بررسی فعالیت آنزیم بتا- ۱ و ۴ اندوگلوکاناز در سه نماتد بیمارگر گیاهی

نوع مقاله: مقاله کامل پژوهشی

نویسندگان

1 دانشجوی دکتری بیماری شناسی گیاهی، دانشکده علوم کشاورزی، دانشگاه گیلان.

2 دانشیار گروه گیاهپزشکی، دانشکده علوم کشاورزی، دانشگاه گیلان.

3 دانشیار گروه بیوتکنولوژی، دانشکده علوم کشاورزی، دانشگاه گیلان.

چکیده

دیواره سلولی گیاه میزبان، سد مکانیکی اصلی برای نفوذ و مهاجرت نماتدهای انگل گیاهی محسوب می­شود. این نماتدها جهت تجزیه دیواره سلولی، انواع مختلفی از آنزیم­های تجزیه کننده دیواره سلولی را ترشح می‌کنند. بتا- 1 و 4 اندوگلوکاناز از اولین آنزیم­های تجزیه کننده شناخته شده در این ارتباط است. در این پژوهش، فعالیت آنزیم مذکور در سه گونه شاملMeloidogyne incognita  ،Pratylenchus loosi  و Aphelenchoides besseyi مورد مطالعه قرار گرفت. ابتدا با انجام نمونه برداری، شناسایی، خالص سازی و تکثیر هر سه گونه نماتد، استخراجDNA  انجام شد. سپس با استفاده از پرایمر دژنره اختصاصی و به کمک واکنش زنجیره­ای پلی­مراز، ژن کد کننده آنزیم بتا- ۱ و ۴ اندوگلوکاناز در نماتدهای مذکور تشخیص داده شد. استخراج RNA  کل از هر سه گونه انجام شد. بعد از آن، cDNA تهیه و با استفاده از ژن مرجع، درستیcDNA ­ ها مورد بررسی قرار گرفت. سپس شدت بیان ژن کد کننده آنزیم بتا- 1 و 4 اندوگلوکاناز برای نمونه­ها بررسی شد. نتایج نشان داد که ژن کدکننده آنزیم بتا- ۱ و ۴ اندوگلوکاناز، بیشترین فعالیت آنزیمی و شدت بیان را در نماتد مولد گره (M. incognita) داشت.نماتد مولد زخم ریشه چای (P. loosi) و نماتد نوک سفیدی برگ برنج (A. besseyi)به ترتیب در رتبه­های بعدی قرار گرفتند.

کلیدواژه‌ها


عنوان مقاله [English]

Study of the activity of β -1,4 endoglucanase enzyme in three plant parasites nematodes*

نویسندگان [English]

  • S.N . Mirghasemi 1
  • S. Jamali 2
  • M.M. Sohani 3
  • A. Zibaei 2
چکیده [English]

Host plant cell wall is the main mechanical barrier to penetration and migration by endoparasitic nematodes. In order to degradation of plant cell walls, the nematodes secrete various types of cell wall degrading enzymes. β‐1, 4‐endoglucanase was the first known cell wall degrading enzyme. In this research, the enzyme activites were studied in three species; Meloidogyne incognita (Kofoid & White1919)Chitwood, 1949, Pratylenchus loosi Loof, 1960 and Aphelenchoides besseyi Christie, 1942. Initially, sampling, identification, purification and multiplication of the nematodes were performed and after extraction of their DNA, the encoding β - 1, 4 endoglucanase gene was detected in the nematodes  by using pairs of specific degenerate primers in polymerase chain reaction. After extraction of total RNA from three species of nematodes, cDNA was synthesized and determination of cDNA goodness was carried out by a reference gene. Gene expression of the β -1,4 endoglucaase was done for all samples. The results showed that the gene encoding β -1 ,4 endoglucanase had the highest activity and expression in the root- knot nematode (M. incognita). Tea root lesion nematode (P. loosi) and Rice white tip nematode (A. besseyi) were classified in  next  step, respectively.

کلیدواژه‌ها [English]

  • Aphelenchoides besseyi
  • Meloidogyne incognita
  • Parasitism
  • Pratylenchus loosi
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