بررسی تولید آنزیم کیتیناز توسط چندین جدایه از قارچ تریکودرما و تاثیر آن بر کنترل بیولوژیک نماتد ریشه گرهی گوجه فرنگی Meloidogyne javanica

نوع مقاله : مقالات پژوهشی

نویسندگان

• مجتبی کواری ¹
• عصمت مهدیخانی مقدم ²
• حمید روحانی ³

¹ دانشگاه فردوسی مشهد

² دانشگاه فردوسی مشهد- گروه گیاهپزشکی

³ دانشگاه فردوسی مشهد-گروه گیاهپزشکی

10.22067/jpp.v29i1.32918

چکیده

کیتین از جمله مهم‌ترین ترکیبات دیواره سلولی قارچ‌های حقیقی به‌شمار می‌رود. این ترکیب به وفور در ساختمان دیواره تخم نماتدها از جمله نماتد مولد گره ریشه گوجه فرنگی Meloidogyne javanica یافت می‌شود. کیتین پس از تجزیه توسط آنزیم‌های کیتیناز، تبدیل به زیر واحد‌های سازنده‌اش یعنی N- استیل گلوکز آمین می‌گردد. گونه‌های قارچ تریکودرما(Trichoderma spp.) به دلیل تولید مقادیر قابل توجهی آنزیم‌های هیدرولیتیک متعدد از جمله کیتیناز، پروتئاز و بتا 1و 3 گلوکاناز، به یکی از عوامل مورد استفاده جهت کنترل بیمارگرهای گیاهی تبدیل شده است. در این پژوهش میزان فعالیت کیتینازی 15 جدایه قارچ تریکودرما در ارتباط با توانایی بیوکنترل نماتد ریشه‌گرهی گوجه فرنگی مورد بررسی قرار گرفت. از بین این جدایه‌ها، جدایه‌های T.BI، T6 و T65 به ترتیب با فعالیت آنزیمی 2/19، 3/18 و 17 واحد بر میلی‌لیتر (U/ml) به عنوان فعال‌ترین و جدایه‌های T16، T12N و T12 به ترتیب با فعالیت آنزیمی 5/5، 4/5 و 7/3 واحد بر میلی‌لیتر به عنوان ضعیف‌ترین جدایه‌ها تعیین شدند. در آزمایشات گلخانه‌ای نیز نتایجی هم‌سو با نتایج آزمایشگاهی به دست آمد و جدایه‌های T.BI ،T65 و T6 به عنوان موثر‌ترین جدایه‌ها در کنترل بیولوژیک این نماتد عمل کردند.

کلیدواژه‌ها

• تریکودرما
• کنترل بیولوژیک
• فعالیت آنزیمی
• نماتد ریشه گرهی گوجه فرنگی

20.1001.1.20084749.1394.29.1.15.8

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عنوان مقاله [English]

Survey on Chitinase Production by Several Isolates of Trichoderama and its Biological Control effect on Tomato Root-knot Nematode Meloidogyne javanica

نویسندگان [English]

• M. Kavari ¹
• E. Mahdikhani Moghadam ²
• H. Rouhani ³

¹ Ferdowsi University of Mashhad

² Ferdowsi University of Mashhad

³ Ferdowsi University of Mashhad

چکیده [English]

Chitin is one of the most important compounds of true fungi cell wall. This compound is used greatly in structural of nematodes egg shell including Meloidogyne javanica. After chitin dissection by chitinase enzyme, convert to its sub units N-Acetyl Glucosamine. Species of Trichoderama genus due to high levels production of many hydrolytic enzymes including Chitinase, Protease and B-1,3-Glucanases, is used as a biocontrol agent of plant pathogens. In the current study, chitinase activity levels of 15 isolates of Trichoderama spp. was investigated to their biocontrol effects on tomato root knot nematode. Among this 15 isolates, T.BI, T6 and T65 respectively with 19.2, 18.3 and 17 U/ml enzyme activity were known as the most active isolates. Also T16, T12N and T12 respectively with 5.5, 5.4 and 3.7 U/ml enzyme activity were known as the weakest isolates. These results was in agreement with the results of greenhouse experiments and T.BI, T65 and T6 isolates were reported as the highest effective in biological control of m.javanica.

کلیدواژه‌ها [English]

• Trichodema
• Biological control
• Enzyme Activity
• Tomato root-knot nematode

مراجع

1- Affokpon A., Coyne D. L., Htay C.C., Agbèdè R.D., Lawouin L., and Coosemans J. 2011. Biocontrol potential of native Trichoderma isolates against root-knot nematodes in West African vegetable production systems. Soil Biology and Biochemistry, 43(3): 600-608.

2- Benitez T., Rincon M.A,. Limon M.C., and Codon A.C. 2004, Biocontrol mechanisms of Trichoderma strains., International Microbiology, 7:249-260.

3- Bird F.A., and McClure M.A. 1976. The tylenchid (Nematoda) egg shell: structure, composition and permeability. Parasitology, 72:19-28.

4- Bonants P.J.M., Fitters P.F.L., Thijis H., Den Belder E., Waalwijk C., and Henfling J.W.D.M. 1995. A basic serin protease from paecilomyces lilacinus with biological activity against Meloidogyne hapla eggs. Microbiology, 141: 775-784.

5- Brants A., Brown C.R., and Earle, A.D. 2000. Trichoderma harzianum Endochitinase Does Not Provide Resistance to Meloidogyne hapla in Transgenic Tobacco. Journal of Nematology, 32(3):289–296.

6- Chahal V.P.S., and Chahal P.P.K. 1991.control of meloidogyne incognita with bacillus thuringensis. Dev. Plant soil Sci., 45:677-680,.

7- De la Veg H., Specht C.A., Liu Y., and Robbins P.W. 1998. Chitinases are a muti-gene family in Aedes, Anopheles and Drosophila. Insect Molecular Biology, 7: 233-239.

8- Dick R.P. 2002. Enzymes in the environment: activity, ecology and applications.. CRC Press. Vol. 86

9- El-Katatny M.H., Somitsch W., Robra K.H., El-Katany M.S., and Gubitz G.M. 2000. production of chitinase and beta-1, 3- glucanase by T. harizianum for control of the phytopathogenic fungus Sclerotinum rolfsii. Food Technology and Biotechnology, 38:173-180.

10- Haran S., Shickler H., Chet I. 1996. Molecular mechanisms of lytic enzymes involved in the biocontrol activity of Trichoderma harzianum. Microbiology, 142: 2321-2331.

11- Harman G.E., Howell C.R., Viterbo A., Chet I., Lorito M. 2004. Trichoderma species-opportunistic, avirulent plant symbionts. Nature Reviews, 2:43-56.( 307).

12- Herrera-Estrella A., and Chet I. 1999. Chitinases in biological control. Express. 87:171- 184.

13- Howell C. R. 2003. Mechanisms employed by Trichoderma species in the biological control of plant diseases: the history and evolution of current concepts. Plant disease, 87(1): 4-10.

14- Jeapson S.B. 1987. Identification of root-knot nematodes Meloidogyne species. Camberian International, 265 pp.

15- Khan A., Williams K.L., and Nevalainen H.K. 2004. Effects of Paecilomyces lilacinus protease and chitinase on the eggshell structures and hatching of Meloidogyne javanica juveniles. Biological Control,31(3): 346-352.

16- Khan A., Williams K., Molloy M.P., and Nevalainenc H., 2003. Purification and characterization of a serine protease and chitinases from Paecilomyces lilacinus and detection of chitinase activity on 2D gels. Protein Expression and Purification, 32: 210–220.

17- Lopez-Ilorca, L.V., Macia-Vicente, J.G. and Jansson, H.B. 2008. Mode of action and interaction of nematophagous fungi. In: Ciancio A. and Mukerji K. G (eds.) Integrated Management and Biocontrol of Vegetable and Grain Crops Nematodes. 51-76.

18- Lunge A.G., and Patil A.S. 2012. Charectization of efficient chitinolytic enzyme producing Trichoderma species: a tool for beter antagonistic approach, International Journal of Science, Environment, 1(5): 377-385.

19- Molan. J., Duram A., and Cabib. E. 1997. A rapid and sensitive assay for chitinase using tritiated chitin., Anal Biochem 1977, 83:648-656.

20- Muzzarelli R.A.A., and Peter M.G. 1997. Chitin handbook. European Chitinase Society, Atec Grottammare, Italy.1:1-10 .

21- Roberts D.P., Lohrke S.M., Meyer S.L.F., Buyer J.S., Bowerrs J.H., Baker C.J., Li W., Souza J.T., Lewis J.A., and Chung S. 2005. Biocontrol agents applied individually and in combination for suppression of soil born diseases of cucumber. Crop Protection, 24(2):141-155.

22- Sahebani N., and Hadavi N. 2008. Biological control of the root-knot nematode Meloidogyne javanica by Trichoderma harzianum. Soil Biology and Biochemistry, 40(8): 2016-2020.

23- Sasser J.N., Carter C.C., and Hartman K.M. 1984. Standardization of host suitability studies and reporting of resistance to root-knot nematodes. Department of Plant Pathology, North Carolina State University.

24- Sharon E., Bar-Eyal M., Chet I., Herrera-Esterella A., Keleifeld O., and Spiegel Y. 2001. Biological control of the root – knot nematode Meloidogyne javanica by Trichoderma harzianum . Phytopathology, 91:687-693.

25- Siddiqui I.A., Amer Zareen M., Javad Zaki M., and Shaukat S.S. 2001. Use of Trichoderma spicies in the control of Meloidogyne javaniva, Roor-rot nematode in the Okra and Mungbean, Pakistan journal of Biological sciences, 4(7): 846-848.

26- Siddiqui I.A., and Shaukat S.S. 2004. Trichoderma harzianum enhances the production of nematicidal compounds in vitro and improves biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato. Letters in applied microbiology, 38(2), 169-175.

27- Singh A., Kirubakaran I., and Sakhtivel N. 2007. Heterologous expression of new antifungal chitinase from wheat. Protein Expression and Purification, 56: 100–109.

28- Spiegel Y., Chet I., and Cohn E. 1987. Use of chitin for controlling plant-parasitic nematodes II. Mode of action. Plant and Soil, 98:337–346.

29- Thiagarajan V., Revathi R., Aparanjini K., Sivamani P., Girilal M., Priya C.S., and Kalaichelvan P.T. 2011. Extracellular chitinase production by Streptomyces sp. PTK19 in submerged fermentation and its lytic activity on Fusarium oxysporum PTK2 cell wall. INT J CURR SCI, 1: 30-44.

30- Verma M., Brar S.K., Tyagi R.D., Surampalli R.Y., and Valero J.R. 2007. Antagonistic fungi, Trichoderma sp. : panoply of biological control. Biochem. Ezymol. J., 37:1-20.

31- Wen C.M., Tseng C.S., Cheng C.Y., and Li Y.K. 2002. Purification, characterization and cloning of a chitinase from Bacillus sp. NCTU2 .Biotechnol. Appl. Biochem, 35’ 213- 219.

32- Xu J., Narabu T., Mizukubo T., and Hibi T. 2001. A molecular marker correlated with selected virulence againts the tomato resistance gene Mi in Meloidogyne incogenita, M.javanica and M. arenaria Phytopathology, 91: 377-382.

33- Zeilinger S., Galhaup C., Payer K., Woo S.L., Mach R.L., Fekete C., Lorito M., Kubicek C.P. 1999. Chitinase gene expression during mycoparasitic interaction of Trichoderma harzianum with its host. Fungal Genetics and Biology, 26: 131-140.