[1]刘洁霞,谭国飞,吴雪君,等.鸭儿芹异黄酮2’-羟化酶基因的克隆及其表达的温度响应[J].江苏农业学报,2016,(03):640.[doi:10.3969/j.issn.1000-4440.2016.03.024]
 LIU Jie-xia,TAN Guo-fei,WU Xue-jun,et al.Cloning of isoflavone 2’-hydroxylase gene from Cryptotaenia japonica Hassk and its expression in response to temperature[J].,2016,(03):640.[doi:10.3969/j.issn.1000-4440.2016.03.024]
点击复制

鸭儿芹异黄酮2’-羟化酶基因的克隆及其表达的温度响应()
分享到:

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2016年03期
页码:
640
栏目:
园 艺
出版日期:
2016-06-30

文章信息/Info

Title:
Cloning of isoflavone 2’-hydroxylase gene from Cryptotaenia japonica Hassk and its expression in response to temperature
作者:
刘洁霞 谭国飞 吴雪君 许瀛之 陈佳凯 熊爱生
南京农业大学园艺学院作物遗传与种质创新国家重点实验室,江苏 南京 210095
Author(s):
LIU Jie-xia TAN Guo-fei WU Xue-jun XU Ying-zhi CHEN Jia-kai XIONG Ai-sheng
State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, China
关键词:
鸭儿芹 CjI2’H基因 克隆 实时荧光定量PCR 温度胁迫 基因表达
Keywords:
Cryptotaenia japonica Hassk CjI2’H gene cloning quantitative real-time PCR temperature stress gene expression
分类号:
S636.3
DOI:
10.3969/j.issn.1000-4440.2016.03.024
文献标志码:
A
摘要:
以贵州野生鸭儿芹为材料,研究了与植物抗逆境胁迫有关的细胞色素P450(CYP450)家族成员异黄酮2’-羟化酶(I2’H)基因对温度胁迫的应答和响应。利用PCR扩增技术克隆得到鸭儿芹异黄酮2’-羟化酶基因CjI2’H,该基因的开放阅读框长为1 527 bp,可编码508个氨基酸,通过DNAMAN软件预测其蛋白质分子质量为 1.270 9×105,等电点(pI)为4.80。进化树分析结果表明CjI2’H编码的氨基酸序列在不同植物间相对保守。实时定量PCR结果显示鸭儿芹在30 ℃处理下,叶柄中CjI2’H基因的相对表达量随处理时间的增加而上升,且在24 h 时达到峰值; 10 ℃处理下,CjI2’H基因的相对表达量明显低于高温处理。说明CjI2’H基因表达与温度胁迫存在关联性,高温胁迫后该基因的表达响应大于低温胁迫。
Abstract:
The CjI2’H gene encoding isoflavone 2’-hydroxylase(I2’H), a member of the cytochrome P450 superfamily related to abiotic stress in plants, was cloned by PCR amplification from wild Cryptotaenia japonica Hassk of Guizhou province. It’s ORF(open reading frame)length is 1 527 bp, encoding 508 amino acids. The enzyme molecular weight predicted by DNAMAN software was 1.270 9×105, and the pI was 4.80. The phylogenetic analysis showed that the CjI2’H-encoded amino acid sequences of I2’H were conservative in different plant species. The expression level of CjI2’H gene in the petiole of C. japonica Hassk at 30 ℃ detected by quantitative real-time PCR were up-regulated and peaked at 24 h. At 10 ℃, the expression level of CjI2’H was lower than that at high temperature. The expression of CjI2’H gene is related to temperature in C. japonica Hassk, greater under high temperature stress.

参考文献/References:

[1] 喻 梅,周守标,吴晓艳,等. 野生鸭儿芹种子休眠特性及破除方法 [J]. 生态学报, 2012, 32(4):1347-1354.
[2] 吴宝成,刘启新.鸭儿芹及其近缘植物地被特性的栽培观察[J].江苏农业科学,2014,42(1):125-128.
[3] 吴宝成,刘启新. 新型蔬菜资源鸭儿芹属植物的栽培技术研究 [J]. 湖北农业科学, 2014, 53(12): 2812-2816.
[4] 孙 兵,娄玉霞,郭水良. 野生蔬菜鸭儿芹在中国栽培的气候适应性研究 [J]. 上海农业学报, 2014, 30(1):85-89.
[5] 杨致荣,毛 雪,杨致芬,等. 细胞色素P450基因及其在植物改良中的应用 [J]. 遗传, 2003, 25(2): 237-240.
[6] 赵 剑,杨文杰,朱蔚华. 细胞色素与植物的次生代谢 [J]. 生命科学, 1999, 11(3):127-131.
[7] AKASHI T, AOKI T, AYABE S. CYP81E1, a cytochrome P450 cDNA of licorice(Glycyrrhiza echinata L.), encodes isoflavone 2’-hydroxylase [J]. Biochemical and Biophysical Research Communications, 1998, 251(1):67-70.
[8] WINKEL-SHIRLEY B. Biosynthesis of flavonoids and effects of stress [J]. Current Opinion in Plant Biology, 2002, 5(3): 218-223.
[8] 陈鸿翰,袁梦求,李双江,等. 苦荞肉桂酸羟化酶基因(FtC4H)的克隆及其UV-B胁迫下的组织表达 [J]. 农业生物技术学报, 2013, 21(2):137-147.
[10] 渠 萌,梁晓薇,张春荣,等. 甘草异黄酮2’-羟化酶cDNA的生物信息学分析 [J]. 广东药学院学报, 2015, 31(2): 243-246.
[11] TAN G F, WANG F, LI M Y, et al. De novo assembly and transcriptome characterization: novel insights into the temperature stress in Cryptotaenia japonica Hassk [J]. Acta Physiologiae Plantarum, 2015, 37(1):1739.
[12] 谭国飞,王 枫,王广龙,等. 鸭儿芹肉桂酸4-羟化酶基因的克隆与不同温度下的表达分析 [J]. 西北植物学报, 2014, 34(7):1298-1304.
[13] AYABE S I, AKASHI T. Cytochrome P450s in flavonoid metabolism [J]. Phytochemistry Reviews, 2006, 5(2):271-282.
[14] CHEN J, YUAN H, ZHANG L, et al. Cloning, expression and purification of isoflavone-2’-hydroxylase from Astragalus membranaceus Bge. var. mongolicus(Bge.)Hsiao [J]. Protein Expression and Purification, 2015, 107: 83-89.
[15] 李成磊,赵海霞,温国琴,等. 苦荞细胞色素CYP81家族同源基因FtP450-R4的克隆、分子鉴定及其功能分析 [J]. 农业生物技术学报, 2015, 23(2):181-192.
[16] 姜晓华,丁 宇,杨光富. 黄烷酮类植物抗毒素的研究简介 [J].华中师范大学学报:自然科学版, 2000, 34(4):427-432.
[17] 钟巍然,柴友荣,张 凯,等. 苯丙烷代谢途径中细胞色素P450的研究 [J]. 安徽农业科学, 2008(13): 5285-5289.
[18] LIU C J, HUHMAN D, SUMNER L W, et al. Regiospecific hydroxylation of isoflavones by cytochrome P450 81E enzymes from Medicago truncatula [J]. Plant Journal, 2003, 36(4):471-484.
[19] 龚凌霄,倪勤学,张 英. 植物抗毒素研究进展及其作为食品功能性成分的应用前景 [J]. 天然产物研究与开发, 2012(24):421-425.
[20] HINDERER W, FLENTJE U, BARZ W. Microsomal isoflavone 2’-and 3’-hydroxylases from chickpea(Cicer arietinum L.)cell suspensions induced for pterocarpan phytoalexin formation [J]. FEBS Letters, 1987, 214(1):101-106.
[21] 方智远,张武男. 中国蔬菜作物图鉴[M].南京:江苏科学技术出版社, 2011:468-469.

备注/Memo

备注/Memo:
收稿日期:2015-09-21
基金项目:国家自然科学基金项目(31272175); 江苏省杰出青年基金项目(BK20130027); 江苏高校优势学科建设项目
作者简介::刘洁霞(1994-),女, 山西晋中人,本科生,主要从事蔬菜分子生物学研究。(E-mail)14112307@njau.edu.cn
通讯作者:熊爱生,(E-mail)xiongaisheng@njau.edu.cn
更新日期/Last Update: 2016-06-30