Wu Wenxian, Cheng Zhiwei, Liu Mengjie, Yang Xiufen, Qiu Dewen
The State Key Laboratory for Biology of Plant Disease and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Science, Beijing, China.
PLoS One. 2014 Jun 5;9(6):e99352. doi: 10.1371/journal.pone.0099352. eCollection 2014.
C3HC4-type RING finger proteins constitute a large family in the plant kingdom and play important roles in various physiological processes of plant life. In this study, a C3HC4-type zinc finger gene was isolated from Nicotiana benthamiana. Sequence analysis indicated that the gene encodes a 24-kDa protein with 191 amino acids containing one typical C3HC4-type zinc finger domain; this gene was named NbZFP1. Transient expression of pGDG-NbZFP1 demonstrated that NbZFP1 was localized to the chloroplast, especially in the chloroplasts of cells surrounding leaf stomata. Virus-induced gene silencing (VIGS) analysis indicated that silencing of NbZFP1 hampered fruit development, although the height of the plants was normal. An overexpression construct was then designed and transferred into Nicotiana benthamiana, and PCR and Southern blot showed that the NbZFP1 gene was successfully integrated into the Nicotiana benthamiana genome. The transgenic lines showed typical compactness, with a short internode length and sturdy stems. This is the first report describing the function of a C3HC4-type RING finger protein in tobacco.
C3HC4型泛素连接酶蛋白在植物界构成一个大家族，并在植物生命的各种生理过程中发挥重要作用。在本研究中，从本氏烟草中分离出一个C3HC4型锌指基因。序列分析表明，该基因编码一个24 kDa的蛋白质，含有191个氨基酸，包含一个典型的C3HC4型锌指结构域；该基因被命名为NbZFP1。pGDG-NbZFP1的瞬时表达表明，NbZFP1定位于叶绿体，尤其是叶片气孔周围细胞的叶绿体中。病毒诱导的基因沉默（VIGS）分析表明，NbZFP1的沉默阻碍了果实发育，尽管植株高度正常。然后设计了一个过表达构建体并转入本氏烟草，PCR和Southern杂交表明NbZFP1基因已成功整合到本氏烟草基因组中。转基因株系表现出典型的紧凑性，节间长度短，茎杆粗壮。这是首次报道C3HC4型泛素连接酶蛋白在烟草中的功能。
