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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (4): 996-1005.doi: 10.3724/SP.J.1006.2023.24062

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

StEFR1 regulates late blight resistance positively in potato (Solanum tuberosum)

ZHANG Wei-Na(), YU Hui-Fang, AN Zhen, LIU Wen-Kai, KANG Yi-Chen, SHI Ming-Fu, YANG Xin-Yu, ZHANG Ru-Yang, WANG Yong, QIN Shu-Hao*()   

  1. College of Horticulture, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2022-03-21 Accepted:2022-07-21 Online:2023-04-12 Published:2022-08-22
  • Contact: *E-mail: qinsh@gsau.edu.cn
  • Supported by:
    Science and Technology Innovation Fund of Gansu Agricultural University (Doctoral Research Start-up Fund Project for Public Recruitment)(GAU-KYQD-2020-10);Natural Science Foundation of Gansu Province(21JR7RA827);National Natural Science Foundation of China(32060441)

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Abstract:

Potato late blight is a devastating oomycete disease, which causes great economic losses to agricultural production. In this study, we analyzed the role and potential regulatory mechanism of StEFR1 in regulating late blight resistance by the relative expression pattern and functional verification combined with the bioinformatics methods. Evolutionary analysis showed that the sequence similarity between StEFR1 and AtEFR was 53.9%. After inoculated with Phytophthora infestans for 3 days and treatment with elf18 for 3 hours, the relative expression level of StEFR1 in isolated leaves of Atlantic was upregulated to 1.87 times and 2.31 times compared with the control, respectively. The late blight resistance significantly increased after the overexpression of StEFR1 in the isolated leaves of Atlantic by agrobacterium infiltration method. Compared with the control, the area of leaf lesion size decreased and the activity of leaf cells increased. And the marker genes of PTI, SA, and JA signaling pathways in overexpressed leaves were significantly up-regulated to varied degrees, while the relative expression levels of ET related-genes did not change significantly. In conclusion, StEFR1 was involved in the PTI resistance and regulated the relative expression levels of SA and JA hormone signaling related genes, suggesting that StEFR1 positively regulated the potato late blight. This study lays a foundation for further revealing the molecular mechanism of StEFR1 in regulating the immune response and provides important reference for the molecular breeding of late blight.

Key words: potato late blight, LRR-RLKs, hormonal signal, marker gene, differential expression

Table 1

Primers for quantitative real-time PCR"

基因名称
Gene name		 上游引物
Forward sequence (5′-3′)		 下游引物
Reverse sequence (5′-3′)
ef1α ATTGGAAACGGATATGCTCCA TCCTTACCTGAACGCCTGTCA
ChtA TTCTGGATGACAGCACAGGATAA GGCGTCCATTGCCCAAT
PR-1b GGCATCCCGAGCACAAAAT CTGCACCGGAATGAATCAAGT
PR-2 GTGAAGCTGGTTTGGGAAATG TTGCCAATCAACGTCATGTCTAC
WRKY7 CCAACTGGAAGCAACAACAA CCTGATTAGAATGATTAGCCAACA
WRKY8 CCTACTGTGACATCTCATCAATCC GGGTGCTCCCATTTCAGAC
ACRE31 CAGGATGAATCGGATCTGAAA CGGCAATCCCAATTTCTCTA
LOX CAGATCAGGCCCCGTTAATG CCTGTAAGTCCACCTTCACTTGTTG
PAL-2 GGTCACTGCCTCGGGTGAT CCTGCCAGTGAGCAAACCA
ERF3 GTGTTGACGTGAAACCAACCAT CCGGTGGAGGAAAGTTAAGGT

Fig. 1

Domain identification, evolutionary analysis, and sequence alignment of StEFR1 A: StEFR1 is identified as a typical member of LRR-RLK family; B: StEFR1 is closely related to AT5G20480 (AtEFR) in Arabidopsis thaliana, which had known functions; C: the similarity between StEFR1 and AtEFR is 53.9%."

Fig. 2

Relative expression pattern of StEFR1 genes in response to pathogen signal * and ** mean significant difference at the 0.05 and 0.01 probability levels, respectively."

Table 2

Putative regulatory cis-elements in StEFR1 promoters"

序号
Serial number		 顺式调控元件
Cis-elements		 数目
Number
1 防御和应激反应元件Defense and stress responsive elements 2
2 脱落酸响应元件Abscisic acid responsive elements 2
3 厌氧诱导相关元件Anaerobic responsive elements 7
4 干旱诱导元件Drought responsive elements 5
5 玉米醇溶蛋白代谢调节元件Regulatory element for zein metabolism 1
6 光照响应元件Light responsive elements 10

Fig. 3

Transient overexpression of StEFR1 leaves enhanced late blight resistance A: the relative expression level of StEFR1 reached the maximum when injected for 2 days; B: the symptoms of WT leaves were more serious on the 4th day; C: the disease spot area in WT and OE; D: a large number of cells in WT group died. WT: the group injected with empty vector pFGC5941; OE: the group injected with StEFR1-PFGC5941. * and ** mean significant difference at the 0.05 and 0.01 probability levels, respectively."

Fig. 4

Relative expression pattern of PTI marker genes (WRKY7, WRKY8, and ACRE31) in WT and OE after inoculation with P. infestans for different times WT: the group injected with empty vector pFGC5941; OE: the group injected with StEFR1-PFGC5941. * and ** mean significant difference at the 0.05 and 0.01 probability levels, respectively."

Fig. 5

Relative expression pattern of SA (ChtA, PR-1B, and PR-2), JA (LOX and PAL-2) and ET (ERF3) related genes in WT and OE after inoculation with P. infestans for different times WT: the group injected with empty vector pFGC5941; OE: the group injected with StEFR1-PFGC5941. * and ** mean significant difference at the 0.05 and 0.01 probability levels, respectively."

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