National Laboratory of Plant Molecular Genetics and National Center for Plant Gene Reserach Shanghai, Institute of Plant Physiology & Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, China.
BMC Genomics. 2012 Feb 10;13:64. doi: 10.1186/1471-2164-13-64.
Cassava is an important tropical root crop adapted to a wide range of environmental stimuli such as drought and acid soils. Nevertheless, it is an extremely cold-sensitive tropical species. Thus far, there is limited information about gene regulation and signalling pathways related to the cold stress response in cassava. The development of microarray technology has accelerated the study of global transcription profiling under certain conditions.
A 60-mer oligonucleotide microarray representing 20,840 genes was used to perform transcriptome profiling in apical shoots of cassava subjected to cold at 7°C for 0, 4 and 9 h. A total of 508 transcripts were identified as early cold-responsive genes in which 319 sequences had functional descriptions when aligned with Arabidopsis proteins. Gene ontology annotation analysis identified many cold-relevant categories, including 'Response to abiotic and biotic stimulus', 'Response to stress', 'Transcription factor activity', and 'Chloroplast'. Various stress-associated genes with a wide range of biological functions were found, such as signal transduction components (e.g., MAP kinase 4), transcription factors (TFs, e.g., RAP2.11), and reactive oxygen species (ROS) scavenging enzymes (e.g., catalase 2), as well as photosynthesis-related genes (e.g., PsaL). Seventeen major TF families including many well-studied members (e.g., AP2-EREBP) were also involved in the early response to cold stress. Meanwhile, KEGG pathway analysis uncovered many important pathways, such as 'Plant hormone signal transduction' and 'Starch and sucrose metabolism'. Furthermore, the expression changes of 32 genes under cold and other abiotic stress conditions were validated by real-time RT-PCR. Importantly, most of the tested stress-responsive genes were primarily expressed in mature leaves, stem cambia, and fibrous roots rather than apical buds and young leaves. As a response to cold stress in cassava, an increase in transcripts and enzyme activities of ROS scavenging genes and the accumulation of total soluble sugars (including sucrose and glucose) were also detected.
The dynamic expression changes reflect the integrative controlling and transcriptome regulation of the networks in the cold stress response of cassava. The biological processes involved in the signal perception and physiological response might shed light on the molecular mechanisms related to cold tolerance in tropical plants and provide useful candidate genes for genetic improvement.
木薯是一种重要的热带块根作物,能适应干旱和酸性土壤等广泛的环境刺激。然而,它是一种对低温极为敏感的热带物种。迄今为止,有关木薯冷胁迫响应相关的基因调控和信号通路的信息十分有限。微阵列技术的发展加速了特定条件下的全转录组谱研究。
使用代表 20840 个基因的 60 聚体寡核苷酸微阵列,对在 7°C 下经历 0、4 和 9 小时冷处理的木薯顶芽进行转录组谱分析。总共鉴定出 508 个转录本为早期冷响应基因,其中 319 个序列与拟南芥蛋白对齐时有功能描述。基因本体论注释分析确定了许多与冷相关的类别,包括“对生物和非生物刺激的反应”、“对胁迫的反应”、“转录因子活性”和“叶绿体”。发现了许多具有广泛生物学功能的各种应激相关基因,例如信号转导成分(例如 MAP 激酶 4)、转录因子(例如 RAP2.11)和活性氧(ROS)清除酶(例如过氧化氢酶 2)以及光合作用相关基因(例如 PsaL)。包括许多研究充分的成员(例如 AP2-EREBP)在内的 17 个主要的 TF 家族也参与了对冷胁迫的早期反应。同时,KEGG 途径分析揭示了许多重要途径,例如“植物激素信号转导”和“淀粉和蔗糖代谢”。此外,通过实时 RT-PCR 验证了 32 个基因在冷和其他非生物胁迫条件下的表达变化。重要的是,大多数测试的应激响应基因主要在成熟叶片、茎形成层和纤维根中表达,而不是在顶芽和嫩叶中表达。作为木薯冷胁迫反应的一部分,还检测到 ROS 清除基因的转录物和酶活性的增加以及总可溶性糖(包括蔗糖和葡萄糖)的积累。
动态表达变化反映了木薯冷胁迫响应网络的综合调控和转录组调控。参与信号感知和生理反应的生物学过程可能揭示与热带植物耐冷性相关的分子机制,并为遗传改良提供有用的候选基因。