Kimber M J, Fleming C C, Bjourson A J, Halton D W, Maule A G
Parasitology Research Group, School of Biology and Biochemistry, Medical Biology Centre, Queen's University Belfast, 97 Lisburn Road, BT9 7BL, Belfast, UK.
Mol Biochem Parasitol. 2001 Sep 3;116(2):199-208. doi: 10.1016/s0166-6851(01)00323-1.
This study presents data demonstrating the presence of FMRFamide-related peptides (FaRPs) in potato cyst nematodes (PCN). Five transcripts of FaRP encoding genes, designated gpflp-1 to gpflp-5, were characterised using RACE. In terms of ORFs, gpflp-1 was 444 base pairs (bp) long and coded for four copies of the FaRP, PF3 (KSAYMRFamide) whilst gpflp-2 was 309 bp long and encoded one copy of the peptide, KNKFEFIRFamide. gpflp-3 (420 bp) Encoded two copies of KHEYLRFamide (AF2) and the genes gpflp-4 and gpflp-5 encoded a total of 11 FaRPs, most of which are novel to PCN. FMRFamide-related peptide (FaRP)-like immunoreactivity was observed in both PCN species, Globodera pallida and Globodera rostochiensis, using an antiserum raised against the invertebrate peptide, FMRFamide. Immunopositive neurones were found throughout the central nervous system in the ventral and dorsal nerve cords and the circumpharyngeal and perianal nerve rings. Reactive neurones were also present peripherally, innervating the highly muscular pharynx with a nerve net and ring-like structures. Positive immunostaining was also observed in neurones running toward the stylet protractor muscles and/or the anterior sensory apparatus. This study implicates a role for FaRPs in feeding, host penetration and sensory function of PCN. This is the first study to characterise FaRP encoding genes from a plant-parasitic nematode using a targeted PCR based RACE approach and further underlines the importance and diversity of this neuropeptide group in the phylum Nematoda.
本研究提供的数据表明，马铃薯胞囊线虫（PCN）中存在FMRF酰胺相关肽（FaRPs）。利用RACE技术对5个FaRP编码基因的转录本进行了鉴定，命名为gpflp-1至gpflp-5。就开放阅读框而言，gpflp-1长444个碱基对（bp），编码4个FaRP拷贝，即PF3（KSAYMRFamide），而gpflp-2长309 bp，编码1个肽拷贝，即KNKFEFIRFamide。gpflp-3（420 bp）编码2个KHEYLRFamide（AF2）拷贝，基因gpflp-4和gpflp-5共编码11个FaRPs，其中大多数对PCN来说是新发现的。使用针对无脊椎动物肽FMRFamide制备的抗血清，在两种PCN物种，即苍白球孢囊线虫和罗斯托克孢囊线虫中均观察到了FMRF酰胺相关肽（FaRP）样免疫反应性。在腹侧和背侧神经索以及咽周和肛周神经环的整个中枢神经系统中均发现了免疫阳性神经元。反应性神经元也存在于外周，通过神经网和环状结构支配高度肌肉化的咽部。在朝向口针牵引肌和/或前部感觉器官的神经元中也观察到了阳性免疫染色。本研究表明FaRPs在PCN的取食、穿透宿主和感觉功能中发挥作用。这是首次使用基于靶向PCR的RACE方法对植物寄生线虫的FaRP编码基因进行鉴定的研究，进一步强调了这一神经肽组在线虫门中的重要性和多样性。
