Welcome to Acta Agronomica Sinica，

Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (8): 1166-1175.doi: 10.3724/SP.J.1006.2019.84166

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

Effects of over-expression of AtDREB1A gene on potato growth and abiotic stress resistance gene expression

JIA Xiao-Xia1,2,QI En-Fang1,2,LIU Shi1,2,WEN Guo-Hong1,2,*(),MA Sheng1,2,LI Jian-Wu1,2,HUANG Wei1,2   

  1. 1 Potato Research Institute, Gansu Academy of Agricultural Sciences/Gansu Engineering Laboratory of Potato Germplasm Resources Innovation, Lanzhou 730070, Gansu, China
    2 The Ministry of Agriculture, Scientific Observation and Experiment Station of Dry Potato in the Northwest, Weiyuan 748201, Gansu, China
  • Received:2018-12-11 Accepted:2019-04-15 Online:2019-08-12 Published:2019-07-16
  • Contact: Guo-Hong WEN E-mail:251580436@qq.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31560412);This study was supported by the National Natural Science Foundation of China(31060200);Science and Technology Support Program of Gansu Academy of Agricultural Sciences(2017GAAS38);China Agriculture Research System(CARS-09-P06)

RichHTML354

11

PDF

268

Abstract:

Longshu 3 (L3) and its AtDREB1A transgenic line T2 were planted in the pot experiment. The water content of the soil in pots was controlled to 45%-50% of the maximum water holding capacity (FWC) during the flowering stage of the potato. The phenotype, MDA content, RWC, activities of SOD and POD, and the gene expression level in leaves were analyzed, showing there were no significant differences between the two lines under normal watering conditions. After 20 d of stress treatment, the phenotype of transgenic plants T2 was significantly better than that of control L3, and RWC was significantly higher than that of L3. The MDA content, SOD and POD activities of leaves in each line increased significantly, while the MDA content of transgenic plants increased less than, and the activities of SOD and POD of transgenic plants increased more than those of the control, indicating that the cell membrane damage and membrane lipid peroxidation of the transgenic plants were lighter than those of the control, and the drought tolerance of transgenic plants was significantly improved. Transcriptome sequencing and bioinformatic analysis showed that compared with L3, there were 430 differentially expressed genes in T2, including 287 up-regulated genes and 143 down-regulated genes. Functional annotation and significance enrichment showed that all differentially expressed genes were involved in three broad categories of GO functional classification systems, that was biological processes, cell components and molecular functions. Most of these differentially expressed genes concentrated on the membrane in cells, and mainly related to signal transduction, redox, biological mediation, stress response, development process, system immune process, nucleic acid and protein binding transcription factor activity, transport activity and catalytic activity. The expression of a large number of abiotic stress-related genes was up-regulated, including PPR protein family, P450 protein family, heat shock protein family and MLO protein family, indicating that these genes play a very important role in drought-stress resistance of AtDREB1A transgenic potato. This study lays a foundation for further understanding AtDREB1A’s regulatory network to improve potato drought resistance.

Key words: potato, AtDREB1Agene, abiotic stress, differentially expressed genes, drought

Table 1

qRT-PCR primers for the selected genes"

基因 ID
Gene ID		 正向引物
Forward primer (5′-3′)		 反向引物
Reverse primer (5′-3′)
EF1α ATTGGAAACGGATATGCTCCA TCCTTACCTGAACGCCTGTCA
AtDREB1A GCCGATCAGCCTGTCTCAAT TCTGCCATATTAGCCAACAAACTC
PGSC0003DMG400016489 AAATGTGTCTTTCAAGCGAA TCCATCTTCCCACATTTACCGTA
PGSC0003DMG400011977 CTTTGCCTGAGAATATTGACT CTTTGGGATTACTACTAGCCTT
PGSC0003DMG400000534 CCGACGATACAAACATATACTGA TGAGTTCCTTCCAGCGATT
PGSC0003DMG402019042 CCAACTCAGAACTGGTACGGACA CGTAAACATCCGTTGGGACCAT
PGSC0003DMG400000242 AACAGCCAGAAGAAGTTGAT ATTCCCATCACAGGCATCTCA
PGSC0003DMG401018057 AGGATCCATTAAAGTTCAAGCCA AATCCTTGAAGAAGCCTAGCA

Fig. 1

Phenotypes of transgenic and no-transgenic potato under drought stress A: 未转基因植株L3; B: 转基因植株T2。A: no-transgenic plant L3; B: transgenic plant T2."

Fig. 2

Changes of SOD and POD activities in potato leaves under drought stress Bars superscripted by different letters are significantly different at P < 0.05 within the same treatment."

Fig. 3

Changes of MDA content and relative water content in potato leaves under drought stress Bars superscripted by different letters are significantly different at P < 0.05 within the same treatment."

Fig. 4

qRT-PCR analysis on AtDREB1A gene in potato leaves Bars superscripted by different letters are significantly different at P < 0.05 within the same treatment."

Fig. 5

Volcano plot of gene expression in potato leaves after drought stress The abscissa shows the log2 (fold change) between the two samples; the ordinate shows the statically significance of the difference. Each gene is represented by one point on the graph. Differential genes with no significant difference were represented by black dots, those left green points and right red points represent significantly different down-regulated and up-regulated genes, respectively."

Fig. 6

Differentially expressed gene GO annotation clustering map The abscissa is the content of each category of GO. The left side of the ordinate is the percentage of the gene number, and the right side is the gene number."

Fig. 7

Cluster diagram of gene expression patterns of PPR protein family under drought stress The lines in graph show the degree of similarity in the variation trend of each gene, and the shorter the line is, the more similar the genes are. Red indicates the up-regulated expression level, while green indicates the down-regulated expression level."

Fig. 8

Cluster diagram of gene expression patterns of HSP protein family under drought stress The lines in graph show the degree of similarity in the variation trend of each gene, and the shorter the line is, the more similar the genes are. Red indicates the up-regulated expression level, while green indicates the down-regulated expression level."

Fig. 9

Cluster diagram of gene expression patterns of P450 protein family under drought stress The lines in graph show the degree of similarity in the variation trend of each gene, and the shorter the line is, the more similar the genes are. Red indicates the up-regulated expression level, while green indicates the down-regulated expression level."

Fig. 10

Cluster diagram of gene expression patterns of MLO protein family under drought stress The lines in graph show the degree of similarity in the variation trend of each gene, and the shorter the line is, the more similar the genes are. Red indicates the up-regulated expression level, while green indicates the down-regulated expression level."

Fig. 11

Verification of DEGs between L3 vs. T2 after drought stress using qRT-PCR"

[1] Seki M, Narusaka M, Abe H, Kasuga M, Shinozaki K, Carninci P, Hayashizaki Y, Shinozaki K . Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA micro array. Plant Cell, 2001,13:61-72.
[2] Fowler S, Thomashow M F . Arabidopsis transcription profiling indicates that multiple regulatory pathways are activated during cold acclimation in addition to the CBF cold response pathway. Plant Cell, 2002,14:1675-1690.
[3] 王萍, 高世庆, 郭永来, 程宪国, 王罡, 马有志, 季静 . 利用农杆菌介导将抗逆相关基因GmDREB导入大豆的研究. 大豆科学, 2008,27:47-51.
Wang P, Gao S Q, Guo Y L, Cheng X G, Wang Z, Ma Y Z, Ji J . Transformation of stress resistance related gene GmDREB into soybean via Agrobacterium-mediation. Soybean Sci, 2008,27:47-51 (in Chinese with English abstract).
[4] Courtois B, McLaren G, Sinha P K, Prasad K, Yadav R, Shen L . Mapping QTLs associated with drought avoidance in upland rice. Mol Breed, 2000,6:55-66.
doi: 10.1023/A:1009652326121
[5] 刘录祥, 赵林姝, 梁欣欣, 郑企成, 刘强, 王晶, 郭会君, 赵世荣, 陈文华 . 基因枪法获得逆境诱导转录因子DREB1A转基因小麦的研究. 中国生物工程杂志, 2003,23(11):53-56.
Liu L X, Zhao L S, Liang X X, Zheng Q C, Liu Q, Wang J, Guo H J, Zhao S R, Chen W H . Study on production of transgenic wheat with a stress-inducible transcription factor gene DREB1A by microprojectile bombardment. China Biotechnol, 2003,23(11):53-56 (in Chinese with English abstract).
[6] Pellegrineschi A, Reynolds M, Pacheco M, Brito R M, Almeraya R, Yamaguchi-Shinozaki K, Hoisington D . Stress-induced expression in wheat of the Arobidopsls thaliana DREB1A gene delays water stress symptoms under greenhouse conditions. Geneme, 2004,47:493-500.
[7] 许昆朋 . 转AtCBF3基因马铃薯提高抗冷性的研究. 山东农业大学硕士毕业论文, 山东泰安, 2011.
Xu K P . Studies on the Mechanisms of Enhanced Chilling Tolerance in Potato Transformed with AtCBF3 Gene. MS Thesis of Shandong Agricultural University, Tai’an, Shandong, China, 2011 (in Chinese with English abstract).
[8] Dou H O, Xu K P, Meng Q W, Li G, Yang X H . Potato plants ectopically expressing Arabidopsis thaliana CBF3 exhibit enhanced tolerance to high-temperature stress. Plant Cell Environ, 2014,38:61-72.
[9] 贾小霞, 齐恩芳, 王一航, 文国宏, 龚成文, 王红梅, 李建武, 马胜, 胡新元 . 转录因子DREB1A基因和Bar基因双价植物表达载体的构建及对马铃薯遗传转化的研究. 草业学报, 2014,23(3):110-114.
doi: 10.11686/cyxb20140312
Jia X X, Qi E F, Wang Y H, Wen G H, Gong C W, Wang H M, Li J W, Ma S, Hu X Y . Construction of a bivalent plant expression vector of DREB1A and Bar genes and studies of genetic transformation of potato. Acta Pratac Sin, 2014,23(3):110-114 (in Chinese with English abstract).
doi: 10.11686/cyxb20140312
[10] 贾小霞, 齐恩芳, 马胜, 胡新元, 王一航, 文国宏, 龚成文, 李建武 . 转DREB1A/Bar双价基因马铃薯的耐旱性及除草剂抗性分析. 草业学报, 2015,24(11):58-64.
Jia X X, Qi E F, Ma S, Hu X Y, Wang Y H, Wen G H, Gong C W, Li J W . Analysis of drought tolerance and herbicide resistance in transgenic potato plants over-expressing DREB1A/Bar. Acta Pratacul Sin, 2015,24(11):58-64 (in Chinese with English abstract).
[11] 张治安, 张美善, 蔚荣海 . 植物生理学实验指导. 北京: 中国农业科学技术出版社, 2004. pp 265-296.
Zhang Z A, Zhang M S, Wei R H. Plant Physiology Experiment Guide. Beijing: China Agricultural Science and Technology Press, 2004. pp 265-296(in Chinese).
[12] 康雯 . 土壤自然失水胁迫对地锦幼苗生理生化特性的影响. 东北林业大学硕士学位论文, 黑龙江哈尔滨, 2010.
Kang W . Effect of Soil Natural Water Stress on Physiological and Biochemical Indexes in Parthenocissus tricuspidata Seedling. MS Thesis of Northeast Forestry University, Harbin, Heilongjiang, China, 2010 (in Chinese with English abstract).
[13] 许冰霞, 尹美强, 温银元, 裴帅帅, 柯贞进, 张彬, 原向阳 . 谷子萌发期响应干旱胁迫的基因表达谱分析. 中国农业科学, 2018,51:1431-1447.
Xu B X, Yin M Q, Wen Y Y, Pei S S, Ke Z J, Zhang B, Yuan X Y . Gene expression profiling of foxtail millet ( Setaria italica L.) under drought stress during germination. Sci Agric Sin, 2018,51:1431-1447 (in Chinese with English abstract).
[14] 李健 . ScCBF1与StCBF1转录因子在马铃薯应答低温、干旱及盐胁迫过程中存在功能差异. 山东农业大学硕士学位论文, 山东泰安, 2018.
Li J . ScCBF1 and StCBF1 have Different Function in Response to Freezing, Drought and Salt Stress in Potato. MS Thesis of Shandong Agricultural University, Tai’an, Shandong, China, 2018 (in Chinese with English abstract).
[15] 杜建雄, 侯向阳, 刘金荣 . 草地早熟禾对干旱及旱后复水的生理响应研究. 草业学报, 2010,19(2):31-38.
doi: 10.11686/cyxb20100205
Du J X, Hou X Y, Liu J R . A study on physiological response to drought and re-watering treatments in Kentucky bluegrass. Acta Pratac Sin, 2010,19(2):31-38 (in Chinese with English abstract).
doi: 10.11686/cyxb20100205
[16] 刘明稀, 卢少云, 郭振飞 . 假俭草抗旱变异体的筛选及其生理鉴定. 草业学报, 2012,21(1):126-132.
doi: 10.11686/cyxb20120116
Liu M X, Lu S Y, Guo Z F . Selection and physiological identification of somaclonal variants for increased drought resistance of centipedegrass. Acta Pratac Sin, 2012,21(1):126-132 (in Chinese with English abstract).
doi: 10.11686/cyxb20120116
[17] 李源, 刘贵波, 高洪文, 孙桂枝, 赵海明, 谢楠 . 紫花苜蓿种质耐盐性综合评价及盐胁迫下的生理反应. 草业学报, 2010,19(4):79-86.
doi: 10.11686/cyxb20100411
Li Y, Liu G B, Gao H W, Sui G Z, Zhao H M, Xie N . A comprehensive evaluation of salt-tolerance and the physiological response of Medicago sativa at the seedling stage. Acta Pratac Sin, 2010,19(4):79-86 (in Chinese with English abstract).
doi: 10.11686/cyxb20100411
[18] 张海娜, 李小娟, 李存东, 肖恺 . 过量表达小麦超氧化物歧化酶(SOD)基因对烟草耐盐能力的影响. 作物学报, 2008,34:1403-1408.
Zhang H N, Li X J, Li C D, Xiao K . Effects of overexpression of wheat superoxide dismutase (SOD) genes on salt tolerant capability in tobacco. Acta Agron Sin, 2008,34:1403-1408 (in Chinese with English abstract).
[19] 范敏, 金黎平, 刘庆昌, 屈冬玉 . 马铃薯PPR蛋白家族基因SoDIPPR的克隆及其在干旱条件下的表达特征分析. 中国农业科学, 2008,41:2249-2257.
Fan M, Jin L P, Liu Q C, Qu D Y . Cloning of SoDIPPR gene of pentatricopeptide repeat (PPR) protein family in potato and analysis of expression characteristics under drought conditions. Sci Agric Sin, 2008,41:2249-2257 (in Chinese with English abstract).
[20] Yan S P, Zhang Q Y, Tang Z C, Su W A, Sun W N . Comparative proteomic analysis provides new insights into chilling stress responses in rice. Mol Cell Proteomics, 2006,5:484-496.
doi: 10.1074/mcp.M500251-MCP200
[21] Baldwin J C, Dombrowski J E . Evaluation of Lolium temulentum as a model grass species for the study of salinity stress by PCR-based subtractive suppression hybridization analysis. Plant Sci, 2006,171:459-469.
[22] Ma S, Gong Q, Bohnert H J . Dissecting salt stress pathways. J Exp Bot, 2006,57:1097-1107.
doi: 10.1093/jxb/erj098
[23] Baxter C J, Redestig H, Schauer N, Repsilber D, Patil K R, Nielsen J, Selbig J, Liu J, Fernie A R, Sweetlove L J . The metabolic response of heterotrophic Arabidopsis cells to oxidative stress. Plant Physiol, 2007,143:312-325.
[24] Umbach A L, Fiorani F, Siedow J N . Characterization of transformed Arabidopsis with altered alternative oxidase levels and analysis of effects on reactive oxygen species in tissue. Plant Physiol, 2005,139:1806-1820.
[25] Hajheidari M, Abdollahian-Noghabi M, Askari H, Heidari M, Sadeghian S Y, Ober E S, Salekdeh G H . Proteome analysis of sugar beet leaves under drought stress. Proteomics, 2005,5:950-960.
doi: 10.1002/(ISSN)1615-9861
[26] Wang W, Vinocur B, Shoseyov O, Altman A . Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends Plant Sci, 2004,9:244-252.
doi: 10.1016/j.tplants.2004.03.006
[27] 崔会婷, 王珍, 张铁军, 蒋旭, 龙瑞才, 杨青川, 康俊梅 . 蒺藜苜蓿MtCYP450基因的克隆及功能分析. 中国草地学报, 2018,40(5):1-10.
Cui H T, Wang Z, Zhang T J, Jiang X, Long R C, Yang Q C, Kang J M . Clong and functional analysis of gene MtCYP450 from Medicago truncatula. Chin J Grassl, 2018,40(5):1-10 (in Chinese with English abstract).
[28] Van Nguyen N T, Vo K T X, Park H, Jeon J S, Jung K H . A systematic view of the MLO family in rice suggests their novel roles in morphological development, diurnal responses, the light- signaling pathway, and various stress responses. Front Plant Sci, 2016,7:1413, doi: 10.3389/fpls.2016.01413.
[29] Wang X, Ma Q, Dou K . Genome-wide characterization and comparative analysis of the MLO gene family in cotton. Plant Physiol Biochem, 2016,103:106-119.
[30] Lim C W, Lee S C . Functional roles of the pepper MLO protein gene, CaMLO2, in abscisic acid signaling and drought sensitivity. Plant Mol Biol, 2013,85:1-10.
[1] CHEN Song-Yu, DING Yi-Juan, SUN Jun-Ming, HUANG Deng-Wen, YANG Nan, DAI Yu-Han, WAN Hua-Fang, QIAN Wei. Genome-wide identification of BnCNGC and the gene expression analysis in Brassica napus challenged with Sclerotinia sclerotiorum and PEG-simulated drought [J]. Acta Agronomica Sinica, 2022, 48(6): 1357-1371.
[2] ZHOU Wen-Qi, QIANG Xiao-Xia, WANG Sen, JIANG Jing-Wen, WEI Wan-Rong. Mechanism of drought and salt tolerance of OsLPL2/PIR gene in rice [J]. Acta Agronomica Sinica, 2022, 48(6): 1401-1415.
[3] LI Yi-Jun, LYU Hou-Quan. Effect of agricultural meteorological disasters on the production corn in the Northeast China [J]. Acta Agronomica Sinica, 2022, 48(6): 1537-1545.
[4] WANG Hai-Bo, YING Jing-Wen, HE Li, YE Wen-Xuan, TU Wei, CAI Xing-Kui, SONG Bo-Tao, LIU Jun. Identification of chromosome loss and rearrangement in potato and eggplant somatic hybrids by rDNA and telomere repeats [J]. Acta Agronomica Sinica, 2022, 48(5): 1273-1278.
[5] WANG Xing-Rong, LI Yue, ZHANG Yan-Jun, LI Yong-Sheng, WANG Jun-Cheng, XU Yin-Ping, QI Xu-Sheng. Drought resistance identification and drought resistance indexes screening of Tibetan hulless barley resources at adult stage [J]. Acta Agronomica Sinica, 2022, 48(5): 1279-1287.
[6] SHI Yan-Yan, MA Zhi-Hua, WU Chun-Hua, ZHOU Yong-Jin, LI Rong. Effects of ridge tillage with film mulching in furrow on photosynthetic characteristics of potato and yield formation in dryland farming [J]. Acta Agronomica Sinica, 2022, 48(5): 1288-1297.
[7] LI A-Li, FENG Ya-Nan, LI Ping, ZHANG Dong-Sheng, ZONG Yu-Zheng, LIN Wen, HAO Xing-Yu. Transcriptome analysis of leaves responses to elevated CO2 concentration, drought and interaction conditions in soybean [Glycine max (Linn.) Merr.] [J]. Acta Agronomica Sinica, 2022, 48(5): 1103-1118.
[8] WANG Xia, YIN Xiao-Yu, Yu Xiao-Ming, LIU Xiao-Dan. Effects of drought hardening on contemporary expression of drought stress memory genes and DNA methylation in promoter of B73 inbred progeny [J]. Acta Agronomica Sinica, 2022, 48(5): 1191-1198.
[9] FENG Ya, ZHU Xi, LUO Hong-Yu, LI Shi-Gui, ZHANG Ning, SI Huai-Jun. Functional analysis of StMAPK4 in response to low temperature stress in potato [J]. Acta Agronomica Sinica, 2022, 48(4): 896-907.
[10] ZHANG Xia, YU Zhuo, JIN Xing-Hong, YU Xiao-Xia, LI Jing-Wei, LI Jia-Qi. Development and characterization analysis of potato SSR primers and the amplification research in colored potato materials [J]. Acta Agronomica Sinica, 2022, 48(4): 920-929.
[11] JIN Rong, JIANG Wei, LIU Ming, ZHAO Peng, ZHANG Qiang-Qiang, LI Tie-Xin, WANG Dan-Feng, FAN Wen-Jing, ZHANG Ai-Jun, TANG Zhong-Hou. Genome-wide characterization and expression analysis of Dof family genes in sweetpotato [J]. Acta Agronomica Sinica, 2022, 48(3): 608-623.
[12] TAN Xue-Lian, GUO Tian-Wen, HU Xin-Yuan, ZHANG Ping-Liang, ZENG Jun, LIU Xiao-Wei. Characteristics of microbial community in the rhizosphere soil of continuous potato cropping in arid regions of the Loess Plateau [J]. Acta Agronomica Sinica, 2022, 48(3): 682-694.
[13] DING Hong, XU Yang, ZHANG Guan-Chu, QIN Fei-Fei, DAI Liang-Xiang, ZHANG Zhi-Meng. Effects of drought at different growth stages and nitrogen application on nitrogen absorption and utilization in peanut [J]. Acta Agronomica Sinica, 2022, 48(3): 695-703.
[14] WU Yan-Fei, HU Qin, ZHOU Qi, DU Xue-Zhu, SHENG Feng. Genome-wide identification and expression analysis of Elongator complex family genes in response to abiotic stresses in rice [J]. Acta Agronomica Sinica, 2022, 48(3): 644-655.
[15] ZHANG Hai-Yan, XIE Bei-Tao, JIANG Chang-Song, FENG Xiang-Yang, ZHANG Qiao, DONG Shun-Xu, WANG Bao-Qing, ZHANG Li-Ming, QIN Zhen, DUAN Wen-Xue. Screening of leaf physiological characteristics and drought-tolerant indexes of sweetpotato cultivars with drought resistance [J]. Acta Agronomica Sinica, 2022, 48(2): 518-528.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No.12 Zhongguancun South Street, Beijing 100081,China Email:xbzw@chinajournal.net.cn
Supported by Beijing Magtech Co.Ltd