College of Plant Protection, Hunan Agricultural University, Changsha, Hunan, 410128, China; Hunan Provincial Engineering & Technology Research Center for Bio-pesticide and Formulation Processing, Changsha, Hunan, 410128, China.
College of Plant Protection, Hunan Agricultural University, Changsha, Hunan, 410128, China.
Biochimie. 2020 Apr-May;171-172:91-102. doi: 10.1016/j.biochi.2020.02.010. Epub 2020 Feb 25.
Voltage-gated calcium channels (VGCCs) mediate the entry of Ca ions into cells in response to membrane depolarization and play fundamental roles in the nervous system, and the α1 subunits are the main subunits of Ca channels. Caenorhabditis elegans possesses genes encoding α1 subunits; however, very few of these genes have been cloned in plant-parasitic nematodes (PPNs). Ditylenchus destructor is a PPN that has been proposed as a new model for studying the biology and control of PPNs. To understand the structure and function of the VGCCs of this PPN, we first cloned and identified three full-length cDNAs of VGCC α1 subunit genes in D. destructor with the defining structural and conserved features of Ca1 (L-type), Ca2 (non-L-type) and Ca3 (T-type). In situ hybridization assays demonstrated that the Ca1 VGCC α1 subunit gene (DdCα1D) was expressed within body wall muscles. The Ca2 VGCC α1 subunit (DdCα1A) was expressed in the oesophageal gland, vulva and vas deferens of the worm, and the Ca3 VGCC α1 subunit (DdCα1G) was localized to the oesophagus and median bulb. In addition, on the basis of the in vitro knockdown of L-, non-L- and T-type genes via RNAi, these genes were predicted to play a key role in the modulation of locomotion, feeding and reproduction. After the silencing of DdCα1G, the median bulb muscle of D. destructor was obviously contracted, and its feeding and reproduction abilities were significantly inhibited. This study provides insight into the structure and function of VGCC α1 subunits in D. destructor.
电压门控钙通道（VGCCs）介导 Ca 离子进入细胞，响应膜去极化，在神经系统中发挥着基本作用，α1 亚基是 Ca 通道的主要亚基。秀丽隐杆线虫（Caenorhabditis elegans）拥有编码α1 亚基的基因；然而，这些基因中只有很少一部分在植物寄生线虫（PPNs）中被克隆。毁灭茎线虫（Ditylenchus destructor）是一种 PPN，已被提议作为研究 PPN 生物学和控制的新模型。为了了解这种 PPN 的 VGCCs 的结构和功能，我们首先使用定义 Ca1（L 型）、Ca2（非 L 型）和 Ca3（T 型）的结构和保守特征，克隆并鉴定了 D. destructor 中三个全长 VGCC α1 亚基基因的 cDNA。原位杂交分析表明，Ca1 VGCC α1 亚基基因（DdCα1D）在体壁肌肉中表达。Ca2 VGCC α1 亚基（DdCα1A）在虫体的食道腺、阴门和输精管中表达，Ca3 VGCC α1 亚基（DdCα1G）定位于食道和中肠球。此外，基于 RNAi 体外敲低 L、非 L 和 T 型基因，预测这些基因在调节运动、摄食和生殖中发挥关键作用。沉默 DdCα1G 后，毁灭茎线虫的中肠球肌肉明显收缩，其摄食和生殖能力明显受到抑制。本研究为了解 D. destructor 中 VGCC α1 亚基的结构和功能提供了线索。
