Molecular Cloning and Real-Time PCR Analysis of Flavone Synthase&#x02161; Gene Full-length cDNA from the Tea Plant

QIAO Xiao-yan; MA Chun-lei; CHEN Liang

doi:10.13305/j.cnki.jts.2009.5.004

Journal of Tea Science >

2009 , Vol. 29 >Issue 5: 347 - 354

DOI: https://doi.org/10.13305/j.cnki.jts.2009.5.004

Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant

QIAO Xiao-yan ,
MA Chun-lei ,
CHEN Liang

Expand

1. Research Center for Tea Germplasm and Improvement, Tea Research Institute of the Chinese Academy of Agricultural Sciences; National Center for Tea Improvement, Hangzhou 310008, China;
2. Tea Research Institute of the Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received date: 2009-02-12

Revised date: 2009-04-09

Online published: 2019-09-09

Fold

Abstract

Using RT-PCR and RACE techniques, flavone synthase Ⅱ (FSⅡ) gene, the key enzyme in synthesize flavone, was cloned in the tea plant. The FS II belongs to cytochrome P450 superfamily. The GenBank Accession No. is FJ169499 in the NCBI. The FSⅡ gene cDNA had 1 824 bp in full-length with 1 605 bp in coding region, encoding 534 amino acids. The putative molecular weight was 60.4 kD. The expression levels of flavone synthaseⅡ gene in spring ‘two and a bud’, mature leaves, petals, stamen and pistils, and seeds of the tea plant were assessed using SYBR Green I Real-time PCR. The expression in mature leaves was significantly higher than that in the other four tissues.

Key words： tea plant; flavone synthaseⅡ; Real-Time PCR; cytochrome P450 superfamily

Cite this article

QIAO Xiao-yan , MA Chun-lei , CHEN Liang . Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant[J]. Journal of Tea Science, 2009 , 29(5) : 347 -354 . DOI: 10.13305/j.cnki.jts.2009.5.004

References

[1] Harborne JB, Williams CA.Advances in flavonoid research since 1992[J]. Phytochemistry, 2000, 55(6): 481~504.
[2] Rice-Evans C.Flavonoid antioxidants[J]. Current Medicinal Chemistry, 2001, 8: 797~807.
[3] Shenouda NS, Zhou C, Browning JD,et al. Phytoestrogens in common herbs regulate prostate cancer cell growthin vitro[J]. Nutrition and Cancer, 2004, 49(2): 200~208.
[4] Zhou Y, Mi MT.Genistein stimulates hematopoiesis and increases survival in irradiated mice[J]. Journal of Radiation Research, 2005, 46(4): 425~433.
[5] Zhu GX, Li CZ, Cao ZG.Inhibitory effect of flavonoid baicalin on degranulation of human polymorphonuclear leukocytes induced by interleukin-8: Potential role in periodontal diseases[J]. Journal of Ethnopharmacology, 2007, 109(2): 325~330.
[6] Chen YC, Shen SC, Chow JM,et al. Flavone inhibition of tumor growth via apoptosisin vitroandin vivo[J]. International Journal of Oncology, 2004, 25(3): 661~670.
[7] Way TD, Kao MC, Lin JK.Apigenin induces apoptosis through proteasomal degradation of HER2/neu in HER2/neuover expressing breast cancer cells via the phosphatidylinositol 3-kinase/Akt-dependent pathway[J]. Journal of Biological Chemistry, 2004, 279(6): 4479~4489.
[8] 刘婵, 陈翔. 芹菜素对中枢神经作用的研究进展[J]. 国外医学: 神经病学·神经外科学分册, 2005, 32(3): 264~267.
[9] Leonard E, Chemler J, Lim KH,et al. Expression of a soluble flavone synthase allows the biosynthesis of phytoestrogen derivatives inEscherichia coli[J]. Applied Microbiology and Biotechnology, 2006, 70(1): 85~91.
[10] Kochs G, Grisebach H.Induction and characterization of a NADPH-dependent flavone synthase from cell cultures ofsoybean[J]. Z Naturforschung, 1987, 42c: 343~348.
[11] Stich K, Ebermann R, Forkmann G.Einfluβ cytochrom P-450-spesifischer inhibitor auf die aktivitat von flavonoid 3’hydroxylase and flavon synthaseⅡ bei verschiedenen Pflanzen[J]. Phyton(Austria), 1988, 28: 237~247.
[12] Kitada C, Gong Z, Tanaka Y,et al. Differential expression of two cytochrome P450s involved in the biosynthesis of flavones and anthocyanins in chemovarietal forms ofPerilla frutescens[J]. Plant and Cell Physiology, 2001, 42(12): 1338~1344.
[13] Martens S, Forkmann G.Genetic control of flavone synthase II activity in flowers ofGerberahybrids[J]. Phytochemistry, 1998, 49(7): 1953~1958.
[14] Britsch L, Heller W, Grisebach H.Conversion of flananone to flavone, and flavonol with an enzyme system from cell cultures of parsley[J]. Z. Naturforschung, 1981, 36c: 742~750.
[15] Martens S, Formann G.Cloning and expression of flavone synthaseⅡfromGerbera hybrids[J]. The Plant Journal, 1999, 20(5): 611~618
[16] Werck-Reichhart D, Feyereisen R. Cytochromes P450: a success story [J]. Genome Biology, 2000, 1(6): reviews3003.1~3003.9.
[17] Nelson DR, Zeldin DC, Hoffman SMG,et al. Comparison of cytochrome P450 (CYP) genes from the mouse and human genomes, including nomenclature recommendations for genes, pseudogenes, and alternative-splice variants[J]. Pharmacogenetics, 2004, 14(1): 1~18.
[18] Holton TA, Brugliera F, Lester DR,et al. Cloning and expression of cytochrome P450 genes controlling flower colour[J]. Nature, 1993, 366(6452): 276~279.
[19] Werck-Reichhart D, Bak S, Paquette S.Cytochrome P450 [J].In CR Somerville, EM Meyerowitz, eds, The Arabidopsis Book. American Society of Plant Biologists, Rockville, MD, doi/10.1199/tab.0028, http://www.aspb.org/publications/Arabidopsis. 2002.
[20] Akashi T, Fukuchi-MizutaniM, Aoki T,et al. Molecular cloning and biochemical characterization of a novel cytochrome P450, flavone synthase II, that catalyzes direct conversion of flavanones to flavones[J]. Plant and Cell Physiology, 1999, 40(11): 1182~1186.
[21] Steele CL, Gijzen M, Qutob D,et al. Molecular characterization of the enzyme catalyzing the aryl migration reaction of isoflavonoid biosynthesis in soybean[J]. Archives of Biochemistry Biophysics, 1999, 367(1): 146~150.
[22] Park JS.Flavonoid biosynthesis-related expression during anthocyanin accumulation in flower. http://www. ncbi.nlm.nih.gov, Accession No. AY945842, 2007.
[23] Okinaka Y, Shimada Y, Nakano-Shimada R,et al. Selective accumulation of delphinidin derivatives in tabacco using a putative flavnoid 3’5’ hydroxylase cDNA fromCampanula nedium[J]. Bioscience, Biotechnology, and Biochemstry, 2003, 67(1): 161~165.
[24] Markham KR.Distribution of flavonoids in the lower plants and its evolutionary significance. In: Harborne, J.B. (Ed.), The Flavonoids: Advances in Research since 1980 [M]. Chapman & Hall, London, 1988, 427~468.
[25] Bohm B A.Introduction to Flavonoids[M]. UK: Harwood Academic Publishers, Reading, 1998.
[26] Niklas KJ, Giannasi DE.Angiosperm paleo-biochemistry ofSuccor Creekflora (Miocene) Oregon, USA[J]. American Journal of Botany, 1978, 65: 943~952.
[27] 张宏杰. 蝗虫细胞色素b基因部分序列的多样性和分子系统学研究[J]. 陕西师范大学学报(自然科学版), 2006, 34(S): 38~44.
[28] 国会艳, 李玲, 马玉堃, 等. 基于线粒体细胞色素b基因序列的雀形目18种鸟系统发育关系[J]. 动物学杂志, 2007, 42(2): 34~40.
[29] Gibson UE, Heid CA, William PM,et al. A novel method for real time quantitative RT-PCR[J]. Genome Research, 1996, 6(10): 995~1001.
[30] 赵丽萍, 陈亮, 王新超, 等. 茶树新梢不同叶片中β-葡萄糖苷酶和β-樱草糖苷酶基因表达的实时定量PCR分析[J]. 茶叶科学, 2006, 26(1): 11~16.
[31] 王开发, 耿越. 花粉中黄酮类研究[J]. 养蜂科技, 1997, 3: 8~12.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References