New Forests

, Volume 48, Issue 4, pp 547–562 | Cite as

Diversity of arbuscular mycorrhizal fungi in Tectona grandis Linn.f. plantations and their effects on growth of micropropagated plantlets

  • Amornrat Chaiyasen
  • David D. Douds
  • Paiboolya Gavinlertvatana
  • Saisamorn Lumyong


Regeneration of stands of valuable tropical hardwood tree species for sustainable harvest requires production of seedlings with high probabilities of survival. One way to enhance the vigor of plants for outplanting is pre-colonization of roots by arbuscular mycorrhizal (AM) fungi. We pursued the strategy that the most promising AM fungus candidates for inoculation would be those associated with the tree of interest in the field. AM fungus communities were assessed in five plantations of Tectona grandis Linn.f. A total of 18 AM fungal morphotypes were found, representing four families: Glomeraceae (49.6%), Acaulosporaceae (24.9%), Claroideoglomeraceae (20.8%), and Gigasporaceae (4.8%). AM fungus spore density was negatively correlated with soil organic carbon. Some of these AM fungi, plus Rhizophagus irregularis, were established in pot culture and in vitro with transformed carrot roots, and subsequently used to inoculate micropropagated plantlets of T. grandis. Tectona grandis plantlets inoculated in vitro were successfully colonized by all AM fungi studied. Plants inoculated with Funneliformis mosseae were taller than uninoculated plants. Tectona grandis plantlets inoculated with the AM fungus Claroideoglomus etunicatum PBT03 were taller than uninoculated controls in ex vitro experiments. This study provides early insight for the targeted use of the AM symbiosis in production of important tree species in future greenhouse studies and reforestation.


Arbuscular mycorrhizal fungi Micropropagated plant Rhizosphere soil Teak 



We would like to acknowledge the financial support of The Royal Golden Jubilee PhD Program (PHD/0150/2550), Thailand Research Fund (TRF), Research-Team Promotion Grant (RTA5880006), Chiang Mai University, and Center of Excellence on Biodiversity (BDC), Office of Higher Education Commission (BDC-PG2-159011).


  1. Abbott LK, Robson AD (1991) Factors influencing the occurrence of vesicular-arbuscular mycorrhizas. Agric Ecosyst Environ 35:121–150CrossRefGoogle Scholar
  2. Aggangan NS, Moon HK, Han SH (2010) Growth response of Acacia mangium Willd. seedlings to arbuscular mycorrhizal fungi and four isolates of the ectomycorrhizal fungus Pisolithus tinctorius (Pers.) Coker and Couch. New Forest 39:215–230CrossRefGoogle Scholar
  3. Bécard G, Fortin JA (1988) Early events of vesicular-arbuscular mycorrhiza formation on Ri T-DNA transformed roots. New Phytol 108:211–218CrossRefGoogle Scholar
  4. Biermann B, Linderman RG (1983) Use of vesicular-arbuscular mycorrhizal roots, intraradical vesicles and extraradical vesicles as inoculum. New Phytol 95:97–105CrossRefGoogle Scholar
  5. Bojarczuk K, Karliński L, Hazubska-Przybył T et al (2015) Influence of mycorrhizal inoculation on growth of micropropagated Populus × canescens lines in metal-contaminated soils. New Forest 46:195–215CrossRefGoogle Scholar
  6. Brundrett MC, Bougher N, Dell B et al (1996) Working with mycorrhizas in forestry and agriculture. Australian Centre for International Agricultural Research Monograph 32, CanberraGoogle Scholar
  7. Brundrett MC, Abbott LK, Jasper DA (1999) Glomalean mycorrhizal fungi from tropical Australia. I. Comparison of the effectiveness and specificity of different isolation procedures. Mycorrhiza 8:305–314CrossRefGoogle Scholar
  8. Chaiyasen A, Young JPW, Teaumroong N et al (2014) Characterization of arbuscular mycorrhizal fungus communities of Aquilaria crassna and Tectona grandis roots and soils in Thailand plantations. PLoS ONE 9(11):e112591. doi: 10.1371/journal.pone.0112591 CrossRefPubMedPubMedCentralGoogle Scholar
  9. Chaiyasen A, Leardwiriyakool C, Douds DD et al (2017) Influence of host plants and soil diluents on arbuscular mycorrhizal fungus propagation for on-farm inoculum production using leaf litter compost and agrowastes. Biol Agric Hortic 33(1):52–62CrossRefGoogle Scholar
  10. Clapp JP, Young JPW, Merryweather JW et al (1995) Diversity of fungal symbionts in arbuscular mycorrhizas from a natural community. New Phytol 130:259–265CrossRefGoogle Scholar
  11. Daniell TJ, Husband R, Fitter AH et al (2001) Molecular diversity of arbuscular mycorrhizal fungi colonising arable crops. FEMS Microbiol Ecol 36:203–209CrossRefPubMedGoogle Scholar
  12. Dhar PP, Mridha MAU (2006) Biodiversity of arbuscular mycorrhizal fungi in different trees of madhupur forest, Bangladesh. J For Res 17(3):201–205CrossRefGoogle Scholar
  13. Dhar PP, Mridha MAU (2012) Arbuscular mycorrhizal associations in different forest tree species of Hazarikhil forest of Chittagong, Bangladesh. J For Res 23(1):115–122CrossRefGoogle Scholar
  14. Douds DD, Galvez L, Janke RR et al (1995) Effect of tillage and farming system upon populations and distribution of vesicular-arbuscular mycorrhizal fungi. Agric Ecosyst Environ 52:111–118CrossRefGoogle Scholar
  15. Dupré de Boulois H, Voets L, Delvaux B et al (2006) Transport of radiocaesium by arbuscular mycorrhizal fungi to Medicago truncatula under in vitro conditions. Environ Microbiol 8:1926–1934. doi: 10.1111/j.1462-2920.2006.01070.x CrossRefGoogle Scholar
  16. Estrada-Luna AA, Davies FT Jr (2003) Arbuscular mycorrhizal fungi influence water relations, gas exchange, abscisic acid and growth of micropropagated chile ancho pepper (Capsicum annuum) plantlets during acclimatization and post-acclimatization. J Plant Physiol 160:1073–1083CrossRefPubMedGoogle Scholar
  17. Estrada-Luna AA, Davies FT FT Jr, Egilla JN (2000) Mycorrhizal fungi enhancement of growth and gas exchange of micropropagated guava plantlets (Psidium guajava L.) during ex vitro acclimatization and plant establishment. Mycorrhiza 10:1–8CrossRefGoogle Scholar
  18. Garland BC, Schroeder-Moreno MS, Fernandez GE et al (2011) Influence of summer cover crops and mycorrhizal fungi on strawberry production in the southeastern United States. HortScience 46:985–992Google Scholar
  19. Gemma JN, Koske RE, Carreiro M (1989) Seasonal dynamics of selected species of V-A mycorrhizal fungi in a sand dune. Mycol Res 92:317–321CrossRefGoogle Scholar
  20. Gerdemann JW, Nicolson TH (1963) Spores of mycorrhizal Endogone species extracted from soil by wet sieving and decanting. Trans Br Mycol Soc 46:235–244CrossRefGoogle Scholar
  21. Ghosh S, Verma NK (2006) Growth and mycorrhizal dependency of Acacia mangium Willd. inoculated with three vesicular arbuscular mycorrhizal fungi in lateritic soil. New Forest 31:75–81CrossRefGoogle Scholar
  22. Giovannetti M, Mosse B (1980) An evaluation of technique for measuring vesicular-arbuscular mycorrhizae infection in roots. New Phytol 84:489–500CrossRefGoogle Scholar
  23. Guadarrama P, Castillo-Arguero S, Ramos-Zapata J et al (2008) Propagules of arbuscular mycorrhizal fungi in a secondary dry forest of Oaxaca, Mexico. Rev Biol Trop 56(1):269–277PubMedGoogle Scholar
  24. Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Circ Calif Agric Exp Stn 347:1–32Google Scholar
  25. Houba VJG, Van Der Lee JJ, Novozamsky I et al (1988) Soil and plant analysis. Part 5: Soil Analysis Procedure. Agricultural University, WageningenGoogle Scholar
  26. IJdo M, Cranenbrouck M, Declerck S (2011) Methods for large-scale production of AM fungi: past, present, and future. Mycorrhiza 21:1–16CrossRefPubMedGoogle Scholar
  27. INVAM Newsletter 3 (1993) Properties of infective propagules at the suborder level (Glomineae versus Gigasporineae). West Virginia University Web. Accessed 9 July 2014
  28. Johnson NC (1993) Can fertilization of soil select less mutualistic mycorrhizas? Ecol Appl 3:749–757CrossRefPubMedGoogle Scholar
  29. Johnson NC, Copeland JP, Crookston RK, Pfleger FL (1992) Mycorrhizae: an explanation for yield decline in continuous corn and soybean. Agron J 84:387–390CrossRefGoogle Scholar
  30. Kapoor R, Sharma D, Bhatnagar AK (2008) Arbuscular mycorrhizae in micropropagation systems and their potential applications. Sci Hortic 116:227–239CrossRefGoogle Scholar
  31. Kobayashi S (2004) Landscape rehabilitation of degraded tropical forest ecosystems, case study of the CIFOR/Japan project in Indonesia and Peru. For Ecol Manag 201:13–22CrossRefGoogle Scholar
  32. Kollert W, Cherubini L (2012) Teak resources and market assessment 2010 (Tectona grandis Linn.f.). Food and Agriculture Organization of the United Nations (FAO) Planted Forests and Trees Working Paper FP/47/E, Rome, pp 8Google Scholar
  33. Lovelock CE, Andersen K, Morton JB (2003) Arbuscular mycorrhizal communities in tropical forests are affected by host tree species and environment. Oecologia 135:268–279CrossRefPubMedGoogle Scholar
  34. Miller MH, McGonigle TP, Addy HD (1995) Functional ecology of vesicular-arbuscular mycorrhizas as influenced by phosphate fertilization and tillage in agricultural ecosystems. Crit Rev Biotechnol 15:241–255CrossRefGoogle Scholar
  35. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–479CrossRefGoogle Scholar
  36. Oehl F, Sieverding E, Ineichen K et al (2009) Distinct sporulation dynamics of arbuscular mycorrhizal fungal communities from different agroecosystems in long-term microcosms. Agric Ecosyst Environ 134:257–268CrossRefGoogle Scholar
  37. Oliveira RS, Vosatka M, Dodd JC et al (2005) Studies on the diversity of arbuscular mycorrhizal fungi and the efficacy of two native isolates in a highly alkaline anthropogenic sediment. Mycorrhiza 16:23–31CrossRefPubMedGoogle Scholar
  38. Peng S-B, Shen C-Y (1990) Seasonal variations of VA mycorrhizae in the rhizospheres of Welsh onion (Allium fistulosum) and corn (Zea mays) in Beijing and their relationship to several environmental factors. Acta Bot Sin 32:141–145Google Scholar
  39. Phillips JM, Hayman DS (1970) Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Trans Br Mycol Soc 55:157–160CrossRefGoogle Scholar
  40. Rajan SK, Reddy BJD, Bagyaraj DJ (2000) Screening of arbuscular mycorrhizal fungi for their symbiotic efficiency with Tectona grandis. For Ecol Manag 126:91–95CrossRefGoogle Scholar
  41. Ramanwong K (1998) Species diversity of vesicular-arbuscular mycorrhizal fungi of teak (Tectona grandis Linn.f.) and their effects on growth of teak seedlings. Dissertation, Kasetsart UniversityGoogle Scholar
  42. Rouphael Y, Franken P, Schneider C et al (2015) Arbuscular mycorrhizal fungi act as biostimulants in horticultural crops. Sci Hortic 196:91–108CrossRefGoogle Scholar
  43. Ruiz-Lozano JM (2003) Arbuscular mycorrhizal symbiosis and alleviation of osmotic stress: New perspectives for molecular studies. Mycorrhiza 13:309–317CrossRefPubMedGoogle Scholar
  44. Schubert A, Lubraco G (2000) Mycorrhizal inoculation enhances growth and nutrient uptake of micropropagated apple rootstocks during weaning in commercial substrates of high nutrient availability. Appl Soil Ecol 15:113–118CrossRefGoogle Scholar
  45. Schüßler A, Walker C (2010) The Glomeromycota: a species list with new families and new genera. Libraries at The Royal Botanic Garden Edinburgh, Edinburgh, UK; The Royal Botanic Garden Kew, Kew, UK; Botanische Staatssammlung Munich, Munich, Germany; and Oregon State University, Corvallis, Oregon, pp 1–56Google Scholar
  46. Singh SS, Tiwari SC, Dkhar MS (2003) Species diversity of vesicular-arbuscular mycorrhizal (VAM) fungi in jhum fallow and natural forest soils of Arunachal Pradesh, North Eastern India. Trop Ecol 44:207–215Google Scholar
  47. Smith SE, Read DJ (2008) Mycorrhizal symbiosis, 3rd edn. Academic Press, New YorkGoogle Scholar
  48. Sparks DL, Page AL, Helmke PA et al (1996) Methods of soil analysis part 3, chemical methods. Soil Science Society of America and American Society of Agronomy, WisconsinGoogle Scholar
  49. Stürmer SL, Filho OK, de Queiroz MH et al (2006) Occurrence of arbuscular mycorrhizal fungi in soils of early stages of a secondary succession of Atlantic Forest in South Brazil. Acta Bot Bras 20(3):513–521CrossRefGoogle Scholar
  50. Swaminathan C, Srinivasan VM (2006) Influence of microbial inoculants on seedling production in teak (Tectona grandis L.f.). J Sustain For 22(3):63–76CrossRefGoogle Scholar
  51. Thomas GV (1988) Vesicular-arbuscular mycorrhizal symbiosis in coconut (Cocos nucifera) in relation to the root (wilt) disease and intercropping or mixed cropping. Indian J Agric Sci 57:145–146Google Scholar
  52. Udaiyan K, Karthikeyan A, Muthukumar T (1996) Influence of edaphic and climatic factors on dynamics of root colonization and spore density of vesicular-arbuscular mycorrhizal fungi in Acacia farnesiana Willd. and A. planifrons Trees 11:65–71Google Scholar
  53. Urgiles N, Strauß A, Loján P et al (2014) Cultured arbuscular mycorrhizal fungi and native soil inocula improve seedling development of two pioneer trees in the Andean region. New Forest 45:859–874CrossRefGoogle Scholar
  54. Verma RK, Jamaluddin (1995) Association and activity of arbuscular mycorrhizae of teak (Tectona grandis) in central India. In Special issue: focus on teak. Indian For 121(6):533–539Google Scholar
  55. Voets L, Dupré de Boulois H, Renard L et al (2005) Development of an autotrophic culture system for the in vitro mycorrhization of potato plantlets. FEMS Microbiol Lett 248:111–118CrossRefPubMedGoogle Scholar
  56. Voets L, de la Providencia IE, Fernandez K et al (2009) Extraradical mycelium network of arbuscular mycorrhizal fungi allows fast colonization of seedlings under in vitro conditions. Mycorrhiza 19:347–356CrossRefPubMedGoogle Scholar
  57. Wright SF, Upadhyaya A (1998) A survey of soils for aggregate stability and glomalin, a glycoprotein produced by hyphae of arbuscular mycorrhizal fungi. Plant Soil 198:97–107CrossRefGoogle Scholar
  58. Zak JC, Danielson RM, Parkinson D (1982) Mycorrhizal fungal spore numbers and species occurrence in two amended mine spoils in Alberta, Canada. Mycologia 74:785–792CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Amornrat Chaiyasen
    • 1
    • 4
  • David D. Douds
    • 2
  • Paiboolya Gavinlertvatana
    • 3
  • Saisamorn Lumyong
    • 1
  1. 1.Department of Biology, Faculty of ScienceChiang Mai UniversityChiang MaiThailand
  2. 2.USDA, Agricultural Research Service, Eastern Regional Research CenterWyndmoorUSA
  3. 3.Thai Orchids Lab Co. Ltd.BangkokThailand
  4. 4.Soil Science Research Group, Agricultural Production Science Research and Development Division, Department of AgricultureMinistry of Agriculture and CooperativesBangkokThailand

Personalised recommendations