Abstract
RNA interference (RNAi) is a powerful tool for the analysis of gene function in nematodes. Fatty acid and retinol binding protein (FAR) is a protein that only exists in nematodes and plays an important role in their life activities. The rice white-tip nematode (RWTN), Aphelenchoides besseyi, is a migratory endoparasitic plant nematode that causes serious damage in agricultural production. In this study, the expression levels of eight RWTN genes were effectively decreased when RWTN was fed Ab-far-n (n: 1–8) hairpin RNA transgenic Botrytis cinerea (ARTBn). These functions of the far gene family were identified to be consistent and diverse through phenotypic changes after any gene was silenced. Such consistency indicates that the body lengths of the females were significantly shortened after silencing any of the eight Ab-far genes. The diversities were mainly manifested as follows: (1) Reproduction of nematodes was clearly inhibited after Ab-far-1 to Ab-far-4 were silenced. In addition, silencing Ab-far-2 could inhibit the pathogenicity of nematodes to Arabidopsis; (2) gonad length of female nematodes was significantly shortened after Ab-far-2 and Ab-far-4 were silenced; (3) proportion of male nematodes significantly increased in the adult population after Ab-far-1, Ab-far-3, and Ab-far-5 were silenced, whereas the proportion of adult nematodes significantly decreased in the nematode population after Ab-far-4 were silenced. (4) Fat storage of nematodes significantly decreased after Ab-far-3, Ab-far-4, and Ab-far-7 were silenced. To our knowledge, this is the first study to demonstrate that Ab-far genes affect sex formation and lipid metabolism in nematodes, which provides valuable data for further study and control of RWTNs.
Highlights
The constructed RNA interference (RNAi) vectors, pBS-Ab-far-n (n = 2, 3, 5, 6, 7, and 8) expressing Ab-far-n hairpin RNA (hpRNA) (Figure 1A), were transferred into B. cinerea mediated by A. tumefaciens
Fed Wild-type B. cinerea (WTB) or GTB (p < 0.05 for both), whereas the latter did not significantly differ from each other (Figure 3A, Table 2). These results indicate that the target gene expression levels of rice white-tip nematode (RWTN) were efficiently silenced when the nematodes were fed the transgenic B
The body length and gonad length of females were not significantly different between the WT and GT treatments (Table 3). These results indicate that each Ab-far (Ab-far-1~Ab-far-8) might affect the growth and development of females in RWTN, and Ab-far-2 and Ab-far-4 might be related to the development female
Summary
RWTN and other plant nematodes cause an economic loss of approximately
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