Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2017, Vol. 43 ›› Issue (06): 839-848.doi: 10.3724/SP.J.1006.2017.00839

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Functional Analysis of 1-Aminocyclopropane-1-carboxylate Oxidase Gene’s Promoter in Mulberry

YU Jian,LIU Chang-Ying,ZHAO Ai-Chun,WANG Chuan-Hong,CAI Yu-Xiang,YU Mao-De*   

  1. College of Biotechnology, Southwest University / State Key Laboratory of Silkworm Genome Biology, Chongqing 400715, China?
  • Received:2016-07-25 Revised:2017-01-21 Online:2017-06-12 Published:2017-02-17
  • Contact: Yu maode, E-mail: yumd@163.com, Tel: 023-68250191 E-mail:yujian1949@163.com
  • Supported by:

    This study was supported by China Special Fund for Agro-scientific Research in the Public Interest (201403064), Graduate Research and Innovation Projects of Chongqing (CYS2015070), and the China Agriculture Research System (CARS-22).

Abstract:

1-Aminocyclopropane-1-carboxylate oxidase (ACO) as a key enzyme catalyzes the reaction from 1-Aminocyclopropane- 1-carboxylate (ACC) to ethylene. In order to explore the function of MnACO in mulberry growth and development and resisting external stresses, we constructed pMnACO::GUS fusion and transformed it into Arabidopsis thaliana, and used GUS histochemical staining to identify GUS activities of transgenic Arabidopsis thaliana treated by different stresses in different growth stages . MnACO1 and MnACO2 promoter fragments obtained by PCR were 1518 bp and 1429 bp, respectively. There were lots of cis-acting elements such as TATA-box, CAAT-box and others responded to external stimuli. MnACO promoter could drive GUS to express in Arabidopsis thaliana; MnACO1 promoter expressed in the root, leaf, flower petal, anther, filament, stigma and silique of Arabidopsis thaliana and their activities were higher than those of MnACO2; but there was no expression of MnACO2 promoter in silique. The pMnACO1::GUS and pMnACO2::GUS transgenetic plants had different activities in different treatments. The GUS activities of the pMnACO1::GUS transgenetic plants were weakened, while those of the pMnACO2::GUS transgenetic plants were strengthened with the elongation of stress treatment time. Detection of MaACO expression in stress treatment of 2-week-old seedlings by qRT-PCRindicated that the trends of expression pattern of MaACO and its GUS activity changes were almost the same. This research indicated that MnACO is inducible promoter, MnACO1 has the characteristic of constitutive promoter, and MnACO2 has the characteristic of tissue-specific promoter. MnACO1 had more ability to respond to stresses in the transgenic plant, indicating it can be used to regulate the target gene of improving mulberry stress resistance. MaACO2 might be relevant to fruit maturation, thus its promoter could be used as fruit-specific promoter to improve the quality of mulberry fruit.

Key words: Mulberry, Ethylene, ACO, Promoter, GUS, Functional Analysis

[1] Bleecker A B, Kende H. Ethylene: a gaseous signal molecule in plants. Annu Rev Cell Dev Biol, 2000, 16: 1–18 [2] Lin Z F, Zhong S L, Grierson D. Recent advances in ethylene research. J Exp Bot, 2009, 60: 3311–3336 [3] Johnson, P R., Ecker J R. The ethylene gas signal transduction pathway: a molecular perspective. Annu Rev Genet, 1998, 32: 227–254 [4] Arc E, Sechet J, Corbineau F, Rajjou L, Marion-Poll1 A. ABA crosstalk with ethylene and nitric oxide in seed dormancy and germination. Front Plant Sci, 2013, 4: 10.3389/fpls.2013.00063 [5] Mattoo, A K, Achilea, O, Fuchs Y, Chalutz E. Membrane association and some characteristics of the ethylene forming enzyme from etiolated pea seedlings. Biochem Biophys Res Commun, 1982, 105: 271–278 [6] Mcgarvey D J, Christoffersen R E. Characterization and kinetic parameters of ethylene-forming enzyme from avocado fruit. J Biol Chem, 1992, 267: 5964–5967 [7] Slater A, Maunders M J, Edwards K, Schuch W, Grierson D. Isolation and characterization of cDNA clones for tomato polygalacturonase and other ripening-related proteins. Plant Mol Biol, 1985, 5: 137–147 [8] Kahana A, Silberstein L, Kessler N, Goldstein R S, Perl-Treves R. Expression of ACC oxidase genes differs among sex genotypes and sex phases in cucumber. Plant Mol Biol, 1999, 41: 517–528 [9] Ruperti B, Bonghi C, Rasori A, Ramina A, Tonutti P. Characterization and expression of two members of the peach 1-aminocyclopropane-1-carboxylate oxidase gene family. Physiol Plant, 2001, 111: 336–344 [10] Ross G S, Knighton M L, Lay-Yee M. An ethylene-related cDNA from ripening apples. Plant Mol Biol, 1992, 19: 231–238 [11] Binnie I E, McManus M T. Characterization of the 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase multigene family of Malus domestica Borkh. Phytochemistry, 2009, 70: 348–60 [12] Shiomi S, Yamamoto M, Nakamura R, Inaba A. Expression of ACC synthase and ACC oxidase genes in melons harvested at different stages of maturity. J Jpn Soc Hort Sci, 1999, 68: 10–17 [13] Barry C S, Blume B, Bouzayen M. Cooper W, Hamilton A J, Grierson D. Differential expression of the 1-aminocyclopropane-1-carboxylic acid oxidase gene family of tomato. Plant J, 1996, 9: 525–535 [14] 朱玉贤. 现代分子生物学. 北京: 高等教育出版社, 2007. pp 75–83 Zhu Y X. Modern Molecular Biology. Beijing: Higher Education Press, 2007, pp 75–83 (in Chinese) [15] Lasserre E, Godard F, Bouquin T, Hernandez J A, Pech J C, Roby D, Balague C. Differential activation of two ACC oxidase gene promoters from melon during plant development and in response to pathogen attack. Mol General Genet, 1997, 256: 211–222 [16] Blume B, Grierson D. Expression of ACC oxidase promoter-GUS fusions in tomato and Nicotiana plumbaginifolia regulated by developmental and environmental stimuli. Plant J, 1997, 12: 731–746 [17] Rasoria A, Bertolasib B, Furini A, Bonghi C, Tonutti P, Ramina A. Functional analysis of peach ACC oxidase promoters in transgenic tomato and in ripening peach fruit. Plant Scie, 2003, 165: 523–530 [18] 毛娟, 陆璐, 陈佰鸿, 禇明宇, 赵长增. 甜瓜CmACOI启动子组织特异性表达研究. 园艺学报, 2013, 40: 1101–1109 Mao J, Lu L, Chen B H, Chu M Y, Zhao C Z. Studies on tissue specific regulation of CmACOI promoter in melon. Acta Hort Sin, 2013, 40: 1101–1109 (in Chinese with English abstract) [19] Checker V G, Khurana P. Molecular and functional characterization of mulberry EST encoding remorin (MiREM) involved in abiotic stress. Plant Cell Reports, 2013, 32(11): 1729-1741 [20] Liu C Y, Lü R H, Zhao A C, Wang X L, Diane U, Wang X H, Wang C H, Yu Y S, Han S M, Lu C, Yu M D. Characterization and expression profiles of MaACS and MaACO genes from mulberry (Morus alba L.). J Zhejiang Univ-Sci B, 2014, 15: 611–623 [21] Liu C Y, Zhao A C, Zhu P P, Li J, Han L, Wang X L, Fan W, Lü R H, Wang C H, Li Z G, Lu C, Yu M D. Characterization and expression of genes involved in the ethylene biosynthesis and signal transduction during ripening of mulberry fruit. PLoS One, 2015, 10: e0122081 [22] Clough S J, Bent A F. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 1998, 6: 735–743 [23] Zheng L, Liu G, Meng X, Li, Y B, Wang, Y C. A versatile agrobacterium mediated transient gene expression system for herbaceous plants and trees. Biochem Genet, 2012, 50: 761–769 [24] Youk E S, Pack I S, Kim Y J, Yoon W K, Kim C G, Ryu S B, Harn C H, Jeong S C, Kim H M. A framework for molecular genetic assessment of a transgenic watermelon rootstock line. Plant Sci, 2009, 176: 805–811 [25] 雷建峰, 李月, 徐新霞, 阿尔祖古丽?塔什, 蒲艳, 张巨松, 刘晓东. 棉花不同GbU6启动子截短克隆及功能鉴定. 作物学报, 2016, 42: 675–683 Lei J F, Li Y, Xu X X, A E Z G L ?Tashi. Cloning and functional analysis of different truncated GbU6 Promoters in Cotton. Acta Agron Sin, 2016, 42: 675–683 (in Chinese with English abstract) [26] 李军, 赵爱春, 王茜龄, 张琼予, 黎其友, 金筱耘, 李镇刚, 余茂德. 三个桑树肌动蛋白基因的克隆与组织表达分析. 作物学报, 2011, 37: 641–649 Li J, Zhao A C, Wang X L, Zhang Q Y, Li Q Y, Jin X Y, Li Z G, Yu M D. Molecular cloning and tissues expression analysis of three actin genes from mulberry (Morus alba). Acta Agron Sin, 2011, 37: 641–649 (in Chinese with English abstract) [27] 吴建, 侯和胜. 高等植物ACC氧化酶基因启动子研究进展. 天津农业科学, 2014, 20(4): 14–16 Wu J, Hou H S. Advances in research of higher plant ACC oxidase promoters. Tianjin Agric Sci, 2014, 20(4): 14–16 (in Chinese with English abstract) [28] Yang S F, Hoffman N E. Ethylene biosynthesis and its regulation in higher plants. Ann Rev Plant Physiol Plant Mol Biol, 1984, 35: 155–189 [29] Carrera E , Holman T, Medhurst A, Dietrich D, Footitt S, Theodoulou F L, Holdsworth M J. Seed after-ripening is a discrete developmental pathway associated with specific gene networks in Arabidopsis. Plant J, 2008, 53: 214–224 [30] Linkies A, Leubner-Metzger G. Beyond gibberellins and abscisic acid: how ethylene and jasmonates control seed germination. Plant Cell Rep, 2012, 31: 253–270 [31] Hermann K, Meinhard J, Dobrev P, Linkies A, Pesek B, Hess B, Machacova I, Fischer U, Leubner-Metzger G. 1-aminocyclopropane-1-carboxylic acid and abscisic acid during the germination of sugar beet (Beta vulgaris L.): a comparative study of fruits and seeds. J Exp Bot, 2007, 58: 3047–3060 [32] Mou W S, Li D D, Bu J W, Jiang Y Y, Khan Z U, Luo Z S, Mao L C, Ying T J. Comprehensive analysis of ABA effects on ethylene biosynthesis and signaling during tomato Fruit ripening. PloS one, 2016, 11: e0154072 [33] Li N, Parsons B L, Liu D R, Mattoo A K. Accumulation of wound-inducible ACC synthase transcript in tomato fruit is inhibited by salicylic acid and polyamines. Plant Mol Biol, 1992, 18: 477–487 [34] Fan X T, Mattheis J P, Fellman J K.Inhibition of apple fruit 1-aminocyclopropane-1-carboxylic activity and respiration by acetylsalicylic acid. J Plant Physiol, 1996, 149: 469–471 [35] Leslie C A, Romani R J. Inhibition of ethylene biosynthesis by salicylic acid. Plant Physiol, 1998, 88: 833–368 [36] Nissen p. Stimulation of somatic embryogenesis in carrot by ethylene effects of modulators of ethylene biosynthesis and action. Physiol Plant, 1994, 92: 397–403 [37] Lasserre E, Bouquin T, Hernandez J A, Bull J, Pech J C, Balague C. Structure and expression of three genes encoding ACC oxidase homologs from melon (Cucumis melo L.). Mol General Genet, 1996, 251: 81–90 [38] 徐忠传. 成熟猕猴桃果实不同组织中ACC氧化酶基因的表达差异研究. 中国科学技术大学学报, 2001, 31: 235–240 Xu Z C. Study on the differences for the expression of 1-aminocyclopropane-1-carboxylic acid oxidase in different tissues of ripening kiwifruit. J China Univ Sci Technol, 200l, 31: 235–240 (in Chinese with English abstract) [39] Hunter D A, Yoo S D, Butcher S M, McManus M T. Expression of l-aminocyclopropane-1-carboxylate oxidase during leaf ontogeny in white clover. Plant Physi1, 1999, 120: 131–141

[1] YANG De-Wei, WANG Xun, ZHENG Xing-Xing, XIANG Xin-Quan, CUI Hai-Tao, LI Sheng-Ping, TANG Ding-Zhong. Functional studies of rice blast resistance related gene OsSAMS1 [J]. Acta Agronomica Sinica, 2022, 48(5): 1119-1128.
[2] ZHOU Yue, ZHAO Zhi-Hua, ZHANG Hong-Ning, KONG You-Bin. Cloning and functional analysis of the promoter of purple acid phosphatase gene GmPAP14 in soybean [J]. Acta Agronomica Sinica, 2022, 48(3): 590-596.
[3] SHI Lei, MIAO Li-Juan, HUANG Bing-Yan, GAO Wei, ZHANG Zong-Xin, QI Fei-Yan, LIU Juan, DONG Wen-Zhao, ZHANG Xin-You. Characterization of the promoter and 5'-UTR intron in AhFAD2-1 genes from peanut and their responses to cold stress [J]. Acta Agronomica Sinica, 2021, 47(9): 1703-1711.
[4] SU Ya-Chun, LI Cong-Na, SU Wei-Hua, YOU Chui-Huai, CEN Guang-Li, ZHANG Chang, REN Yong-Juan, QUE You-Xiong. Identification of thaumatin-like protein family in Saccharum spontaneum and functional analysis of its homologous gene in sugarcane cultivar [J]. Acta Agronomica Sinica, 2021, 47(7): 1275-1296.
[5] WANG Xiao-Chun, WANG Lu-Lu, ZHANG Zhi-Yong, QIN Bu-Tan, YU Mei-Qin, WEI Yi-Hao, MA Xin-Ming. Transcription characteristics of wheat glutamine synthetase isoforms and the sequence analysis of their promoters [J]. Acta Agronomica Sinica, 2021, 47(4): 761-769.
[6] YANG Yang, LI Huai-Lin, HU Li-Min, FAN Chu-Chuan, ZHOU Yong-Ming. Genetic analysis and molecular characterization of multilocular trait in the srb mutant of Brassica rapa L. [J]. Acta Agronomica Sinica, 2021, 47(3): 385-393.
[7] LI Lan-Lan, MU Dan, YAN Xue, YANG Lu-Ke, LIN Wen-Xiong, FANG Chang-Xun. Effect of OsPAL2;3 in regulation of rice allopathic inhibition on barnyardgrass (Echinochloa crusgalli L.) [J]. Acta Agronomica Sinica, 2021, 47(2): 197-209.
[8] WANG Zhen, ZHANG Xiao-Li, MENG Xiao-Jing, YAO Meng-Nan, MIU Wen-Jie, YUAN Da-Shuang, ZHU Dong-Ming, QU Cun-Min, LU Kun, LI Jia-Na, LIANG Ying. Identification of upstream regulators for mitogen-activated protein kinase 7 gene (BnMAPK7) in rapeseed (Brassica napus L.) [J]. Acta Agronomica Sinica, 2021, 47(12): 2379-2393.
[9] LI Na-Na, LIU Ying, ZHANG Hao-Jie, WANG Lu, HAO Xin-Yuan, ZHANG Wei-Fu, WANG Yu-Chun, XIONG Fei, YANG Ya-Jun, WANG Xin-Chao. Promoter cloning and expression analysis of the hexokinase gene CsHXK2 in tea plant (Camellia sinensis) [J]. Acta Agronomica Sinica, 2020, 46(10): 1628-1638.
[10] Mao-Ni CHAO,Hai-Yan HU,Run-Hao WANG,Yu CHEN,Li-Na FU,Qing-Qing LIU,Qing-Lian WANG. Cloning and functional analysis of promoter of potassium transporter gene GhHAK5 in upland cotton (Gossypium hirsutum L.) [J]. Acta Agronomica Sinica, 2020, 46(01): 40-51.
[11] CHANG Jian-Zhong,DONG Chun-Lin,ZHANG Zheng,QIAO Lin-Yi,YANG Rui,JIANG Dan,ZHANG Yan-Qin,YANG Li-Li,WU Jia-Jie,JING Rui-Lian. Function analysis of 5′ untranslated region introns in drought-resistance gene TaSAP1 [J]. Acta Agronomica Sinica, 2019, 45(9): 1311-1318.
[12] SONG Song-Quan,LIU Jun,XU Heng-Heng,ZHANG Qi,HUANG Hui,WU Xian-Jin. Biosynthesis and signaling of ethylene and their regulation on seed germination and dormancy [J]. Acta Agronomica Sinica, 2019, 45(7): 969-981.
[13] Xiao-Hong ZHANG,Gen-Hai HU,Han-Tao WANG,Cong-Cong WANG,Heng-Ling WEI,Yuan-Zhi FU,Shu-Xun YU. Expression and promoter activity of GhTFL1a and GhTFL1c in Upland cotton [J]. Acta Agronomica Sinica, 2019, 45(3): 469-476.
[14] Rui-Juan YANG,Jian-Rong BAI,Lei YAN,Liang SU,Xiu-Hong WANG,Rui LI,Cong-Zhuo ZHANG. Cloning and Expression Analysis of Strong Inducible Promoter P1502-ZmPHR1 Responding to Low Phosphorus Stress in Maize [J]. Acta Agronomica Sinica, 2018, 44(7): 1000-1009.
[15] Bo JIAO, Feng BAI, Yan-Yan LI, Jia LU, Xiao ZHANG, Yi-Ru CAO, Rong-Chao GE, Bao-Cun ZHAO. Cloning and Regulation Function Analysis of TaSC Promoter from Salt Tolerant Wheat [J]. Acta Agronomica Sinica, 2018, 44(04): 620-626.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!