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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)

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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"

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