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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (5): 1191-1198.doi: 10.3724/SP.J.1006.2022.13027

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

Effects of drought hardening on contemporary expression of drought stress memory genes and DNA methylation in promoter of B73 inbred progeny

WANG Xia*(), YIN Xiao-Yu, Yu Xiao-Ming, LIU Xiao-Dan   

  1. College of Jilin Agriculture Science and Technology, Jilin 132101, Jilin, China
  • Received:2021-03-24 Accepted:2021-09-09 Online:2022-05-12 Published:2021-10-14
  • Contact: WANG Xia E-mail:wangxhangj@126.com
  • Supported by:
    “13th five-year plan” Science and Technology Project of Jilin Province(JJKH20190972KJ);Science Technology Foundation for Young Scientists of College of Jilin Agriculture Science and Technology (Ji Nong Yuan He Zi [2018] 1001).

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Abstract:

Plants may remember the existing stresses, and this memory can be passed down across generations. To study the effects of drought hardening in G0 generation on the contemporary drought stress memory genes’ expression and DNA methylation in promoter region in G1 generation, 20% PEG-6000 was used to simulate drought condition, qRT-PCR was used to analyze the expression level changes of six genes with drought-stressed memory function in maize and Arabidopsis. The changes of DNA methylation rates in the promoter regions of the most differentially expressed gene was analyzed by bisulfite sequencing technology. The results showed that the expression of six genes were up-regulated under drought stress in a +/+ mode, and the expression level of G1 generation was significantly higher than that of G0 generation. Drought stress reduced the DNA methylation’ level in the promoter region of GRMZM2G088396 gene. The reduction of methylation in the detection region 1 of two generations was mainly caused by the reduction of CHG and CHH methylation level, the decrease of methylation in the detection region 2 was mainly caused by the decrease of CG and CHH methylation level. The total cytosine methylation rate of G1 generation was significantly lower than that of G0 generation, suggesting that drought exercise in G0 generation resulted in heritable variation of DNA methylation in the promoter region of GRMZM2G088396 gene in G1 generation, which may be directly involved in the expression of GRMZM2G088396 gene.

Key words: drought hardening, B73, drought stress memory gene, transgenerational memory, qRT-PCR, bisulfite sequencing PCR

Table 1

Primers for qRT-PCR and BSP"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 引物名称
Primer name		 引物序列
Primer sequence (5′-3′)
GAPDH-F CCCTTCATCACCACGGACTAC GRMZM2G059836-R ACGCTCGCAACCTTCGCAGTC
GAPDH-R TCCCACCACGGTTCTTCCAA GRMZM2G088396-F TGAGAAGGGGCAAGAATACCA
GRMZM5G851862-F TGTTTTCTGCTGGGAGTGACA GRMZM2G088396-R CTCGTCTCCAAGCACTGTCCT
GRMZM5G851862-R GAACCTTCGGGAATCACGTAG GRMZM2G079440-F GCACTTGCGAGTGGCTTTACTTG
GRMZM2G126505-F CCACCGCAACAAGCAGAGT GRMZM2G079440-R ACCGCTGGAGGTAATATCGACAC
GRMZM2G126505-R CGGTTTTTCGCGTTCCTGG GRMZM2G088396-BSP1-F TTTATAGTAGTAAAAGAGGTATTTTTAAT
GRMZM2G028535-F CAAAGGCGAAAAGATCGGTAC GRMZM2G088396-BSP1-R TCCCTATACTAAATTTTAAACTCTAC
GRMZM2G028535-R TTGCGTTCCTCTGATGACAAC GRMZM2G088396-BSP2-F TAGTATAGGGAATTAAATAATTGTTA
GRMZM2G059836-F TTGCGTTCCTCTGATGACAAC GRMZM2G088396-BSP2-R CATCCCTAACACCACTAAAAAATATC

Fig. 1

Phenotypes of B73 G0 and G1 generation under drought stress"

Table 2

Drought stress memory genes’ information"

基因编号Gene ID 基因描述Gene description 位置Position 染色体Chr.
GRMZM5G851862 cytochrome P450 family 76 subfamily C polypeptide 7 21,883,592-21,886,533 5
GRMZM2G126505 abscisic acid 8’-hydroxylase2 162,654,597-162,657,413 4
GRMZM2G028535 Deltal-pyrroline-5-carboxylate synthetase 173,415,159-173,423,316 8
GRMZM2G059836 Farnesylated protein 2 669,554-670,477 6
GRMZM2G088396 4-hydroxyphenylpyruvate dioxygenase 1 86,084,655-86,086,755 5
GRMZM2G079440 dehydrin DHN1 141,323,718-141,325,549 6

Fig. 2

Relative expression profiles of drought stress memory genes between G0 and G1 generations Different lowercase letters indicate significant differences at P < 0.05."

Fig. 3

Prediction of CG loci distribution in promoter region of GRMZM2G088396 gene The blue areas are CG enrichment areas."

Fig. 4

BSP amplified products of GRMZM2G088396 promoter M: 100 bp DNA marker; 1-4: the stress time of G0 generations was 0, 1, 3, and 5 hour(s); 5-8: the stress time of G1 generations was 0, 1, 3, and 5 hour(s)."

Fig. 5

Distribution of CG, CHG, and CHH sites in the promoter region of GRMZM2G088396 gene"

Fig. 6

Effects of drought training on DNA methylation in promoter region of G1 GRMZM2G088396 gene Different letters indicate significant difference at P < 0.05;The DNA methylation rate was the ratio of methylated cytosines to all cytosines."

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