نخستین گزارش گونه‌ی Meloidogyne javanica (Treub, 1885) Chitwood, 1949 از نارنگی در ایران

نوع مقاله : علمی پژوهشی-فارسی

نویسندگان

1 دانشجوی کارشناسی ارشد دانشگاه شهید چمران اهواز

2 استاد دانشگاه شهید چمران اهواز

3 استادیار دانشگاه شهید چمران اهواز

چکیده

     نماتد‌ ریشه‌گرهی انگل اجباری گیاهان است که تعداد زیادی از گیاهان آوندی را در سراسر جهان مورد حمله قرار می‌دهد و از نظر اقتصادی بسیار مهم می‌باشد. در طی بررسی تنوع زیستی نماتدهای انگل گیاهی در باغ‌های مرکبات استان خوزستان درسال 97-1396، یک جمعیت نماتد ریشه‌گرهی از نمونه ‌خاک وریشه نارنگی جمع‌آوری شد. ماده‌های بالغ و لارو سن دو از ریشه‌های آلوده استخراج گردید. بعد ازکشتن و ثابت کردن نماتد‌ها،اسلاید دائم تهیه شد. شناسایی گونه با استفاده از کلید‌های معتبر صورت گرفت. DNA ژنومی از لارو سن دو و ماده‌های بالغ استخراج گردید. DNA استخراج شده توسط جفت آغازگر ناحیه D2/D3 از ژن 28S و جفت آغازگرهای ناحیه 18S تکثیر و توالی‌یابی شد. بر اساس خصوصیات ریخت‌شناختی، ریخت‌سنجی و تجزیه و تحلیل فیلوژنی نواحی تکثیر شده، گونه M. javanica شناسایی گردید. تفاوت‌های ریخت‌شناختی جمعیت به‌دست‌آمده با سایر جمعیت‌های این گونه مورد بحث قرار گرفت. در این مطالعه دو قطعه 799 جفت بازی توسط آغازگرهای ناحیه 28S و دو قطعه 1678 جفت بازی توسط آغازگرهای ناحیه 18S تولید شد. بر اساس منابع موجود، این اولین گزارش از آلودگی نارنگی به نماتد ریشه ‏گرهی M. javanica در ایران می‌باشد.

کلیدواژه‌ها


عنوان مقاله [English]

First report of Meloidogyne javanica (Treub, 1885) Chitwood, 1949 from tangerine tree in Iran

نویسندگان [English]

  • P. Eisvand 1
  • R. Farrokhi Nejad 2
  • S. Azimi 3
1 M.Sc. student of Plant Pathology,, Shahid Chamran University of Ahvaz
2 Professor, , Shahid Chamran University of Ahvaz
3 Assistant Professor, Shahid Chamran University of Ahvaz
چکیده [English]

Background and Objectives: The root-knot nematode is an obligatory plant parasite that attacks a large number of vascular plants worldwide and is economically important. The present study aimed to identify the Meloidogyne javanica species based on morphological, morphometric data and molecular characterization.
Materials and Methods: During a survey on the biodiversity of plant-parasitic nematodes in citrus orchards of Khuzestan province from 2017 to 2018, a root-knot nematode population was collected from soil and root samples of tangerine. Adults and the second-stage juveniles were extracted from the infected roots. After killing and fixing the nematodes, permanent slides were prepared. Species identification was performed using valid keys. Genomic DNA was extracted from both juvenile and adult females. The extracted DNA was amplified and sequenced by the primers of the D2-D3 expansion segments of the 28S rRNA gene and the 18S rRNA gene. The newly obtained sequences of the D2-D3 fragments of 28S and 18S rRNA gene sequences and additional sequences of relevant species were selected after a BlastN search and were aligned by Clustal X using the default parameters. The both phylogenetic analyses were evaluated under the GTR + G + I model with two million generations.
Results: M. javanica was identified based on morphological, morphometric data and phylogenetic analysis of amplified regions. The morphological differences of the recovered population in comparison with other populations were discussed. In this study, two 799 bp fragments were generated by the 28S rDNA gene region primers and two 1678 bp fragments were generated by the 18S rDNA gene region primers.
Discussion and conclusion: The perineal pattern of the studied female nematode is distinguished from the other species of this genus by having two parallel lines at the lateral surfaces of the body so that these lines clearly divide the perineal pattern into the dorsal and ventral parts. To the best of our knowledge, this is the first report of M. javanica in association with tangerine in Iran.

کلیدواژه‌ها [English]

  • Citrus
  • Root-knot nematode
  • Morphometric data
  • Morphology
  • Phylogeny
Adam, M. A. M., Phillips, M. S., and Blok, V. C. 2007. Molecular diagnostic key for identification of single juveniles of seven common and economically important species of root-knot nematode (Meloidogyne spp.). Plant Pathology, 56: 190-197.
Ahmadi, K., Abadzadeh, H. R., Hatami, F., Hosseinpour, R., and Abdullah, E. H. 2019. Agricultural Statistics 2018. Tehran: Horticultural Products. Ministry of Jihad-e-Agriculture, Deputy of Planning and Economics, ICT Center. P. 241. (in Farsi)
Askarian, H., Sharifnabi, B., Olia, M., Mahdikhani-Moghadam, E., and Akhavan A. 2009. Identification of Meloidogyne javanica using morphological and morphometrical characters and species specific primers. Journal of Science and Technology of Agriculture and Natural Resources, 13: 279-290. (In Farsi with English abstract).
Blok, V. C., and Powers, T. O. 2009. Biochemical and molecular identification. In: R. N. Perry, M. Moens, and J. L. Starr (eds). Root-knot nematodes. CAB International, Wallingford. pp. 98-118.  
Coyne, D. L., Nicol, J. M., and Claudius-Cole, B. 2014. Practical plant nematology: A field and laboratory guide. 2nd edition. SP-IPM Secretariat, International Institute of Tropical Agriculture (IITA), Cotonou, Benin. P. 96.
De Grisse, A. T. 1969. Redescription and modification of some techniques used in the study of nematodes phytoparasitaires. Mededelingen Rijksfacultiet Landbouw Wetenschappe Gent, 34: 351-369.
De Ley, I. T., Karssen, G., De Ley, P., Vierstraete, A., Waeyenberge, L., Moens, M., and Vanfleteren, J. 1999. Phylogenetic analyses of internal transcribed spacer region sequences within Meloidogyne. Journal of Nematology, 31: 530-531.
Eisenback, J. D., and Triantaphyllou, H. H. 1991. Root-knot nematodes: Meloidogyne species and races. In: W. R. Nickle (ed). Manual of Agricultural Nematology. Marcell Dekker, New York. pp. 191-274.
Garcia, L. E., and Sanchez-Puerta, M. V. 2012. Characterization of a root-knot nematode population of Meloidogyne arenaria from Tupungato (Mendoza, Argentina). Journal of Nematology, 44: 291-301.
Ghaderi R., Kashi L., and Karegar A. 2018. Plant-parasitic nematodes in Iran. Tehran, Iran: Science Reference in collaboration with the Iranian Society of Nematology.
Harris, T. S., Sandall, L. J., and Powers, T. O. 1990. Identification of single Meloidogyne juveniles by polymerase chain reaction amplification of mitochondrial DNA. Journal ofNematology, 22: 518-524.
Hartman, K. M., and Sasser, J. N. 1985. Identification of Meloidogyne species on the basis of differential host test and perineal pattern morphology. In: K. R. Barker, C. C. Carter, and J. N. Sasser (eds). An advanced treatise onMeloidogyne. Volume II:Methodology. A cooperative publication of the Department of Plant Pathology and the United States Agency for International Development, North Carolina State University Graphics, North Carolina. pp. 69-77.
Hesar, A. M., Moghaddam, E. M., and Maafi, Z. T. 2011. Morphometrical and genetic diversity of Meloidogyne javanica isolates from the north east of Iran. Journal of nematode morphology and systematics, 14: 1-11.
Holterman, M., van der Wurff, A., van den Elsen, S., van Megen, H., Holovachov, T. M. O., Bakker, J., and Helder, J. 2006. Phylum wide analysis of SSU rDNA reveals deep phylogenetic relationships among nematodes and accelerated evolution toward crown clades. Molecular Biology and Evolution, 23: 1792-1800.
Hunt, D. J., and Handoo, Z. A. 2009. Taxonomy, identification and principal species. In: R. N. Perry, M. Moens, and J. L. Starr (eds). Root-knot nematodes. CAB International, Wallingford. pp. 55-97.
Hyman, B. C. 1990. Molecular diagnosis of Meloidogyne species. Journal of Nematology, 22: 24-30.
Janssen, T., Karssen, G., Verhaeven, M., Coyne, D., and Bert, W. 2016. Mitochondrial coding genome analysis of tropical root-knot nematodes (Meloidogyne) supports haplotype based diagnostics and reveals evidence of recent reticulate evolution. Scientific Reports, 6: 22591.
Jones, J. T., Haegeman, A., Danchin, E. G. J., Gaur, H. R., Helder, J., Jones, M. G. K., Kikuchi, T., Manzanilla-Lopez, R., Palomares-Rius, J. E., Wesemael, W. M. L., and Perry, R. N. 2013. Top 10 plant-parasitic nematodes in molecular plant pathology. Molecular Plant pathology, 14: 946-961.
Karssen, G., and Moens, M. 2006. Root-knot nematodes. In:R. N. Perry, and M. Moens, (eds). Plant nematology. CAB International. Wallingford. pp. 59-90.
Kiewnick, S., Holterman, M., van den Elsen, S., van Megen, H., Frey, J. E., and Helder, J. 2014. Comparison of two short DNA barcoding loci (COI and COII) and two longer ribosomal DNA genes (SSU & LSU rRNA) for specimen identification among quarantine root-knot nematodes (Meloidogyne spp.) and their close relatives. European Journal of Plant Pathology, 140(1): 97-110.
Kiewnick, S., Wolf, S., Willareth, M., and Frey, J. E. 2013. Identification of the tropical root-knot nematode species Meloidogyne incognita, M. javanica and M. arenaria using a multiplex PCR assay. Nematology, 15 (7): 891-894. 
Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thompson, J. D., Gibson, T. J., and Higgins, D. G. 2007. Clustal W and Clustal X version 2.0. Bioinformatics, 23: 2947-2948.
Mahdikhani-Moghadam, E., Kheiri, A., and Mohammadi, M. 2007. Molecular comparison of Iranian populations of Meloidogyne javanica and Meloidogyne incognita with PCR-RFLP. Journal of Science and Technology of Agriculture and Natural Resources, 10: 405-410. (in Farsi with English abstract).
Nunn, G. B. 1992. Nematode molecular evolution. PhD. Thesis, University of Nottingham, Nottingham, U.K.
Onkendi, E. M., Kariuki, G. M., Marais, M. and Moleleki, L. N. 2014. The threat of root-knot nematodes (Meloidogyne spp.) in Africa: A review. Plant Pathology, 68: 727-737.
Pedram, M., Pourjam, E., Ye, W., Atighi, M. R., Robbins, R. T. and Ryss, A. 2011. Description of Bursaphelenchus mazandaranense sp. n. (Nematoda: Parasitaphelenchidae) from Iran. Russian Journal of Nematology, 19: 121-129.
Powers, T. O., and Harris, T. S. 1993. A polymerase chain reaction method for identification of five major Meloidogyne species. Journal of Nematology, 25: 1-6.
Powers, T. O., Todd, T. C., Burnell, A. M., Murray, P. C. B., Flemming, C. C., Szalanski, A. L., Adams, B.A., and Harris, T. S. 1997. The rDNA internal transcribed spacer region as a taxonomic marker for nematodes. Journal of Nematology, 29: 441-450. 
Rafiee, H., Mahdikhani-Moghaddam, E., and Najafinia, M. 2010. Isolation of Meloidogyne cruciani from cucumber greenhouses in Jiroft region and evaluation of intra-species genetic diversity using RAPD-PCR marker. Journal of Plant Protection, 24 (1): 43-52. (in Farsi with English abstract).
Salalia, R., Walia, R. K., Somvanshi, V. S., Kumar, P., and Kumar, A. 2017. Morphological, morphometric, and molecular characterization of intraspecific variations within Indian populations of Meloidogyne graminicola. Journal of Nematology, 49 (3): 254-267.
Skantar, A. M., Carta, L. K., and Handoo, Z. A. 2008. Molecular and morphological characterization of an unusual Meloidogyne arenaria population from traveler’s tree, Ravenala madagascariensis. Journal of Nematology, 40: 179-189.
Tamura, K., Stecher, G., Peterson, D., Filipski, A., and Kumar, S. 2013. MEGA6: Molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution, 30: 2725-2729.
Verdejo-Lucas, S., and McKenry, M. V. 2004. Management of the citrus nematode, Tylenchulus semipenetrans. Journal of Nematology, 36 (4): 424-432.
 Wu, G. A., Prochnik, S., Jenkins, J., Salse, J., Hellsten, U., Murat, F., … Rokhsar, D.  2014. Sequencing of diverse mandarin, pummelo and orange genomes reveals complex history of admixture during citrus domestication. Nature Biotechnology, 32 (7): 656-662.