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کنترل Phytophthora sojae و Macrophomina Phaseolina در سویا توسط عوامل زیستی

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه بیوتکنولوژی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران

2 گروه تنوع زیستی، پژوهشکده علوم محیطی، پژوهشگاه علوم و تکنولوژی پیشرفته و علوم محیطی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته، کرمان، ایران

3 دانشجوی سابق رشته بیماری شناسی گیاهی، دانشگاه زابل، زابل، ایران

چکیده

سویا یکی از مهم‌ترین دانه‎‌های روغنی در جهان است که نیمی از پروتئین و روغن گیاهی مصرفی انسان را تامین می‌کند. بیماری پوسیدگی زغالی و پوسیدگی طوقه و ریشه سویا ناشی از Macrophomina phaseolina و Phytophthora sojae از مهم‌ترین بیماری‌های سویا هستند که باعث کاهش کمی و کیفی عملکرد محصول می‌شوند. راهبرد مهار زیستی با استفاده از خانواده Actinobacteria، به ویژه اعضای جنس Streptomyces و در خانواده Hypocreaceae، جنس Trichoderma، به عنوان یک روش امیدوارکننده برای مدیریت بیماری‌ها و آفات در نظر گرفته می‌شود. در این مطالعه فعالیت مهار زیستی یک جدایه استرپتومایسس و یک جدایه تریکودرما‌ی جداسازی شده از خاک باغات استان کرمان در برابر قارچ‌های بیمارگرM. phaseolina و P. sojae مورد ارزیابی قرار گرفت. آزمایش‌ها در آزمایشگاه بر پایه طرح کاملاتصادفی با سه تکرار انجام شد. تجزیه و تحلیل توالی 16s rDNA و ناحیه ITS نشان داد که  Streptomyces sp.جدایه ۲۳ وTrichoderma sp. جدایه۱ به ترتیب متعلق به S. bacillaris وT. Longibrachiatum  می‌باشند. نتایج روش کشت دوگانه و آزمون متابولیت‌های فرار نشان داد که S. bacillaris جدایه23 و T. longibrachiatum جدایه۱ بطور معنی‌داری از رشد M. phaseolina و P. sojae جلوگیری کردند. در هر دو آزمون، دو عامل بیوکنترل مورد استفاده اثر بازدارندگی بیشتری بر روی رشد میسلیومیP. sojae در مقایسه با M. phaseolina از خود نشان دادند.

کلیدواژه‌ها

عنوان مقاله [English]

Control of Phytophthora sojae and Macrophomina phaseolina in soybean by biological agents

نویسندگان [English]

  • saeid mirzaei 1
  • Masoud Ahmadi -Afzadi 1
  • Mohammadreza Lashkari 2
  • Batul Sadeghi 3
1 Department of Biotechnology, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, kerman. Iran.
2 Department of Biodiversity, Institute of Science and High Technology and Environmental Sciences, Graduate University of Advanced Technology, Kerman, Kerman, Iran
3 Former student of plant protection, Agricultural Faculty, Zabol University, Zabol, Iran.
چکیده [English]

Soybean is one of the most important oilseed crops, supplying half of the protein and vegetable oil by humans. Charcoal rot disease and soybean crown and root rot caused by Macrophomina phaseolina and Phytophthora sojae, respectively, are among the most important soybean pathogens that cause a reduction in crop yield. A biocontrol strategy using Actinobacteria, especially different species of Streptomyces, and Hypocreaceae especially different species of Trichoderma, is considered a method for the management of disease and pests. In this study, biological control of these pathogens was investigated using Streptomyces sp. isolate 23 and Trichoderma sp. isolates 1 and their volatile metabolites. The activity of Streptomyces isolates and a Trichoderma isolate taken from the soil of Kerman was evaluated against pathogenic fungi, M. phaseolin, and P. sojae. Experiments in the laboratory were conducted based on a completely randomized design with three replications. Sequence analysis of 16s rDNA and ITS region showed Streptomyces sp. isolate 23 and Trichoderma sp. isolate 1 belong to S. bacillaris and T. longibrachiatum, respectively. The results of the double culture method and volatile metabolites test showed that S. bacillaris isolate 23 and T. longibrachiatum isolate 1, significantly inhibited the growth of M. phaseolina and P. sojae. In both tests, the two biocontrol agents used showed more inhibitory effects on P. sojae compared to M. phaseolina. The results of the dual culture and volatile metabolites tests showed that S. bacillaris isolate 23 and T. longibrachiatum isolate 1 had a high inhibitory effect on M. phaseolina and P. sojae.

کلیدواژه‌ها [English]

  • Actinobacteria
  • Charcoal rot disease
  • Soybean
  • Biocontrol

Extended Abstract

Introduction

    Soybean is one of the most important oilseed crops in the world, supplying half of the protein and vegetable oil consumed by humans. It is a nitrogen-fixing legume that is usually cultivated in alternation with cereals. Charcoal rot disease and soybean crown and root rot caused by Macrophomina phaseolina and Phytophthora sojae, respectively, are among the most important soybean pathogens that cause quantitative and qualitative reduction of crop yield. The common method of using chemical fungicides to control these diseases poses negative side effects on humans and the environment.

 Biological control offers the best alternative for controlling plant diseases. Actinobacteria, especially different species of Streptomyces, and Hypocreaceae especially members of the genus Trichoderma, are considered as the promising method for the management of diseases and pests. Furthermore, Actinobacteria produce antifungal antibiotics, cell wall degrading enzymes, and plant growth promoters which protect plants from a wide range of phytopathogenic fungi. Trichoderma is of great importance in controlling plant pathogens due to its rapid growth, high ability to produce spores, and the ability of myco-parasitism against a wide range of fungal pathogens.

 

Materials and Methods

    In this study, the activity of Streptomyces isolates and Trichoderma isolates taken from the soil of Kerman was evaluated against pathogenic fungi, M. phaseolina, and P. sojae. Experiments in the laboratory were conducted based on a completely randomized design with three replications. To confirm the antagonistic activity of Trichoderma sp. isolate 1 and Streptomyces sp. isolate 23, a dual culture assay and Sandwiched Petri dish method were used. Streptomyces sp. isolate 23 and Trichoderma sp. isolate 1 were identified based on morphological and molecular characteristics (16s rDNA and ITS regions).

 

Results and Discussion

    Sequence analysis of 16s rDNA and ITS region showed Streptomyces sp. isolate 23 and Trichoderma sp. isolate 1 belong to S. bacillaris and T. longibrachiatum respectively.

Results of dual culture showed that the hyphal growth of M. phaseolina and P. sojae stopped by T. longibrachiatum with an average of 58% and 97.14%, respectively. In addition, the T. longibrachiatum was able to advance the inhibition zone and cover the pathogenic fungal colonies, and exert intense and rapid sporadic activity at various locations of hyphae, including on the pathogenic fungal colonies. This physiological feature is the most important stage in parasitic activity. Moreover, S. bacillaris significantly inhibited the hyphal growth of M. phaseolina and P. sojae in dual culture.

Streptomyces species have been identified as one of the promising sources for the biological control of plant diseases. They produce active antifungal and antibacterial substances. In this study was observed that volatile compounds of S. bacillaris have inhibitory effects on the hyphal growth of phytopathogenic fungi M. phaseolina and P. sojae. Furthermore, it was observed that volatile compounds of S. bacillaris stopped the production of microsclerotia in M. phaseolina and the radial growth of P. sojae. Also, Streptomyces generates antifungal volatile compounds. Furthermore, our results revealed that T. longibrachiatum volatile compound significantly decreased the rate of microsclerotia production in M. phaseolina and the density of the colony over passing time.

 

Conclusion

    The results of the present study showed that two biocontrol agents T. longibrachiatum and S. bacillaris and their volatile compounds are highly efficient in the biocontrol of pathogenic fungi of M. phaseolina and P. sojae in vitro. Both T. longibrachiatum and S. bacillaris could decrease the production of microsclerotium of M. phaseolina. Microsclerotia are living structures that persist in the soil for many years and play an important role in the disease cycle. Studies have shown that inhibition of germination of the fungus's survival organs can disrupt the disease cycle and protect the plant. T. longibrachiatum and S. bacillaris can be good candidates for controlling M. phaseolina and P. sojae in soybean plants. Further investigation is needed in the greenhouse and field conditions.

مراجع

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دانش گیاهپزشکی ایران
دوره 54، شماره 1
شهریور 1402
صفحه 77-100

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  • تاریخ دریافت: 14 اسفند 1401
  • تاریخ بازنگری: 08 خرداد 1402
  • تاریخ پذیرش: 10 خرداد 1402

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