Identification of cold tolerance genes from leaves of mangrove plant Kandelia obovata by suppression subtractive hybridization
Identification of cold tolerance genes from leaves of mangrove plant Kandelia obovata by suppression subtractive hybridization
复制标题
抑制消减杂交鉴定红树植物秋茄叶片耐寒基因
DOI：
10.1007/s10646-015-1486-9
复制
发表时间：
2015-10-01
期刊：
影响因子：
2.7
通讯作者：
Zhang, Jian-Dong
中科院分区：
文献类型：
--
作者：
Fei, Jiao;Wang, You-Shao;Zhang, Jian-Dong
低温是严重限制红树林生产力和分布的主要非生物胁迫，目前对红树林耐寒性的分子机制还知之甚少。利用抑制消减杂交技术对秋茄低温相关基因进行了筛选。从670个克隆中分离出334个冷相关表达序列标签（EST）并进行测序。在这些EST序列中，共鉴定出143个独立的cDNA序列，根据NCBI blast将其分为代谢、能量、细胞拯救和防御、转录和光合作用等10个大类。生物信息学分析表明，这些EST主要与刺激反应和代谢过程相关，共包含72条KEGG通路。两个选择的基因（例如，水通道蛋白基因和锌家族蛋白基因）进行定量实时PCR分析。这两个基因在低温胁迫下均表达上调，部分证实了消减cDNA文库的构建。这些基因功能的多样性表明，在低温胁迫下，青冈草对冷胁迫的反应是复杂的。深入研究这些基因的功能和可能的作用途径，将有助于理解红树林植物抗寒性的分子适应机制。
Low temperature is a major abiotic stress that seriously limits mangrove productivity and distribution, the molecular mechanisms of cold tolerance involved in mangroves are still poorly understood at present. It was used to identify the potential cold-related genes in Kandelia obovata (K. obovata) by suppression subtractive hybridization. 334 cold-related expressed sequence tags (ESTs) out of 670 clones were isolated and sequenced. Among these ESTs, 143 unique cDNAs were identified and classified into ten groups, such as metabolism, energy, cell rescue and defense, transcription and photosynthesis according to NCBI blast. Based on bioinformatics analysis, these ESTs were mainly related to response to stimulus and metabolic process, and were included to 72 KEGG pathways. Two selected genes (e.g., aquaporin gene and zinc family protein gene) from the library were further analyzed by quantitative real-time PCR analysis. Both the two genes were found to be transcriptionally up-regulated under cold stress, which partly approve the construction of the subtractive cDNA library. The diversity of the putative functions of these genes indicated that cold stress resulted in a complex response in K. obovata. Further investigation on the functions and potential pathways of these genes will facilitate the understanding of the molecular adaptations to cold tolerance in mangrove plants.