American Journal of Microbiological Research

ISSN (Print): 2328-4129

ISSN (Online): 2328-4137

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Website: http://www.sciepub.com/journal/AJMR

   

Article

Prevalence of GBV-C and Its Impacts among Patients with Hepatitis B and C Viruses in Addis Ababa, Ethiopia

1Department of Microbial Cellular & Molecular Biology, Addis Ababa University, Addis Ababa, Ethiopia

2Schools of Medicine, Addis Ababa University, Addis Ababa, Ethiopia

3Infectious and Non-infectious Diseases Research Directorate, Ethiopian Health & Nutrition Research


American Journal of Microbiological Research. 2017, 5(1), 1-6
doi: 10.12691/ajmr-5-1-1
Copyright © 2017 Science and Education Publishing

Cite this paper:
Mohamed Farouk, Abate Bane Shewaye, Desta Kassa, Mekuria Lakew. Prevalence of GBV-C and Its Impacts among Patients with Hepatitis B and C Viruses in Addis Ababa, Ethiopia. American Journal of Microbiological Research. 2017; 5(1):1-6. doi: 10.12691/ajmr-5-1-1.

Correspondence to: Mohamed  Farouk, Department of Microbial Cellular & Molecular Biology, Addis Ababa University, Addis Ababa, Ethiopia. Email: mohamed.f.mirghani@gmail.com

Abstract

Background: Hepatocellular carcinoma (HCC) caused by hepatitis B virus (HBV) and hepatitis C virus (HCV) is currently one of the most common neoplasms worldwide. GB virus C/hepatitis G virus (HGV/GBV-C) is a virus in the Flaviviridae family isolated from patients with liver disease. It has the same mode of transmission with HBV and HCV and is common in high risk group. The impact of HGV/GBV-C in clinical outcome among HBV and HCV is controversy. Therefore, this study was conducted to determine the prevalence and the association of (HGV/GBV-C) in the clinical outcome among HCV and HBV patients. Materials: This case-control study was performed in Addis Ababa University, Ethiopia. The cases were 101 patients with viral hepatitis collected from Adera internal medical specialty center. The control group consisted of 50 healthy individuals collected from the Ethiopian Public Health and Research Institutes. The serological analysis and liver enzyme levels were determined for each of the participants. RNA was extracted, reversed transcribed, and amplified by Real Time polymerase chain reaction (PCR), using primers for 5- untranslated region (5-UTR) of the GBV-C. Results: Analysis of the 101 samples of the hepatitis patients showed that; 83(82.2%) were positive for HBV while only18 (17.8%) for HCV. The prevalence of (HGV/GBV-C) RNA was 11(13.2%) in HBV, 2 (11.1%) in HCV and rests of the control group were negative. There was no significant difference (P > 0.05) in the liver enzymes level among (HGV/GBV-C) negative and positive individuals. Conclusion: Our study showed that the co-infection rate of (HGV/GBV-C) RNA among hepatitis patients was significantly higher (P <0.05) in HBV than in HCV patients, and the virus has no association in the course of the disease.

Keywords

References

[1]  Hu DJ, Bower WA, Ward JW. Viral hepatitis. In: Morse S, Moreland AA, Holmes KK, eds. Atlas of sexually transmitted diseases and AIDS. London: Elsevier; 2010: 203-29.
 
[2]  CDC. Recommendations for prevention and control of hepatitis C virus (HCV) infection and HCV-related chronic disease. MMWR 1998; 47.
 
[3]  Armstrong GL, Wasley A, Simard EP, McQuillan GM, Kuhnert WL, Alter MJ. The prevalence of hepatitis C virus infection in the United States, 1999 through 2002. Ann Intern Med 2006; 144: 705-4.
 
[4]  Peters MG. Special populations with hepatitis B virus infection. Hepatology 2009; 49(Suppl 5): S146-S155.
 
[5]  Koopmans M., Duizer E. (2004): Foodborne viruses: an emerging problem. International Journal of Food Microbiology, 90, 23-41.
 
Show More References
[6]  Vasickova P., Dvorska L., Lorencova A., Pavlik I. (2005): Viruses as a cause of foodborne diseases: a review of the literature. Veterinarni Medicina 50, 89-104.
 
[7]  Ashbolt N.J. (2004): Microbial contamination of drinking water and disease outcomes in developing regions. Toxicology, 198, 229-238.
 
[8]  Global policy report on the prevention and control of viral hepatitis (2013).World Health organization; Geneva, Switzerland.
 
[9]  Nassal M.HBV cccDNA: viral persistence reservoir and key obstacle for a cure of chronic hepatitis B. Gut 2015; 64: 1972-84.
 
[10]  Penin F. structural biology of hepatitis C virus. Clin Liver Dis 2003; 7: 1-21.
 
[11]  Stapleton JT, Foung S, Muerhoff AS, Bukh J, Simmonds P. The GB viruses: a review and proposed classification of GBV-A, GBV-C (HGV), and GBV-D in genus Pegivirus within the family Flaviviridae. J Gen Virol 2011; 92: 233-46.
 
[12]  Lavanchy D. Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures. J Viral Hepat. 2004; 11(2): 97-107.
 
[13]  Fattovich G. Natural history and prognosis of hepatitis B. Semin Liver Dis 2003; 23: 47-58.
 
[14]  McMahon BJ. The natural history of chronic hepatitis B virus infection. Semin Liver Dis 2004; 24: 17-21.
 
[15]  World Health Organization. Hepatitis C Fact Sheet N°164. Available from: Http: //www. who. int/ mediacentre /factsheets /fs164/en/. Last accessed: March 2016.
 
[16]  Sultan MT, Rahman MM, Begum S. Epidemiology of hepatitis C virus (HCV) infection. J Bangladesh Coll Phy Surg. 2009; 27: 160-2.
 
[17]  Lee MH, Yang HI, Yuan Y, Lltalien G, Chen CJ (2014). Epidemiology and natural history of hepatitis C virus infection. World J. Gastroenterol. 20(28): 9270-9280.
 
[18]  Lavanchy D. Evolving epidemiology of hepatitis C virus. Clin Microbiol Infect 2011;17: 107-115
 
[19]  Simons, J. N., T. P. Leary, G. J. Dawson, T. J. Pilot-Matias, A. S. Muerhoff, G. G. Schlauder, S. M. Desai, and I. K. Mushahwar. 1995a. Isolation of novelvirus-like sequences associated with human hepatitis. Nat Med 1: 564-569.
 
[20]  Linnen, J., J. Wages, Jr., Z. Y. Zhang-Keck, K. E. Fry, K. Z. Krawczynski, H. Alter, E. Koonin, M. Gallagher, M. Alter, S. Hadziyannis, P. Karayiannis, K. Fung, Y. Nakatsuji, J. W. Shih, L. Young, M. Piatak, Jr., C. Hoover, J. Fernandez, S. Chen, J. C. Zou, T. Morris, K. C. Hyams, S. Ismay, J. D. Lifson, G. Hess, S. K. Foung, H. Thomas, D. Bradley, H. Margolis, and J. P. Kim. 1996. Molecular cloning and disease association of hepatitis G virus: a transfusion-transmissible agent. Science 271: 505-508.
 
[21]  Leary, T. P., A. S. Muerhoff, J. N. Simons, T. J. Pilot-Matias, J. C. Erker, M. L. Chalmers, G. G. Schlauder, G. J. Dawson, S. M. Desai, and I. K. Mushahwar. 1996. Sequence and genomic organization of GBV-C: a novel member of the flaviviridae associated with human non-A-E hepatitis. J Med Virol 48: 60-67.
 
[22]  Cheung RC, Keeffe EB, and Greenberg HB, Hepatitis G virus: Is it a hepatitis virus? West J Med 167: 23-33. 1997.
 
[23]  Pavesi A, Detection of signature sequences in overlapping genes and prediction of a novel overlapping gene in hepatitis G virus. J Mol Evol 50: 284-295. 2000
 
[24]  Feng Y, Zhao W, Dai J, Li Z, Zhang X, Liu L, Bai J, Zhang H, Lu L, and Xia X, A novel genotype of GB virus C: its identification and predominance among injecting drug users in Yunnan, China, PLoS One 6: e21151. 2011.
 
[25]  Sathar MA, York DF, Group 5: GBV-C/HGV isolates from South Africa, J Med Virol 65: 121-122. 2011.
 
[26]  Scallan MF, Clutterbuck D, Jarvis LM, et al., Sexual transmission of GB virus C/hepatitis G virus, J Med Virol, 1998; 55: 203-8.
 
[27]  Nerurkar VR, Chua PK, Hoffmann PR, et al., High prevalence of GB virus C/hepatitis G virus infection among homosexual men infected with human immunodeficiency virus type 1: evidence for sexual transmission, J Med Virol, 1998; 56: 123-7.
 
[28]  Fischler B, Lara C, Chen M, et al., Genetic evidence for mother-to-infant transmission of hepatitis G virus, J Infect Dis, 1997; 176: 281-5.
 
[29]  Dawson,G.J. et al. Prevalence studies of GB virus-C infection using reverse transcriptase-polymerase chain reaction. J Med V irol 50, 97-103 (1996).
 
[30]  Liu,H.F., Muyembe-Tamfum,J.J., Dahan,K., Desmyter,J. & Goubau,P. High prevalence of GB virus C/hepatitis G virus in Kinshasa. J Med Virol 60, 159-165 (2000).
 
[31]  Tucker,T.J., Louw,S.J., Robson,S.C., Isaacs,S. & Kirsch,R.E. High prevalence of GBV-C hepatitis G virus infection in a rural South African population. J Med Virol 53, 225-228 (1997).
 
[32]  Bhattarai N, Stapleton JT, GB virus C: the good boy virus? Trends Microbiol, 20(3): 124-30. 2012.
 
[33]  Toyoda H, Fukuda Y, Hayakawa T, Takamatsu J, and Saito H, Effect of GB virus C/hepatitis G virus co-infection on the course of HIV infection in hemophilia patients in Japan. Journal Acquired Immune Deficiency Syndromes.17: 209-13. 1998.
 
[34]  Heringlake S, Ockenga J, Tillmann HL, Trautwein C, Meissner D, Stoll M, Hunt J, Jou C, Solomon N, Schmidt RE, and Manns MP, GB virus C/hepatitis G virus infection: a favorable prognostic factor in human immunodeficiency virus-infected patients? Journal of Infectious Diseases.177: 1723-6. 1998.
 
[35]  Bjorkman P, Flamholc L, Molnegren V, Marshall A, Guner N, Widell A (2007). Enhanced and resumed GB virus C replication in HIV-1-infected individuals receiving HAART. AIDS. 21(12): 1641-1643.
 
[36]  Yoshiba M, Okamoto H, Mishiro S. Detection of the GBV-C hepatitis virus genome in serum from patients with fulminant hepatitis of unkown etiology. Lancet 1995; 346: 1131-2.
 
[37]  Alter J. A reassessment of the literature on the hepatitis G virus. Transfusion 1997; 37: 569-72.
 
[38]  Chams V, Fournier-Wirth C, Chabanel A, et al. Is GB virus-C alias “hepatitis G” involved in human pathology. Trans Clin Bioi 2003; 10: 292 306.
 
[39]  Birgit Kallinowski, Christine Buhrmann, Stefanie Seipp, Tobias Goeser, Wolfgang Stremmel, Gerd Otto, and Lorenz Theilmann (1998). Incidence, Prevalence, and Clinical Outcome of Hepatitis GB-C Virus Infection in Liver Transplant Patients. Liver Transplantation and Surgery, (4)1: 28-33.
 
[40]  Yue Feng, Li Liu, Yue-Mei Feng, Wenhua Zhao, Zheng Li, A-Mei Zhang, Yuzhu Song, and Xueshan Xia (2014). GB Virus C infection in Patients With HIV/Hepatitis C Virus Coinfection: Improvement of the Liver Function in Chronic Hepatitis C. Hepat Mon. 14(3): e14169.
 
[41]  Berg T, Naumann U, Fukumoto T, Lobeck W, Bechstein P, Neuhaus P, et al. GB virus C infection in patients with chronic hepatitis B and C before and after liver transplantation. Transplantation 1996;62: 711-714
 
[42]  Alvarado-Mora MV, Botelho L, Nishiya A, Neto RA, Gomes-Gouvea MS, Gutierrez MF, Carrilho FJ, Pinho JR (2011) Frequency and genotypic distribution of GB virus C (GBV-C) among Colombian population with Hepatitis B (HBV) or Hepatitis C (HCV) infection. Virol J 8: 345.
 
[43]  Tanaka E, Alter HJ, Nakatsuji Y, et al. Effect of hepatitis G virus infection on chronic hepatitis C. Ann Intern Med. 1996; 125(9): 740-3.
 
[44]  Ruiz V, Espinola L, Mathet VL, Perandones CE and Oubina JR, Design, development and evaluation of a competitive RT-PCR for quantitation of GBV-C RNA, J Virol Methods, 2006; 136: 58-64.
 
[45]  Thomas DL, Vlahov D, Alter HI, Hunt, Marshall R, Astemborski I and Nelson KE. (1998) Association of antibody to GB Virus C (Hepatitis G Virus) with viral clearance and protection from reinfection. J. Infect. Dis. 177, 539-542.
 
[46]  Masaru Enomoto, Shuhei,Shiguchi, Katsuhiko Fukuda, Tetsuo Kuroki, Motoharu Tanaka, Shuzo Otani, Masayuki Ogami, and Takeyuki Monna (1998). Characteristics of Patients with Hepatitis C Virus With and Without GB Virus C/Hepatitis G Virus Co-infection and Efficacy of Interferon Alfa. HEPATOLOGY, (27)5.
 
[47]  Tanaka Y, Mizokami M, Orito E, Ohba K, Nakano T, Kato T, Kondo Y, Ding X, Ueda R, Sonoda S, Tajima K, Miura T, Hayami M: GB virus C/hepatitis G virus infection among Colombian native Indians. Am J Trop Med Hyg 1998, 59: 462-467.
 
[48]  Berzsenyi MD, Bowden DS, Kelly HA, Watson KM, Mijch AM, Ham¬mond RA, et al. Reduction in hepatitis C-related liver disease associated with GB virus C in human immunodeficiency virus coinfection. Gastroenterology. 2007; 133(6): 1821-30.
 
[49]  El-Zayadi AR, Abe K, Selim O, Naito H, Hess G, Ahdy A: Prevalence of GBVC/ hepatitis G virus viraemia among blood donors, health care personnel, chronic non-B non-C hepatitis, chronic hepatitis C and hemodialysis patients in Egypt. J Virol Methods 1999, 80: 53-58.
 
[50]  Monica, V., Alvarado, M., Livia, B., Anna, N., Raymundo, A., Neto3, M. and Maria, F. (2011). Frequency and genotypic distribution of GB virus C (GBV-C) among Colombian population with Hepatitis B (HBV) or Hepatitis C (HCV) infection. Virol J. 8: 345.
 
[51]  Abu Odeh RO, Al‐Moslih MI, Al‐Jokhdar MW, Ezzeddine SA. Detection and genotyping of GBV‐C virus in the United Arab Emirates. J Med Virol 2005; 76: 534-40.
 
[52]  Akcali S, Sanlidag T, Ozbakkaloglu B. Prevalence of GBV-C/ hepatitis G virus viremia among chronic hepatitis B, chronic hepatitis C and hemodialysis patients in Turkey. Ann Saudi Med 2006; 26: 68-9.
 
[53]  Grassi M, Raffa S, Traditi F, Ferrazzi M, Cioschi S, Fontana M, et al. Detection and clinical evaluation of GBV-C/HGV in plasma from patients with chronic hepatitis of unknown etiology. Clin Ter. 2000 Jul-Aug; 151(4): 241-5.
 
[54]  Ramezani A, Gachkar L, Eslamifar A, et al. Detection of hepatitis G virus envelope protein E2 antibody in blood donors. Int J Infect Dis. 2008; 12: 57-61.
 
Show Less References

Article

Antagonistic Fungi, Soil Amendment and Soil Solarization as an Integrated Tactics for Controlling Fusarium Root Rot of Lupine (Lupinus termis)

1Department of Botany and Microbiology, Faculty of Science, University of Port Said, Port Said, Egypt

2Department of Biological sciences, University of Calgary, Alberta, Canada


American Journal of Microbiological Research. 2017, 5(1), 7-14
doi: 10.12691/ajmr-5-1-2
Copyright © 2017 Science and Education Publishing

Cite this paper:
Mohsen E. Ibrahim, Ahmed E. M. Abdelaziz. Antagonistic Fungi, Soil Amendment and Soil Solarization as an Integrated Tactics for Controlling Fusarium Root Rot of Lupine (Lupinus termis). American Journal of Microbiological Research. 2017; 5(1):7-14. doi: 10.12691/ajmr-5-1-2.

Correspondence to: Mohsen  E. Ibrahim, Department of Botany and Microbiology, Faculty of Science, University of Port Said, Port Said, Egypt. Email: mohsenhbrahim@yahoo.com

Abstract

The efficient control of soil-borne pathogens while avoiding environmental hazards and degradation of natural resources is the paramount challenge in crop protection sciences. Lupinus termis (lupine) is a fabaceous crop grown in Egypt for food, medical and industrial uses. The management of Fusarium root rot pathogens, which are responsible for serious losses on a number of economically important crops, including lupine, is being investigated. This study aimed to control Fusarium lupine root rot, under field conditions with no or low environmental impact; by using different control measures such as: antagonistic fungi, soil amendment and solarization singly or as an integrated disease management strategy. The integration of pest management methods is not only merely worthy, but also the more powerful practical solution for controlling soil-borne pests. Results showed that the solarized treated soil as well as the fungal antagonists, as single treatment methods in controlling Fusarium root rot revealed 80% and 81% healthy plants compared with their control that showed only 41%. Integrating soil solarization with mixed fungal inocula and soil organic amendment have been improving efficacy of controlling of lupine-fusarium-root rot by increasing the percentage number of healthy plants from 81 %, 79 % in treated unsolarized soil to 86 %, 82 % respectively in treated solarized soil. Using tactics such as solarization antagonistic fungi, and organic amendment as an integrated method, proved to be efficient for controlling the Fusarium root rot pathogen in Lupinus termis.

Keywords

References

[1]  Ibrahim, D., Khafagy, M. and El-Kheer, A. A. (1990). Some growth substances affecting the growth, chemical composition and alkaloidal content of Lupinus termis, L. Egypt. J. Applied Sci, 5, 367-381.
 
[2]  Van Bruggen, A. H. (1995). Plant disease severity in high-input compared to reduced-input and organic farming systems. Plant disease, 79, 976-984.
 
[3]  Kennedy, A. and Smith, K. (1995). Soil microbial diversity and the sustainability of agricultural soils. Plant and soil, 170, 75-86.
 
[4]  Sturz, A. and Christie, B. (2003). Beneficial microbial allelopathies in the root zone: the management of soil quality and plant disease with rhizobacteria. Soil and Tillage Research, 72, 107-123.
 
[5]  Gullino, M., Clini, C. and Garibaldi, A. (2004). Life without methyl bromide: the Italian experience in replacing the fumigant. Communications in agricultural and applied biological sciences, 70, 13-25.
 
Show More References
[6]  Katan, J. Soil disinfestation: one minute before methyl bromide phase out. VI International Symposium on Chemical and non-Chemical Soil and Substrate Disinfestation-SD2004 698, 2004. 19-26.
 
[7]  Baker, C., Henis, J., Baker, R. and Dunn, P. Commercial production and formulation of microbial biocontrol agents. New directions in biological control. Alternatives for suppressing agricultural pests and diseases. Proceedings of a UCLA Colloquium held at Frisco, Colorado, January 20-27, 1989., 1990. Alan R. Liss, Inc., 333-344.
 
[8]  Raupach, G. S. and Kloepper, J. W. (1998). Mixtures of plant growth-promoting rhizobacteria enhance biological control of multiple cucumber pathogens. Phytopathology, 88, 1158-1164.
 
[9]  Guetsky, R., Shtienberg, D., Elad, Y. and Dinoor, A. (2001). Combining biocontrol agents to reduce the variability of biological control. Phytopathology, 91, 621-627.
 
[10]  Roberts, D. P., Lohrke, S. M., Meyer, S. L., Buyer, J. S., Bowers, J. H., Baker, C. J., Li, W., De souza, J. T., Lewis, J. A. and Chung, S. (2005). Biocontrol agents applied individually and in combination for suppression of soilborne diseases of cucumber. Crop Protection, 24, 141-155.
 
[11]  Hubbard, J., Harman, G. and Hadar, Y. (1983). Effect of soilborne Pseudomonas spp. on the biological control agent. Trkhoderma hamatum, on pea seeds. Ecology and Epidemiology, 73, 655-659.
 
[12]  Dandurand, L. and Knudsen, G. (1993). Influence of Pseudomonas fluorescens on hyphal growth and biocontrol activity of Trichoderma harzianum in the spermosphere and rhizosphere of pea. Phytopathology, 83, 265-270.
 
[13]  Baker, R. and Hornby, D. An overview of current and future strategies and models for biological control. Biological control of soil-borne plant pathogens., 1990. CAB International, 375-388.
 
[14]  Harman, G. E., Howell, C. R., Viterbo, A., Chet, I. and Lorito, M. (2004). Trichoderma species—opportunistic, avirulent plant symbionts. Nature reviews microbiology, 2, 43-56.
 
[15]  Vinale, F., Marra, R., Scala, F., Ghisalberti, E., Lorito, M. and Sivasithamparam, K. (2006). Major secondary metabolites produced by two commercial Trichoderma strains active against different phytopathogens. Letters in Applied Microbiology, 43, 143-148.
 
[16]  Viterbo, A., Inbar, J., Hadar, Y. and Chet, I. (2007). Plant disease biocontrol and induced resistance via fungal mycoparasites. The mycota IV: environmental and microbial relationships. Springer Verlag, Berlin, Germany, 325-340.
 
[17]  Punja, Z. K., Utkhede, R. S. and Arora, D. (2004). Biological control of fungal diseases on vegetable crops with fungi and yeasts. Fungal biotechnology in agricultural, food, and environmental applications, 157-171.
 
[18]  Marwah, R. G., Fatope, M. O., Deadman, M. L., Al-Maqbali, Y. M. and Husband, J. (2007). Musanahol: a new aureonitol-related metabolite from a Chaetomium sp. Tetrahedron, 63, 8174-8180.
 
[19]  Zhang, H. and Yang, Q. (2007). Expressed sequence tags-based identification of genes in the biocontrol agent Chaetomium cupreum. Applied microbiology and biotechnology, 74, 650-658.
 
[20]  Tomilova, O. and Shternshis, M. (2006). The effect of a preparation from Chaetomium fungi on the growth of phytopathogenic fungi. Applied Biochemistry and Microbiology, 42, 67-71.
 
[21]  Borrego‐Benjumea, A., Basallote‐Ureba, M. J., Abbasi, P. A., Lazarovits, G. and Melero‐Vara, J. M. (2014). Effects of incubation temperature on the organic amendment‐mediated control of Fusarium wilt of tomato. Annals of applied biology, 164, 453-463.
 
[22]  Borrego-Benjumea, A., Basallote-Ureba, M., Melero-Vara, J. and Abbasi, P. (2014). Characterization of Fusarium isolates from asparagus fields and influence of soil organic amendments on Fusarium crown and root rot. Phytopathology, 104.
 
[23]  Motisi, N., Montfort, F., Dore, T., Romillac, N. and Lucas, P. (2009). Duration of control of two soilborne pathogens following incorporation of above‐and below‐ground residues of Brassica juncea into soil. Plant pathology, 58, 470-478.
 
[24]  Hansen, Z. and Keinath, A. (2013). Increased pepper yields following incorporation of biofumigation cover crops and the effects on soilborne pathogen populations and pepper diseases. Applied soil ecology, 63, 67-77.
 
[25]  Katan, J. (1981). Solar heating (solarization) of soil for control of soilborne pests. Annual Review of Phytopathology, 19, 211-236.
 
[26]  Katan, J. and Devay, J. E. (1991). Soil solarization: historical perspectives, principles, and uses. Soil solarization, 23-37.
 
[27]  Rubin, B. and Benjamin, A. (1984). Solar heating of the soil: involvement of environmental factors in the weed control process. Weed Science, 138-142.
 
[28]  Spadaro, D. and Gullino, M. L. (2005). Improving the efficacy of biocontrol agents against soilborne pathogens. Crop Protection, 24, 601-613.
 
[29]  Whipps, J., Van Elsas, J., Trevors, J. and Wellington, E. (1997). Ecological considerations involved in commercial development of biological control agents for soil-borne diseases. Modern soil microbiology., 525-546.
 
[30]  Ramirez-Villapudua, J. and Munnecke, D. E. (1987). Control of cabbage yellows (Fusarium oxysporum f. sp. conglutinans) by solar heating of field soils amended with dry cabbage residues. Plant disease, 71, 217-221.
 
[31]  Gamliel, A. and Stapleton, J. (1993b). Effect of chicken compost or ammonium phosphate and solarization on pathogen control, rhizosphere microorganisms, and lettuce growth. Plant Disease, 77, 886-891.
 
[32]  Gamliel, A. and Stapleton, J. (1993a). Characterization of antifungal volatile compounds evolved from solarized soil amended with cabbage residues. Phytopathology, 83, 899-905.
 
[33]  Chellemi, D., Olson, S. M., Mitchell, D., Secker, I. and Mcsorley, R. (1997). Adaptation of soil solarization to the integrated management of soilborne pests of tomato under humid conditions. Phytopathology, 87, 250-258.
 
[34]  Stapleton, J. J. and Devay, J. E. (1995). Soil solarization: a natural mechanism of integrated pest management. Novel approaches to integrated pest management, 309-322.
 
[35]  Jackson, M. L. (1958). Soil chemical analysis.
 
[36]  Klein, E., Katan, J. and Gamliel, A. (2011a). Combining residues of herb crops with soil heating for control of soilborne pathogens in a controlled laboratory system. Crop Protection, 30, 368-374.
 
[37]  Gullino, M. L., Katan, J. and Garibaldi, A. (2012). Fusarium wilts of greenhouse vegetable and ornamental crops, American Phytopathological Society.
 
[38]  Garibaldi, A., Gilardi, G. and Gullino, M. Critical aspects in disease management as a consequence of the evolution of soil-borne pathogens. VIII International Symposium on Chemical and Non-Chemical Soil and Substrate Disinfestation 1044, 2014. 43-50.
 
[39]  Gilardi, G., Demarchi, S., Gullino, M. L. and Garibaldi, A. (2014). Effect of simulated soil solarization and organic amendments on Fusarium wilt of rocket and basil under controlled conditions. Journal of Phytopathology, 162, 557-566.
 
[40]  Gilardi, G., Pugliese, M., Gullino, M. and Garibaldi, A. (2015). Effect of different organic amendments on lettuce fusarium wilt and on selected soilborne microorganisms. Plant Pathology.
 
[41]  Bonanomi, G., Antignani, V., Pane, C. and Scala, F. (2007). Suppression of soilborne fungal diseases with organic amendments. Journal of Plant Pathology, 311-324.
 
[42]  Klein, E., Katan, J. and Gamliel, A. (2011b). Soil suppressiveness to Fusarium disease following organic amendments and solarization. Plant Disease, 95, 1116-1123.
 
[43]  Chen, Y., Gamliel, A., STapleton, J. J. and Aviad, T. (1991). Chemical, physical, and microbial changes related to plant growth in disinfested soils. Soil solarization, 103-129.
 
[44]  Devay, J. E., Stapleton, J. J. and Elmore, C. L. (1991). Soil Solarization. FAO Plant Production and Protection Paper.
 
[45]  Stapleton, J. J. Modes of action of solarization and biofumigation. Second International Conference on Soil Solarization and Integrated Management of Soilborne Pests, Aleppo (Syria), 16-21 Mar 1997, 1997. ICARDA.
 
[46]  Stapleton, J. J. (2000). Soil solarization in various agricultural production systems. Crop protection, 19, 837-841.
 
[47]  Abdel-Rahim, M., Satour, M., Mickail, K., El-Eraki, S., Grinstein, A., Chen, Y. and Katan, J. (1988). Effectiveness of soil solarization in furrow-irrigated Egyptian soils. Plant Dis, 72, 143-146.
 
[48]  El-Shami, M., Salem, D., Fadl, F., Ashour, W. and El-Zayat, M. (1990). Soil solarization and plant disease management: II. Effect of soil solarization in comparison with soil fumigation on the management of Fusarium wilt of tomato. Agricultural Research Review, 68, 601-611.
 
[49]  Chellemi, D. (2006). Effect of urban plant debris and soil management practices on plant parasitic nematodes, Phytophthora blight and Pythium root rot of bell pepper. Crop Protection, 25, 1109-1116.
 
[50]  Wang, K.-H. and Sipes, B. S. (2009). Solarization and Cover Cropping as Alternatives to Soil Fumigants for Nematode Management in Hawai ‘i’s Pineapple Fields. CTAHR Cooperative Extension Publication SCM-29.
 
[51]  Gamliel, A. and Katan, J. (2012). Soil solarization: Theory and practice, APS press St. Paul.
 
[52]  Mahrer, Y. and Shilo, E. (2012). Physical principles of solar heating of soils. Soil solarization, theory and practice, 147-152.
 
[53]  Goswami, B. K., Singh, N. and Bhattacharya, C. (2013). Solar Assisted Integrated Approach for the Management of Soil Borne Fungus and Root-Knot Nematode Diseases on Tomato at Nursery Level. Int. J. Pure Appl. Sci. Technol, 19, 75-81.
 
[54]  Pane, C., Spaccini, R., Piccolo, A., Scala, F. and Bonanomi, G. (2011). Compost amendments enhance peat suppressiveness to Pythium ultimum, Rhizoctonia solani and Sclerotinia minor. Biological Control, 56, 115-124.
 
[55]  Navrozidis, E., Zartaloudis, Z., Thomidis, T., Karagiannidis, N., Roubos, K. and Michailides, Z. (2007). Effect of Soil Plowing and Fertilization on the Susceptibility of Four Olive Cultivars to the InsectBactrocera oleae and the FungiSphaeropsis dalmatica andSpilocaea oleagina. Phytoparasitica, 35, 429-432.
 
[56]  Singh, K. (2011). Organic amendments to soil inoculated arbuscular mycorrhizal fungi and Pseudomonas fluorescens treatments reduce the development of root-rot disease and enhance the yield of Phaseolus vulgaris L. European Journal of Soil Biology, 47, 288-295.
 
[57]  Hamel, C., Vujanovic, V., Jeannotte, R., Nakano-Hylander, A. and St-Arnaud, M. (2005). Negative feedback on a perennial crop: Fusarium crown and root rot of asparagus is related to changes in soil microbial community structure. Plant and Soil, 268, 75-87.
 
[58]  Sabet, K. K., Saber, M. M., El-Naggar, M. A.-A., El-Mougy, N. S., El-Deeb, H. M. and El-Shahawy, I. E.-S. (2013). Using commercial compost as control measures against cucumber root-rot disease. Journal of Mycology, 2013.
 
[59]  Bernard, E., Larkin, R. P., Tavantzis, S., Erich, M. S., Alyokhin, A., Sewell, G., Lannan, A. and Gross, S. D. (2012). Compost, rapeseed rotation, and biocontrol agents significantly impact soil microbial communities in organic and conventional potato production systems. Applied Soil Ecology, 52, 29-41.
 
[60]  Taylor, F. I. (2013). Control of soil borne potato pathogens using Brassica spp. mediated biofumigation. University of Glasgow.
 
[61]  Bonanomi, G., Giorgi, V., Neri, D. and Scala, F. (2006). Olive mill residues affect saprophytic growth and disease incidence of foliar and soilborne plant fungal pathogens. Agriculture, ecosystems & environment, 115, 194-200.
 
[62]  Matthiessen, J. N. and Kirkegaard, J. A. 2006. Biofumigation and enhanced biodegradation: opportunity and challenge in soilborne pest and disease management. Critical Reviews in Plant Sciences, 25, 235-265.
 
[63]  Mazzola, M., Brown, J., Izzo, A. D. and Cohen, M. F. (2007). Mechanism of action and efficacy of seed meal-induced pathogen suppression differ in a Brassicaceae species and time-dependent manner. Phytopathology, 97, 454-460.
 
[64]  Stapleton, J. and Banuelos, G. (2009). Biomass crops can be used for biological disinfestation and remediation of soils and water. California agriculture, 63, 41-46.
 
[65]  Larkin, R. P. and Griffin, T. S. (2007). Control of soilborne potato diseases using Brassica green manures. Crop protection, 26, 1067-1077.
 
[66]  Janisiewicz, W. (1988). Biocontrol of postharvest diseases of apples with antagonist mixtures. Phytopathology, 78, 194-198.
 
[67]  Alabouvette, C., Hoeper, H., Lemanceau, P. and Steinberg, C. (1996). Soil suppressiveness to diseases induced by soilborne plant pathogens. Soil biochemistry, 9, 371-413.
 
[68]  Alabouvette, C., Schippers, B., Lemanceau, P. and Bakker, P. (1998). Biological control of Fusarium wilts. Plant-Microbe Interactions and Biological Control. GJ Boland and LD Kuykendall, eds. Marcel Dekker, New York, 15-36.
 
[69]  Ibrahim, M. (1994). Soil fungi as biocontrol agent of tomato fusarial-wilt. MSc thesis, Faculty of Science, Suez Canal University, Ismailia.
 
[70]  Ibrahim, M. (1999). Management of tomato fusarial-wilt through integrated control. Ph. D Thesis, Faculty of Science, Suez Canal University.
 
[71]  Wahid, O. A. (2006). Improving control of Fusarium wilt of leguminous plants by combined application of biocontrol agents. Phytopathologia Mediterranea, 45, 231-237.
 
[72]  Bonaterra, A., Badosa, E., Cabrefiga, J., Frances, J. and Montesinos, E. (2012). Prospects and limitations of microbial pesticides for control of bacterial and fungal pomefruit tree diseases. Trees, 26, 215-226.
 
[73]  Gil, S. V., Pedelini, R., Oddino, C., Zuza, M., Marinelli, A. and March, G. J. (2008). The role of potential biocontrol agents in the management of peanut root rot in Argentina. Journal of plant pathology, 35-41.
 
[74]  El-Mehalawy, A. A., Hassanein, N. M., Khater, H. M., El-Din, E. K. and Youssef, Y. A. (2004). Influence of maize root colonization by the rhizosphere actinomycetes and yeast fungi on plant growth and on the biological control of late wilt disease. Int. J. Agric. Biol, 6, 599-605.
 
[75]  Hernández-Montiel, L. G., Rueda-Puente, E. O., Cordoba-Matson, M. V., Holguín-Peña, J. R. and Zulueta-Rodríguez, R. (2013). Mutualistic interaction of rhizobacteria with arbuscular mycorrhizal fungi and its antagonistic effect on Fusarium oxysporum in Carica papaya seedlings. Crop Protection, 47, 61-66.
 
[76]  Martínez-Medina, A., Roldán, A. and Pascual, J. A. (2011). Interaction between arbuscular mycorrhizal fungi and Trichoderma harzianum under conventional and low input fertilization field condition in melon crops: Growth response and Fusarium wilt biocontrol. Applied Soil Ecology, 47, 98-105.
 
[77]  Akhtar, M. and Siddiqui, Z. (2007). Biocontrol of a chickpea root-rot disease complex with Glomus intraradices, Pseudomonas putida and Paenibacillus polymyxa. Australasian Plant Pathology, 36, 175-180.
 
[78]  Saldajeno, M. and Hyakumachi, M. (2011). The plant growth‐promoting fungus Fusarium equiseti and the arbuscular mycorrhizal fungus Glomus mosseae stimulate plant growth and reduce severity of anthracnose and damping‐off diseases in cucumber (Cucumis sativus) seedlings. Annals of Applied Biology, 159, 28-40.
 
[79]  Arriaga, H., Núñez-Zofio, M., Larregla, S. and Merino, P. (2011). Gaseous emissions from soil biodisinfestation by animal manure on a greenhouse pepper crop. Crop Protection, 30, 412-419.
 
[80]  Núñez-Zofío, M., Larregla, S. and Garbisu, C. (2011). Application of organic amendments followed by soil plastic mulching reduces the incidence of Phytophthora capsici in pepper crops under temperate climate. Crop Protection, 30, 1563-1572.
 
[81]  Blok, W. J., Lamers, J. G., Termorshuizen, A. J. and Bollen, G. J. (2000). Control of soilborne plant pathogens by incorporating fresh organic amendments followed by tarping. Phytopathology, 90, 253-259.
 
Show Less References

Article

Phenotyping Groundnut Rhizobia Native to Phosphorus Deficient Soils in Western Kenya

1Department of Biological Sciences, University of Eldoret, P.O. Box 1125-30100, Eldoret, Kenya


American Journal of Microbiological Research. 2017, 5(1), 15-24
doi: 10.12691/ajmr-5-1-3
Copyright © 2017 Science and Education Publishing

Cite this paper:
Velma Okaron, Beatrice A. Were, Benson O. Nyongesa. Phenotyping Groundnut Rhizobia Native to Phosphorus Deficient Soils in Western Kenya. American Journal of Microbiological Research. 2017; 5(1):15-24. doi: 10.12691/ajmr-5-1-3.

Correspondence to: Velma  Okaron, Department of Biological Sciences, University of Eldoret, P.O. Box 1125-30100, Eldoret, Kenya. Email: vokaron@yahoo.com

Abstract

Most soils in western Kenya are characterized by high acidity level and phosphorus (P) deficiency, which affect nodulation and nitrogen fixation of groundnut (Arachis hypogaea L.). This study aimed at characterising rhizobia capable of nodulating groundnut in P deficient soils in Western Kenya. Sixty four isolates out of the 68 were confirmed to be rhizobia due to their ability to nodulate groundnut. Ninety six percent of the isolates exhibited semi-globose to globose colony shape on yeast extract mannitol agar (YEMA). Groundnut was nodulated by both fast and slow growing rhizobia isolates with 81% being fast growers. Fifty one isolates representing 75% produced acid on YEMA medium supplemented with bromothymol blue (BTB). The isolates varied in their response to pH with 39 and 61 growing at pH 4.0 and 5.5, respectively. All the isolates grew at pH 7.0 and 8.5. YEMA medium containing glucose, sucrose, starch and citrate had 64, 61, 56 and 5 isolates growing, respectively. Sixty four isolates exhibited clear zone of solubilization on medium containing dicalcium phosphate as source of inorganic phosphate. Solubilization index (SI) varied from 1.1 to 6.8. Fast-growing rhizobia isolates N01, B02, I06, Q01, F05, C02, E01, Q03, I01 and B01 recorded the highest solubilization index of 3.8, 4.5, 4.6, 4.6, 4.7, 5.0, 5.1, 6.1, 6.1 and 6.8, respectively. Groundnut rhizobia showed variation in their potential to solubilize inorganic phosphate and effectively nodulate the host. The most promising isolates from this study would be used as bio-fertilizer upon further validation in the greenhouse and field.

Keywords

References

[1]  Mokgehle, S. N., Dakora, F. D., & Mathews, C. Variation in N2 fixation and N contribution by 25 groundnut (Arachis hypogaea L.) varieties grown in different agro-ecologies, measured using 15 N natural abundance. Agriculture, Ecosystems & Environment, 195, 161-172. 2014.
 
[2]  Taurian, T., Ibañez, F., Fabra, A., & Aguilar, O. M. Genetic diversity of rhizobia nodulating Arachis hypogaea L. in central Argentinean soils. Plant and soil, 282(1-2), 41-52.2006.
 
[3]  Nekesa, P., Maritim, H. K., Okalebo, J. R., & Woomer, P. L. Economic analysis of maize-bean production using a soil fertility replenishment product (PREP-PAC) in western Kenya. African Crop Science Journal, 1999; 7(4): 585-590.
 
[4]  Owino, C. O., Owuor, P. O., & Sigunga, D. O. Elucidating the causes of low phosphorus levels in ferralsols of Siaya County, Western Kenya. Journal of Soil Science and Environmental Management, 2015; 6(9): 260-267.
 
[5]  Kisinyo, P. O., Othieno, C. O., Gudu, S. O., Okalebo, J. R., Opala, P. A., Ng'etich, W. K., & Opile, W. R. Immediate and residual effects of lime and phosphorus fertilizer on soil acidity and maize production in western Kenya. Experimental Agriculture, 2014; 50(1): 128-143.
 
Show More References
[6]  Khan, M. S., Zaidi, A., Ahemad, M., Oves, M., & Wani, P. A. Plant growth promotion by phosphate solubilizing fungi–current perspective. Archives of Agronomy and Soil Science, 2010.; 56(1): 73-98.
 
[7]  Zhang, D., Zhang, C., Tang, X., Li, H., Zhang, F., Rengel, Z., & Shen, J. Increased soil phosphorus availability induced by faba bean root exudation stimulates root growth and phosphorus uptake in neighbouring maize. New Phytologist, 2016; 209(2): 823-831.
 
[8]  Marra, L. M., Oliveira, S. M., Soares, C. R. F. S., & Moreira, F. M. S. Solubilisation of inorganic phosphates by inoculant strains from tropical legumes. Science Agriculture (Piracicaba, Braz.), 2011; 68(5):.603-609.
 
[9]  Divito, G. A., & Sadras, V. O. How do phosphorus, potassium and sulphur affect plant growth and biological nitrogen fixation in crop and pasture legumes. A meta-analysis. Field Crops Research, 2014; 156:161-171.
 
[10]  Mathu, S., Herrmann, L., Pypers, P., Matiru, V., Mwirichia, R., & Lesueur, D. Potential of indigenous bradyrhizobia versus commercial inoculants to improve cowpea (Vigna unguiculata L. walp.) and green gram (Vigna radiata L. wilczek.) yields in Kenya. Soil science and plant nutrition, 2012; 58 (6):750-763.
 
[11]  Benson, O., Beatrice, A., Regina, N., Koech, P. K., Skilton, R. A., & Francesca, S. Morphological, genetic and symbiotic characterization of root nodule bacteria isolated from Bambara groundnuts (Vigna subterranea L. Verdc) from soils of Lake Victoria basin, western Kenya. Journal of Applied Biology and Biotechnology; 2015; 3(01): 1-10.
 
[12]  Jaetzold, R., Schmidt, H., Hornetz, B., & Shisanya, C.. Ministry of Agriculture Farm Management Handbook of Kenya VOL. II-Part C Subpart C1. Nairobi, Kenya: Ministry of Agriculture, 2006.
 
[13]  Muhati, S. I., Shepherd, K. D., Gachene, C. K., Mburu, M. W., Jones, R., Kironchi, G. O., & Sila, A. Diagnosis of soil nutrient constraints in small-scale groundnut (Arachis hypogaea L.) production systems of Western Kenya using infrared spectroscopy. Journal of Agricultural Science and Technology 2011; 111-127.
 
[14]  Okalebo, J. R., Gathua, K. W., & Woomer, P. L. Laboratory methods of plant and soil analysis:a working manual. Tropical Soil Biology and Fertility Programme, Nairobi. 2002
 
[15]  Hue, Andrade DS, Chueira LM. Isolation and characterization of new efficient and competitive bean (Phaseolus vulgaris L.) rhizobia in Brazil. Soil Biology and Biochemistry, 2000; 32: 1515-1528.
 
[16]  Vincent, J. M. A manual for the practical study of the root-nodule bacteria. (IBP Handbuch No. 15 des International Biology Program, London . XI u. 164 S., 10 Abb., 17 Tab., 7 Taf. Oxford-Edinburgh: Blackwell Scientific Publ., 45 s.1970.
 
[17]  Gupta, A., Gupta, A. K., Mahajan, R., Singh, D., Khosla, K., Lal, R., & Gupta, V. Protocol for isolation and identification of Agrobacterium Isolates from Stone Fruit plants and sensitivity of native A. tumefaciens isolates against agrocin produced by A. radiobacter strain K84. National Academy Science Letters, 2013; 36(1): 79-84.
 
[18]  Huang, B,., Lü, C., Wu, B. & Fan, L. A rhizobia strain isolated from root nodule of gymnosperm Podocarpus macrophyllus. Science China Serie. C: Life Science,2007; 50:1-6.
 
[19]  Mehta, S., & Nautiyal, C. S. An efficient method for qualitative screening of phosphate-solubilizing bacteria. Current microbiology, 2001; 43(1): 51-56.
 
[20]  Alikhani, H. A., Saleh-Rastin, N., & Antoun, H. Phosphate solubilization activity of rhizobia native to Iranian soils. In First international Meeting on microbial phosphate solubilization , 35-41. Springer Netherlands. 2006.
 
[21]  Laurette, N. N., Maxémilienne, N. B., Henri, F., Souleymanou, A., Kamdem, K., Albert, N. & François-Xavier, E. Isolation and Screening of Indigenous Bambara Groundnut (Vigna Subterranea) Nodulating Bacteria for their Tolerance to Some Environmental Stresses. American Journal of Microbiological Research, 2015; 3(2): 65-75.
 
[22]  Somasegaran, P., & Hoben, H. J. (1994). Collecting nodules and isolating rhizobia. In Handbook for Rhizobia (pp. 7-23). Springer New York
 
[23]  Team, R. C. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. 2015. URL h ttp. www. R-project. org. Accessed September .13, 2016
 
[24]  Muthini, M., Maingi, J. M., Muoma, J. O., Amoding, A., Mukaminega, D., Osoro, N. & Ombori, O.Morphological assessment and effectiveness of indigenous rhizobia isolates that nodulate P. vulgaris in water hyacinth compost testing field in Lake.
 
[25]  Sharma, M. P., Srivastava, K., & Sharma, S. K. Biochemical characterization and metabolic diversity of soybean rhizobia isolated from Malwa region of central India. Plant Soil Environ, 56(8), 375-383.2010Victoria Basin. British Journal of Applied Science & Technology,2014; 4(5): 718.
 
[26]  Sanginga, N., Danso, S. K. A., & Bowen, G. D. Nodulation and growth response of Allocasuarina and Casuarina species to phosphorus fertilization. Plant and Soil, 1989; 118 (1-2): 125-132.
 
[27]  Teixeira, F.C.P., Borges, W. L., Xavier, G. R, Rumjanek, N. G. Characterization of indigenous rhizobia from Caatinga. Brazilian Journal of Microbiology, 2010; 41: 201-208.
 
[28]  Boakye, E. Y., Lawson, I. Y. D., & Danso, S. K. A. Characterization and diversity of rhizobia nodulating selected tree legumes in Ghana. Symbiosis, 69 (2), 89-99. 2016
 
[29]  Kapembwa, R., Mweetwa, A. M., Ngulube, M., & Yengwe, J. Morphological and Biochemical Characterization of Soybean Nodulating Rhizobia Indigenous to Zambia. Journal of Agricultural Research, 2016; 5: 84.
 
[30]  Sprent, J.I. Evolution and diversity in the legume-rhizobium symbiosis: chaos theory? Plant Soil, 1994; 161:1-10.
 
[31]  Sayyed, R. Z., Jamadar, D. D., & Patel, P. R. Production of Exo-polysaccharide by Rhizobium sp. Indian journal of microbiology, 2011; 51(3): 294-300.
 
[32]  Batista, J. S. S., Hungria, M., Barcellos, F. G., Ferreira, M. C., & Mendes, I. C. Variability in Bradyrhizobium japonicum and B. elkanii seven years after introduction of both the exotic microsymbiont and the soybean host in a Cerrados soil. Microbial ecology, 2007; 53(2): 270-284.
 
[33]  Wahab, A. A., Zahran, H. H., & Abd-Alla, M. H. Root-hair infection and nodulation of four grain legumes as affected by the form and the application time of nitrogen fertilizer. Folia microbiologica, 199 41(4), 303-308. 1996.
 
[34]  Sobczak, I., & Lolkema, J. S. The 2-hydroxycarboxylate transporter family: physiology, structure, and mechanism. Microbiology and Molecular Biology Review, 2005; 69: 665-695.
 
[35]  Küçük, Ç., Kivanç, M., & Kinaci, E. Characterization of Rhizobium sp. isolated from bean. Turkish Journal of Biology, 2000; 30(3): 127-132.
 
[36]  Hungria, M., de S Andrade, D., de O Chueire, L. M., Probanza, A., Guttierrez-Mañero, F. J., & Megı́as, M. Isolation and characterization of new efficient and competitive bean (Phaseolus vulgaris L.) rhizobia from Brazil. Soil Biology and Biochemistry, 2000; 32, 1515-1528.
 
[37]  Brígido, C., & Oliveira, S. Most acid-tolerant chickpea mesorhizobia show induction of major chaperone genes upon acid shock. Microbial Ecology, 2013; 65: 145-153.
 
[38]  Rodrigues, C. S., Laranjo, M., & Oliveira, S. Effect of heat and pH stress in the growth of chickpea mesorhizobia. Current Microbiology, 2006; 53(1): 1-7.
 
[39]  Kumari, B. S., Ram, M. R., & Mallaiah, K. V. Studies on exopolysaccharide and indole acetic acid production by Rhizobium strains from Indigofera. African Journal of Microbiology Research, 2009; 3(1): 10-14.
 
[40]  Kumar, G. K., & Ram, M. R. Phosphate solubilizing rhizobia isolated from Vigna trilobata. American Journal of Microbiological Research, 2014; 2(3): 105-109.
 
[41]  Sujatha, E., Grisham, S., Reddy, S. M. Phosphate solubilization by thermophillic microorganisms. Indian Journal of Microbiology, 2004; 44: 101–104.
 
[42]  Harun, M. Sattar, M. A., Uddin, M. I. and Young, J. P. W. Molecular characterization of symbiotic root nodulating rhizobia isolated ntifrom lentil (Lens culinaris Medik.). Journal of Environmental Agriculture and Food Chemistry, 2009; 8: 602-612.
 
Show Less References