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非宿主幼虫会对昆虫病原真菌球孢白僵菌在土壤中的持久性产生负面影响。

Non-host larvae negatively impact persistence of the entomopathogen Beauveria bassiana in soil.

机构信息

Faculty of Agriculture and Life Sciences, PO Box 85084, Lincoln University, Lincoln, Canterbury, New Zealand.

Plant & Food Research Ltd, Lincoln, Canterbury, New Zealand; Previously Faculty of Agriculture and Life Sciences, PO Box 85084, Lincoln University, Lincoln, Canterbury, New Zealand.

出版信息

J Invertebr Pathol. 2018 Jul;156:19-28. doi: 10.1016/j.jip.2018.07.001. Epub 2018 Jul 9.

Abstract

A better understanding of the ecology of the insect pathogenic fungus, Beauveria bassiana, in soil is needed to identify reasons behind the variable efficacy often seen after field application. A transformed strain of a candidate commercial strain of B. bassiana (F418 gfp tr3), expressing the green fluorescent protein and the hygromycin B resistance gene, was used to assess the effects of the larvae of a host insect, Tenebrio molitor L. (Coleoptera: Tenebrionidae), a non-host, Costelytra zealandica (Coleoptera: Scarabaeidae) and the absence of larvae on the persistence of F418 gfp tr3 in pasteurised and non-sterile soil over 4 months. In the presence of a T. molitor larvae, F418 gfp tr3 populations increased significantly in pasteurised and non-sterile soil; however, populations increased less in non-sterile soil than in pasteurised soil. Lower populations of F418 gfp tr3 were recovered in pasteurised soil in the presence of C. zealandica larvae than in pasteurised soil without larvae. No difference was observed between F418 gfp tr3 populations in non-sterile soil with a non-host larvae or without larvae. Accompanying studies showed that F418 gfp tr3 conidia germinated and produced appressoria on live and excised cuticle of non-host (C. zealandica) larvae but infection did not occur, leading to a net loss of viable conidia in the soil. Conidia administrated orally to C. zealandica larvae were viable on recovery from faecal samples, suggesting that ingestion of the fungus by the larvae had little impact on the viable fungal population. Soil bacterial and fungal community patterns were analysed using Single-Strand Conformation Polymorphism (SSCP) and showed a correlation between changes in F418 gfp tr3 persistence in pasteurised and non-sterile soil and changes in soil communities in the presence of a host insect, non-host insect or in the absence of insect. In pasteurised soil, non-specific germination of F418 gfp tr3 conidia on the non-host larval cuticle and the presence of antagonistic bacteria introduced with the field-collected larvae are most likely responsible for the differences observed. The more complex microbial community structures in non-sterile soil could lead to fungistasis, preventing potentially antagonistic bacteria degrading conidia or inhibiting attachment and germination on the non-host larval cuticle, resulting in the observed lack of difference between non-host and no larval treatments.

摘要

为了确定田间应用后效果变化的原因,我们需要更好地了解昆虫病原真菌绿僵菌在土壤中的生态学。我们使用一株经转化的候选商业绿僵菌(F418 gfp tr3)菌株,该菌株表达绿色荧光蛋白和潮霉素 B 抗性基因,以评估宿主昆虫黄粉虫幼虫、非宿主新西兰大坎皮幼虫和无幼虫对 F418 gfp tr3 在巴氏灭菌和非无菌土壤中 4 个月内持续存在的影响。在有黄粉虫幼虫存在的情况下,F418 gfp tr3 在巴氏灭菌和非无菌土壤中的种群显著增加;然而,非无菌土壤中的种群增加量小于巴氏灭菌土壤。在有新西兰大坎皮幼虫存在的巴氏灭菌土壤中,F418 gfp tr3 的回收量低于无幼虫存在的巴氏灭菌土壤。在非无菌土壤中,非宿主幼虫或无幼虫存在时,F418 gfp tr3 的种群没有差异。在无菌土壤中,非宿主幼虫或无幼虫存在时,F418 gfp tr3 的种群没有差异。伴随的研究表明,F418 gfp tr3 分生孢子在非宿主(新西兰大坎皮幼虫)幼虫的活体和离体表皮上萌发并产生附着胞,但未发生感染,导致土壤中活分生孢子净损失。从粪便样本中回收的新西兰大坎皮幼虫口服给予的分生孢子在恢复后具有活力,这表明幼虫摄入真菌对真菌种群的活力影响不大。使用单链构象多态性(SSCP)分析了土壤细菌和真菌群落模式,结果表明,在有宿主昆虫、非宿主昆虫或无昆虫存在的情况下,F418 gfp tr3 在巴氏灭菌和非无菌土壤中的持久性变化与土壤群落的变化之间存在相关性。在巴氏灭菌土壤中,F418 gfp tr3 分生孢子在非宿主幼虫表皮上的非特异性萌发和与田间收集的幼虫一起引入的拮抗细菌的存在可能是造成这种差异的原因。非无菌土壤中更复杂的微生物群落结构可能导致真菌抑制,阻止潜在的拮抗细菌降解分生孢子或抑制在非宿主幼虫表皮上的附着和萌发,导致非宿主和无幼虫处理之间没有差异。

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