Molecular characterization and functions of fatty acid and retinoid binding protein gene (Ab-far-1) in Aphelenchoides besseyi.
Molecular characterization and functions of fatty acid and retinoid binding protein gene (Ab-far-1) in Aphelenchoides besseyi.
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贝氏滑刃藻脂肪酸和类视黄醇结合蛋白基因 (Ab-far-1) 的分子特征和功能
DOI：
10.1371/journal.pone.0066011
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Li Y
中科院分区：
文献类型：
--
作者：
Cheng X;Xiang Y;Xie H;Xu CL;Xie TF;Zhang C;Li Y
水稻白色尖线虫是一种植物寄生线虫，在水稻等多种作物上造成严重损失。脂肪酸和维甲酸结合蛋白（Fatty acid and retinoid binding protein，FAR）是线虫体内的特异性蛋白，与线虫的发育、繁殖、侵染寄主以及破坏植物防御反应等有关，因此抑制FAR功能是防治线虫病的潜在途径。besseyi。Ab-far-1cDNA全长805 bp，开放阅读框546 bp，编码181个氨基酸。软件分析表明，该蛋白富含α-螺旋结构，含有预测的酪蛋白激酶II磷酸化位点和疏水性分泌信号肽，但不含糖基化位点。Ab-FAR-1与Gp-FAR-1蛋白同源性为52%。系统发育树显示，Ab-FAR-1和Gp-FAR-1属于同一分支。配体结合分析证实，重组抗体FAR 1（rAb-FAR 1）与荧光类似物11-（（5-二甲氨基萘-1-磺酰基）氨基）十一烷酸（DAUDA）、顺式帕林酸和视黄醇结合，但油酸会与结合位点竞争。采用定量PCR（qPCR）技术检测了Ab-far-1在不同发育阶段的表达水平。Besseyi，在雌性中发现最高表达，其次是卵，幼体和雄性。利用原位杂交技术，Ab-far-1 mRNA存在于幼鱼和成年鱼的皮下组织，雌性的卵巢和雄性的睾丸。当A. Ab-far-1 dsRNA处理besseyi 48 h后，Ab-far-1的沉默效果最好，胡萝卜上的线虫数量最少。因此，FAR在线虫的发育和繁殖中起着重要的作用。本研究为植物寄生线虫的防治提供了一条新的途径。
Rice white tip nematode, Aphelenchoides besseyi, is a kind of plant parasitic nematodes that cause serious losses in rice and many other crops. Fatty acid and retinoid binding protein (FAR) is a specific protein in nematodes and is related to development, reproduction, infection to the host, and disruption of plant defense reactions, so the inhibition of FAR function is the potential approach to control A. besseyi. The full-length of Ab-far-1 cDNA is 805 bp, including 546 bp of ORF that encodes 181 amino acids. Software analysis revealed that the Ab-FAR-1 was rich in α-helix structure, contained a predicted consensus casein kinase II phosphorylation site and a hydrophobic secretory signal peptide, but did not have glycosylation sites. The Ab-FAR-1 had 52% homology to Gp-FAR-1 protein. The Ab-FAR-1 and Gp-FAR-1 were grouped in the same branch according to the phylogenetic tree. Fluorescence-based ligand binding analysis confirmed that the recombinant Ab-FAR-1 (rAb-FAR-1) bound with the fluorescent analogues 11-((5-dimethylaminonaphthalene-1-sulfonyl) amino) undecannoic acid (DAUDA), cis-parinaric acid and retinol, but the oleic acid would compete with the binding site. Quantitative PCR (qPCR) was used to assess the expression level of Ab-far-1 at different development stages of A. besseyi, the highest expression was found in the females, followed by eggs, juveniles and males. Using in situ hybridization technique, Ab-far-1 mRNA was present in the hypodermis of juveniles and adults, the ovaries of females and the testes of males. When A. besseyi was treated with Ab-far-1 dsRNA for 48 h, the silencing efficiency of Ab-far-1 was the best and the number of nematodes on the carrot was the least. Thus FAR plays important roles in the development and reproduction of nematodes. This study is useful and helpful to figure out a new way to control the plant parasitic nematodes.
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作者：
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DOI：
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