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Acta Agronomica Sinica ›› 2026, Vol. 52 ›› Issue (1): 131-147.doi: 10.3724/SP.J.1006.2026.53032

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

Screening and identification of candidate resistance genes to gibberella ear rot caused by Fusarium graminearum in maize

Dong Li-Hua(), Dong Cheng-Yan, Li Zheng-Nan, Yu Jing, Ye Liang, Liu Fang, Tan Jing*()   

  1. School of Agriculture, Yunnan University, Kunming 650500, Yunnan, China
  • Received:2025-05-29 Accepted:2025-10-30 Online:2026-01-12 Published:2025-11-05
  • Contact: *E-mail: tanjingli@sina.com
  • Supported by:
    National Natural Science Foundation of China(32260516)

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Abstract:

Gibberella ear rot (GER) of maize (Zea mays L.), caused by Fusarium graminearum (Fg), is a major factor contributing to yield losses in southwestern China. In this study, two highly resistant inbred lines (4019 and NMJT) and two highly susceptible lines (Huangzaosi and GEMS61) were selected. Kernels at 15 days after pollination were inoculated with Fg, and samples were collected at three post-infection time points for transcriptome sequencing. Candidate resistance genes were identified, cloned, and preliminarily validated through a combination of differentially expressed gene (DEG) analysis, previous genome-wide association study (GWAS) results, qRT-PCR validation, cloning and sequence alignment, and expression pattern analysis. The main findings were as follows: (1) Transcriptome analysis revealed a large number of DEGs between resistant and susceptible lines at all three infection stages. The two resistant lines shared common defense responses, with DEGs significantly enriched in pathways such as plant secondary metabolism, plant hormone signal transduction, calcium signaling, and the antioxidant system. In addition, each resistant line exhibited specific defense mechanisms by regulating unique gene expression patterns. (2) Integration of DEG and GWAS data identified 24 co-localized genes. Based on gene annotation and literature reports, 12 genes were predicted as potential candidates for GER resistance. Among these, two genes—Zm00001eb104020 and Zm00001eb195780—were successfully cloned and validated by qRT-PCR. (3) Protein sequence analysis revealed shared mutations and deletions between resistant and susceptible lines. The two candidate genes also showed distinct spatiotemporal expression patterns, with significantly higher expression levels in resistant lines. Both genes were strongly induced and upregulated in kernels following Fg inoculation. These results provide a theoretical foundation for further functional characterization of GER resistance mechanisms and the breeding of resistant maize germplasm.

Key words: maize, gibberella ear rot, candidate resistance genes, functional site identification, spatio-temporal expression characteristics

Table 1

Information on maize inbred lines with high resistance and high susceptibility to GER"

抗性
Resistance		 代号
No.		 自交系
Inbred lines		 系谱
Pedigree		 来源/类群
Source/group
高抗
High resistance		 R1 4019 G108×G172 中国/温带非硬秆种China / temperate non-stiff stalk group
R2 NMJT 地方种质
Local germplasm		 中国/温热混合China / temperate & tropical mixed group
高感
High susceptibility		 S1 黄早四Huangzaosi 黄改Huanggai 中国/唐四平头China / Tangsipingtou
S2 GEMS61 ARI16026:S17-10-1-B-B 美国/热带USA / tropical group

Table 2

DEGs between highly resistant inbred R1 and highly susceptible inbreds S1 and S2 under the same treatment at three sampling time points"

组合
Paired samples		 处理
Treatment		 0 hpi 1.5 hpi 6 hpi
上调Up 下调Down 上调Up 下调Down 上调Up 下调Down
R1 vs S1 CK 7461 7959 6487 6514 7105 7111
Fg 6593 6624 6896 6709 7161 6789
Fg-CK 1875 1554 2305 2323 2382 2328
R1 vs S2 CK 7047 7552 5918 6826 5451 5752
Fg 5110 5872 4727 5162 6380 6375
Fg-CK 1956 1731 1724 1464 2525 2195

Table 3

DEGs between highly resistant inbred R2 and highly susceptible inbreds S1 and S2 under the same treatment at three sampling time points"

组合
Paired samples		 处理
Treatment		 0 hpi 1.5 hpi 6 hpi
上调Up 下调Down 上调Up 下调Down 上调Up 下调Down
R2 vs S1 CK 7683 7630 5677 5207 7497 7672
Fg 5660 5824 6460 6001 4037 4074
Fg-CK 1175 939 2476 2335 1051 1176
R2 vs S2 CK 7634 7869 5782 6321 5933 6361
Fg 5529 6193 6221 5911 4903 5458
Fg-CK 1687 1517 2575 1695 1753 1654

Fig. 1

Venn diagrams of common DEGs between R1 vs S1 & S2 and R2 vs S1 & S2 at three sampling time points A: Venn diagrams of Up DEGs; B: Venn diagrams of Down DEGs. 0 hpi, 1.5 hpi, and 6 hpi indicated the grains cultured on PDA plate for 0 h, 1.5 h and 6 h, respectively."

Fig. 2

GO and KEGG enrichment analysis of common DEGs between R1 vs S1 & S2 and R2 vs S1 & S2 at three sampling time points A: GO analysis of up-regulated DEGs; B: GO analysis of down-regulated DEGs; C: KEGG analysis of up-regulated DEGs; D: KEGG analysis of down-regulated DEGs. 0 hpi, 1.5 hpi and 6 hpi indicated the grains cultured on PDA plate for 0 h, 1.5 h and 6 h, respectively. BP: biological process; CC: cellular component; MF: molecular function."

Table 4

Co-localization of transcriptome DEGs with previous GWAS results"

基因
Gene		 描述
Description
Zm00001eb240340 含ABC转运蛋白结构域的蛋白质ABC transporter domain-containing protein
Zm00001eb309360 MYB-CC型转录因子LHEQLE含结构域蛋白
MYB-CC type transcription factor LHEQLE-containing domain-containing protein
Zm00001eb309010 磷酸甘油酸变位酶家族蛋白Phosphoglycerate mutase family protein
Zm00001eb070740 四肽重复序列(TPR)样超家族蛋白Tetratricopeptide repeat (TPR)-like superfamily protein
Zm00001eb070690 DNA聚合酶DNA polymerase eta
Zm00001eb070770 光系统Ⅰ的积累Accumulation of photosystem Ⅰ
Zm00001eb354690 植物血红素过氧化物酶家族结构域内含蛋白Plant heme peroxidase family profile domain-containing protein
Zm00001eb070760 蛋白质阿戈核酸酶2 Protein argonaute 2
Zm00001eb248650 细胞周期蛋白-T1-3 Cyclin-T1-3
Zm00001eb195760 抗病蛋白RPM1 Disease resistance protein RPM1
Zm00001eb197630 谷氨酰tRNA (Gln)酰胺转移酶亚基B叶绿体/线粒体
Glutamyl-tRNA (Gln) amidotransferase subunit B chloroplastic / mitochondrial
Zm00001eb070720 富含亮氨酸的重复跨膜蛋白激酶Leucine-rich repeat transmembrane protein kinase
Zm00001eb239920 蛋白质丝氨酸/苏氨酸磷酸酶Protein-serine/threonine phosphatase
Zm00001eb195720 未鉴定蛋白Uncharacterized protein
Zm00001eb195790 TRAF样超家族蛋白TRAF-like superfamily protein
Zm00001eb195710 假定的无酶家族蛋白Putative apyrase family protein
Zm00001eb104020 磷脂酶D Phospholipase D
Zm00001eb070750 OSJNBa0008M17.14样蛋白OSJNBa0008M17.14-like protein
Zm00001eb195780 V型质子ATP酶亚单位S1/VOA1跨膜结构域含蛋白
V-type proton ATPase subunit S1/VOA1 transmembrane domain-containing protein
Zm00001eb106040 聚腺苷酸结合蛋白RBP45C Polyadenylate-binding protein RBP45C
Zm00001eb070700 LRR受体样丝氨酸/苏氨酸蛋白激酶LRR receptor-like serine/threonine-protein kinase
Zm00001eb248680 含NAC结构蛋白质NAC domain-containing protein
Zm00001eb070730 富含亮氨酸的重复跨膜蛋白激酶Leucine-rich repeat transmembrane protein kinase
Zm00001eb354680 过氧化物酶Peroxidase

Fig. 3

qRT-PCR verification of candidate genes R1: highly resistant material 4019; R2: highly resistant material NMJT; S1: highly susceptible material Huangzaosi; S2: highly susceptible material GEMS61. CK: sterile water treatment; Fg: inoculation treatment. 0 hpi, 1.5 hpi, and 6 hpi indicated the grains cultured on PDA plate for 0 h, 1.5 h, and 6 h respectively."

Fig. 4

Amino acid sequence alignment of candidate genes Zm00001eb104020 (A) and Zm00001eb195780 (B) between high resistant and high susceptibility inbreds R1: highly resistant material 4019; R2: highly resistant material NMJT; S1: highly susceptible material Huangzaosi; S2: highly susceptible material GEMS61."

Fig. 5

Expression profiles of candidate genes Zm00001eb104020 and Zm00001eb195780 in different tissues of four inbreds across four periods R1: highly resistant material 4019; R2: highly resistant material NMJT; S1: highly susceptible material Huangzaosi; S2: highly susceptible material GEMS61."

Fig. 6

Expression profiles of candidate genes Zm00001eb104020 and Zm00001eb195780 in different tissues of four inbreds at two periods under Fg infection R1: highly resistant material 4019; R2: highly resistant material NMJT; S1: highly susceptible material Huangzaosi; S2: highly susceptible material GEMS61. CK: sterile water treatment; Fg: inoculation treatment. R: root; S: stalk; L: leaves; MF: male flower; FF: female flower; C: corn grian. *: P < 0.05; **: P < 0.01; ***: P < 0.001."

Fig. S1

GO and KEGG enrichment analysis of DEGs between highly resistant inbred R1 and highly susceptible inbreds S1, S2 at three sampling time points A: GO analysis of up-regulated DEGs; B: GO analysis of down-regulated DEGs; C: KEGG analysis of up-regulated DEGs; D: KEGG analysis of down-regulated DEGs. 0 hpi, 1.5 hpi, and 6 hpi indicated the grains cultured on PDA plate for 0 h, 1.5 h, and 6 h, respectively. BP: biological process; CC: cellular component; MF: molecular function."

Fig. S2

GO and KEGG enrichment analysis of DEGs between highly resistant inbred R2 and highly susceptible inbreds S1, S2 at three sampling time points A: GO analysis of up-regulated DEGs; B: GO analysis of down-regulated DEGs; C: KEGG analysis of up-regulated DEGs; D: KEGG analysis of down-regulated DEGs. 0 hpi, 1.5 hpi, and 6 hpi indicated the grains cultured on PDA plate for 0 h, 1.5 h, and 6 h, respectively. BP: biological process; CC: cellular component; MF: molecular function."

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