diversity of AMF associated with Ammophila arenaria ssp. arundinacea in Portuguese sand dunes[J]. Mycorrhiza, 2006, 16(8):543-552.
    [33] CHAIYASEN A, CHAIYA L, DOUDS DD, LUMYONG S. Influence of host plants and soil diluents on arbuscular mycorrhizal fungus propagation for on-farm inoculum production using leaf litter compost and agrowastes[J]. Biological Agriculture & Horticulture, 2017, 33(1):52-62.
    [34] 张琳. 三种土壤长期不同施肥措施对丛枝菌根真菌的影响[D]. 南京:南京农业大学硕士学位论文, 2017. ZHANG L. Response of arbuscular mycorrhizal fungi to long-term chemical and organic nutrient inputs in three distinct soils[D]. Nanjing:Master' Thesis of Nanjing Agricultural University, 2017(in Chinese).
    [35] 陈梅梅, 陈保冬, 王新军, 朱永官, 王幼珊. 不同磷水平土壤接种丛枝菌根真菌对植物生长和养分吸收的影响[J]. 生态学报, 2009, 29(4):1980-1986. CHEN MM, CHEN BD, WANG XJ, ZHU YG, WANG YS. Influences of arbuscular mycorrhizal fungi (AMF) on the growth and ecological stoichiometry of clover and ryegrass grown in monoculture or in mixture at different phosphorus (P) levels[J]. Acta Ecologica Sinica, 2009, 29(4):1980-1986(in Chinese).
    [36] 孙金华, 毕银丽, 裘浪, 江彬. 土壤中丛枝菌根真菌对宿主植物磷吸收作用机制综述[J]. 土壤通报, 2016, 47(2):499-504. SUN JH, BI YL, QIU L, JIANG B. A review about the effect of AMF on uptaking phosphorus by host plants in soil[J]. Chinese Journal of Soil Science, 2016, 47(2):499-504(in Chinese).
    [37] LIN CY, WANG YX, LIU MH, LI Q, XIAO WF, SONG XZ. Effects of nitrogen deposition and phosphorus addition on arbuscular mycorrhizal fungi of Chinese fir (Cunninghamia lanceolata)[J]. Scientific Reports, 2020, 10:12260.
    [38] GAMPER H, PETER M, JANSA J, LÜSCHER A, HARTWIG UA, LEUCHTMANN A. Arbuscular mycorrhizal fungi benefit from 7 years of free air CO2 enrichment in well-fertilized grass and legume monocultures[J]. Global Change Biology, 2004, 10(2):189-199.
    [39] MA N, YOKOYAMA K, MARUMOTO T. Promotion of host plant growth and infection of roots with arbuscular mycorrhizal fungus Gigaspora margarita by the application of peat[J]. Soil Science and Plant Nutrition, 2006, 52(2):162-167.
    [40] 马俊卿, 侯宁, 孙晨瑜, 杨怡森, 覃圣峰, 王勇, 刘璐, 廖虹霖, 黄京华. 宿主不同对丛枝菌根真菌扩繁效应的影响[J]. 中国农学通报, 2022, 38(1):7-14. MA JQ, HOU N, SUN CY, YANG YS, QIN SF, WANG Y, LIU L, LIAO HL, HUANG JH. Effects of different hosts on propagation of arbuscular mycorrhizal fungi[J]. Chinese Agricultural Science Bulletin, 2022, 38(1):7-14(in Chinese).
    引证文献
    网友评论
    网友评论
    分享到微博
    发 布
引用本文

黄玉丹,张淑彬,李琳,梁斌,高雪,武亚芬,李敏,向丹. 丛枝菌根真菌繁殖体的高效扩繁[J]. 微生物学通报, 2023, 50(2): 503-513

复制
分享
文章指标
  • 点击次数:558
  • 下载次数: 1093
  • HTML阅读次数: 970
  • 引用次数: 0
历史
  • 收稿日期:2022-04-29
  • 录用日期:2022-06-28
  • 在线发布日期: 2023-02-03
  • 出版日期: 2023-02-20
文章二维码