DNA sequence analysis, morphology and pathogenicity of Fusarium oxysporum f. sp. lentis isolates inciting lentil wilt in Pakistan

Paper Details

Research Paper 01/12/2015
Views (333) Download (8)
current_issue_feature_image
publication_file

DNA sequence analysis, morphology and pathogenicity of Fusarium oxysporum f. sp. lentis isolates inciting lentil wilt in Pakistan

Khola Rafique, Chaudhary Abdul Rauf, Farah Naz, Ghulam Shabbir
Int. J. Biosci.7( 6), 74-91, December 2015.
Certificate: IJB 2015 [Generate Certificate]

Abstract

Fusarium wilt of lentil (Lens culinaris Medikus) incited by F. oxysporum f. sp. lentis causes huge lentil yield losses worldwide. Understanding morpho-molecular and pathogenic variation of F. oxysporum isolates is of utmost importance, which has not been previously studied in Pakistan. In this study, extensive two year (2011-12 and 2012-13) wilt surveys of seven districts viz. Chakwal, Jhelum, Gujrat, Sialkot, Layyah, Bhakkar and Khushab of Punjab, Pakistan were conducted, which revealed 100% mean disease prevalence and 25.7% mean incidence. Morphology assessment of recovered 105 isolates showed unique identification characters and variability in morphological measurements that differentiates F. oxysporum from other fusaria. Furthermore, sequencing of TEF-1α gene supported the morphological study and confirmed the associated fungi at species level. Phylogenetic analysis grouped all the selected type 31 isolates, obtained from different districts, under a single lineage within F. oxysporum species complex. Characterized isolates screened for pathogenicity revealed considerable pathogenic variability on two lentil genotypes (NARC-08-1 and Masoor-93). The inoculation of lentils confirmed the pathogenicity of 30 (96.77%) isolates on NARC-08-1 with 0 to 100% disease severity index and incidence with 19.33 to 100% yield reduction. On Masoor-93, 22 (70.97%) isolates were pathogenic with 0 to 66.66% severity index, 0 to 100% incidence and 6.47 to 53.68% yield reduction. Eight (25.81%) isolates viz. FWC15, FWJ35, FWG1, FWL6, FWL9, FWL12, FWB10 and FWK2 proved the most pathogenic resulting in highly virulent disease reaction (94.07% mean severity index and 100% mean incidence and yield reduction) and complete death of the NARC-08-1 seedlings.

VIEWS 10

Abd-Elsalam KA, Schnieder F, Guo JR. 2003. A modified  DNA  extraction  minipreparation  protocol for Fusarium isolates. Journal of Rapid Methods and Automation in Microbiology 11(1), 75-79. http://dx.doi.org/10.1111/j.17454581.2003.tb00410.x

Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. 1997. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Research 25(17), 3389-3402. http://dx.doi.org/10.1093/nar/25.17.3389

Aoki T, O’Donnell K, Homma Y, Lattanzi AR. 2003. Sudden-death syndrome of soybean is caused by two morphologically and phylogenetically distinct species within the Fusarium solani species complex-F. virguliforme in North America and F. tucumaniae in South America. Mycologia 95(4), 660-684. http://dx.doi.org/10.2307/3761942

Arif M, Chawla S, Zaidi NW, Rayar JK, Variar M, Singh US. 2012. Development of specific primers for genus Fusarium and F. solani using rDNA sub-unit and transcription elongation factor (TEF-1α) gene. African Journal of Biotechnology 11(2), 444-447. http://dx.doi.org/10.5897/AJB10.489

Ayub K, Rahim M, Khan A. 2001. Performance of exotic lentil varieties under rainfed conditions in Mingora (NWFP). Online Journal of Biological Sciences 1(5), 343-344.

Baayen RP, O’Donnell K, Bonants PJM, Cigelnik E, Kroon LPNM, Roebroeck EJA. 2000. Gene genealogies and AFLP analyses in the Fusarium oxysporum complex identify monophyletic and non monophyletic formae speciales causing wilt and rot disease. Phytopathology 90(8), 891-900. http://dx.doi.org/10.1094/PHYTO.2000.90.8.891

Bayaa B, Erskine W, Abbas A. 1994. Evaluating different methods for screening lentil germplasm for resistance to lentil wilt caused by Fusarium oxysporum f. sp. lentis. Arab Journal of Plant Protection 12(2), 83-91.

Bayaa B, Erskine W, Hamdi A. 1995. Evaluation of a wild lentil collection for resistance to vascular wilt. Genetic Resources and Crop Evolution 42(3), 231-235.

Bayaa BO, Erskine W, Khoury L. 1986. Survey of wilt damage on lentil in Northern Syria. Arab Journal of Plant Protection 4(2), 118-119. http://dx.doi.org/10.1007/BF02431257

Belabid L, Baum M, Fortas Z, Bouznad Z, Eujayl I. 2004. Pathogenic and genetic characterization of Algerian isolates of Fusarium oxysporum f. sp. lentis by RAPD and AFLP analysis. African Journal of Biotechnology 3(1), 25-31. http://dx.doi.org/10.5897/AJB2004.000-2005

Belabid L, Fortas Z, Dalli D, Khiare M, Amdjad D. 2000. Wilt root rot of lentils in North-Western Algeria. Cahiers Agricultures 9(6), 515-518.

Belabid L, Fortas Z. 2002. Virulence and vegetative compatibility of Algerian isolates of Fusarium oxysporum f. sp. lentis (Lens culinaris Medik.). Phytopathologia Mediterranea 41(3), 179-187.

Bogale M, Wingfield BD, Wingfield MJ, Steenkamp ET. 2007. Species-specific primers for Fusarium redolens and  a  PCR-RFLP  technique  to distinguish among three clades of Fusarium oxysporum. FEMS Microbiology Letters 271(1), 27-32. http://dx.doi.org/10.1111/j.1574-6968.2007.00687.x

Boonham N, Glover R, Tomlinson J, Mumford R. 2008. Exploiting generic platform technologies for the detection and identification of plant pathogens. European Journal of Plant Pathology 121, 355-363. http://dx.doi.org/10.1007/s10658-008-9284-3

Booth C. 1977. Fusarium: Laboratory guide to the identification of the major species. Commonwealth Mycological Institute, Kew Surrey, England, 31.

Bowers JH, Locke JC. 2000. Effect of botanical extracts on the population density of Fusarium oxysporum in soil and control of Fusarium wilt in the greenhouse. Plant Disease 84(3), 300-305. http://dx.doi.org/10.1094/PDIS.2000.84.3.300

Burgess LW, Nelson PE, Summerell BA. 1989. Variability and stability of morphological characters in Fusarium oxysporum. Mycologia 81(6), 818-822. http://dx.doi.org/10.2307/3759891

Cenis JL. 1992. Rapid extraction of fungal DNA for PCR amplification. Nucleic Acids Research 20(9), 238.

Chaudhary RG, Amarjit K. 2002. Wilt disease as a cause of shift from lentil cultivation in Sangod Tehsil of Kota, Rajasthan. Indian Journal of Pulses Research 15(2), 193-194.

Chaudhary RG, Saxena DR, Dhar V, Singh RK, Namdev JK. 2010. Prevalence of wilt-root rot and their associated pathogens at reproductive phase in lentil. Archives of Phytopathology and Plant Protection 43(10), 996-100. http://dx.doi.org/10.1080/03235400802214851

Chaudhry M, Ilyas M, Ghazanfar M. 2008. Sources of resistance from lentil international Fusarium wilt nursery 2006-7. Pakistan Journal of Phytopathology 20, 122-124.

Choi YW, Hyde KD, Ho WH. 1999. Single spore isolation of fungi. Fungal Diversity 3(1), 29-38.

Erskine W, Muehlbauer F, Sarker A, Sharma B. 2009. The Lentil: Botany, production and uses. CAB International, Wallingford, UK, 457. http://dx.doi.org/10.1079/9781845934873.0000

Geiser DM, Jimenez-Gasco MM, Kang S, Makalowska I, Veeraraghavan N, Ward TJ, Zhang N, Kuldau GA, O’Donnell K. 2004. FUSARIUM-ID v.1.00: A DNA sequence database for identifying Fusarium. European Journal of Plant Pathology 110, 473-479. http://dx.doi.org/10.1023/B:EJPP.0000032386.75915.a0

Gupta OM, Khare MN, Kotasthane SR. 1986. Variability among six isolates of F. oxysporum f. sp ciceris causing vascular wilt of chickpea. Indian Phytopathology 39(2), 279-281.

Hamdi A, Hassanein AM. 1996. Survey of fungal diseases of lentil in North Egypt. LENS Newsletter 23(1/2), 52-53.

Haqqani AM, Zahid MA, Malik MR. 2000. Legumes in Pakistan. In: Johansen C, Duxburv JM, Virmani SM, Gowda CLL, Pande S, Joshi PK, Eds. Legumes in rice and wheat cropping systems of the Indo-Gangetic Plain-constraints and opportunities. International Crops Research Institute for the Semi-Arid Tropics, Cornell University, Ithaca, New York, USA, 98-128.

Haware M, Nene Y. 1982. Races of Fusarium oxysporum f. sp. ciceri. Plant Disease 66, 809-810. http://dx.doi.org/10.1094/PD-66-809

JimenezGasco M, Milgroom M, JimenezDiaz R. 2002. Gene genealogies support Fusarium oxysporum f. sp. ciceris as a monophyletic group. Plant Pathology 51, 72-77. http://dx.doi.org/10.1046/j.00320862.2001.00610.x -i1

Katoh     K, Standley D. 2013.  MAFFT  multiple sequence alignment software version 7: Improvements in performance and usability. Molecular Biology and Evolution 30, 772-780. http://dx.doi.org/10.1093/molbev/mst010

Khare M, Agrawal S, Jain A. 1979. Lentil diseases and their control. Technical Bulletin, Jabalpur, India, 29.

Khare MN. 1981. Diseases of lentil. In: Webb C, Hawtin G, Eds. Lentils. CAB-ICARDA, Farnham Royal, UK, 163-172.

Kranz J. 1988. Measuring plant disease. In: Kranz J, Rotem J, Eds. Experimental techniques in plant disease epidemiology. Springer, The Netherlands, 35-50. http://dx.doi.org/10.1007/978-3-642-95534-14

Leslie JF, Summerell BA. 2006. The Fusarium laboratory manual. Wiley-Blackwell Publishing Professional, Ames, IA, USA, 212. http://dx.doi.org/10.1002/9780470278376

Mandhare VK, Deshmukh GP, Patil JV, Kale AA, Chavan UD. 2011. Mophological, pathogenic and molecular characterization of Fusarium oxysporum f. sp. Cicero isolates from Maharashtra,India. Indonesian Journal of Agricultural Science 12(2), 47-56.

Martin C, Roberts D, Weide M, Rossau R, Jannes G, Smith T, Maher M. 2000. Development of a PCR-based line probe assay for identification of fungal pathogens. Journal of Clinical Microbiology 38, 3735-3742.

Mohammadi N, Puralibaba H, Goltapeh EM, Ahari AB, Sardrood BP. 2012. Advanced lentil lines screened for resistance to Fusarium oxysporum f. sp. lentis under greenhouse and field conditions. Phytoparasitica 40(1), 69-76. http://dx.doi.org/10.1007/s12600-011-0201-5

Naimuddin, Chaudhary RG. 2009. Pathogenic variability in isolates of Fusarium oxysporum f. sp. lentis. Trends in Biosciences 2(1), 50-52.

Nelson PE, Toussoun TA, Marasas WFO. 1983. Fusarium species: An illustrated manual for identification. Pennsylvania State University Press, University Park, Pennsylvania, USA, 193.

Nicolaisen M, Supronienb S, Nielsena LK, Lazzaroa I, Spliid NH, Justesen AF. 2009. Real-time PCR for quantification of eleven individual Fusarium species in cereals. Journal of Microbiological Methods 76(3), 234-240. http://dx.doi.org/10.1016/j.mimet.2008.10.016

O’Donnell K, Nirenberg HI, Aoki T, Cigelnik E. 2000. A multigene phylogeny of the Gibberella fujikuroi  species  complex:  Detection  of  additional phylogenetically distinct species. Mycoscience 41(1), 61-78. http://dx.doi.org/10.1007/BF02464387

O’Donnell K, Rooney A, Proctor R, Brown D, McCormick S, Ward T, Frandsen R, Lysoe E, Rehner SA, Aoki T. 2013. Phylogenetic analyses of RPB1 and RPB2 support a middle cretaceous origin for a clade comprising all agriculturally and medically important fusaria. Fungal Genetics and Biology 52, 20-31. http://dx.doi.org/10.1016/j.fgb.2012.12.004

Rahman T, Ahmed AU, Islam MR, Hosen MI. 2010. Physiological study and both in vitro and in vivo antifungal activities against Stemphylium botryosum  causing  Stemphylium  blight  disease  in lentil (Lens culinaris). Plant Pathology Journal 9(4), 179-187. http://dx.doi.org/10.3923/ppj.2010.179.187

Saleem MT. 2013. Pakistan agricultural economy. In: Saleem MT, Akhtar ME, Tahir E, Eds. Farming Outlook: A quarterly educational magazine on policy and developments of progressive agriculture. Pakistan, 9-12.

Seifert KA. 2001. Fusarium and anamorph generic concepts. In: Summerell BA, Leslie JF, Backhouse D, Bryden WL, Burgess LW, Eds. Fusarium: Paul E. Nelson memorial symposium. American Phytopathological Society Press, St. Paul, Minnesota, 15-28.

Skovgaard K, Rosendahl S, O’Donnell K, Nirenberg HI. 2003. Fusarium commune is a new species identified by morphological and molecular phylogenetic data. Mycologia 95(4), 630-636. http://dx.doi.org/10.2307/3761939

Summerell BA, Salleh B, Leslie JF. 2003. A utilitarian approach to Fusarium identification. Plant Disease 87(2), 117-128.

Taheri N, Rastegar MF, Jafarpour B, Bagheri AR, Jahanbaghsh V. 2010. Pathogenic and genetic characterization of Fusarium oxysporum f. sp. lentis by RAPD and IGS analysis in Khorasan province. World Applied Sciences Journal 9(3), 239-244.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. 2013.  MEGA6:  Molecular  evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30, 2725-2729. http://dx.doi.org/10.1093/molbev/mst197

Toussoun TA, Nelson PE. 1976. A pictorial guide to the identification of Fusarium species. 2nd edn. The Pennsylvania State University Press, University Park, USA, 43.

Williams JGK, Kubelik AR, Livak KJ, Rafalski JA, Tingey SV. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 18(22), 6531-6535. http://dx.doi.org/10.1093/nar/18.22.6531

Wulff EG, Sorensen JS, Lubeck M, Nielsen KF, Thrane U, Torp J. 2010. Fusarium spp. associated with rice Bakanae: Ecology, genetic diversity, pathogenicity and toxigenicity. Environmental Microbiology 12(3), 649-657. http://dx.doi.org/10.1111/j.1462-2920.2009.02105.x