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شناسایی مولکولی باکتری‌های اندوفیت‌ مرکبات در شرق استان گیلان

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه گیاهپزشکی، دانشکده تولید‌گیاهی، دانشگاه علوم‌کشاورزی و منابع‌طبیعی گرگان، گرگان، ایران.

2 دانشکده تولید گیاهی گروه گیاه پزشکی دانشگاه علوم کشاورزی منابع طبیعی گرگان

3 ریس پژوهشکده مرکبات و میوه های نیمه گرمسیری

4 گروه گیاهپزشکی- دانشگاه علوم کشاورزی و منابع طبیعی گرگان

چکیده

این تحقیق به منظور شناسایی اندوفیت‌های باکتریایی درخت‌‌های مرکبات بالای پنج سال در شرق استان گیلان انجام شد. تعداد 63 باکتری اندوفیت‌ از برگ‌های سالم نارنج، پرتقال محلی و تامسون ناول، نارنگی‌ انشو، لیموشیرین، کامکوات و بالنگ جداسازی گردید. جدایه‌های بدست آمده براساس شکل و رنگ‌ پرگنه گروه‌بندی شدند، سپس خصوصیات ریخت-شناسی و بیوشیمیایی شش جدایه از آن‌ها به عنوان نماینده گروه‌های مختلف مورد ارزیابی قرار گرفت. براساس ارزیابی-های مولکولی و مطالعات تبارزایی ناحیه 16S rRNA مشخص گردید که این جدایه‌ها به گونه‌های Bacillus megaterium،Bacillus cereus، ‌Arthrobacter agilis، Acinetobacter junii و Pseudomonas azotoformans تعلق دارند. که از این میان سه گونه‌ A. agilis، A. junii و P. azotoformans برای اولین بار به عنوان باکتری‌های اندوفیت مرکبات از ایران و جهان گزارش می‌گردند. ویژگی‌های ریخت‌شناسی، بیوشیمیایی‌، مولکولی و تجزیه و تحلیل‌های تبارزایی به خوبی قادر به تمایز جنس‌ها و گونه‌ها‌ی اندوفیت جدا شده می باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Molecular identification of citrus endophytic bacteria in the east of guilan provience

نویسندگان [English]

  • S.leila Akbari kiarood 1
  • kamran Rahnama 2
  • morteza Golmohammadi 3
  • Saeed Nasrollanejad 4
1 Department of Plant Protection, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
2 Faculty member
3 Assistant Professor Horticultural Science Research Institute, Citrus and Subtropical Fruits Research Center
4 Department of Plant Protection, Gorgon University of Agricultural Sciences & Natural Resources
چکیده [English]

This research was conducted to identify endophytic bacteria over five-year old citrus trees in the east of Guilan province. A total of 63 endophytic bacteria were isolated from symptomless leaves of Citrus aurantium, Citrus sinensis, Citrus sinensis var. thomson, Citrus unshiu, Citrus limon, Fortunella margarita and Citrus medica. The isolates were grouped based on colony type and color. Then the morphological and biochemical characteristics of the six isolates were assessed as candidate of different groups. According to the molecular studies and phylogenetic analyses of 16srRNA region these isolates were identified as Bacillus megaterium, Bacillus cereus, Arthrobacter agilis, Acinetobacter junii and Pseudomonas azotoformans species. . Among them, three species including A. agilis, A. junii and P. azotoformans are reported as citrus endophytic bacteria from Iran and the world for the first time. The morphological, biochemical, molecular and phylogenetical analyses are able to differentiate genus and species of the endophyte isolates.

کلیدواژه‌ها [English]

  • Bacterial endophyte
  • Citrus
  • 16SrRNA
  • phylogeny
  1. Akbari kiarood, S. L., Rahnama, K., Golmohammadi, M. & Nasrollahnejad, S. (2018). Identification of one species of Xylaria sp. as endophytic fungi on Thomson Novel sweet Orange In: Proceedings of 23rd Iranian Plant Protection Congress, 27-30 August, Gorgan, Iran. (In Farsi).
  2. Akbari‌ kiarood, S. L., Rahnama, K., Golmohammadi, M. & Nasrollahnejad, S. (2019). Epicoccum nigrum, from Citrus and Kiwi trees common endophytes in the North of Iran. In: Proceedings of 4thIranian Mycological Congress, 26-28 August, Sari Agricultural Sciences and Natural Resources University, Iran. (In Farsi).
  3. Ali, S., Charles, T. C. & Glick, B. R. (2014). Amelioration of high salinity stress damage by plant growth-promoting bacterial endophytes that contain ACC deaminase. Plant Physiology and Biochemistry, 80, 160-167.
  4. Alizadeh, M. (2017). First Report of Arthrobacter agilis Associated with Elm Trees in Iran. Journal of Plant Chemistry and Ecophysiology, 2(1), 1013.
  5. Araújo, W. L., Maccheroni, W. ‏J., Aguilar-Vildoso, C. I., Barroso, P. A., Saridakis, H. O. & Azevedo, J. L. (2001). Variability and interactions between endophytic bacteria and fungi isolated from leaf tissues of citrus rootstocks. Canadian Journal of Microbiology, 47(3), 229-236.
  6. Ashfaq, M., Haider, M. S., Ali, A., Ali, M., Saleem, I. & Mubashar, U. (2016). Morphological characterization of endophytic bacterial strains isolated from discolored rice grain. Pakistan Journal of Phytopathology, 28(1), 01-08.
  7. Azevedo, J. L., Araújo, W. L., Luiz, W. & Lacava, P. T. (2016). The diversity of citrus endophytic bacteria and their interactions with Xylella fastidiosa and host plants. Genetics and Molecular Biology, 39(4), 476-491.
  8. Bashan, Y. & Holguin, G. (1998). Proposal for the division of plant growth promoting rhizobacteria into two classifications: biocontrol-PGPB (plant growth-promoting bacteria) and PGPB. Soil Biology Biocheimistry, 30, 1225-1228.
  9. Chen, C., Bauske, E. M., Musson, G., Rodriguez- kabana, R. & Kloepper, J. W. (1995). Biological control of Fusarium wilt on cotton by use of endophytic bacteria. Biological Control, 5, 83-91.

10. Douanla-Meli, C. & Langer, E. (2013). Diversity and molecular phylogeny of fungal endophytes associated with Diospyros crassiflora. Mycology, An International Journal on Fungal Biology, 3, 175-187.

11. Ek-Ramos, M.J., Gomez-Flores, R., Orozco-Flores, A. A., Rodríguez-Padilla. C., González-Ochoa. G. & Tamez-Guerra. P. (2019). Bioactive products from plant-endophytic gram-positive bacteria. Frontiers Microbiology, 10, 463.

12. Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39, 783-791.

13. Forchetti, G., Masciarelli, O., Alemano, S., Alvarez, D. & Abdala, G. (2007). Endophytic bacteria in sunflower (Helianthus annuus L.): isolation, characterization, and production of jasmonates and abscisic acid in culture medium. Applied Microbiology and Biotechnology, 76, 1145-1152.

14. Golinska, P., Wypij, M., Agarkar, G., Rathod, D., Dahm, H. & Rai, M. (2015). Endophytic actinobacteria of medical plants: diversity and bioactivity. Antonie van Leeuwenhoek, 108, 267-28.

15. Goryluk-Salmonowicz, A., Orzeszko-Rywka, A., Piórek, M., Rekosz-Burlaga, H., Otłowska, A., Gozdowski, D. & Błaszczyk, M. (2018). Plant growth promoting bacterial endophytes isolated from Polish herbal plants. Acta Scientiarum Polonorum. Hortorum Cultus, 17(5), 37-46.

16. Gupta, R. S. (2011). Origin of diderm (Gram-negative) bacteria: antibiotic selection pressure rather than endosymbiosis likely led to the evolution of bacterial cells with two membranes. Antonie Van Leeuwenhoek, 100(2), 171-182.

17. Hallmann, J., Quadt-Hallmann, A., Mahaffee, W. F. & Kloepper, J. W. (1997). Bacterial endophytes in agricultural crops. Canadin Journal of Microbiology, 43, 895-914.

18. Herrera, H., Sanhueza, T., Novotná, A., Charles T. C. & Arriagada, C. (2020). Isolation and identification of endophytic bacteria from mycorrhizal tissues of terrestrial orchids from southern chile. Diversity, 12 (2), 55.

19. Jannah, M., Agustien, A., Zam, S. I., Lalfari, R. S., Aldi, Y., Dewi, A. P. & Djamaan, A. (2018). Isolation and characterization of antibiotic-producing endophytic bacteria from citrus aurantifolia swingle. Journal of Pure and Applied Microbiology, 12(3), 1473-1481.

20. Kántor, A., Mareček, J., Ivanišová, E., Terentjeva, M. & Kačániová, M. (2017). Microorganisms of grape berries. Proceedings of the Latvian Academy of Sciences. Section B. Natural, Exact, and Applied Sciences, 6 (711), 502-508.

21. Kobayashi, D. Y. & Palumbo, J.  D. (2000).  Bacterial endophytes and their effects on plants and uses in agriculture. In: C. W. Bacon & J. F. White (Ed), Microbial Endophytes. (pp. 99-233). Marcel Dekker, New York.

22. Kuklinsky-Sobral, J., Araujo, W. L., Mendes, R., Pizzirani-Kleiner, A. A. & Azevedo, J. L. (2005). Isolation and characterization of endophytic bacteria from soybean (Glycine max) grown in soil treated with glyphosate herbicide. Plant and soil, 273(1-2), 91-99.

23. Lewin, G. R., Carlos, C., Chevrette, M. G., Horn, H. A., McDonald, B. R., Stankey, R. J., Fox, B. G.  & Currie, C. R. (2016). Evolution and ecology of Actinobacteria and their bioenergy applications. Annual review of microbiology, 70, 235-254.

24. Li, Y., Wu, C., Xing, Z., Gao, B. & Zhang, L. (2017).  Engineering the bacterial endophyte Burkholderia pyrrocinia JK-SH007 for the control of lepidoptera larvae by introducing the cry218 genes of Bacillus thuringiensis. Biotechnology & Biotechnological Equipment, 31, 1167-1172.

25. Liop, P., Caruso, P., Cubero, J., Morente, C. & López, M. M. (1999). A simple extraction procedure for efficient routine detection of pathogenic bacteria in plant material by polymerase chain reaction. Journal of Microbiological Methods, 37, 23-31.

26. Liu, H., Carvalhais, L. C., Crawford, M., Singh, E., Dennis, P. G., Pieterse, C. M. & Schenk, P. M. (2017). Inner plant values: diversity, colonization and benefits from endophytic bacteria. Frontiers in Microbiology, 8, 2552.

27. Liu, X. L., Liu, S. L., Liu, M., Kong, B. H., Liu, L. & Li, Y. H. (2014). A primary assessment of the endophytic bacterial community in a xerophilous moss (Grimmia montana) using molecular method and cultivated isolates. Brazilian Journal of Microbiology, 45(1), 163-173.

28. Ma, Y., Rajkumar, M., Moreno, A., Zhang, C. & Freitas, H. (2017). Serpentine endophytic bacterium Pseudomonas azotoformans ASS1 accelerates phytoremediation of soil metals under drought stress. Chemosphere, 185, 75-85.

29. Mahmoudi, E., Ahmadi, A., Sayed-Tabatabaei, B.E., Ghobadi, C., Akhavan, A., Hasanzadeh, N. & Venturi, V. (2011). A novel AHL- degrading rhizobacterium quenches the virulence of Pectobacterium atrosepticum on potato plant. Journal Plant Pathology, 93 (3), 587-594.

30. Misaghi, I. J. & Donndelinger, C. R. (1990). Endophytic bacteria in symptom-free cotton plants. Phytopathology, 80, 808-811.

31. Mushtaq, S., Shafiq, M., Asim, M. & Haider, M. S. (2018).  Effect of bacterial endophytes isolated from citrus on the physiology of Brassica OleraceaInternational Journal of Biosciences, 12(6), 225-234.

32. Qin, S., Zhang, Y. J., Yuan, B., Xu, P. Y.,  Xing, K., Wang,  J. & Jiang, J. H. (2014) Isolation of ACC deaminase-producing habitat-adapted symbiotic bacteria associated with halophyte Limonium sinense (Girard) Kuntze and evaluating their plant growth-promoting  activity under salt stress. Plant Soil, 374, 753-766.

33. Quambusch, M., Pirttilä, A. M., Tejesvi, M. V., Winkelmann, T. & Bartsch, M. (2014). Endophytic bacteria in plant tissue culture: differences between easy-and difficult-to-propagate Prunus avium genotypes. Tree Physiology, 34(5), 524-533.

34. Rabbee, M. F., Ali, M. & Baek, K. H. (2019). Endophyte Bacillus velezensis isolated from citrus spp. controls Streptomycin-Resistant Xanthomonas citri subsp. citri that causes citrus bacterial canker. Agronomy, 9(8), 470.

35. Rolli, E., Marasco, R., Vigani, G., Ettoumi, B., Mapelli, F., Deangelis, M. L., Gandolfi, C., Casati, E., Previtali, F., Gerbino, R., Pierotti, C. F., Borin, S., Sorlini, C., Zocchi, G. & Daffonchio, D. (2015). Improved plant resistance to drought is promoted by the root-associated microbiome as a water stress-dependent trait. Environmental Microbiology,17, 316-331.

36. Sambrook, J. & Russell, D. W. (2001) Molecular Cloning, a Laboratory Manual (3rd Ed.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

37. Santoyo, G., Moreno-Hagelsieb, G., Orozco-Mosqueda, M. C. & Glick, B. R. (2016). Plant growth-promoting bacterial endophytes. Microbiological Research, 183, 92-99.

38. Schaad, N. W., Jones, J. B. & Chun, W. (2001). Laboratory Guide for Identification of Plant Pathogenic Bacteria. APS Press. St. Paul, Minnesota, USA.

39. Schulz, B. & Boyle, C. (2006). What are endophytes? In:Schulz B, Boyle C and Sieber T.N (eds). Microbial Root Endophytes, Springer-Verlag, Berlin, pp. 1-13.

40. Shi, Y., Lou, K & Li, C.  (2011). Growth promotion effects of the endophyte Acinetobacter johnsonii strain 3-1 on sugar beet. Journal of Genetic Resources, 54(3), 159-166.

41. Shuai, Z., Na, Z., Zheng-Yong, Z., Ke, Z., Guo-Hua, W & Chang-Yan, T. (2016). Isolation of endophytic plant growth-promoting bacteria associated with the halophyte Salicornia europaea and evaluation of their promoting activity under salt stress. Current Microbiology, 73(4), 574-81.

42. Shutsrirung A., Chromkaew Y., Pathom-Aree W., Choonluchanon S. & Boonkerd N. (2013). Diversity of endophytic actinomycetes in mandarin grown in northern Thailand, their phytohormone production potential and plant growth promoting activity. Soil Science and Plant Nutrition, 59(3), 322-330.

43. Siala, R., Chobba, I. B., Vallaeys, T., Triki, M. A., Jrad, M., Cheffi, M., Ayedi, I., Elleuch, A., Nemsi, A., Cerqueira, F., Gdoura, R., Drira, N. and & Gharsallah, N. (2016). Analysis of the cultivable endophytic bacterial diversity in the date palm (Phoenix dactylifera L.) and evaluation of its antagonistic potential against pathogenic Fusarium species that cause date palm bayound disease. Journal of Applied and Environmental Microbiology, 4(5), 93-104.

44. Su, F., Jacquard, C., Villaume, S., Michel, J., Rabenoelina, F., Clément, C., Barka, E. A., Dhondt-Cordelier, S. & Vaillant-Gaveau, N. (2015). Burkholderia phytofirmans PsJN reduces impact of freezing temperatures on photosynthesis in Arabidopsis thaliana. Frontiers Plant Science, 6, 810.

45. Tamura, K. & Nei, M. (1993). Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution, 10, 512-526.

46. Tran, P. N., Tan, N. E. H., Lee, Y.P., Gan, H. M., Polter, S. J., Dailey, L. K., Hudson, A. O. & Savka, M. A. (2015). Whole-genome sequence and classification of 11 endophytic bacteria from poisonivy (Toxicodendron radicans). Genome Announcement 3(6):e01319-15.

  1. Vega, F. E., Pava-Ripoll, M., Posada, F. & Buyer, J. S. (2005). Endophytic bacteria in Coffea arabica L. Journal Basic Microbiology, 45, 371-380.