Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (07): 1253-1263.doi: 10.3724/SP.J.1006.2012.01253

• RESEARCH NOTES • Previous Articles     Next Articles

Molecular Cloning and Information Analysis of ANS Genes Encoding Anthocyanin Synthases from Mulberry (Morus alba)

ZHANG Qiong-Yu,LI Jun,ZHAO Ai-Chun,WANG Xi-Ling,JIN Xiao-Yun,Li Zhen-Gang,YU Mao-De*   

  1. State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei 400715, China
  • Received:2011-06-13 Revised:2012-04-28 Online:2012-07-12 Published:2012-05-11
  • Contact: 余茂德, E-mail: yumd@163.com, Tel: 023-68250191 E-mail:noscjoey@yahoo.cn

Abstract: Anthocyanidin synthase (ANS, leucoanthocyanidin oxygenase) is one of the critical enzymes in the biosynthesis of the anthocyanin. Anthocyanidin synthase gene fragment (designated as MaANS) was isolated from mulberry fruit (Moru alba) by RT-PCR based on homology cloning and genome walking technology. MaANS with the 5′ and 3′ was cloned by genome-walking. The full-length genomic sequence of MaANS is 1 535 bp, which consists of two exons and one intron. The coding region length was
1 077 bp, and their deduced protein consisted of 358 amino acid residues. Multiple alignments revealed that the nucleic acid of MaANS shared above 80% identity with that of Fragaria × ananassa, Ipomoea batatas, Malus domestica, and Pyrus pyrifolia. Structural analysis showed that the MaANS protein might belong to the 2 OG and Fe(II)-dependent oxygenase family. Phylogenetic tree analysis revealed that MaANS was the most close with Fragaria × ananassa, then Malus domestica and Pyrus pyrifolia. Reverse transcription-PCR (RT-PCR) analyses of MaANS transcripts showed that it was abundantly expressed in the young leaves and ripened fruit. All research made an essential foundation for pathway and regulation of gene expression in anthocyanin biosynthesis in mulberry fruit.

Key words: Mulberry, Anthocyanin synthase, Cloning, Information analysis, Tissues expression

[1]Schaefer H M, Wilkinson D M, Red leaves, insects and coevolution: a red herring. Trends Ecol Evol, 2004, 19: 616-618

[2]Tsuda T, Horio F, Uchida K, Aoki H, Osawa T. Dietary cyanidin 3-O-beta-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia in mice. J Nutr, 2003, 133: 2125-2130

[3]Wang S Y, Jiao H. Scavenging capacity of berry crops on superoxide radicals, hydrogen peroxide, hydroxyl radicals, and singlet oxygen. J Agric Food Chem, 2000, 48: 5677-5684

[4]Matsumoto H, Inaba H, Kishi M, Tominaga S, Hirayama M, Tsuda T. Orally administered delphinidin 3-rutinoside and cyanidin 3-rutinoside are directly absorbed in rats and humans and appear in the blood as the intact forms. J Agric Food Chem, 2001, 49: 1546-1551

[5]Nakaishi H, Matsumoto H, Tominaga S, Hirayama M. Effects of black current anthocyanoside intake on dark adaptation and VDT work-induced transient refractive alteration in healthy humans. Altern Med Rev, 2000, 5: 553-562

[6]Muth E R, Laurent J M, Jasper P. The effect of bilberry nutritional supplementation on night visual acuity and contrast sensitivity. Altern Med Rev, 2000, 5: 164-173

[7]Mazza G, Kay C D, Cottrell T, Holub B J. Absorption of anthocyanins from blueberries and serum antioxidant status in human subjects. J Agric Food Chem, 2002, 50: 7731-7737

[8]Wang X-L(王茜龄). Studies on Induction of Polyploid and Breeding of Good New Variety in Mulberry. PhD Dissertation of Southwest University, 2009 (in Chinese with English abstract)

[9]Turnbull J J, Sobey W J, Aplin R T, Hassan A, Firmin J L, Schofield C J, Prescott A G. Are anthocyanidins the immediate products of anthocyanidin synthase. Chem Commun, 2000, 24: 2473-2474

[10]Menssen A, Höhmann S, Martin W, Schnable P S, Peterson P A, Saedler H, Gierl A. The En/Spm transposable element of Zea mays contains splice sites at the termini generating a novel intron from a dSpm element in the A2 gene. EMBO J, 1990, 9: 3051-3057

[11]Martin C, Prescott A, Mackay S, Bartlett J, Vrijlandt E. Control of anthocyanin biosynthesis in flowers of Antirrhinum majus. Plant J, 1991, 1: 37-49

[12]Weiss D, van der Luit A H, Kroon J T, Mol J N, Kooter J M. The petunia homologue of the Antirrhinum majus candi and Zea mays A2 flavonoid genes; homology to flavanone 3-hydroxylase and ethylene-forming enzyme. Plant Mol Biol, 1993, 22: 893-897

[13]Gong Z, Yamazaki M, Sugiyama M, Tanaka Y, Saito K. Cloning and molecular analysis of structural genes involved in anthocyanin biosynthesis and expressed in a forma-specific manner in Perilla frutescens. Plant Mol Biol, 1997, 35: 915-927

[14]Szankowski I, Flachowsky H, Li H, Halbwirth H, Treutter D, Regos I, Hanke MV, Stich K, Fischer T C. Shift in polyphenol profile and sublethal phenotype caused by silencing of anthocyanidin synthase in apple (Malus sp.). Planta, 2009, 229: 681-692

[15]Xu F, Cheng H, Cai R, Li L L, Chang J, Zhu J, Zhang F X, Chen L J, Wang Y, Cheng S H, Cheng S Y. Molecular cloning and function analysis of an anthocyanidin synthase gene from Ginkgo biloba, and its expression in abiotic stress responses. Mol Cells, 2008, 26: 536-547

[16]Jaakola L, Määttä K, Pirttilä Maria A, Törrönen R, Kärenlampi S and Hohtola A. Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin, proanthocyanidin, and flavonol levels during bilberry fruit development. Plant Physiol, 2002, 130: 729-739

[17]Mano H, Ogasawara F, Sato K, Higo H, Minobe Y. Isolation of a regulatory gene of anthocyanin biosynthesis in tuberous roots of purple-fleshed sweet potato. Plant Physiol, 2007, 143: 1252-1268

[18]Xiao Y H, Zhang Z S, Yin M H, Luo M, Li X B, Hou L, Pei Y. Cotton flavonoid structural genes related to the pigmentation in brown fibers. Biochem Biophys Res Commun, 2007, 358: 73-78

[19]Park J S, Choung M G, Kim J B, Hahn B S, Kim J B, Bae S C, Roh K H, Kim Y H, Cheon C I, Sung M K, Cho K J. Genes up-regulated during red coloration in UV-B irradiated lettuce leaves. Plant Cell Rep, 2007, 26: 507-516

[20]Kim S, Binzel M L, Yoo K S, Park S, Pike L M. Pink (P), a new locus responsible for a pink trait in onions (Allium cepa) resulting from natural mutations of anthocyanidin synthase. Mol Genet Genomics, 2004, 272: 18-27

[21]Wilmouth R C, Turnbull J J, Welford R W, Clifton I J, Prescott A G, Schofield C J. Structure and mechanism of anthocyanidin synthase from Arabidopsis thaliana. Structure, 2002, 10: 93-103

[22]Jiang Z, Wang H, Ma Y, Wei D. Characterization of two novel lipase genes isolated directly from environmental sample. Appl Microbiol Biotechnol, 2006, 70: 327-332

[23]Sambrook J, Russell D W, eds. Huang P-T(黄培堂) trans. Molecular Cloning: a Laboratory Manual (分子克隆实验指南). Beijing: Science Press, 2003. pp 28-30

[24]Aljanabi S M , Martinez I. Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucl Acids Res, 1997, 25: 4692-4693

[25]Geourjon C, Deléage G. SOPMA: significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments. Comput Appl Biosci, 1995, 11: 681-684

[26]Benkert P, Tosatto S C, Schomburg D. QMEAN: a comprehensive scoring function for model quality assessment. Proteins, 2008, 71: 261-77

[27]Benkert P, Künzli M, Schwede T. QMEAN server for protein model quality estimation. Nucl Acids Res, 2009, 37: W510-W 514

[28]Li J(李军), Zhao A-C(赵爱春), Wang X-L(王茜龄), Zhang Q-Y(张琼予), Li Q-Y(黎其友), Jin X-Y(金筱耘), Li Z-G(李镇刚), Yu M-D(余茂德). Molecular cloning and tissues expression analysis of three Actin genes from mulberry (Morus alba). Acta Agron Sin (作物学报), 2011, 37(4): 641-649 (in Chinese with English abstract)

[29]Almeida J R, D’Amico E, Preuss A, Carbone F, de Vos C H, Deiml B, Mourgues F, Perrotta G, Fischer T C, Bovy A G, Martens S, Rosati C. Characterization of major enzymes and genes involved in flavonoid and proanthocyanidin biosynthesis during fruit development in strawberry (Fragaria × ananassa). Arch Biochem Biophys, 2007, 465: 61-71

[30]Rausher M D, Lu Y, Meyer K. Variation in constraint versus positive selection as an explanation for evolutionary rate variation among anthocyanin genes. J Mol Evol, 2008, 67: 137-144

[31]Lukacin R, Britsch L. Identification of strictly conserved histidine and arginine residues as part of the active site in Petunia hybrida flavanone 3beta-hydroxylase. Eur J Biochem, 1997, 249: 748-757

[32]Rosati C, Cadic A, Duron M, Ingouff M, Simoneau P. Molecular characterization of the anthocyanidin synthase gene in Forsythia × intermedia reveals organ-specific expression during flower development. Plant Sci, 1999, 149: 73-77

[33]Matsuda J, Okabe S, Hashimoto T, Yamada Y. Molecular cloning of hyoscyamine 6 beta-hydroxylase, a 2-oxoglutarate-dependent dioxygenase, from cultured roots of Hyoscyamus niger. J Biol Chem, 1991, 266: 9460-9464

[34]Shimada S, Inoue Y T, Sakuta M. Anthocyanidin synthase in non-anthocyanin-producing Caryophyllales species. Plant J, 2005, 44: 950-959

[35]Chalker-Scott L. Environmental significance of anthocyanins in plant stress responses. Photochem Photobiol, 1999, 70: 1-9

[36]Jose A M, Schäfer E. Distorted phytochrome action spectra in green plants. Planta, 1978, 138: 25

[37]Li Y(李跃), Liu Y-J(刘延吉). Research on metabolism mechanism of fruit anthocyanin and control technique. J Anhui Agric Sci (安徽农业科学), 2007, 35(16): 4755-4756 (in Chinese with English abstract)

[38]Zhao Z-F(赵宗方), Xie J-B(谢嘉宝), Wu G-F(吴桂法), Wang C-L(王成良). Correlation analysis on development of anthocyanidin in pericarp of fuji apple. J Fruit Sci (果树科学), 1992, 9(3): 134-237 (in Chinese with English abstract)

[39]Ramirez-Tortosa C, Andersen Ø M, Gardner P T, Morrice P C, Wood S G, Duthie S J, Collins A R, Duthie G G. Anthocyanin-rich extract decreases indices of lipid peroxidation and DNA damage in vitamin E-depleted rats. Free Radic Biol Med, 2001, 31: 1033-1037

[40]Cheng L Q, Xu Y J, Grotewold E, Jin Z P, Wu F Y, Fu C X, Zhao D X. Characterization of anthocyanidin synthase (ANS) gene and anthocyanidin in rare medicinal plant-Saussurea medusa. Plant Cell Tissue Organ Culture, 2007, 89: 63-73

[41]Pang Y, Peel G J, Wright E, Wang Z, Dixon R A. Early steps in proanthocyanidin biosynthesis in the model legume Medicago truncatula. Plant Physiol, 2007, 145: 601-615

[42]Reddy A M, Reddy V S, Scheffler B E, Wienand U, Reddy A R. Novel transgenic rice overexpressing anthocyanidin synthase accumulates a mixture of flavonoids leading to an increased antioxidant potential. Metab Eng, 2007, 9: 95-111

[43]Angiosperm Phylogeny Group. An update of the angiosperm phylogeny group classi?cation for the orders and families of ?owering plants: APG III. Bot J Linnean Soc, 161: 105-121

[44]Brouillard R, Dangles O. Flavonoids and Flower Colour in the Flavonoids. In: Harborne J B ed. The Flavonoids: Advances in Research Since 1986. London: Chapman & Hall Ltd., 1993. pp 565-588

[45]Holton T A, Cornish E C. Genetics and biochemistry of anthocyanin biosynthesis. Plant Cell, 1995, 7: 1071-1083
[1] Zuo Tong-Hong, Zhang He-Cui, Zeng Jing, Zhu Li-Quan. Molecular cloning and expression analysis of BoPUB3L associated with self-incompatibility in Brasscia oleracea [J]. Acta Agronomica Sinica, 2026, 52(6): 1698-1710.
[2] XUE Xiao-Fei, DAI Yun-Jing, LI Xi-Lin, DING Yan-Yan, WANG Xiang, LEI Zhang-Ying, HAN Huan-Yong, HE Dao-Hua. Characterization of GhCDN10 encoding cadinene synthase and its involvement in gossypol biosynthesis pathway in Gossypium hirsutum [J]. Acta Agronomica Sinica, 2025, 51(8): 2060-2076.
[3] WANG Lin, CHEN Xiao-Yu, ZHANG Wen-Meng-Long, WANG Si-Qi, CHENG Bing-Yun, CHENG Jing-Qiu, PAN Rui, ZHANG Wen-Ying. Molecular characteristics and functional analysis of HvMYB2 in response to drought stress in barley [J]. Acta Agronomica Sinica, 2025, 51(4): 873-887.
[4] ZHANG Zheng-Kang, SU Yan-Hong, RUAN Sun-Mei, ZHANG Min, ZHANG Pan, ZHANG Hui, ZENG Qian-Chun, LUO Qiong. Cloning and functional study of OgXa13 in Oryza meyeriana [J]. Acta Agronomica Sinica, 2025, 51(2): 334-346.
[5] LI Chun-Mei, CHEN Jie, LANG Xing-Xuan, ZHUANG Hai-Min, ZHU Jing, DU Zi-Jun, FENG Hao-Tian, JIN Han, ZHU Guo-Lin, LIU Kai. Map-based cloning and functional analysis of Dwarf and Tillering 1 (DT1) gene in rice [J]. Acta Agronomica Sinica, 2025, 51(2): 347-357.
[6] WANG Yu-Jiao, WANG Yong-Le, TIAN Chang-Jiu, YU Chun-Wang, LYU Jia-Bin, ZHU Jia-Bao. Cloning of the VQ4 gene and preliminary analysis of its role in salt tolerance in Coix lacryma-jobi L. [J]. Acta Agronomica Sinica, 2025, 51(12): 3198-3210.
[7] LI Shi-Kuan, HONG Hui-Long, FU Jia-Qi, GU Yong-Zhe, SUN Ru-Jian, QIU Li-Juan. Mine the genes of premature yellowing and aging in soybean leaves by BSA-seq combined with RNA-seq technology [J]. Acta Agronomica Sinica, 2024, 50(2): 294-309.
[8] ZHU Xiao-Tong, YE Ya-Feng, GUO Jun-Yao, YANG Hui-Jie, WANG Zi-Yao, ZHAN Yue, WU Yue-Jin, TAO Liang-Zhi, MA Bo-Jun, CHEN Xi-Feng, LIU Bin-Mei. Heredity and fine mapping of an early-senescence leaf gene ESL8 in rice [J]. Acta Agronomica Sinica, 2023, 49(3): 662-671.
[9] LI Meng-Yuan, ZHANG Wen-Cheng, GAO Yong, QIN Yong-Tian, BO Shi-Rong, SONG Kun-Yang, TANG Ji-Hua, FU Zhi-Yuan. Map-based cloning and allelic analysis of gene controlling maize kernel mutant crk4 [J]. Acta Agronomica Sinica, 2023, 49(10): 2613-2620.
[10] WANG Mu-Mu, HE Yan-Fan, ZHENG Yong-Sheng, WANG Hui, WANG Li-Yuan, WANG Dong-Jian, ZHANG Han, LI Ru-Yu. Fine mapping and cloning of a seed shattering gene SH8 in rice (Oryza sativa L.) [J]. Acta Agronomica Sinica, 2022, 48(8): 1948-1956.
[11] CUI Lian-Hua, ZHAN Wei-Min, YANG Lu-Hao, WANG Shao-Ci, MA Wen-Qi, JIANG Liang-Liang, ZHANG Yan-Pei, YANG Jian-Ping, YANG Qing-Hua. Molecular cloning of two maize (Zea mays) ZmCOP1 genes and their transcription abundances in response to different light treatments [J]. Acta Agronomica Sinica, 2022, 48(6): 1312-1324.
[12] ZHOU Hui-Wen, QIU Li-Hang, HUANG Xing, LI Qiang, CHEN Rong-Fa, FAN Ye-Geng, LUO Han-Min, YAN Hai-Feng, WENG Meng-Ling, ZHOU Zhong-Feng, WU Jian-Ming. Cloning and functional analysis of ScGA20ox1 gibberellin oxidase gene in sugarcane [J]. Acta Agronomica Sinica, 2022, 48(4): 1017-1026.
[13] YANG Lu-Hao, WANG Li-Jian, SUN Guang-Hua, WANG Shao-Ci, CUI Lian-Hua, CHEN Chang, SONG Mei-Fang, ZHANG Yan-Pei, JIANG Liang-Liang, YANG Jian-Ping, WANG Chen-Yang. Transcription abundances of PHYA, PHYB, and PHYC genes in response to different light treatments in Secale cereale [J]. Acta Agronomica Sinica, 2022, 48(12): 3057-3070.
[14] XIE Qin-Qin, ZUO Tong-Hong, HU Deng-Ke, LIU Qian-Ying, ZHANG Yi-Zhong, ZHANG He-Cui, ZENG Wen-Yi, YUAN Chong-Mo, ZHU Li-Quan. Molecular cloning and expression analysis of BoPUB9 in self-incompatibility Brassica oleracea [J]. Acta Agronomica Sinica, 2022, 48(1): 108-120.
[15] TANG Rui-Min, JIA Xiao-Yun, ZHU Wen-Jiao, YIN Jing-Ming, YANG Qing. Cloning of potato heat shock transcription factor StHsfA3 gene and its functional analysis in heat tolerance [J]. Acta Agronomica Sinica, 2021, 47(4): 672-683.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!