Molecular cloning and characterization of a chlorophyll degradation regulatory gene from bamboo

Leaf senescence constituted the final stage of leaf development and it is always accompanied by the leaf yellowing. The non-yellowing gene (NYE1), initially identified from Arabidopsis in our laboratory, is a key regulatory gene responsible for chlorophyll degradation during senescence. In this study, an orthologue of AtNYE1 was isolated from the bamboo (Bambusa emeiensis cv. Viridiflavus) and tentatively named BeNYE1. The full length sequence of 1 386 bp contains an open reading frame of 801 bp. The protein encoded by BeNYE1 consists of 266 amino acids. Sequence analysis revealed that BeNYE1 had high similarity with other NYE/SGR proteins from various monocotyledon species. BeNYE1 was strongly induced by natural senescence and dark-induced senescence in bamboo. Driven by a 1.5 kb upstream fragment of AtNYE1, BeNYE1 could rescue the stay-green phenotype of nye1-1. The constitutive over-expression of BeNYE1 could accelerate the chlorophyll degradation. These results indicated that BeNYE1 might play an important role in the regulation of chlorophyll degradation during leaf senescence in bamboo.     

金鱼藤中ApNAP基因的克隆和表达模式分析

金鱼藤(Asarina procumbens)是玄参科金鱼藤属多年生藤本植物,在园林中常用于垂直绿化,是良好的观赏植物,但金鱼藤叶片衰老后很快干枯并脱落,仅留光秃的茎蔓,使其观赏价值大大降低。AtNAP是拟南芥NAC基因家族中与衰老相关的转录因子,在调控叶片衰老过程中有重要的作用。本研究以金鱼藤衰老叶片为材料,运用RT-PCR和RACE技术克隆金鱼藤中ApNAP基因cDNA序列。结果显示,该基因位于细胞核中,cDNA全长为1341 bp,开放阅读框为936 bp,编码311个氨基酸,相对分子量为35.21 kD,等电点为9.13;ApNAP基因组全长1526 bp,含有2个内含子和3个外显子。多重序列比对表明,ApNAP属于NAC转录因子家族。系统进化树分析显示,ApNAP属于NAP亚组,与AtNAM的同源关系最近。实时荧光定量PCR结果表明,受自然状态下衰老和黑暗处理的诱导,ApNAP基因表达量增加,这进一步说明ApNAP基因参与调控金鱼藤叶片衰老的过程。该基因的分离对利用转基因技术延缓金鱼藤叶片衰老,提高金鱼藤在园林绿化中的观赏价值具有实际意义。