RNA干扰（RNA interference，RNAi）是一种广泛应用于基因功能分析的研究技术。在最新的研究中，入侵生物松材线虫可经取食表达其靶基因dsRNA的丝状真菌引发，即Feeding RNAi。该技术可为线虫提供持续的干扰环境，但其作用效率在不同实验时存在较大差异，因而通过技术探索优化并稳定Feeding RNAi技术十分有必要。同时，RNAi的非靶效应长期以来便被研究者所广泛关注。本研究计划在优化后的方案中，针对非内源性靶标（绿色荧光蛋白基因，gfp）以及内源性靶标（松材线虫Bx-dpy-4l基因）进行feeding RNAi的非靶效应分析，探索gfp在RNAi对照实验中的实际价值以及靶标基因Bx-dpy-4l干扰后所引起的基因调控表达网路。通过此类分析，可以对Feeding RNAi的效用进行评估，以增加其在基因功能性分析中的应用性，并成为松材线虫研究中一项便捷、高效的技术。

RNA interference (RNAi) is a valuable and widely used tool for gene functional study. In the latest study, RNAi could be triggerred by feeding worm fungi of transgenic expression double strand RNA (dsRNA) for the invasive species, Bursaphelenchus xylophilus. This RNAi technology was also known as “Feeding RNAi”, which offered a continuous interference environment for nematode. However, the RNAi efficiency varies between different experiments. So it is necessary to optimize the operating procedures of Feeding RNAi in order to maintain the relative high RNAi efficiency. Meantime, nontarget effect of RNAi has been widely concerned by the researchers since it was developmented. In this study, we planned to analyze the non-target effect by using a nonendogenous target (green fluorescent protein gene, gfp) and an endogenous target (Bx-dpy-4l) under the optimized Feeding RNAi standards. According to the results, we could explore the practical application value of gfp in RNAi control experiment and also the regulation network of Bx-dpy-4l gene after RNAi. Through such analysis, utility of Feeding RNAi could be evaluated to increase its applicability and the possibility in gene functional analysis of pinewood nematode with convenience and efficiency.