Tea Science Research Institute, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China.
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China.
Int J Mol Sci. 2022 Aug 18;23(16):9294. doi: 10.3390/ijms23169294.
Nitrate nitrogen is an important nitrogen source for tea plants' growth and development. LBD transcription factors play important roles in response to the presence of nitrate in plants. The functional study of LBD transcription factors in tea plants remains limited. In this study, the LBD family gene was isolated and characterized from tea plants. Sequence analysis indicated that CsLBD39 contained a highly conserved CXCXCXCX domain. The phylogenetic tree assay showed that CsLBD39 belonged to class II subfamily of the LBD family. was highly expressed in flowers and root; we determined that its expression could be induced by nitrate treatment. The CsLBD39 protein was located in the nucleus and has transcriptional activation activity in yeast. Compared with the wild type, overexpression of gene in resulted in smaller rosettes, shorter main roots, reduced lateral roots and lower plant weights. The nitrate content and the expression levels of genes related to nitrate transport and regulation were decreased in transgenic hosting gene. Compared with the wild type, overexpression in transgenic had smaller cell structure of leaves, shorter diameter of stem cross section, and slender and compact cell of stem longitudinal section. Under KNO treatment, the contents of nitrate, anthocyanins, and chlorophyll in leaves, and the content of nitrate in roots of overexpressing were reduced, the expression levels of nitrate transport and regulation related genes were decreased. The results revealed that CsLBD39 may be involved in nitrate signal transduction in tea plants as a negative regulator and laid the groundwork for future studies into the mechanism of nitrate response.
硝酸盐氮是茶树生长发育的重要氮源。LBD 转录因子在植物对硝酸盐存在的响应中发挥重要作用。茶树 LBD 转录因子的功能研究仍然有限。本研究从茶树中分离并鉴定了 LBD 家族基因。序列分析表明,CsLBD39 含有一个高度保守的 CXCXCXCX 结构域。系统进化树分析表明,CsLBD39 属于 LBD 家族的 II 类亚家族。在根和花中高表达;我们确定其表达可被硝酸盐处理诱导。CsLBD39 蛋白定位于细胞核中,在酵母中具有转录激活活性。与野生型相比,过表达 基因的 株系的莲座叶较小,主根较短,侧根减少,植株重量降低。过表达 基因的 株系的硝酸盐含量和硝酸盐转运及调控相关基因的表达水平降低。与野生型相比,过表达 基因的 株系叶片的细胞结构较小,茎横切直径较短,茎纵切的细胞细长而紧凑。在 KNO3 处理下,过表达 基因的 株系叶片的硝酸盐、花青素和叶绿素含量以及根中的硝酸盐含量降低,硝酸盐转运和调控相关基因的表达水平降低。结果表明,CsLBD39 可能作为负调节剂参与茶树硝酸盐信号转导,并为进一步研究硝酸盐响应机制奠定了基础。