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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (9): 1303-1311.doi: 10.3724/SP.J.1006.2020.03004

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

Exploration of specific gene(s) for ear rot resistance to Fusarium verticilloides in maize

WEN Jing1, SHEN Yan-Qi1, HAN Si-Ping1, XING Yue-Xian2, ZHANG Ye1, WANG Zi-Yu1, LI Shi-Jie1, YANG Xiao-Hong3, HAO Dong-Yun1, ZHANG Yan1,*()   

  1. 1 Agro-Biotechnology Research Institute, Jilin Academy of Agricultural Sciences, Changchun 130033, Jilin, China
    2 Maize Research Institute, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, Jilin, China
    3 College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China
  • Received:2020-01-18 Accepted:2020-04-15 Online:2020-09-12 Published:2020-04-26
  • Contact: Yan ZHANG E-mail:zhangyan4023@163.com
  • Supported by:
    National Natural Science Foundation of China(31701504)

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Abstract:

Ear rot in maize, caused by fungal pathogens, poses a grave threat to maize production, and current inbred lines in use generally lack resistance to ear rot. It is essential to explore the resistant loci and corresponding resistance genes for improvement of the resistance to ear rot in maize by molecular marker-assisted breeding and biotechnology breeding. The purpose of this study was to explore the resistance loci of ear rot caused by Fusarium verticilloides and preliminary identify candidate genes by transcriptome analysis and genome-wide association. The result of transcriptome analysis showed that there were 10,761 differentially expressed genes between inbred lines Fa A and Ye 81162 at seven days after artificial inoculation with Fusarium verticilloides. A total of five SNPs significantly associated with ear rot resistance were detected by genome-wide association analysis, and these SNPs were distributed on chromosomes 1 and 9. By comparing B73 RefGen_v3 and annotating, it was found that the genes involved near the SNP site included acyl activating enzyme 1 peroxisome, protein phosphatase 2C 48, magnesium transporter, receptor protein kinase CRINKLY4 and zinc finger CCCH domain protein 19. The zinc finger CCCH domain protein 19 detected in genome-wide association just was the differentially expressed gene from transcriptome analysis, indicating that Zinc finger CCCH domain protein 19 probably is related to resistance to Fusarium verticilloides ear rot in maize. The results not only provide theoretical basis and important genetic resources for resistance gene cloning and molecular breeding of maize, but also lay a foundation for analysing the interaction between maize and pathogen.

Key words: maize, Fusarium verticilloides, ear rot, resistance genes

Table 1

Classification criteria of Fusarium verticillioides ear rot in maize"

病情评级Disease severity 发病百分比Percentage of symptom
1 发病面积占雌穗总面积0~1% Percentages of lesion area: 0-1%
3 发病面积占雌穗总面积2%~10% Percentages of lesion area: 2%-10%
5 发病面积占雌穗总面积11%~25% Percentages of lesion area: 11%-25%
7 发病面积占雌穗总面积26%~50% Percentages of lesion area: 26%-50%
9 发病面积占雌穗总面积51%~100% Percentages of lesion area: 51%-100%

Table 2

Sequences of primers"

基因名称
Gene name		 上游引物序列
Forward primer sequence (5'-3')		 下游引物序列
Reverse primer sequence (5'-3')		 产物长度
Product length (bp)
actin CCTTCATTGGCATGGAATCT GCAACCACCTTCACCTTCAT 128
β-tubulin2 AGACCGGTCAGTGCGGTAAC CGTGCTCGCCAGAGAGATGGT 70
CCCH domain protein 19 CAGGTGAGTTGTGGACATGG CTCCAAGCACAAGCAAACAA 197

Fig. 1

RT-PCR analysis of β-tubulin2 of Ye 81162 M: DL2000 marker; 1-6: Grain samples from ears before inoculation and at 1 d, 2 d, 3 d, 4 d, 7 d after inoculation."

Fig. 2

Relative expression of β-tubulin2 gene after artificial inoculation with Fusarium verticillium"

Fig. 3

Analysis of differentially expressed genes of Fa A and Ye 81162 after inoculation with Fusarium verticillium R: resistant; S: susceptible; R-0: Fa A before inoculation; R-7d: Fa A at seven days after inoculation; S-0: Ye 81162 before inoculation; S-7d: Ye 81162 at seven days after inoculation."

Fig. 4

Manhattan plot and QQ plot for genome-wide association analysis of resistance to ear rot caused by Fusarium verticillium in maize"

Table 3

Relative expression of β-tubulin2 gene after artificial inoculation with Fusarium verticillium"

自交系
Inbred line		 β-tubulin2基因的相对表达量 Relative expression of β-tubulin2 gene
接菌前
Before inoculation		 1 d 2 d 3 d 4 d 7 d
法A Fa A 0±0.00 0.0004±0.00 0.0002±0.00 0.0003±0.00 0.0002±0.00 1.006E-07±0.00
掖81162 Ye 81162 0±0.00 2.3894±0.93 0.6537±0.14 3.1675±0.38 3.0244±0.65 4.2477±0.26

Table 4

Gene information related to SNP loci associated with Fusarium verticillium ear rot resistance"

标记
Marker		 注释
Annotation		 染色体
Chromosome		 描述
Description
chr1.S_7569254 GRMZM2G110616 1 酰基激活酶1过氧化物酶体
Probable acyl-activating enzyme 1 peroxisomal
chr1.S_8288770 GRMZM5G891266 1 蛋白磷酸酶2C 48 Probable protein phosphatase 2C 48
chr1.S_8353697 GRMZM2G018706 1 镁转运蛋白10 Magnesium transporter 10
chr9.S_154383692 GRMZM2G092776 9 受体蛋白激酶CRINKLY4 Receptor protein kinase CRINKLY4
chr9.S_154384544 GRMZM2G393471 9 锌指CCCH域蛋白19 Zinc finger CCCH domain-containing protein 19

Fig. 5

Relative expression of zinc finger CCCH domain-containing protein 19 gene after artificial inoculation with Fusarium verticillium"

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