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鉴定与粉虱 Perkinsiella saccharicida 相关的酵母:斐济叶瘿控制的潜在应用。

Identification of yeast associated with the planthopper, Perkinsiella saccharicida: potential applications for Fiji leaf gall control.

机构信息

School of Biological Sciences, The University of Queensland, Brisbane, QLD 4072, Australia.

出版信息

Curr Microbiol. 2011 Oct;63(4):392-401. doi: 10.1007/s00284-011-9990-5. Epub 2011 Aug 18.

Abstract

Yeasts associate with numerous insects, and they can assist the metabolic processes within their hosts. Two distinct yeasts were identified by PCR within the planthopper Perkinsiella saccharicida, the vector of Fiji disease virus to sugarcane. The utility of both microbes for potential paratransgenic approaches to control Fiji leaf gall (FLG) was assessed. Phylogenetic analysis showed one of the microbes is related to yeast-like symbionts from the planthoppers: Laodelphax striatellus, Nilaparvata lugens, and Sogetella furcifera. The second yeast was a member of the Candida genus, a group that has been identified in beetles and recently described in planthoppers. Microscopy revealed the presence of yeast in the fat body of P. saccharicida. The Candida yeast was cultured, and transformation was accomplished by electroporation of Candida albicans codon optimized plasmids, designed to integrate into the genome via homologous recombination. Transgenic lines conferred resistance to the antibiotic nourseothricin and expression of green fluorescent protein was observed in a proportion of the yeast cells. Stably transformed yeast lines could not be isolated as the integrative plasmids presumably replicated within the yeast without integration into the genome. If stable transformation can be achieved, then this yeast may be useful as an agent for a paratransgenic control of FLG.

摘要

酵母与许多昆虫有关联,并且它们可以帮助其宿主进行代谢过程。在斐济病病毒的介体甘蔗叶蝉 Perkinsiella saccharicida 中,通过 PCR 鉴定出了两种不同的酵母。评估了这两种微生物在潜在的共生转化方法控制斐济叶瘤 (FLG) 方面的应用。系统发育分析表明,其中一种微生物与稻飞虱 Laodelphax striatellus、褐飞虱 Nilaparvata lugens 和红尾白蜡蚧 Sogetella furcifera 中的酵母样共生体有关。第二种酵母是假丝酵母属的成员,该属已在甲虫中被鉴定,并在最近的研究中在叶蝉中被描述。显微镜观察显示酵母存在于 P. saccharicida 的脂肪体中。假丝酵母被培养,并通过电穿孔转化了经过密码子优化的 Candida albicans 质粒,这些质粒设计通过同源重组整合到基因组中。转基因株系对抗生素潮霉素表现出抗性,并且在一部分酵母细胞中观察到绿色荧光蛋白的表达。由于整合质粒可能在酵母内复制而没有整合到基因组中,因此无法分离出稳定转化的酵母株系。如果能够实现稳定转化,那么这种酵母可能作为控制 FLG 的共生转化剂有用。

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