Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (14): 2889-2896.doi: 10.3864/j.issn.0578-1752.2014.14.019

• AGRICULTURE INFORMATION TECHNOLOGY • Previous Articles     Next Articles

Isolation of a Maize Wound-Induced Gene Promoter and Characterization of the Gene Expression in Maize

 LIAN  Yun-1, 2 , LIU  Yun-Jun-1, WANG  Guo-Ying-1   

  1. 1、Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081;
    2、Institute of Industrial Crops, Henan Academy of Agricultural Sciences, Zhengzhou 450002
  • Received:2014-01-14 Online:2014-07-15 Published:2014-03-27

RichHTML

4

PDF

765

Cited

1

Abstract: 【Objective】 Protease inhibitor is an important defense protein in plant for resistance to infection or feeding by pathogen and animals. Maize Wip1(wound-induced protein) is the member of Bowman-Birk inhibitor(BBI) gene family. To provide new information for the application of resistance gene to insect in plant, this experiment was designated to understand the activity of Wip1 promoter, assay spatial and temporal expression of Wip1 gene and its response to phytohormones and abiotic stresses.【Method】A clone containing the upstream region of Wip1 was isolated and sequenced from maize leaf cDNA. The promoter sequences of Wip1 gene was inserted into binary expression vector p1300 to produce recombinant plasmids p1300-Wip1-GUS, which was transferred into Agrobacterium LBA4404 by freeze thawing method. Functional analysis of the promoter was conducted through heterologous expression in maize callus by agro-infiltration method. Total RNA was isolated from maize tissues using hot phenol method. Purified RNA was subjected to formaldehyde-containing-agarose gel, blotted onto a nylon membrane, and Northern blotting was performed with [α-32P] dCTP-labeled probe to characterized tissue-speci?c location, phytohormones and abiotic stresses analysis of Wip1. 【Result】 Promoter region was 1 737 bp and coding sequence was 512 bp. The result showed that GUS activity driven by the Wip1 promoter was detected in maize callus tissues after infection by Agrobacterium haboring recombinant plasmids p1300-Wip1-GUS. The expression of Wip1 was assayed at various developmental stages. Northern blotting showed that Wip1 gene abundantly expressed in injured coleoptile, root, stem, leaf, ear, tassel, silk and husk, while expressed little in uninjured coleoptile and not expressed in other uninjured detection tissues. The results also showed that Wip1 was not affected by submergence, cold, dehydration, PEG, ABA, NaCl and IAA treatments. The dynamic expression of Wip1 gene under injure stress from 0 h to 24 h with 8 time points, was analyzed in maize leaf tissue. Wip1 gene was detected after stressing for 2 h, the expression level increased rapidly, and with a continually increasing trend from 4 h to 24 h. 【Conclusion】 Wip1 gene is induced by injure-specific stress. Wip1 promoter is an effective injure-specific promoter to express the desired genes in plants.

Key words: maize , wound induced , Wip1 , serine protease inhibitor , bowman-birk

[1]Kim T H, Barrera L O, Qu C, Van Calcar S, Trinklein N D, Cooper S J, Luna R M, Glass C K, Rosenfeld M G, Myers R M, Ren B. Direct isolation and identification of promoters in the human genome. Genome Research, 2005, 15(6): 830-839.

[2]Sandelin A, Carninci P, Lenhard B, Ponjavic J, Hayashizaki Y,  Hume D A. Mammalian RNA polymerase II core promoters: Insights from genome-wide studies. Nature Reviews Genetics, 2007, 8(6): 424-436.

[3]Hutchison W D, Burkness E C, Mitchell P D, Moon R D, Leslie T W, Fleischer S J, Abrahamson M, Hamilton K L, Steffey K L, Gray M E, Hellmich R L, Kaster L V, Hunt T E, Wright R J, Pecinovsky K, Rabaey T L, Flood B R, Raun E S. Areawide suppression of European corn borer with Bt maize reaps savings to non-Bt maize growers. Science, 2010, 330(6001): 222-225.

[4]Lu M G, Rui C H, Zhao J Z, Jian G L, Fan X L, Gao X W. Selection and heritability of resistance to Bacillus thuringiensis subsp kurstaki and transgenic cotton in Helicoverpa armigera (Lepidoptera: Noctuidae). Pest Management Science, 2004, 60(9): 887-893.

[5]Kathage J, Qaim M. Economic impacts and impact dynamics of Bt (Bacillus thuringiensis) cotton in India. Proceedings of the National Academy of Sciences of the USA, 2012, 109(29): 11652-11656.

[6]Fang R X, Nagy F, Sivasubramaniam S, Chua N H. Multiple cis regulatory elements for maximal expression of the cauliflower mosaic virus 35S promoter in transgenic plants. The Plant Cell, 1989, 1(1): 141-150.

[7]Lian Y, Jia Z, He K, Liu Y, Song F, Wang B, Wang G. Transgenic tobacco plants expressing synthetic Cry1Ac and Cry1Ie genes are more toxic to cotton bollworm than those containing one gene. Chinese Science Bulletin, 2008, 53(9): 1381-1387.

[8]Christensen A H, Sharrock R A, Quail P H. Maize polyubiquitin genes: structure, thermal perturbation of expression and transcript splicing, and promoter activity following transfer to protoplasts by electroporation. Plant Molecular Biology, 1992, 18(4): 675-689.

[9]Zhang W, McElroy D, Wu R. Analysis of rice Act1 5′ region activity in transgenic rice plants. The Plant Cell, 1991, 3(11): 1155-1165.

[10]Cazzonelli C I, McCallum E J, Lee R, Botella J R. Characterization of a strong, constitutive mung bean (Vigna radiata L.) promoter with a complex mode of regulation in planta. Transgenic Research, 2005, 14(6): 941-967.

[11]Stavolone L, Kononova M, Pauli S, Ragozzino A, de Haan P, Milligan S, Lawton K, Hohn T. Cestrum yellow leaf curling virus (CmYLCV) promoter: A new strong constitutive promoter for heterologous gene expression in a wide variety of crops. Plant Molecular Biology, 2003, 53(5): 663-673.

[12]Xie Y, Liu Y, Meng M, Chen L, Zhu Z. Isolation and identification of a super strong plant promoter from cotton leaf curl Multan virus. Plant Molecular Biology, 2003, 53(1/2): 1-14.

[13]Bates S L, Zhao J Z, Roush R T, Shelton A M. Insect resistance management in GM crops: Past, present and future. Nature Biotechnology, 2005, 23(1): 57-62.

[14]Hamilton K A, Pyla P D, Breeze M, Olson T, Li M, Robinson E, Gallagher S P, Sorbet R, Chen Y. Bollgard II cotton: compositional analysis and feeding studies of cottonseed from insect-protected cotton (Gossypium hirsutum L.) producing the Cry1Ac and Cry2Ab2 proteins. Journal of Agricultural and Food Chemistry, 2004, 52(23): 6969-6976.

[15]Qi R F, Song Z W, Chi C W. Structural features and molecular evolution of Bowman-Birk protease inhibitors and their potential application. Acta Biochimica et Biophysica Sinica, 2005, 37(5): 283-292.

[16]Hilder V A, Gatehouse A M R, Sheerman S E, Barker R F, Boulter D. A novel mechanism of insect resistance engineered into tobacco. Nature, 1987, 330(6144): 160-163.

[17]Shitan N, Horiuchi K, Sato F, Yazaki K. Bowman-birk proteinase inhibitor confers heavy metal and multiple drug tolerance in yeast. Plant & Cell Physiology, 2007, 48(1): 193-197.

[18]Clemente A, Sonnante G, Domoney C. Bowman-Birk inhibitors  from legumes and human gastrointestinal health: Current status and perspectives. Current Protein & Peptide Science, 2011, 12(5): 358-373.

[19]Clementea A, Domoney C. Biological significance of polymorphism in legume protease inhibitors from the Bowman-Birk family. Current Protein & Peptide Science, 2006, 7(3): 201-216.

[20]Clemente A, Carmen Marin-Manzano M, Jimenez E, Carmen Arques M, Domoney C. The anti-proliferative effect of TI1B, a major Bowman-Birk isoinhibitor from pea (Pisum sativum L.), on HT29 colon cancer cells is mediated through protease inhibition. The British Journal of Nutrition, 2012, 108( Suppl 1): 135-144.

[21]Qi R F, Song Z W, Chi C W. Structural features and molecular evolution of Bowman-Birk protease inhibitors and their potential application. Acta Biochimica et Biophysica Sinica, 2005, 37(5): 283-292.

[22]Shitan N, Horiuchi K, Sato F, Yazaki K. Bowman-birk proteinase inhibitor confers heavy metal and multiple drug tolerance in yeast. Plant & Cell Physiology, 2007, 48(1): 193-197.

[23]Qu L J, Chen J, Liu M, Pan N, Okamoto H, Lin Z, Li C, Li D, Wang J, Zhu G, Zhao X, Chen X, Gu H, Chen Z. Molecular cloning and functional analysis of a novel type of Bowman-Birk inhibitor gene family in rice. Plant Physiology, 2003, 133(2): 560-570.

[24]Che F-S, Entani T, Marumoto T, Taniguchi M, Takayama S, Isogai A. Identification of novel genes differentially expressed in compatible and incompatible interactions between rice and Pseudomonas avenae. Plant Science, 2002, 162(3): 449-458.

[25]陈军, 刘静, 郭蕾, 瞿礼嘉, 陈章良, 顾红雅. 水稻Bowman-Birk蛋白酶抑制剂基因OsWIP1-2的诱导表达及其抑制活性. 科学通报, 2004, 49(7): 657-661.

Chen J, Liu J, Guo L, Qu L J, Chen Z L, Gu H Y. Inducible expression pattern of rice Bowman-Birk inhibitor gene OsWIP1-2 and its protease inhibitory activity. Chinese Science Bulletin, 2004, 49(7): 657-661. (in Chinese)

[26]Rohrmeier T, Lehle L. WIP1, a wound-inducible gene from maize with homology to Bowman-Birk proteinase inhibitors. Plant Molecular Biology, 1993, 22(5): 783-792.

[27]Eckelkamp C, Ehmann B, Schopfer P. Wound-induced systemic accumulation of a transcript coding for a Bowman-Birk trypsin inhibitor-related protein in maize (Zea mays L.) seedlings. Febs Letters, 1993, 323: 73-76.

[28]Zhang S, Lian Y, Liu Y, Wang X, Liu Y, Wang G. Characterization of a maize wip1 promoter in transgenic plants. International Journal of Molecular Sciences, 2013, 14(12): 23872-23892.

[29]练云. 人工改造的cry1Ac、cry1Ie基因在转基因烟草和玉米中的共表达[D]. 北京: 中国农业大学, 2008: 26.

Lian Y. Co-expression of the modified cry1Ac, cry1Ie genes in transgenic tobacco and maize plants (Dr.). BeiJing: China Agriculture University, 2008: 26. (in Chinese)

[30]Jefferson R A, Kavanagh T A, Bevan M W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. The European Molecular Biology Organization Journal, 1987, 6(13): 3901-3907.

[31]Kay R, Chan A, Daly M, McPherson J. Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science, 1987, 236(4806): 1299-1302.

[32]Zheng J, Zhao J, Tao Y, Wang J, Liu Y, Fu J, Jin Y, Gao P, Zhang J, Bai Y, Wang G. Isolation and analysis of water stress induced genes in maize seedlings by subtractive PCR and cDNA macroarray. Plant Molecular Biology, 2004, 55(6): 807-823.
[1] WANG YaFei, YAN Peng, XUE JinTao, DONG XueRui, MENG FanQi, GUO LiNa, LUO Yi, ZHANG Juan, DONG ZhiQiang, LU Lin. Effects of Ethephon-Glycine Betaine-Salicylic Acid Mixture on Root System Architecture, Physiological Function and Yield of Maize Under Heat Stress [J]. Scientia Agricultura Sinica, 2026, 59(7): 1439-1455.
[2] WANG JiaNuo, CHEN GuiPing, LI Pan, WANG LiPing, NAN YunYou, HE Wei, FAN ZhiLong, HU FaLong, CHAI Qiang, YIN Wen, ZHAO LiaoHao. Photo-Physiological Mechanism at Grain Filling Stage of No-Tillage with Plastic Re-Mulching to Increase Maize Yield in Oasis Irrigation Areas [J]. Scientia Agricultura Sinica, 2026, 59(6): 1189-1202.
[3] ZHOU XinJie, REN Hao, CHEN YingLong, ZHANG JiWang, ZHAO Bin, REN BaiZhao, LIU Peng, WANG HongZhang. Effects of Calcium Peroxide on Root Morphology and Yield Formation of Summer Maize in Waterlogging Farmland [J]. Scientia Agricultura Sinica, 2026, 59(6): 1203-1216.
[4] HE JiHang, ZHANG Qing, LÜ XiangYue, XUE JiQuan, XU ShuTu, LIU JianChao. Evaluation of Nitrogen Efficiency of Different Stay-Green Maize Hybrids [J]. Scientia Agricultura Sinica, 2026, 59(6): 1217-1230.
[5] LI YongJuan, ZHANG YueTong, WANG YiBo, ZHAO ChangJiang, SONG Jie, CHEN XueLi, YAO Qin. Effects of Biochar Application on the Abundance and Community Composition of Nitrogen-Fixing Microbial nifH Gene in Soybean Rotation and Continuous Cropping Systems [J]. Scientia Agricultura Sinica, 2026, 59(6): 1272-1285.
[6] LI SiYuan, LI HongPing, CHANG HongQing, ZHANG SenYan, LI SiJia, CUI XinFei, QIAO Po, ZENG Bo, LIU GuiZhen, LIU TianXue, TANG JiHua, LI ChaoHai. Effects of Density Increase on Dynamic Change of Yield and Agronomic Traits of Maize Cultivars with Different Plant Heights [J]. Scientia Agricultura Sinica, 2026, 59(5): 967-984.
[7] DONG JinLong, ZHAO Ying, YU HaiBing, LÜ JianYe, QIN JiaQi, LIANG Chen, MING Bo, LI ShaoKun. Multi-Model Elucidating of Nutritional Quality Contributions to Maize Kernel Test Weight and Regional Heterogeneity [J]. Scientia Agricultura Sinica, 2026, 59(5): 985-995.
[8] CHEN GuiPing, WEI JinGui, GUO Yao, LI Pan, WANG FeiEr, QIU HaiLong, FENG FuXue, YIN Wen. Synergistic Effects of Wide-Narrow Row and Density Enhancement on the Photosynthetic Characteristics and Resource Utilization of Maize in Oasis Irrigation Areas [J]. Scientia Agricultura Sinica, 2026, 59(2): 278-291.
[9] ZHANG ZhiYong, TAN ShiChao, XIONG ShuPing, MA XinMing, WEI YiHao, WANG XiaoChun. Effects of Annual Water and Nitrogen Optimization on Yield and Nitrogen Migration of Wheat-Maize Rotation System in Irrigation Area of Northern Henan [J]. Scientia Agricultura Sinica, 2026, 59(2): 336-353.
[10] WEI WenHua, LI Pan, SHAO GuanGui, FAN ZhiLong, HU FaLong, FAN Hong, HE Wei, CHAI Qiang, YIN Wen, ZHAO LianHao. Response of Silage Maize Yield and Quality to Reduced Irrigation and Combined Organic-Inorganic Fertilizer in Northwest Irrigation Areas [J]. Scientia Agricultura Sinica, 2025, 58(8): 1521-1534.
[11] XUE YuQi, ZHAO JiYu, SUN WangSheng, REN BaiZhao, ZHAO Bin, LIU Peng, ZHANG JiWang. Effects of Different Nitrogen Forms on Yield and Quality of Summer Maize [J]. Scientia Agricultura Sinica, 2025, 58(8): 1535-1549.
[12] CHEN GuiPing, LI Pan, SHAO GuanGui, WU XiaYu, YIN Wen, ZHAO LianHao, FAN ZhiLong, HU FaLong. The Regulatory Effect of Reduced Irrigation and Combined Organic- Inorganic Fertilizer Application on Stay-Green Characteristics in Silage Maize Leaves After Tasseling Stage [J]. Scientia Agricultura Sinica, 2025, 58(7): 1381-1396.
[13] YUE RunQing, LI WenLan, DING ZhaoHua, MENG ZhaoDong. Molecular Characteristics and Resistance Evaluation of Transgenic Maize LD05 with Stacked Insect and Herbicide Resistance Traits [J]. Scientia Agricultura Sinica, 2025, 58(7): 1269-1283.
[14] ZHAO Yao, CHENG Qian, XU TianJun, LIU Zheng, WANG RongHuan, ZHAO JiuRan, LU DaLei, LI CongFeng. Effects of Plant Type Improvement on Root-Canopy Characteristics and Grain Yield of Spring Maize Under High Density Condition [J]. Scientia Agricultura Sinica, 2025, 58(7): 1296-1310.
[15] ZOU XiaoWei, XIA Lei, ZHU XiaoMin, SUN Hui, ZHOU Qi, QI Ji, ZHANG YaFeng, ZHENG Yan, JIANG ZhaoYuan. Analysis of Disease Resistance Induced by Ustilago maydis Strain with Overexpressed UM01240 Based on Transcriptome Sequencing [J]. Scientia Agricultura Sinica, 2025, 58(6): 1116-1130.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd