鸭儿芹羟基肉桂酸转移酶基因的克隆及其对不同温度的响应
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国家自然科学基金(31272175);教育部新世纪优秀人才支持计划(NCET-11-0670);江苏省杰出青年基金(BK20130027);江苏高校优势学科建设项目


Isolation and Expression of Shikimate/Quinate Hydroxycinnamoyltransferase Gene under Different Temperatures in Cryptotaenia japonica Hassk
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    摘要:

    以贵州省野生鸭儿芹为研究对象,用RT-PCR克隆获得与木质素合成有关的羟基肉桂酸转移酶基因(CjHCT),并分析CjHCT基因对不同温度的响应,探索人工栽培鸭儿芹的适宜温度。结果表明:(1)CjHCT基因开放阅读框长度为1 290 bp,编码429个氨基酸;蛋白质分子质量为47.58 kD,等电点为6.67;进化树分析表明,CjHCT与茄科植物的HCT亲缘关系最近。(2)荧光定量PCR分析显示,CjHCT基因在贵州鸭儿芹根、叶柄、叶及花不同组织中的表达具有组织特异性,且在根中CjHCT基因的表达量最高,而在叶中表达量最低,花与叶中该基因表达量相近。(3)对鸭儿芹生长过程中主要面临的4种不同温度(10 ℃、18 ℃、30 ℃和38 ℃)分别处理0、0.5、1、2、4、8、12、24 h,荧光定量PCR检测显示,CjHCT基因的表达量(以0 h处理作为对照,表达量为1)在高温(30 ℃和38 ℃)处理下于0.5 h 时最高,其中30 ℃处理的CjHCT基因表达量高于38 ℃处理;在低温(10 ℃和18 ℃)下鸭儿芹CjHCT基因表达量于12 h时最高,其中10 ℃处理0.5 h时CjHCT基因较对照表现为下调;高温(30 ℃和38 ℃)下CjHCT基因表达量峰值比低温(10 ℃和18 ℃)下表达量峰值高,而且峰值出现的时间也较早。该实验意义在于初步研究发现低温栽培鸭儿芹能抑制CjHCT基因的表达,为人工栽培鸭儿芹温度调控提供了参考。

    Abstract:

    The shikimate/quinate hydroxycinnamoyltransferase (HCT) gene involved into lignin synthesis was cloned by RT-PCR from wild Cryptotaenia japonica Hassk from Guizhou Province.In this paper,the expression profiles of CjHCT under different temperature treatments have been analyzed and explore suitable temperature of artificial cultivation of C.japonica Hassk.The results showed as follows:(1)CjHCT gene contains 1 290 bp nucleotide acids,which encodes 429 amino acids.Using DNAMAN software prediction,the molecular weight and pI of CjHCT were 47.58 kD and 6.67,respectively.Phylogenetic analysis demonstrated that CjHCT showed the highest identity with HCTs from Apiaceae family plants.(2)Quantitative real-time PCR analysis showed that the expression of CjHCT gene was different in root,petiole,leaf,and flower of C.japonica Hassk.The expression level of CjHCT gene was the highest in root and the lowest in leaf.The expression levels of CjHCT gene were similar in leaf and flower.(3)The expression profiles of CjHCT gene was analyzed under 10 ℃,18 ℃,30 ℃ and 38 ℃ temperature treatments for 0,0.5,1,2,4,8,12,and 24 h.The expression level of 0 h treatment which was used as control was treated as 1.Under high temperatures (30 ℃ and 38 ℃) treatments,the CjHCT gene showed the highest expression levels at 0.5 h.The expression level of CjHCT under 30 ℃ treatments is higher than that under 38 ℃ treatments at this time.Under low temperature (10 ℃ and 18 ℃) treatments,the CjHCT showed the highest expression level at 12 h.The expression level of CjHCT under 10 ℃ treatment for 0.5 h is lower than that of control.The highest expression levels of CjHCT under high temperature (30 ℃ and 38 ℃) treatments are higher than that under low temperature (10 ℃ and 18 ℃) treatments.The CjHCT under high temperature (30 ℃ and 38 ℃) treatments showed the highest expression levels earlier than that under low temperature (10 ℃ and 18 ℃) treatments.C.japonica Hassk is an edible vegetable.In order to improve the taste,the lignin contents in C.japonica Hassk should be reduced.This study aims to choose the optimum temperature for C.japonica Hassk cultivation by cloning CjHCT and analyzing its expression profiles under the different temperatures.And at the same time,this paper provides novel insights into the condition of C.japonica Hassk cultivation.The experiment is a preliminary study that low temperature treatment inhibits the expression of CjHCT gene and provides the reference for suitable temperature of artificial cultivation for C.japonica Hassk.

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吴雪君,谭国飞,徐志胜,等.鸭儿芹羟基肉桂酸转移酶基因的克隆及其对不同温度的响应[J].西北植物学报,2015,35(7):1289-1296

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  • 在线发布日期: 2015-08-29
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