Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (12): 2829-2842.doi: 10.16420/j.issn.0513-353x.2024-0015
• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles
YANG Xue1, HU Jinhong1, LI Jingjing1, ZHANG Yingcai1, WANG Lingxia1,2, LIANG Wenyu1,2()
Received:
2024-04-02
Revised:
2024-09-03
Online:
2024-12-25
Published:
2024-12-13
Contact:
LIANG Wenyu
YANG Xue, HU Jinhong, LI Jingjing, ZHANG Yingcai, WANG Lingxia, LIANG Wenyu. Genetic Transformation of Lycium Barbarum TGA2 Gene in Arabidopsis Enhances Its Salt Tolerance[J]. Acta Horticulturae Sinica, 2024, 51(12): 2829-2842.
Add to citation manager EndNote|Ris|BibTeX
URL: https://www.ahs.ac.cn/EN/10.16420/j.issn.0513-353x.2024-0015
用途 Purpose | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence |
---|---|---|
载体构建和表达模式 Vector construction and expression mode | TGA2-F | ATGGAGAGTCAAAGAATTGG |
TGA2-R | TCACATATTTGAGAAATGG | |
3′GSP1 | CAAGGTGCTTCATGGTGATAGGGGAAT | |
3′GSP2 | CCCTATGGGCATCACGTCCTAGAGAGA | |
5′GSP1 | GCTGGTGGTTCTTGAT | |
5′GSP2 | GGAGGATTTAGGCCACGAA | |
5′GSP3 | AAGGCATGTTGTGATCAGC | |
3′CDS | AAGCAGTGGTATCAACGCAGAGTAC(T)30 V N | |
基因启动子克隆 Gene promoter cloning | TGA2-R1 | TCTCCTCCTCCTGATTTTGAGCTTCC |
TGA2-R2 | TCATCATTGCTCATCTTAACTCCTTGCAG | |
TGA2-R3 | AGAAGTGCATAAGGCATGTTGTGATC | |
目的基因扩增引物 Target gene amplification primers | TGA2-2300-EGFP-F | ACGGGGGACGAGCTCGGTACC ATGGAGAGTCAAAGAATTGGAGAC |
TGA2-2300-EGFP-F | CACCATGGTGTCGACTCTAGA CATATTTGAGAAATGGTGTTGGGATG | |
基因定量分析 Quantitative analysis of genes | TGA2-T-F | CCACTATCCTTCGCAAGACC |
TGA2-T-R | GCTGAACCTGAATCACTACTCT | |
TGA2-F-Y | CCAAGAACAGGAGTGTCACATA | |
TGA2-R-Y | GTCTTAGCATCAGTCACCCTATC | |
目的基因扩增引物 Target gene amplification primers | TGA2-QRT-F | GTCTTAGCATCAGTCACCCTATC |
TGA2-QRT-R | CCAAGAACAGGAGTGTCACATA | |
内参引物 Internal reference primer | Actin-F | CCTAAGGCCAACAGAGAGAAA |
Actin-R | CGACCACTAGCATACAAGGAA |
Table 1 Primers sequence
用途 Purpose | 引物名称 Primer name | 引物序列(5′-3′) Primer sequence |
---|---|---|
载体构建和表达模式 Vector construction and expression mode | TGA2-F | ATGGAGAGTCAAAGAATTGG |
TGA2-R | TCACATATTTGAGAAATGG | |
3′GSP1 | CAAGGTGCTTCATGGTGATAGGGGAAT | |
3′GSP2 | CCCTATGGGCATCACGTCCTAGAGAGA | |
5′GSP1 | GCTGGTGGTTCTTGAT | |
5′GSP2 | GGAGGATTTAGGCCACGAA | |
5′GSP3 | AAGGCATGTTGTGATCAGC | |
3′CDS | AAGCAGTGGTATCAACGCAGAGTAC(T)30 V N | |
基因启动子克隆 Gene promoter cloning | TGA2-R1 | TCTCCTCCTCCTGATTTTGAGCTTCC |
TGA2-R2 | TCATCATTGCTCATCTTAACTCCTTGCAG | |
TGA2-R3 | AGAAGTGCATAAGGCATGTTGTGATC | |
目的基因扩增引物 Target gene amplification primers | TGA2-2300-EGFP-F | ACGGGGGACGAGCTCGGTACC ATGGAGAGTCAAAGAATTGGAGAC |
TGA2-2300-EGFP-F | CACCATGGTGTCGACTCTAGA CATATTTGAGAAATGGTGTTGGGATG | |
基因定量分析 Quantitative analysis of genes | TGA2-T-F | CCACTATCCTTCGCAAGACC |
TGA2-T-R | GCTGAACCTGAATCACTACTCT | |
TGA2-F-Y | CCAAGAACAGGAGTGTCACATA | |
TGA2-R-Y | GTCTTAGCATCAGTCACCCTATC | |
目的基因扩增引物 Target gene amplification primers | TGA2-QRT-F | GTCTTAGCATCAGTCACCCTATC |
TGA2-QRT-R | CCAAGAACAGGAGTGTCACATA | |
内参引物 Internal reference primer | Actin-F | CCTAAGGCCAACAGAGAGAAA |
Actin-R | CGACCACTAGCATACAAGGAA |
名称 Name | 特征序列 Characteristic sequence | 元件功能 Component function | 位置/bp Site |
---|---|---|---|
ABRE | ACGTG | ABA响应 ABA response | -5 |
ACE | CTAACGTATT | 光响应 Photoresponse | + 9 |
AE-box | AGAAACAA | 光响应 Photoresponse | -8 |
Box4 | ATTAAT | 光响应 Photoresponse | -6 |
G-box | CACGTT | 光响应 Photoresponse | + 6 |
P-box | CCTTTTG | 赤霉素响应 Gibberellin response | + 7、-7 |
TCA-element | CCATCTTTTT | 水杨酸响应 Salicylic acid response | + 9 |
TGACG-motif | TGACG | 茉莉酸甲酯响应 Response of methyl jasmonate | + 5、-5 |
TCT-motif | TCTTAC | 光响应 Photoresponse | -953 |
TC-rich repeats | GTTTTCTTAC | 防御和应激反应响应 Defense and stress response | -622、+ 213 |
MYB | CAACCA | MYB结合位点;干旱响应;ABA响应 MYB binding site;Drought response;ABA response | -676、+ 1702、-1125、 -2263 |
MYC | CATTTG | MYC结合位点;干旱响应;ABA响应 MYC binding site;Drought response;ABA response | -81、-1587 |
as-1 | TGACG | + 1、-131 |
Table 2 cis-Acting elements of TGA2 promoters
名称 Name | 特征序列 Characteristic sequence | 元件功能 Component function | 位置/bp Site |
---|---|---|---|
ABRE | ACGTG | ABA响应 ABA response | -5 |
ACE | CTAACGTATT | 光响应 Photoresponse | + 9 |
AE-box | AGAAACAA | 光响应 Photoresponse | -8 |
Box4 | ATTAAT | 光响应 Photoresponse | -6 |
G-box | CACGTT | 光响应 Photoresponse | + 6 |
P-box | CCTTTTG | 赤霉素响应 Gibberellin response | + 7、-7 |
TCA-element | CCATCTTTTT | 水杨酸响应 Salicylic acid response | + 9 |
TGACG-motif | TGACG | 茉莉酸甲酯响应 Response of methyl jasmonate | + 5、-5 |
TCT-motif | TCTTAC | 光响应 Photoresponse | -953 |
TC-rich repeats | GTTTTCTTAC | 防御和应激反应响应 Defense and stress response | -622、+ 213 |
MYB | CAACCA | MYB结合位点;干旱响应;ABA响应 MYB binding site;Drought response;ABA response | -676、+ 1702、-1125、 -2263 |
MYC | CATTTG | MYC结合位点;干旱响应;ABA响应 MYC binding site;Drought response;ABA response | -81、-1587 |
as-1 | TGACG | + 1、-131 |
Fig. 5 Construction and transformation of TGA2 gene expression vector A:Target gene amplification;B:Identification of E. coli by PCR;1-10:TGA2 gene PCR product.
Fig. 6 Overexpression of TGA2 in Arabidopsis thaliana T1(A)and T2(B)DNA identification M:DL2000;1-14:Transgenic A. thaliana;WT:Negative control(wild type);CK:Blank control(water);P:Positive control(Agrobacterium tumefaciens).
Fig. 7 Phenotypic changes and survival rate of wild-type and TGA2 transgenic Arabidopsis thaliana under different NaCl concentrations The different lowercase letters between groups indicate significant differences(P < 0.05). The same below.
Fig. 8 The expression levels of TGA2 in Lycium barbarum seedlings(A)and TGA2 in wild-type and transgenic(TGA2-OE)Arabidopsis thaliana(B)under different NaCl concentrations stress
[1] |
|
[2] |
|
[3] |
|
[4] |
|
[5] |
|
[6] |
|
高俊凤. 2006. 植物生理学实验指导. 北京: 高等教育出版社:210-217.
|
|
[7] |
|
[8] |
doi: 10.1093/jxb/eraa534 pmid: 33188435 |
[9] |
doi: 10.16420/j.issn.0513-353x.2020-1000 URL |
黎春红, 汪开拓, 雷长毅, 许凤, 季娜娜, 蒋永波. 2022. 桃TGA家族鉴定及BABA诱导的抗病表达分析. 园艺学报, 49 (2):265-280.
doi: 10.16420/j.issn.0513-353x.2020-1000 URL |
|
[10] |
doi: 10.16420/j.issn.0513-353x.2021-0732 URL |
李丽仙, 王烁, 陈莹, 邬滢涛, 王雅倩, 房月, 陈学森, 田长平, 冯守千. 2022. 甜樱桃PavMYB10.1促进PavRiant表达和花青苷积累. 园艺学报, 49 (5):1023-1030.
doi: 10.16420/j.issn.0513-353x.2021-0732 URL |
|
[11] |
|
[12] |
doi: 10.16420/j.issn.0513-353x.2014-0359 |
李涛,
doi: 10.16420/j.issn.0513-353x.2014-0359 |
|
[13] |
|
梁斌, 谢琦. 2016. GST融合蛋白活性测定条件的优化. 承德医学院学报, 33 (5):413-415.
|
|
[14] |
|
梁晓婕, 何昕孺, 何军, 田建文, 戴国礼, 秦垦. 2023. 不同篱架树形及采摘期对宁夏枸杞新品系果实品质的影响. 北方园艺,(2):104-111.
|
|
[15] |
|
梁晓婕, 李越鲲, 王亚军, 段淋渊, 安巍. 2019. 枸杞主栽品种耐盐性评价. 宁夏农林科技, 60 (6):17-21.
|
|
[16] |
|
[17] |
|
[18] |
|
[19] |
|
罗秀云, 李园园, 周赓, 田云, 卢向阳. 2015. TGA转录因子在植物氧化胁迫应答中的调控作用. 化学与生物工程, 32 (3):1-5.
|
|
[20] |
|
秦垦, 张波, 王丹青, 高燕, 戴国礼, 黄婷, 何昕孺, 周旋, 段淋渊, 焦恩宁. 2024a. 宁夏枸杞新品种‘宁农杞15号’. 园艺学报, 51 (S1):235-236.
|
|
[21] |
|
秦垦, 张波, 王丹青, 高燕, 戴国礼, 黄婷, 何昕孺, 周旋, 段淋渊, 焦恩宁. 2024b. 宁夏枸杞新品种‘科杞6082’. 园艺学报, 51 (S1):237-238.
|
|
[22] |
|
[23] |
doi: 10.13560/j.cnki.biotech.bull.1985.2022-0570 |
史光珍, 王兆晔, 孙琦, 朱新霞. 2022. 雪莲SikCDPK1启动子的克隆和活性分析. 生物技术通报, 38 (9):191-197.
doi: 10.13560/j.cnki.biotech.bull.1985.2022-0570 |
|
[24] |
|
[25] |
|
[26] |
|
田露辰. 2020. SmTGA2与丹酚酸B合成关键酶基因启动子的互作分析[硕士论文]. 杨凌: 西北农林科技大学.
|
|
[27] |
|
王艳红, 席克勇, 田野, 刘德麒, 周克贵, 尹军良, 刘奕清, 朱永兴. 2024. 姜GST基因家族成员鉴定与表达分析. 园艺学报, 51 (8):1803-1822.
|
|
[28] |
doi: 10.1023/a:1005873500238 pmid: 9225852 |
[29] |
|
[30] |
|
朱熙. 2021. 马铃薯中与胁迫相关StMAPKs基因的分离及功能研究[硕士论文]. 兰州: 甘肃农业大学.
|
[1] | LI Min, LI Siyu, SHI Zihan, CHEN Shuang, XU Yan, and LIU Guotian. Studies on the Efficiency of GRFs/GIFs for Genetic Transformation and Regeneration in Grapevine [J]. Acta Horticulturae Sinica, 2025, 52(1): 51-65. |
[2] | WANG Shanshan, GUO Rui, HE Ling, WU Chunhong, CHEN Chanyou, WAN Heping, and ZHAO Huixia. Identification of Long Cowpea Lhc Gene Family and Its Expression Analysis Under Salt Stress [J]. Acta Horticulturae Sinica, 2025, 52(1): 111-122. |
[3] | HAN Qin, GAO Huihui, MA Xiang, SU Jianyu, XU Chunyan. Screening and Identification of Antagonistic Rhizosphere Bacteria and Their Biological Control Effect on Root Rot of Lycium barbarum [J]. Acta Horticulturae Sinica, 2024, 51(5): 1162-1172. |
[4] | YI Qian, ZHANG Manman, WANG Xiaoli, FENG Jipeng, ZHU Shiping, WANG Fusheng, ZHAO Xiaochun. CclSAUR49 Affects Growth and Limonoids Metabolism in Citrus [J]. Acta Horticulturae Sinica, 2024, 51(3): 479-494. |
[5] | FENG Zhijuan, LIU Na, ZHANG Guwen, BU Yuanpeng, WANG Bin, GONG Yaming. Promoter of Vegetable Soybean GmDi19-3 Responds to Salt Stress and Exogenous ABA and MeJA [J]. Acta Horticulturae Sinica, 2024, 51(12): 2791-2799. |
[6] | YANG Juanbo, GUO Lili, LU Shixiong, GOU Huimin, WANG Shuaiting, ZENG Baozhen, MAO Juan. Cloning and Functional Analysis of FaGH3.17 Gene in Strawberry [J]. Acta Horticulturae Sinica, 2024, 51(11): 2483-2494. |
[7] | FENG Zhijuan, LIU Na, BU Yuanpeng, ZHANG Guwen, WANG Bin, GONG Yaming. Establishment of Agrobacterium rhizogenes-Mediated Genetic Transformation System in Vegetable Pea [J]. Acta Horticulturae Sinica, 2024, 51(10): 2439-2448. |
[8] | PENG Ling, FENG Lu, SONG Aiyun, DONG Linshui, LI Qingjun, LIU Jingtao. Interaction of Different Concentrations of Salt and Nitrogen Treatment on Fruit Quality Formation of Ziziphus jujuba‘Dongzao’ [J]. Acta Horticulturae Sinica, 2023, 50(10): 2192-2206. |
[9] | CHEN Sijie, ZHANG Tao, JIA Baosen, DU Juan, YAN Siyuan, GU Peiwen. Study on Antibacterial Activity of Dark Septate Endophytes Against Fusarium oxysporum in Chinese Wolfberry [J]. Acta Horticulturae Sinica, 2022, 49(7): 1519-1531. |
[10] | LIU Zhongjie, ZHENG Ting, ZHAO Fanggui, FU Weihong, ZHUGE Yaxian, ZHANG Zhichang, FANG Jinggui. Resistance Difference and Physiological Response Mechanism of Grape Rootstocks to Osmotic Stress [J]. Acta Horticulturae Sinica, 2022, 49(5): 984-994. |
[11] | LI Yamei, MA Fuli, ZHANG Shanqi, HUANG Jinqiu, CHEN Mengting, ZHOU Junyong, SUN Qibao, SUN Jun. Optimization of Jujube Callus Transformation System and Application of ZjBRC1 in Regulating ZjYUCCA Expression [J]. Acta Horticulturae Sinica, 2022, 49(4): 749-757. |
[12] | LIU Xiaowei, XIA Bin, LI Ziwei, YANG Yujia, CHEN Bin, ZHOU Yunwei, HE Miao. Overexpression of Chrysanthemum indicum miR396a Gene in Arabidopsis Enhances Its Salt Tolerance [J]. Acta Horticulturae Sinica, 2022, 49(4): 816-826. |
[13] | XIANG Li, ZHAO Lei, WANG Mei, LÜ Yi, WANG Yanfang, SHEN Xiang, CHEN Xuesen, YIN Chengmiao, MAO Zhiquan. Cloning and Functional Analysis of MdWRKY74 in Apple [J]. Acta Horticulturae Sinica, 2022, 49(3): 482-492. |
[14] | YANG Ni, WAN Qiwen, LI Yimin, HAN Miaohua, TENG Ruimin, LIU Jiexia, ZHUANG Jing. Effects of Exogenous Spermidine on Photosynthetic Characteristics and Gene Expression of Key Enzymes Under Salt Stress in Tea Plant [J]. Acta Horticulturae Sinica, 2022, 49(2): 378-394. |
[15] | SU Liyao, WANG Peiyu, JIANG Mengqi, HUANG Shuqi, XUE Xiaodong, LIU Mengyu, XIAO Xuechen, LAI Chunwang, ZHANG Zihao, CHEN Yukun, LAI Zhongxiong, LIN Yuling. The Activity Verification of pri-miR319a Encode Regulatory Peptide of Dimocarpus longan [J]. Acta Horticulturae Sinica, 2021, 48(5): 908-920. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||
Copyright © 2012 Acta Horticulturae Sinica 京ICP备10030308号-2 国际联网备案号 11010802023439
Tel: 010-82109523 E-Mail: yuanyixuebao@126.com
Support by: Beijing Magtech Co.Ltd