通过转录和拮抗分析测定土耳其木霉对镰刀菌的生物防治能力

IF 2.1 Q3 MYCOLOGY Frontiers in fungal biology Pub Date : 2023-11-13 DOI:10.3389/ffunb.2023.1278525
Özlem Sefer, Esma Özsoy, Emre Yörük, Evrim Özkale
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引用次数: 0

摘要

本研究旨在探索潜在的真菌生物防治剂,通过转录分析对几种木霉菌株的拮抗作用进行了评价。首先从土耳其马尼萨的天然木屑和林区土壤中分离得到21株单孢子木霉。通过tef1-α测序,分离的木霉属4个不同种(T. atroviride、T. harzianum、T. koningii和T. breviccompacactum)。计算各菌种的线性生长率(LGR)为13.22±0.71 mm/d (T. atroviride TR2) ~ 25.06±1.45 mm/d (T. harzianum K30)。简单序列重复(ISSR)基因分型通过在树形图中呈现两个亚簇来验证tef1-α测序结果。我们确定了基因上最相似的(TR1 &TR2;97.77%)和不相似(K9 &K17;40.40%),分别属于同一种和不同种。双夹心培养试验(可用于拮抗研究)表明,抑菌力最小的哈氏T. harzianum K21和抑菌力最大的T. brevicompactum K26分别以3%和46%的增长率抑制了F. culmorum。利用实时荧光定量聚合酶链反应(qRT-PCR)检测了此前与支原体-植物保护-次生代谢相关的基因(nag1、tgf-1和tmk-1)的表达。在K21中,这三个基因的表达量没有显著差异(p>0.05),但在K26中,这三个基因的表达量上调了两倍,分别为2.69±0.26 (p<0.001)、2.23±0.16 (p<0.001)和5.38±2.01 (p<0.05),这意味着真菌的生理过程发生了显著变化。同时,对感染了F. culmorum FcUK99菌株的小麦进行了分枝寄生试验。试验结果,包括特定的植物生长参数(植株的重量或长度),证实了该分离物的分枝寄生潜力。结果表明:(1)nag1、tgf-1和tmk-1基因可作为评价木霉BCA能力的可靠标记;(2)短孔霉K26菌株可作为防治霉霉病的有希望的候选菌株,在确保其无害和安全的必要程序下。
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Determining the biocontrol capacities of Trichoderma spp. originating from Turkey on Fusarium culmorum by transcriptional and antagonistic analyses
In this study aiming to investigate potential fungal biocontrol agents for Fusarium culmorum , several isolates of Trichoderma spp. were evaluated for their antagonistic effects by means of transcriptional analyses. At first, 21 monosporic Trichoderma spp. isolates were obtained from natural wood debris and wood area soils in Manisa, Turkey. Trichoderma spp. Isolates were identified as belonging to four different species ( T. atroviride, T. harzianum, T. koningii , and T. brevicompactum ) by tef1-α sequencing. Then, the linear growth rate (LGR) of each species was calculated and determined to be in a range between 13.22 ± 0.71 mm/day ( T. atroviride TR2) and 25.06 ± 1.45 mm/day ( T. harzianum K30). Inter-simple sequence repeat (ISSR) genotyping validated the tef1-α sequencing results by presenting two sub-clusters in the dendrogram. We determined the genetically most similar (TR1 &amp; TR2; 97.77%) and dissimilar (K9 &amp; K17; 40.40%) individuals belonging to the same and different species, respectively. Dual sandwich culture tests (which are useful for antagonism studies) revealed that T. harzianum K21 (the least suppressive) and T. brevicompactum K26 (the most suppressive) isolates suppressed F. culmorum with growth rates of 3% and 46%, respectively. Expressions of genes previously associated with mycoparasitism-plant protection-secondary metabolism ( nag1 , tgf-1 , and tmk-1 ) were tested by quantitative real-time polymerase chain reaction (qRT-PCR) in both those isolates. While there were no significant differences (p&gt;0.05) in expression that were present in the K21 isolate, those three genes were upregulated with fold change values of 2.69 ± 0.26 (p&lt;0.001), 2.23 ± 0.16 (p&lt;0.001), and 5.38 ± 2.01 (p&lt;0.05) in K26, meaning that the presence of significant alteration in the physiological processes of the fungus. Also, its mycoparasitism potential was tested on Triticum aestivum L. cv Basribey in planta , which was infected with the F. culmorum FcUK99 strain. Results of the trials, including specific plant growth parameters (weight or length of plantlets), confirmed the mycoparasitic potential of the isolate. It can be concluded that (i) nag1 , tgf-1 , and tmk-1 genes could be approved as reliable markers for evaluation of BCA capacities of Trichoderma spp. and (ii) the T. brevicompactum K26 strain can be suggested as a promising candidate for combating in F. culmorum diseases following the necessary procedures to ensure it is non-hazardous and safe.
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