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Characterization and Functional Importance of Two Glycoside Hydrolase Family 16 Genes from the Rice White Tip Nematode Aphelenchoides besseyi

文献类型： 外文期刊

作者： Feng, Hui 1 ; Zhou, Dongmei 1 ; Daly, Paul 1 ; Wang, Xiaoyu 1 ; Wei, Lihui 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing 210014, Peoples R China

关键词： glycoside hydrolase family 16; Aphelenchoides besseyi; fungal feeding; RNA interference

期刊名称：ANIMALS （ 影响因子：2.752； 五年影响因子：2.942 ）

ISSN： 2076-2615

年卷期： 2021 年 11 卷 2 期

页码：

收录情况： SCI

摘要： Simple Summary The rice white tip nematode Aphelenchoides besseyi is a plant parasite but can also feed on fungi if this alternative nutrient source is available. Glucans are a major nutrient source found in fungi, and beta-linked glucans from fungi can be hydrolyzed by beta-glucanases from the glycoside hydrolase family 16 (GH16). The GH16 family is abundant in A. besseyi, but their functions have not been well studied, prompting the analysis of two GH16 members (AbGH16-1 and AbGH16-2). AbGH16-1 and AbGH16-2 are most similar to GH16s from fungi and probably originated from fungi via a horizontal gene transfer event. These two genes are important for feeding on fungi: transcript levels increased when cultured with the fungus Botrytis cinerea, and the purified AbGH16-1 and AbGH16-2 proteins inhibited the growth of B. cinerea. When AbGH16-1 and AbGH16-2 expression was silenced, the reproduction ability of A. besseyi was reduced. These findings have proved for the first time that GH16s contribute to the feeding and reproduction of A. besseyi, which thus provides novel insights into how plant-parasitic nematodes can obtain nutrition from sources other than their plant hosts. The glycoside hydrolase family 16 (GH16) is widely found in prokaryotes and eukaryotes, and hydrolyzes the beta-1,3(4)-linkages in polysaccharides. Notably, the rice white tip nematode Aphelenchoides besseyi harbors a higher number of GH16s compared with other plant-parasitic nematodes. In this work, two GH16 genes, namely AbGH16-1 and AbGH16-2, were isolated and characterized from A. besseyi. The deduced amino acid sequences of AbGH16-1 and AbGH16-2 contained an N-terminal signal peptide and a fungal Lam16A glucanase domain. Phylogenetic analysis revealed that AbGH16-1 and AbGH16-2 clustered with ascomycete GH16s, suggesting AbGH16-1 and AbGH16-2 were acquired by horizontal gene transfer from fungi. In situ hybridization showed that both AbGH16-1 and AbGH16-2 were specifically expressed in the nematode gonads, correlating with qPCR analysis that showed the high transcript levels of the two genes in the female nematodes. AbGH16-1 and AbGH16-2 were also significantly induced in nematodes feeding on Botrytis cinerea. Characterization of the recombinant protein showed AbGH16-1 and AbGH16-2 displayed pronounced inhibition of both conidial germination and germ tube elongation of B. cinerea. In addition, silencing of AbGH16-1 and AbGH16-2 by RNA interference significantly decreased the reproduction ability of A. besseyi and had a profound impact on the development process of offspring in this nematode. These findings have firstly proved that GH16s may play important roles in A.besseyi feeding and reproduction on fungi, which thus provides novel insights into the function of GH16s in plant-parasitic nematodes.

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