Low-Density Co-Inoculation of Myanmar Bradyrhizobium yuanmingense MAS34 and Streptomyces griseoflavus P4 to Enhance Symbiosis and Seed Yield in Soybean Varieties


This study examined whether low-density co-inoculation of Myanmar Bradyrhizobium yuanmingense strain MAS34 and Streptomyces griseoflavus P4 would enhance nodulation, N2 fixation, and seed yield in two soybean varieties. A field experiment was conducted during the July to November 2012 growing season at Kyushu University Farm, Japan, using a split-plot design with three replications and the following four treatments: T1, an uninoculated treatment with peat moss (uninoculated); T2, a single inoculation with S. griseoflavus P4 (P4); T3, a single inoculation of B. yuanmingense MAS34 (MAS34); and T4, a dual inoculation of P4 with MAS34 (P4 + MAS34). Two varieties of soybean, Yezin-3 (Rj4) and Yezin-6 (non-Rj), were used. The N2 fixation activity of soybean was evaluated by the relative ureide method using xylem solute from root bled sap at the early pod-fill stage (R3.5). Dry matter production, N2 fixation, and seed yield were significantly (P < 0.01) different between the inoculated treatments. The effect of variety was also significant (P < 0.05) for nodule dry weight at the V6 stage, percentage of N derived from the atmosphere at the R3.5 stage, and seed yield at the maturity stage. The number of nodules on the tap roots was significantly higher in Yezin-3 than in Yezin-6. The single inoculation of P4 did not have a significant effect on dry matter production, N2 fixation, and seed yield in either soybean variety. The dry matter production, relative ureide index, percentage of N derived from the atmosphere, and seed yield were significantly (P < 0.01) enhanced by a single inoculation of MAS34 in Yezin-3 and by dual inoculation of P4 + MAS34 in Yezin-6. These results indicate that low inoculum concentrations (105 cells seed-1) increase N2 fixation and seed yield in these soybean varieties under open field conditions. Myanmar B. yuanmingense MAS34 and S. griseoflavus P4 are expected to be useful biofertilizers for soybean production.

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K. Soe and T. Yamakawa, "Low-Density Co-Inoculation of Myanmar Bradyrhizobium yuanmingense MAS34 and Streptomyces griseoflavus P4 to Enhance Symbiosis and Seed Yield in Soybean Varieties," American Journal of Plant Sciences, Vol. 4 No. 9, 2013, pp. 1879-1892. doi: 10.4236/ajps.2013.49231.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] M. Aslam, S. Mirza, M. Shjah, S. M. Jayed and N. Naeemullah, “New Early Maturing and High Yielding Soybean Varieties,” Crop Production Bulletin, Vol. 4, 1995, pp. 1-11.
[2] FAO, “Legume Inoculants and Their Use,” NifTAL Project, USA Food and Agriculture Organization of the United Nations, Via delle Terme di Caracalla, Rome, Italy, 1984.
[3] CSO, “Myanmar Agricultural Statistics (1992-1993 to 2004-2005),” Central Statistical Organization, Ministry of National Planning and Economic Development, Yangon, Republic of the Union of Myanmar, 2006.
[4] N. Boonkerd and P. Singleton, “Production of Rhizobium Biofertilizer,” In: S. Kannaiyan, Ed., Biotechnology of Biofertilizers, Narosa Publishing House, New Delhi, 2002, pp. 122-128.
[5] G. Stacey, M. Libault, L. Brechenmacher, J. Wan and G. D. May, “Genetics and Functional Genomics of Legume Nodulation,” Current Opinion in Plant Biology, Vol. 9, No. 2, 2006, pp. 110-121. doi:10.1016/j.pbi.2006.01.005
[6] P. H. Graham, “Nitrogen Transformations,” In: M. E. Sumner, Ed., Hand Book of Soil Science, CRC Press, London, 2000, pp. 139-147.
[7] G. U. Okereke, C. Onochie and E. Onyeagba, “Effectiveness of Foreign Bradyrhizobia Strains in Enhancing Nodulation, Dry Matter and Seed Yield of Soybean (Glycine max L.) Cultivars in Nigeria,” Biology and Fertility of Soils, Vol. 33, No. 1, 2001, pp. 3-9. doi:10.1007/s003740000264
[8] H. H. Tien, T. M. Hien, M. T. Son and D. Herridge, “Rhizobial Inoculation and N2 Fixation of Soybean and Mungbean in the Eastern Region of South Vietnam,” Proceedings of a Workshop Held in Hanoi, Vietnam, 2002, pp. 29-36.
[9] S. K. Mahna, “Individual Partner Report,” INCO-DEV Research Project on Soybean BNF and Mycorrhization for Improved Production in South Asia, Department of Botany, Maharshi Dayanand Saraswati University, Ajmer, 2006.
[10] R. W. Weaver and L. R. Frederick, “Effects of Inoculum Rate on Competitive Nodulation of Glycine max L. Merril. II. Field Studies,” Agronomy Journal, Vol. 66, No. 2, 1974, pp. 233-236. https://www.soils.org/publications/aj/abstracts/66/2/AJ0660020233
[11] S. M. Lohrke, C. J. Madrzak, H. Hur, A. K. Judd, J. H. Orf and M. J. Sadowsky, “Inoculum Density-Dependent Restriction of Nodulation in the Soybean-Bradyrhizobium japonicum Symbiosis,” Symbiosis, Vol. 29, No. 1, 2000, pp. 59-70. http://eurekamag.com/research/003/479/inoculum-density-dependent-restriction-nodulation-soybean-bradyrhizobium-japonicum-symbiosis.php
[12] T. Yamakawa and Y. Saeki, “Inoculation Methods of Bradyrhizobium japonicum on Soybean in South-West Area of Japan,” In: J. E. Board, Ed., A Comprehensive Survey of International Soybean Research—Genetics, Physiology, Agronomy and Nitrogen Relationships, InTech, Rijeka, 2013, pp. 83-114.
[13] J. Tang-um and H. Niamsup, “Chitinase Production and Antifungal Potential of Endophytic Streptomyces Strain P4,” Maejo International Journal of Science and Technology, Vol. 6, No. 1, 2012, pp. 95-104. http://www.mijst.mju.ac.th/vol6/95-104.pdf
[14] P. Thapanapongworakul, “Characterization of Endophytic Actinomycetes Capable of Controlling Sweet Pea Root Rot Diseases and Effects on Root Nodule Bacteria,” M.Sc. Dissertation, Chiang Mai University, Chiang Mai, 2003.
[15] K. M. Soe and T. Yamakawa, “Evaluation of Effective Myanmar Bradyrhizobium Strains Isolated from Myanmar Soybean and Effects of Coinoculation with Streptomyces griseoflavus P4 for Nitrogen Fixation,” Soil Science and Plant Nutrition, Vol. 59, No. 3, 2013, pp. 361-370. doi:10.1080/00380768.2013.794437
[16] S. K. Dansoc and J. D. Owiredu, “Competitiveness of Introduce and Indigenous Cowpea in Three Soil,” Soil Biology and Biochemistry, Vol. 20, No. 3, 1988, pp. 305-310. doi:10.1016/0038-0717(88)90008-9
[17] M. Yahya-Abadi, “Evaluation of Nitrogen Fixation Potential and Nutrients Uptaking in Some Common Bean Symbiosis Bacteria,” Proceeding of 10th Iranian Congress of Crop Production and Plant Breeding, Karaj, 19-21 August 2008, pp. 1-75.
[18] M. Mehrpouyan, “Nitrogen Fixation Efficiency in Native Strains Compared with Non-Native Strains of Rhizobium leguminosarum,” In: International Conference on Environment Science and Engineering IPCBEE, Vol. 8, IACSIT Press, Singapore, 2011.
[19] J. Muramoto, I. Goto and M. Ninaki, “Rapid Analysis of Exchangeable Cation and Cation Exchange Capacity (CEC) of Soils by Shaking Extraction Method,” Japanese Soil Science and Plant Nutrition, Vol. 63, No. 2, 1992, pp. 210-215 (in Japanese).
[20] D. A. Cataldo, L. E. Schrader and V. L. Youngs, “Analysis by Digestion and Colrimetric Assay of Total Nitrogen in Plant Tissues High in Nitrate,” Crop Science, Vol. 14, No. 6, 1974, pp. 854-856. doi:10.2135/cropsci1974.0011183X001400060024x
[21] J. Murphy and J. P. Riley, “A Modified Single Solution for the Determination of Phosphate in Natural Waters,” Analytica Chimica Acta, Vol. 27, No. 1, 1962, pp. 31-36. doi:10.1016/S0003-2670(00)88444-5
[22] E. Truog, “The Determination of the Readily Available Phosphorus in Soils,” Agronomy Journal, Vol. 22, No. 10, 1930, pp. 874-882. doi:10.2134/agronj1930.00021962002200100008x
[23] P. Somasegaran and H. J. Hoben, “Handbook for Rhizobia,” In: R. C. Garber, Ed., Methods in Legumes-Rhizobium Technology, Springer-Verlag, Inc., New York, 1994, pp. 58-64.
[24] K. M. Soe, T. Yamakawa, S. Hashimoto and P. Sarr, “Phylogenetic Diversity of Indigenous Soybean Bradyrhizobia from Different Agro-Climatic Regions in Myanmar,” ScienceAsia, 2013, in Press.
[25] T. E. Devine and L. D. Kuykendall, “Host Genetic Control of Symbiosis in Soybean (Glycine max L.),” Plant and Soil, Vol. 186, No. 1, 1996, pp. 173-187. doi:10.1007/BF00035072
[26] J. Ishizuka, Y. Suemasu and K. Mizogami, “Preference of Rj-Soybean Cultivars for Bradyrhizobium japonicum for Nodulation,” Japanese Soil Science and Plant Nutrition, Vol. 37, No. 1, 1991, pp. 15-21. http://ci.nii.ac.jp/naid/110001720006
[27] Y. Saeki, I. Akagi, H. Takaki and Y. Nagatomo, “Diversity of Indigenous Bradyrhizobium Strains Isolated from Three Different Rj-Soybean Cultivars in Terms of Randomly Amplified Polymorphic DNA and Intrinsic Antibiotic Resistance,” Japanese Soil Science and Plant Nutrition, Vol. 46, No. 4, 2000, pp. 917-926. http://ci.nii.ac.jp/ naid/110001717919
[28] G. Vest and B. E. Caldwell, “Rj4-A Gene Conditioning Ineffective Nodulation in Soybean,” Crop Science, Vol. 12, No. 5, 1972, pp. 692-693.
[29] L. D. Kuykendall, “Isolation and Identification of Genetically Marked Strains of Nitrogen-Fixing Microsymbionts of Soybeans,” In: G. H. Elkan, Ed., Practical Symbiotic Nitrogen Fixation Methodology, Marcel Dekker, New York, 1987, pp. 205-220.
[30] M. Shimizu, Y. Nakagawa, Y. Sato, T. Furumai, Y. Igarashi, H. Onaka, R. Yoshida and H. Kunoh, “Studies on Endophytic Actinomycetes (I) Streptomyces sp. Isolate from Rhododendron and Its Antifungal Activity,” Journal of General Plant Pathology, Vol. 66, No. 4, 2000, pp. 360-366. doi:10.1007/PL00012978
[31] T. Ohyama, M. Ito, K. Kobayashi, S. Araki, S. Yasuyoshi, O. Sasaki, T. Yamazaki, K. Soyama, R. Tanemura, Y. Mizuno and T. Ikarashi, “Analytical Procedures of N, P, K Contents in Plant and Manure Materials Using H2SO4-H2O2 Kjeldahl Digestion Method,” Bulletin of the Faculty of Agriculture, Niigata University, Vol. 43, 1991, pp. 110-120 (in Japanese).
[32] M. B. Peoples, A. W. Faizah, B. Rerkasem and D. F. Herridge, “Methods for Evaluating Nitrogen Fixation by Nodulated Legumes in the Field,” Australian Centre for International Agricultural Research Monograph series, Vol. 11, No. 7, 1989, pp. 39-40.
[33] S. Moore and W. H. Stein, “A Modified Ninhydrin Reagent for the Photometric Determination of Amino Acids and Realted Compounds,” The Journal of Biological Chemistry, Vol. 211, No. 2, 1954, pp. 907-913. http://www.jbc.org/content/211/2/907.citation
[34] D. A. Cataldo, M. Haroon, L. E. Schrader and V. L. Youngs, “Rapid Colorimetric Determination of Nitrate in Plant Tissue by Nitration of Salicylic Acid,” Communications in Soil Science and Plant Analysis, Vol. 6, No. 1, 1975, pp. 71-80. doi:10.1080/00103627509366547
[35] E. G. Young and C. F. Conway, “On the Estimation of Allantoin by the Rimini-Schryver Reaction,” The Journal of Biological Chemistry, Vol. 142, No. 2, 1942, pp. 839-853. http://www.jbc.org/content/142/2/839.citation
[36] D. F. Herridge and M. B. Peoples, “Ureide Assay for Measuring Nitrogen Fixation by Nodulated Soybean Calibrated by 15N Methods,” Plant Physiology, Vol. 93, No. 2, 1990, pp. 495-503. doi:10.1104/pp.93.2.495
[37] D. F. Herridge and M. B. Peoples, “Timing of Xylem Sampling for Ureide Analysis of Nitrogen Fixation,” Plant and Soil, Vol. 238, No. 1, 2002, pp. 57-67.
[38] H. M. A. Purcino, P. M. Festin and G. H. Elkan, “Identification of Effective Strains of Bradyrhizobium for Arachis pintoi,” Tropical Agriculture, Vol. 77, No. 4, 2000, pp. 226-231.
[39] B. D. Pant and B. N. Prasad, “Effectiveness of Bradyrhizobium Isolates on Seedling Growth and Nitrogen Content in Soybean [Glycine max (L.) Merr.],” Botanica Orientalis, Vol. 4, No. 1, 2004, pp. 1-3. http://nepjol.info/index.php/BOTOR/article/view/493
[40] K. M. Soe, A. Bhromsiri and D. Karladee, “Effect of Selected Endophytic Actinomycetes (Steptomyces sp.) and Bradyrhizobia from Myanmar on Growth, Nodulation, Nitrogen Fixation and Yield of Different Soybean Varieties,” Chiang Mai University Journal of Natural Sciences, Vol. 9, No. 1, 2010, pp. 95-109.
[41] S. T. Takats, “Suppression of Nodulation in Soybeans by Superoptimal Inoculation with Bradyrhizobium japonicum,” Physiologia Plantarum, Vol. 66, No. 4, 1986, pp. 669-673. doi:10.1111/j.1399-3054.1986.tb05597.x
[42] R. S. Smith, “Legume Inoculant Formulation and Application,” Canadian Journal of Microbiology, Vol. 38, No. 6, 1992, pp. 485-492. doi:10.1139/m92-080
[43] S. Jitacksorn and M. J. Sadowsky, “Nodulation Gene Regulation and Quorum Sensing Control Density-Dependent Suppression and Restriction of Nodulation in the Bradyrhizobium japonicum-Soybean Symbiosis,” Applied and Environmental Microbiology, Vol. 74, No. 12, 2008, pp. 3749-3756. doi:10.1128/AEM.02939-07
[44] J. E. Thies, P. W. Singleton and B. B. Bohlool, “Influence of Size of Indiogenous Rhizobial Populations on Establishment and Symbiotic Performance of Introduced Rhizobia on Field-grown Legumes,” Applied and Environmental Microbiology, Vol. 57, No. 1, 1991, pp. 19-28.
[45] T. Sato, N. Onoma, H. Fujikake, N. Ohtake, K. Sueyoshi and T. Ohyama, “Changes in Four Leghemoglobin Components in Nodules of Hypernodulating Soybean (Glycine max [L] Merr.) Mutant and Its Parent in the Early Nodule Developmental Stage,” Plant and Soil, Vol. 237, No. 1, 2001, pp. 129-135. doi:10.1023/A:1013317219871
[46] A. Souleimanov, B. Prithiviraj and D. L. Smith, “The Major Nod Factor of Bradyrhizobium japonicum Promotes Early Growth of Soybean and Corn,” Journal of Experimental Botany, Vol. 53, No. 376, 2002, pp. 1929-1934. doi:10.1093/jxb/erf034
[47] H. C. Huang and R. S. Erickson, “Effect of Seed Treatment with Rhizobium leguminosarum on Pythium Dumping-Off, Seedling Height, Root Nodulation, Root Biomass, Shoot Biomass, and Seed Yield of Pea and Lentil,” Journal of Phytopathology, Vol. 155, No. 1, 2006, pp. 31-37. doi:10.1111/j.1439-0434.2006.01189.x
[48] S. P. Wani, O. P. Rupela and K. K. Lee, “Sustainable Agriculture in a Semiarid Tropic through Biological Nitrogen Fixation in Grain Legume,” Plant and Soil, Vol. 174, No. 2, 1995, pp. 29-49. doi:10.1007/BF00032240
[49] D. Egamberdiyeva, D. Qarshieva and K. Davranov, “The Use of Bradyrhizobium to Enhance Growth and Yield of Soybean in Calcareous Soil in Uzbekistan,” Journal of Plant Growth Regulation, Vol. 23, No. 1, 2004, pp. 54-57. doi:10.1007/s00344-004-0069-4
[50] V. Milic, N. Mrkovacki, M. Popovic and D. Malencic, “Nodule Efficiency of Three Soybean Genotypes Inoculated by Different Methods,” Rostlinna Vyroba, Vol. 48, No. 8, 2002, pp. 356-360. http://www.agriculturejournals.cz/publicFiles/53851.pdf
[51] K. M. Soe, A. Bhromsiri, D. Karladee and T. Yamakawa, “Effects of Endophytic Actinomycetes and Bradyrhizobium japonicum Strains on Growth, Nodulation, Nitrogen Fixation and Seed Weight of Different Soybean Varieties,” Soil Science and Plant Nutrition, Vol. 58, No. 3, 2012, pp. 319-325.
[52] A. Akarapisan, A. Bhromsiri and P. Sangmanee, “Selection of Suitable Isolates of Endophytic Actinomycetes and Rhizobia for Improvement of N2 Fixation and Disease Control of Various Pisum sativum on the Highland Area,” Final Report for Highland Research and Development Institute, Thailand, 2008, pp. 297-306.

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