Cloning and Expression Analysis of IiCYP79B2 from Isatis indigotica Fort.

doi:10.7668/hbnxb.201751719

Abstract

Abstract: In order to explore the structure, characteristics and functions of IiCYP79B2 gene in Isatis indigotica, the IiCYP79B2 gene was cloned and the bioinformatics analysis and expression patterns under different conditions were carried out. The results showed that the full length of the gene was 1 827 bp with an intron, and its ORF was 1 629 bp, encoding 542 amino acids. IiCYP79B2 protein contained two transmembrane domains without signal peptide. The protein was localized in the chloroplast thylakoid membrane and mainly consisted of random coils and α-helix structures, having the closest relationship with Sinapis alba. The qRT-PCR analysis showed that the expression levels of IiCYP79B2 in different tissues were leaf > stem > fruit > flower > root, and the expression levels at different development stages were vegetative growth > flowering > seedling > germination. Moreover, IiCYP79B2 could be induced significantly by MJ and Glu, and repressed by SA and cold. The obtained results provided some experimental evidences for further studying the functions of IiCYP79B2 gene.

Key words: Isatis indigotica Fort., IiCYP79B2, Gene cloning, Expression analysis

CLC Number:

S567.03
Q78

HU Xiangyang, QU Xinyun, WU Gege, QIN Miaomiao, LIU Rui, GAO Tian, LI Tao. Cloning and Expression Analysis of IiCYP79B2 from Isatis indigotica Fort.[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(5): 23-29. doi: 10.7668/hbnxb.201751719.

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References

[1] 国家药典委员会. 中华人民共和国药典[M]. 2015版.一部.北京:中国医药科技出版社,2015.
Sate pharmacopoeia commission. Pharmacopoeia of the People's Republic of China[M]. 2015 Edition. Part I. Beijing:China Pharmacopoeia Science and Technology Press, 2015.
[2] 刘倩倩, 王康才, 罗春红, 邹立思. 不同品种类型菘蓝根中表告依春累积规律研究[J]. 中药材, 2013, 36(2):199-201. doi:10.13863/j.issn1001-4454.2013.02.003.
Lu Q Q, Wang K C, Luo C H, Zou L S. Accumulation law of epigoitrin in roots of Isatis indigotica of different breed type[J]. Journal of Chinese Medicinal Materials, 2013,36(2):199-201.
[3] 周孟清, 张慧茹, 贾峰, 赵银丽, 黄进, 齐仁立, 罗宇. 黄芪和板蓝根多糖的抗菌抗病毒作用研究[J]. 饲料工业, 2014, 35(21):58-64.doi:10.13302/j.cnki.fi.2014.21.015.
Zou M Q, Zhang H R, Jia F, Zhao Y L, Huang J, Qi R L, Luo Y. The effects of anti-bacterial and anti-viral and the impactions on immune system of astragalus polysaccharide and radix isatidis polysaccharide[J]. Feed Industry, 2014,35(21):58-64.
[4] 袁高峰, 陈思学, 汪俏梅. 芥子油苷及其代谢产物的生物学效应研究与应用[J]. 核农学报, 2009, 23(4):664-668, 716.
Yan G F, Chen S X, Wang Q M. Biological functions and application of glucosinolates and their degradation products[J]. Acta Agriculturae Nucleatae Sinica, 2009,23(4):664-668, 716.
[5] 李友, 马莉, 沈芃, 龚慕辛. 板蓝根三氯甲烷提取部位抗内毒素作用的"谱效"关系研究[J]. 中国药学杂志, 2011, 46(10):741-744.
L Y, Ma L, Shen P, Gong M X. Spectrum-effect relationships between HPLC fingerprints and anti-endotoxin activity of chloroform from Isatidis radix[J]. Chinese Pharmaceutical Journal, 2011,46(10):741-744.
[6] Halkier B A, Gershenzon J. Biology and biochemistry of glucosinolates[J].Annual Review of Plant Biology,2006,57(1):303-333.doi:10.1146/annurev.arplant.57.032905.105228.
[7] Hu K L, Zhu Z J, Zang Y X, Hussain S A. Accumulation of glucosinolates and nutrients in pakchoi (Brassica campestris L. ssp. chinensis var. communis) two cultivar plants exposed to sulfur deficiency[J]. Horticulture Environment and Biotechnology, 2011, 52(2):121-127. doi:10.1007/s13580-011-0097-5.
[8] 陈亚州, 阎秀峰. 芥子油苷在植物-生物环境关系中的作用[J]. 生态学报, 2007, 27(6):2584-2593. doi:10.3321/j.issn:1000-0933.2007.06.053.
Cen Y Z, Yan X F. The role of glucosinolates in plant biotic environment interaction[J]. Acta Ecologica Sinica, 2007,27(6):2584-2593.
[9] Hopkins R H,van Dam N M,van Loon J J A. Role of glucosinolates in Insect-plant relationships and multitrophic interactions[J].Annual Review of Entomology, 2009,54(1):57-83. doi:10.1146/annurev.ento.54.110807.090623.
[10] Clarke D B. Glucosinolates, structures and analysis in food[J]. Analytical Methods, 2010,2(4):310-325. doi:10.1039/b9ay00280d.
[11] 宋傲男, 程文财, 刘彩霞, 赵云, 王茂林. 甘蓝型油菜BnCYP79B1 基因的克隆与表达[J]. 西北植物学报, 2013, 33(6):1085-1090. doi:10.3969/j.issn.1000-4025.2013.06.003.
Sng A N, Cheng W C, Liu C X, Zhao Y, Wang M L. Molecular cloning and expression analysis of BnCYP79B1 gene from Brassica napus L.[J]. Acta Botanica Boreali-Occidentalia Sinica, 2013,33(6):1085-1090.
[12] Chen S X, Glawischnig E, Jørgensen K, Naur P, Jørgensen B, Olsen C E, Hansen C H, Rasmussen H, Pickett J A, Halkier B A. CYP79F1 and CYP79F2 have distinct functions in the biosynthesis of aliphatic glucosinolates in Arabidopsis[J]. Plant Journal, 2003,33(5):923-937. doi:10.1046/j.1365-313X.2003.01679.x.
[13] Mostafa I, Zhu N, Yoo M J, Balmant K M, Misra B B,Dufresne C, Abou-Hashem M, Chen S X, El-Domiaty M. New nodes and edges in the glucosinolate molecular network revealed by proteomics and metabolomics of Arabidopsis myb28/29 and cyp79B2/B3 glucosinolate mutants[J]. Journal of Proteomics,2016,138(1):1-19. doi:10.1016/j.jprot.2016.02.012.
[14] Mikkelsen M D, Petersen B L, Olsen C E, Halkier B A. Biosynthesis and metabolic engineering of glucosinolates[J]. Amino Acids, 2002, 22(3):279-295. doi:10.1007/s007260200014.
[15] Wittstock U, Halkier B A. Cytochrome P450 CYP79A2 from Arabidopsis thaliana L.catalyzes the conversion of L-phenylalanine to phenylacetaldoxime in the biosynthesis of benzylglucosinolate[J]. Journal of Biological Chemistry, 2000, 275(19):14659. doi:10.1074/jbc.275.19.14659.
[16] Banerjee A, Rai A N, Penna S A,Variyar P S. Aliphatic glucosinolate synthesis and gene expression changes in gamma-irradiated cabbage[J]. Food Chemistry,2016,209:99-103. doi:10.1016/j.foodchem.2016.04.022.
[17] Hemm M R, Ruegger M O, Chapple C. The Arabidopsis ref2 mutant is defective in the gene encoding CYP83A1 and shows both phenylpropanoid and glucosinolate phenotypes[J].The Plant Cell,2003,15(1):179-194. doi:10.1105/tpc.006544.
[18] Mikkelsen M D, Hansen C H, Wittstock U, Halkier B A. Cytochrome P450 CYP79B2 from Arabidopsis catalyzes the conversion of tryptophan to indole-3-acetaldoxime,a precursor of indole glucosinolates and indole-3-acetic acid[J]. Journal of Biological Chemistry, 2000, 275(43):33712-33717. doi:10.1074/jbc.M001667200.
[19] Meenu, Augustine R, Majee M, Pradhan A K,Bisht N C. Erratum to:Genomic origin, expression differentiation and regulation of multiple genes encoding CYP83A1, a key enzyme for core glucosinolate biosynthesis, from the allotetraploid Brassica juncea[J].Planta,2015,241(3):667.doi:10.1007/s00425-014-2210-3.
[20] Mikkelsen M D, Peter B L, Glawischnig E, Jensen A B, Andreasson E, Halkier B A. Modulation of CYP79 genes and glucosinolate profiles in Arabidopsis by defense signaling pathways[J]. Plant Physiology,2003,131(1):298-308. doi:10.1104/pp.011015.
[21] 高灿红,董丽丽,关晓弯,赵良侠,林俊城,徐福乐,张水明.青花菜萝卜硫素合成相关基因BoCYP83A1 的克隆与表达分析[J].西北植物学报,2016,36(7):1302-1307.doi:10.7606/j.issn.1000-4025.2016.07.1302.
Go C H, Dong L L, Guan X W, Zhao L X, Lin J C, Xu F L, Zhang S M. Cloning and expression analysis of sulforaphane synthesis related gene BoCYP83A1 in Broccoli[J]. Acta Botanica Boreali-Occidentalia Sinica, 2016,36(7):1302-1307.
[22] 杜海, 冉凤, 刘静, 文婧, 马珊珊, 柯蕴倬, 孙丽萍, 李加纳. 拟南芥硫苷生物合成相关基因的组织和胁迫诱导表达谱的全基因组分析[J]. 中国农业科学, 2016,49(15):2879-2897. doi:10.3864/j.issn.0578-1752.2016.15.003.
D H, Ran F, Liu J, Wen J, Ma S S, Ke Y Z, Sun L P, Li J N. Genome-wide expression analysis of glucosinolate biosynthetic genes in Arabidopsis across diverse tissues and stresses induction[J]. Scientia Agricultura Sinica, 2016,49(15):2879-2897.

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