大豆 CYP707A 基因家族的鉴定和表达分析对干旱和盐胁迫的响应。
Identification and expression analysis of the Glycine max CYP707A gene family in response to drought and salt stresses.
机构信息
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Root Biology Center, South China Agricultural University, Guangzhou 510642, China.
出版信息
Ann Bot. 2012 Aug;110(3):743-56. doi: 10.1093/aob/mcs133. Epub 2012 Jul 1.
BACKGROUND AND AIMS
Abscisic acid (ABA) plays crucial roles in plants' responses to abiotic stresses. ABA 8'-hydroxylation controlled by CYP707A genes has been well studied in Arabidopsis and rice, but not in legumes. The aims of the present study were to identify and functionally analyse the soybean CYP707A gene family, and to explore their expression patterns under dehydration and salt stresses.
METHODS
A complementation experiment was employed to verify the function of soybean CYP707A1a in ABA catabolism. Genomic and cDNA sequences of other soybean CYP707A genes were isolated from the Phytozome database based on soybean CYP707A1a. The structure and phylogenetic relationship of this gene family was further analysed. The expression patterns of soybean CYP707A genes under dehydration and salt stress were analysed via quantitative real-time PCR.
KEY RESULTS
Over-expression of GmCYP707A1a in the atcyp707a2 T-DNA insertion mutant decreased its sensitivity to ABA, indicating that GmCYP707A1a indeed functions as an ABA 8'-hydroxylase in higher plants. The soybean genome contains ten CYP707A genes. Gene structure and phylogenetic analysis showed high conservation of ten GmCYP707A genes to the other CYP707A genes from monocots and dicots. Seed imbibition induced expression of A1a, A1b, A2a, A2b, A2c, A3a and A5 in embryo, and expression of A1a, A1b, A2a and A2b in cotyledon. Dehydration induced expression of A1a, A1b, A2b, A2c, A3a, A3b, A4a, A4b and A5 both in roots and in leaves, whereas rehydration stimulated transcription of A2a, A2b, A3b, A4a and A5 in roots, and only A3b and A5 in leaves. Expression of all soybean CYP707A genes was induced either by short- or by long-term salt stress.
CONCLUSIONS
The first biological evidence is provided that GmCYP7071a encodes an ABA 8'-hydroxylase through transgenic studies. Ten soybean GmCYP707A genes were identified, most of them expressed in multiple soybean tissues, and were induced by imbibition, dehydration and salinity.
背景与目的
脱落酸（ABA）在植物应对非生物胁迫的反应中起着至关重要的作用。CYP707A 基因控制的 ABA 8'-羟化作用在拟南芥和水稻中已有深入研究，但在豆科植物中尚未研究。本研究的目的是鉴定和功能分析大豆 CYP707A 基因家族，并探讨其在脱水和盐胁迫下的表达模式。
方法
采用互补实验验证大豆 CYP707A1a 在 ABA 分解代谢中的作用。根据大豆 CYP707A1a，从 Phytozome 数据库中分离出其他大豆 CYP707A 基因的基因组和 cDNA 序列。进一步分析该基因家族的结构和系统进化关系。通过定量实时 PCR 分析大豆 CYP707A 基因在脱水和盐胁迫下的表达模式。
结果
在 atcyp707a2 T-DNA 插入突变体中过表达 GmCYP707A1a 降低了其对 ABA 的敏感性，表明 GmCYP707A1a 确实在高等植物中作为 ABA 8'-羟化酶发挥作用。大豆基因组包含十个 CYP707A 基因。基因结构和系统进化分析表明，十个 GmCYP707A 基因与单子叶和双子叶植物的其他 CYP707A 基因高度保守。种子吸胀诱导胚中 A1a、A1b、A2a、A2b、A2c、A3a 和 A5 的表达，以及子叶中 A1a、A1b、A2a 和 A2b 的表达。脱水诱导根和叶中 A1a、A1b、A2b、A2c、A3a、A3b、A4a、A4b 和 A5 的转录，而再水合仅刺激根中 A2a、A2b、A3b、A4a 和 A5 的转录，叶中仅 A3b 和 A5 的转录。所有大豆 CYP707A 基因都被短期或长期盐胁迫诱导表达。
结论
通过转基因研究提供了第一个生物学证据，表明 GmCYP7071a 通过编码 ABA 8'-羟化酶。鉴定出十个大豆 GmCYP707A 基因，其中大多数在多种大豆组织中表达，并被吸胀、脱水和盐胁迫诱导。
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