کنترل زیستی بیماری پاخوره گندم با استفاده از دو قارچ اندوفیت Penicillium canescens و Penicillium hordei

نوع مقاله : مقاله پژوهشی

نویسندگان

• معصومه غلامی ¹
• جهانشیر امینی ¹
• جعفر عبداله زاده ¹
• مراحم آشنگرف ²

¹ گروه گیاهپزشکی، دانشکده کشاورزی، دانشگاه کردستان، سنندج، ایران

² گروه علوم زیستی، دانشکده علوم پایه، دانشگاه کردستان، سنندج، ایران

چکیده

کلیدواژه‌ها

• پاخوره گندم
• ژن کالمودولین
• Ggt
• گلخانه
• شدت بیماری

عنوان مقاله [English]

Biological control of Take-all disease of wheat using two fungal endophytes, Penicillium canescens and Penicillium hordei

نویسندگان [English]

• Masumeh gholami ¹
• Jahanshir Amini ¹
• Jafar abdollahzadeh ¹
• Morahem Ashengroph ²

کلیدواژه‌ها [English]

• Calmodulin gene
• disease severity
• greenhouse
• Ggt
• Wheat Take-all

Extended Abstract

Introduction

Gaeumannomyces graminis var. tritici Walker (Ggt) cause wheat take-all disease and it is the most important disease of wheat and other temperate small-grain cereal such as barley and rye (Cook., 1994; Yu et al., 2009; Cook, 2003). The pathogen survives parasitically in the roots during the growing season and saprophytically on plant debris after crop harvest (Bailey et al., 1999). The pathogen damages cereal plants by rotting the root, crown, and base of the stems and the disease can spread through different ways such as soil, diseased plants, and plant debris ((Asher and Shipton, 1981). Management of the disease by current methods such as crop rotation, use of chemical fungicides, resistant varieties, tillage to hasten infected residue decomposition has been reported to be ineffective (Ghahfarokhy et al., 2011; Liu et al., 2009). Biocontrol of wheat take-all disease by means of different microorganisms could be used as an effective and sustainable approach to control take-all disease of wheat (Lirong Yang et al., 2015; Youn-Sig Kwak et al., 2012) and the use of biocontrol agents (BCAs) to management of wheat take-all can be a more efficient and environmentally friendly alternative (Lirong Yang et al., 2015). For example, Gholami et al. (2019) found that soil drenches of Coprinopsis urticicola and Rhizoctonia endophytica decreased disease severity of wheat take-all comparable to fungicide application, and Sari et al. (2006) used a seed soaking treatment with Pseudomonas fluorescens to control wheat take-all in the greenhouse and field conditions. The aim of this research using two endophyte isolates for their ability to suppress Ggt causing take-all disease of wheat in vitro and greenhouse conditions.

Materials and Methods

The pathogen Ggt was obtained from the collection of plant pathology department, university of Kurdistan (GenBank accession number: MH488732). Two fungal endophytes were isolated from healthy cereal plants and were identified under a microscope (Olympus BX51) based on the morphology (colonies, mycelia and spores) and molecular phylogenetic analysis. Pathogenicity test of Ggt was demonstrated on wheat (Triticum aestivum L.) seeds of cultivars Pishtaz susceptible to take-all disease. Antagonistic abilities of fungal endophytes against the pathogen were investigated through in vitro assays (dual culture, volatile metabolite and non-volatile metabolite) and greenhouse conditions in a completely randomized design with four replications. For greenhouse tests, pathogen and fungal endophytes inocula were prepared on sterilized wheat seed and incubated for 4 weeks. The wheat cultivars Pishtaz susceptible to wheat take-all disease was planted in sterilized soil -sand-peat mix (1:1:1, w/w/w) inoculated with pathogen (20 g/kg soil, in each pot) and endophytes (40 g/kg soil in each pot) in greenhouse conditions. After 35 days, the disease severity and growth parameters of the wheat plant were evaluated (Weller and Cook, 1983). Data were statistically analyzed by standard analysis of variance (ANOVA) using SAS software (Version 8.2; SAS Institute, Cary, NC, USA, 2013). Differences among different treatments were analyzed using Duncan’s Multiple Range Test (DMRT) at P≤0.05. Data were reported as mean values ± standard error (SE).

Results and Discussion

Two fungal endophytes were identified as Penicillium canescens Ag1T5 and Penicillium hordei TB4L1 based on phenotypic characteristics and molecular phylogenetic analysis of the calmodulin gene (Cam) sequences. The results of this research showed that two endophytes were highly capable of inhibition the mycelial colony growth of Ggt with inhibition 52.65-70.69% and 29.44-38.11% respectively by dual culture and non-volatile metabolite assays as compared to untreated control treatments after 7 days of inoculation, while volatile of P. canescens and P. hordei had no significant inhibitory effect on Ggt. The fungal endophytes such as Fomes fomentarius and Coprinopsis urticicola showed in vitro antifungal activity against Ggt by dual culture assay on PDA medium with 39.1% and 43.6% inhibition, respectively. Two fungal endophytes produced antifungal metabolites including chitinase, cellulase, siderophore, protease and pectinase. Furthermore, both isolates produced IAA and gibberellin, which had the most effect on increasing of growth parameters of wheat plant (Gholami et al., 2019). Isolate of C. urticicola suppressed the mycelia growth and caused change in morphology of Ggt so that it may be correlated with production metabolites or lytic enzymes such as protease and pectinase that caused abnormalities hyphae morphology (Gholami et al., 2019). In greenhouse conditions, both endophytes P. canescens and P. hordei were reduced disease severity 83.57% and 38.74% respectively. Furthermore, both fungal endophytes improved plant growth parameters of wheat plant. Several biocontrol agents such as of Bacillus subtilis (Lirong Yang et al., 2015; Lirong Yang et al., 2018), Pseudomonas fluorescens (Youn-Sig Kwak et al., 2012; Mohamed and Grossmann, 1994), Streptomyces, Microbispora and Nocardioides (Justin et al., 2004) were evaluated to control wheat take-all disease and reduced disease severity of disease. Also, C. urticicola M2 and Rhizoctonia zeae M32 effectively reduced wilt disease and increased the plant growth parameters of wheat plant in greenhouse conditions (P ≤0.05) in comparison with the inoculated control (Gholami et al., 2019).

Conclusion

The results of this research indicated P. canescens Ag1T5 had a great potential as biocontrol agent against take-all of wheat caused by Ggt in vitro and greenhouse conditions. In addition, P. canescens Ag1T5 was improved wheat plant growth in greenhouse conditions and it can be used as a safe and suitable alternative to chemical fungicides and could be a useful biocontrol agents BCA) for take-all management.

Author Contributions

 MG and JA conceptualized the project. MG and JA performed the experiments and wrote the manuscript. JA and MA edited the manuscript. All authors read and approved the manuscript.

Data Availability Statement

Data are available upon request from the authors.

Acknowledgements

This research was supported by a grant from the Plant Protection Department of the University of Kurdistan (Grant No: 96/19/32668).

Ethical considerations

The authors avoided data fabrication, falsification, plagiarism, and misconduct.

Conflict of interest

The authors declare no conflict of interest.