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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (11): 2923-2932.doi: 10.3724/SP.J.1006.2025.41080

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

Identification and agronomic characterization of imazamox-resistant dwarf mutants in Ningmai 36

GUI Ling-Xing1,2(), LING Xi-Tie2, TANG Zhao-Cheng2, LUO Wen-Zhen1,2, ZHU Pan-Zhen1,2, QIU Ze-Yu2,*(), ZHANG Bao-Long1,2,*()   

  1. 1 College of Life Sciences, Nanjing Agricultural University, Nanjing 210014, Jiangsu, China
    2 Institute of Germplasm Resources and Biotechnology, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu, China
  • Received:2024-11-15 Accepted:2025-08-13 Online:2025-11-12 Published:2025-08-26
  • Contact: *E-mail: qzy910525@163.com; E-mail: zhbl2248@hotmail.com
  • Supported by:
    Jiangsu Provincial Independent Innovation Project((CX (23) 3094)

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Abstract:

The frequent occurrence of herbicide-resistant volunteer weed plants in wheat fields, combined with poor agronomic performance due to lodging, poses a significant challenge to achieving high yields and yield stability. To address this issue, we applied ethyl methanesulfonate (EMS) mutagenesis to the widely cultivated winter wheat variety Ningmai 36 and generated an M2 population screened for both resistance to the acetolactate synthase (ALS) inhibitor imazamox and reduced plant height. Two stable mutants, WK120 and WK121, were identified that combined both traits. These mutants survived imazamox doses lethal to the wild type and exhibited height reductions of 37% and 29%, respectively. Genotyping revealed a G-to-A point mutation in the ALS gene on the D subgenome, resulting in a serine-to-asparagine substitution at position 627. Comprehensive evaluation of agronomic and yield-related traits showed that WK120 and WK121 outperformed the wild type in several key metrics: thousand-grain weight increased by 4.29% and 5.17%, grain number per spike by 1.9% and 3.1%, and spike length by 1.8% and 2.7%, respectively. Gibberellin (GA3) application experiments confirmed that both mutants remain responsive to GA, indicating that their dwarf phenotype is not due to GA insensitivity. Furthermore, molecular marker assays and Sanger sequencing ruled out the presence of known GA-sensitive dwarfing alleles (Rht4, Rht8, Rht9, Rht11, and Rht22) in both the wild type and mutant lines, suggesting the presence of a previously uncharacterized dwarfing locus in WK120 and WK121. These herbicide-resistant, dwarf derivatives of Ningmai 36 demonstrate strong tolerance to ALS-inhibiting herbicides along with improved agronomic performance, representing valuable genetic resources for future wheat breeding programs.

Key words: wheat, dwarf mutant, imazamox, gibberellin, agronomic traits

Table 1

Selected dwarf genes in gibberellin-sensitive wheat"

基因
Gene		 引物名称
Primer name		 引物序列
Primer sequence
(5'-3')		 片段长度
Fragment length (bp)		 退火温度
Annealing temperature
(℃)		 染色体
Chromosome
Rht4 WMC317 F
WMC317 R		 TGCTAGCAATGCTCCGGGTAAC
TCACGAAACCTTTTCCTCCTCC		 170 58 2BL
Rht8 WMC503 F
WMC503 R		 GCAATAGTTCCCGCAAGAAAAG
ATCAACTACCTCCAGATCCCGT		 275 55 2DS
Rht9 BARC151 F
BARC151 R		 TGAGGAAAATGTCTCTATAGCATCC
CGCATAAACACCTTCGCTCTTCCACTC		 220 55 5AL
Rht11 WR3 F
WR3 R		 GGTAGGGAGGCGAGAGGCGAG
GGCCATCTCCAGCTGCTCCAGCTA		 220 65 4B
Rht22 Xgwm471 F
Xgwm471 R		 CGGCCCTATCATGGCTG
GCTTGCAAGTTCCATTTTGC		 149 60 7AS

Fig. 1

ALS gene sequences from the genomes of imazamox-resistant wheat lines A: WK120 mutation site; B: WK121 mutation site. The red box represents a single point mutation from G-A."

Fig. 2

Resistance levels to imazamox in wild type (WT) Ningmai 36 and dwarf mutants WK120 and WK121 The numbers below the picture indicate the concentration, and the unit of concentration: g a.i. hm-2. Scale bar: 10 cm."

Table 2

Comparison of major agronomic traits between wild type Ningmai 36 and dwarf mutants WK120 and WK121"

材料
Material		 宁麦36
Ningmai 36		 WK120 WK121
株高Plant height (cm) 80.45±3.29 a 50.60±2.06 c 57.00±3.00 b
旗叶长度Flag leaf length (cm) 18.87±2.32 a 20.17±3.42 a 19.60±3.27 a
旗叶宽度Flag leaf width (cm) 1.69±0.14 a 1.44±0.22 b 1.46±0.32 b
每穗粒数Number of grains per panicle 40.66±7.75 a 41.47±4.36 a 41.93±4.76 a
穗长Spike length (cm) 8.77±0.78 a 8.92±1.08 a 9.01±0.81 a
节间数 Number of internodes 5.90±0.32 a 4.80±0.42 b 4.70±0.48 b
平均节间长Average internode length (cm) 14.28±3.15 a 10.67±3.56 b 13.05±4.88 ab
第4节间充实度Intersegmental fullness of fourth internode (mg cm-1) 24.54±3.46 a 23.18±2.30 a 20.36±5.23 a

Table 3

Comparison of grain morphology and thousand-grain weight between wild type Ningmai 36 and dwarf mutants WK120 and WK121"

材料 Material 宁麦36 Ningmai 36 WK120 WK121
粒长Grain length (cm) 0.66 a 0.64 a 0.66 a
粒宽Grain width (cm) 0.33 a 0.34 a 0.34 a
千粒重Thousand-seed weight (g) 40.82 a 42.58 a 42.93 a

Fig. 3

Major phenotypic characteristics of wild type (WT) Ningmai 36 and mutants WK120 and WK121 A: spike length; B: grain length; C: cross-section of the stem; D: grain width; E: plant height. Scale bars in A-E: 5 cm, 1 cm, 10 mm, 1 cm, and 5 cm."

Fig. 4

Morphological responses of wild type (WT) Ningmai 36 and mutant lines WK120 and WK121 to exogenous gibberellin (GA3) treatment during seedling stage A and B were the effects of gibberellin on the length of the first leaf and the elongation of the coleoplasty at the seedling stage of wheat Ningmai 36 and mutants WK120 and WK121, respectively. C shows the phenotypic comparison of all genotypes under GA3 and MOCK treatments. GA: gibberellin. Different lowercase and uppercase letters represent significant difference at the 0.05 and 0.01 levels, respectively. Scale bar in C: 1 cm."

Fig. 5

Molecular marker detection and Sanger sequencing of wheat dwarfing-related genes A and B represent molecular marker detection and Sanger sequencing, respectively. M: marker, 100-5000 bp. Lanes 1-3 were indicated: wild type Ningmai 36, mutant WK120 and mutant WK121. The target fragment size of each gene has been noted. In figure B, the red box indicates that wild type (WT), WK120, and WK121 all have CG bases in the target region, with no T-type mutations present in the mutants."

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