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Acta Agronomica Sinica ›› 2026, Vol. 52 ›› Issue (10): 2898-2911.doi: 10.3724/SP.J.1006.2026.63030

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

Evaluation of maize germplasm for resistance to common smut and identification of related genes

Yang Wen-Yan1,2(), Qu Jian-Zhou1, Liu Geng-Yu1, Lu Yu1, Liu Jian-Zhuo1, Zhang Hong-Wei2, Du Wan-Li1,*()   

  1. 1 College of Agronomy, Shenyang Agricultural University, Shenyang 110161, Liaoning, China
    2 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2026-03-08 Accepted:2026-07-15 Online:2026-10-12 Published:2026-07-22
  • Contact: Du Wan-Li, E-mail: dwl2014@syau.edu.cn
  • Supported by:
    Basic Research Project of Liaoning Provincial Department of Education(JYTXS2024004);National Natural Science Foundation of China(32272152)

Abstract:

Maize common smut, caused by Ustilago maydis, is a widespread disease that seriously threatens maize yield and quality worldwide. Breeding resistant varieties is the most economical and effective strategy for controlling this disease. To identify elite germplasm resistant to common smut and uncover key resistance genes, this study systematically evaluated the resistance levels of 425 maize inbred lines, aiming to dissect the genetic basis of resistance and screen for resistance-related genes. Artificial inoculation and resistance evaluation were conducted in the field at two locations over two years, based on disease index (DI), incidence rate (IR), and area under the disease progress curve (AUDPC). A genome-wide association study (GWAS) was performed using 970,761 high-quality SNPs, and candidate resistance genes were further screened by integrating spatiotemporal transcriptomic data, followed by functional enrichment analysis. Among the 425 inbred lines, 6 were identified as highly resistant, 27 as resistant, 67 as moderately resistant, 154 as susceptible, and 171 as highly susceptible. The broad-sense heritabilities of DI, IR, and AUDPC were all above 75%. GWAS detected a total of 1665 significantly associated SNP loci, leading to the identification of 376 candidate genes, among which 79 were differentially expressed in response to pathogen infection and 19 were significantly upregulated. GO enrichment analysis indicated that the candidate genes were mainly involved in processes such as defense response, cell wall organization, and hormone signaling. This study confirms that resistance to maize common smut is highly heritable and controlled by multiple genes. The resistance-associated loci and candidate genes identified here provide important germplasm resources and genetic targets for the molecular breeding of maize varieties resistant to common smut.

Key words: maize, common smut, genome-wide association analysis, transcriptome, disease resistance candidate genes

Fig. 1

Correlation analysis of repeated measurements and BLUE values across locations for maize common smut resistance traits A-C: heatmaps of correlation coefficients for disease index (DI), incidence rate (IR), and area under the disease progress curve (AUDPC) across three replicates at two locations, Shenyang and Jinzhou, in two years, 2023 and 2024. 23SY, 24SY, 23JZ, and 24JZ represent Shenyang in 2023, Shenyang in 2024, Jinzhou in 2023, and Jinzhou in 2024, respectively; 1, 2, and 3 indicate the three replicates. D: SY-DI, JZ-DI, All-DI, SY-IR, JZ-IR, All-IR, SY-AUDPC, JZ-AUDPC, and All-AUDPC denote the best linear unbiased estimation (BLUE) values for disease index in Shenyang across two years, disease index in Jinzhou across two years, disease index across two years and two locations, incidence rate in Shenyang across two years, incidence rate in Jinzhou across two years, incidence rate across two years and two locations, AUDPC in Shenyang across two years, AUDPC in Jinzhou across two years, and AUDPC across two years and two locations, respectively. BLUE: best linear unbiased estimation; *, **, and *** indicate P < 0.05, P < 0.01, and P < 0.001, respectively. NS indicates no significant difference."

Fig. 2

Frequency distribution of incidence rate, disease index, and area under the disease progress curve for maize common smut 23SY, 24SY, 23JZ, 24JZ, and BLUE represent Shenyang in 2023, Shenyang in 2024, Jinzhou in 2023, Jinzhou in 2024, and the overall BLUE value across two years and two locations, respectively. BLUE: best linear unbiased estimation."

Fig. 3

Comprehensive assessment of maize inbred lines for resistance to common smut HS, S, MR, R, and HR represent highly susceptible, susceptible, moderately resistant, resistant, and highly resistant, respectively."

Fig. 4

Analysis of critical environmental time windows affecting maize common smut resistance traits A: identification of critical environmental time windows for disease index (DI). B: identification of critical environmental time windows for incidence rate (IR). C: identification of critical environmental time windows for area under the disease progress curve (AUDPC). Tmax, Tmin, Tmean, PTT, RH and SD represent maximum temperature, minimum temperature, mean temperature, precipitation, relative humidity, and sunshine duration, respectively."

Table 1

Grey relational grades between maize common smut resistance traits and meteorological factors"

气象因子
Meteorological factor
病情指数DI 发病率IR 病程曲线下面积AUDPC
关联度
Correlation
排名
Rank
关联度
Correlation
排名
Rank
关联度
Correlation
排名
Rank
最高气温Maximum temperature (Tmax) 0.639 5 0.641 5 0.585 5
最低气温Minimum temperature (Tmin) 0.702 3 0.705 3 0.655 3
平均气温Mean temperature (Tmean) 0.670 4 0.673 4 0.620 4
降水量Precipitation (PTT) 0.935 1 0.933 1 0.909 1
相对湿度Relative humidity (RH) 0.405 6 0.405 6 0.347 6
日照时数Sunshine duration (SD) 0.893 2 0.898 2 0.875 2

Fig. 5

Genetic structure analysis of the maize association panel A: scatter plot of principal component analysis (PCA) based on genome-wide SNPs. B: heatmap of the kinship matrix among individuals; diagonal elements approaching 1 indicate complete genotypes and high homozygosity of the inbred lines; the off-diagonal region is predominantly light blue, with generally low kinship coefficients, indicating substantial genetic divergence and relatively distant relationships among individuals; the absence of distinct high-kinship blocks is consistent with the PCA results, suggesting that this panel does not contain strong subpopulation-driven kinship clusters and has a relatively dispersed genetic structure. C: population structure analysis, with different colors representing the proportion of each accession assigned to different ancestral subpopulations (K = 6). The x-axis represents 421 maize inbred lines, and the y-axis indicates the probability of each inbred line being assigned to different subpopulations."

Fig. 6

Manhattan plots from genome-wide association analysis of maize common smut resistance traits A: disease index (DI); B: incidence rate (IR); C: area under the disease progress curve (AUDPC). The red horizontal dashed line indicates the significance threshold (P = 2.87×10-6)."

Fig. 7

Venn diagram of SNP loci associated with disease index, incidence rate, area under the disease progress curve, and their combined maize common smut resistance index"

Fig. 8

Integrative analysis of GWAS candidate genes and transcriptome differentially expressed genes Heatmap of expression patterns for 79 candidate genes at different time points (1, 2, 4, 6, 8, and 10 days) after inoculation with the standard Ustilago maydis strain SG200. Blue to red indicates expression values from low to high. Inf1 d, Inf2 d, Inf4 d, Inf6 d, Inf8 d, and Inf10 d represent 1, 2, 4, 6, 8, and 10 days post-inoculation, respectively; Mock1 d, Mock2 d, Mock4 d, Mock6 d, Mock8 d, and Mock10 d represent the corresponding mock-inoculated controls at 1, 2, 4, 6, 8, and 10 days post-inoculation, respectively. FPKM: fragments per kilobase of transcript per million mapped reads."

Table S1

List of important candidate genes for common smut resistance in maize and their functional annotations"

基因编号
Gene ID
染色体
Chromosome
起始位置
Start position
终止位置
End position
描述
Description
Zm00001eb363720 8 159,957,159 159,961,745 Aminopeptidase
Zm00001eb425600 10 127,370,239 127,371,829 Endochitinase B
Zm00001eb298310 7 1,229,176 1,233,458 Monosaccharide transporter 1
Zm00001eb325320 7 160,143,290 160,148,271 Pheophytinase, chloroplastic
Zm00001eb400350 9 147,292,024 147,294,766 Putative MYB DNA-binding domain superfamily protein
Zm00001eb220250 5 22,100,009 22,104,260 Trihelix transcription factor GT-2
Zm00001eb342970 8 65,781,256 65,785,083 Unknown
Zm00001eb325790 7 161,346,807 161,349,645 Unknown
Zm00001eb152660 3 195,940,555 195,943,584 Unknown
Zm00001eb136360 3 125,744,774 125,746,538 Unknown
Zm00001eb296740 6 165,988,425 165,989,350 Calmodulin
Zm00001eb154300 3 201,227,822 201,232,263 Subtilisin-like protease SBT5.6
Zm00001eb029990 1 161,025,196 161,028,265 Unknown
Zm00001eb169930 4 20,398,975 20,399,475 Unknown
Zm00001eb116370 2 232,357,624 232,362,179 Unknown
Zm00001eb325340 7 160,215,143 160,216,461 Stress-induced protein 1
Zm00001eb404840 10 1,146,198 1,148,897 Unknown
Zm00001eb233720 5 89,152,269 89,156,855 4-coumarate-CoA ligase 1
Zm00001eb325300 7 160,098,845 160,101,078 Putative lectin-like receptor protein kinase family protein

Fig. 9

Gene ontology (GO) enrichment analysis of differentially expressed candidate genes for common smut resistance in maize FDR (false discovery rate) is a statistical error control metric used in multiple hypothesis testing, defined as the expected proportion of incorrectly rejected null hypotheses among all rejected null hypotheses."

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