Screening of Trichoderma strains by optimized conditions in test plate media for siderophore production and measuring of cellulytic enzyme

Document Type : Research Paper

Authors

1 Ph.D. Student, Department of Plant Protection, Faculty of Agriculture, Vali-e-Asr University of Rafsanjan, Kerman, Iran

2 Associate Professor, Department of Plant Protection, Faculty of Agriculture, Vali-e-Asr University of Rafsanjan, Kerman, Iran

3 Professor, Department of Plant Protection, Faculty of Agriculture, Valiasr University, Rafsanjan, Rafsanjan, Iran.

4 Assistant Professor, Department of Soil Science, Faculty of Agriculture, Vali-e-Asr University of Rafsanjan, Kerman, Iran

Abstract

Objective
The production of siderophore and cellulase by Trichoderma species is one of the effective indexs in plant growth, effective control and management of pathogens as well as application in industry. Therefore, primary screening of best isolates is very important based on the defined indicators.
Materials and methods
In this study, the optimal conditions for the qualitative and quantitative production of siderophore and cellulase were investigated using test plate method for the initial screening of 27 Trichoderma strains. For measuring cellulase enzymes (Endoglucanase, exoglucanase and beta glucosidase), the reactive dyes phenol red, congo red (0.5, 0.05 and 0.015%) were used at pH levels of 4, 5, 7 and 9 with carbon sources including Avicel, CMC and cellobiose. Quantitative measurement of cellulase was investigated using the dinitrosalicylic acid method. Screening of Trichoderma strains for siderophore production was studied by CAS agar method as mycelial culture and crude extract in a well method.
Results
The results of this research showed phenol red 0.015% under pH5 is the best reagent for detection of cellulase, Because of there is no inhibition on mycelial growth and the purple zone diameter is clear. In quantitative cellulase assay, the highest enzyme specific activity is related to T. harzianum CT-763 with 3 U/mg. T. atroviride CT-482, CT-473 and CT-865 were known as active strains in siderophore production, which were able to produce two types of siderophore, hydroximate and catechol. The results of siderophore production in liquid medium using the well method showed that T. afroharzianum CT-891 has the highest siderophore production by creating a halo zone of 1.2 cm, while T. harzianum CT566 produced the lowest amount of siderophore without creating halo.
Conclusions
Considering the application efficiency using of siderophore and cellulase in sustainable agriculture and importance of the role of Trichoderma species in production these metabolites, the well method for siderophore and dinitrosalicylic acid method for cellulase detection suggested. In this study T. atroviride and T. harzianum species had the highest production in siderophore and cellulase, respectively

Keywords


اسدی فرزانه، علایی حسین، صابری ریسه روح الله، زین الدینی ریسه اعظم (1397) بررسی تاثیر سویه های Trichoderma جدا شده از خاک های شور و سدیمی در کنترل بیماری پوسیدگی فوزاریومی ریشه خیار (Fusarium solani). مهار زیستی در گیاه پزشکی 6 (2)، 43-45.
باقری خدیجه، شهبازی سمیرا، عسکری حامد، موجرلو شیده، امیرلو فرنگیس (1396) افزایش فعالیت آنزیم سلولاز در قارچ تریکودرما ویریده با پرتو گاما. علوم زیستی 4(4)، 329-336.
براهویی نسیبه، علایی حسین، صابری روح­الله، صداقتی ابراهیم (1401) ارزیابی اثرآنتاگونیستی گونه­های Trichoderma روی Phytophthora citrophthora، عامل پوسیدگی ریشه و طوقه­ مرکبات. دانش گیاهپزشکی ایران 53، 295-311.
جمالی سکینه، پنجه­که ناصر، محمدی امیرحسین (1395) مطالعه اثر بازدارندگی گونه های مختلف Trichoderma بر رشد رویشی و تولید زئوسپورهای Phytophthora drechsleri و بررسی نقش آنزیم های هیدرولیتیک. مجله بین­المللی خشکبار (7)2، 148-137.
خدادادی-پورارپنایی صدیقه، مهرابی کوشکی مهدی، فرخی­نژاد رضا (1399) بررسی اثرات ضد قارچی چند سویه Trichoderma علیه چهار بیمارگر ریشه و طوقه لوبیا چشم­بلبلی. فصلنامه علمی زیست­شناسی میکروارگانیسم­ها 10(39)، 51-63.

زواری فاطمه، صاحبانی نوازالل، اعتباریان حسن رضا (1391) اندازه­گیری فعالیت آنزیمی بتا 1و3 گلوکاناز برون یاخته­ای در جدایه­های Trichoderma virens  و انتخاب جدایه­های برتر در کنترل بوته­میری خیار. دانش کشاورزی و تولید پایدار (22) 4، 150-159.

مرید مائده، مظفری دوست­مراد (1392) بررسی تاثیر ریزمغذی ها روی میزان تولید و فعالیت آنزیم کیتیناز گونه هایی از تریکودرما. مجله بیمارهای گیاهی 49(3)، 341-335.
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