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作物学报 ›› 2025, Vol. 51 ›› Issue (11): 2923-2932.doi: 10.3724/SP.J.1006.2025.41080

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

宁麦36抗甲氧咪草烟矮秆突变体的鉴定及农艺性状分析

桂灵星1,2(), 凌溪铁2, 唐兆成2, 罗文臻1,2, 朱盼珍1,2, 仇泽宇2,*(), 张保龙1,2,*()   

  1. 1 南京农业大学生命科学学院, 江苏南京 210014
    2 江苏省农业科学院种质资源与生物技术研究所, 江苏南京 210014
  • 收稿日期:2024-11-15 接受日期:2025-08-13 出版日期:2025-11-12 网络出版日期:2025-08-26
  • 通讯作者: *仇泽宇, E-mail: qzy910525@163.com; 张保龙, E-mail: zhbl2248@hotmail.com
  • 作者简介:E-mail: 2023816136@stu.njau.edu.com
  • 基金资助:
    江苏省自主创新项目((CX (23) 3094)

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 Published:2025-11-12 Published online: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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摘要：

麦田除草剂抗性杂草频发与小麦倒伏风险已成为制约小麦高产稳产的双重挑战。本研究基于甲基磺酸乙酯(EMS)诱变技术创制新型抗除草剂矮秆小麦种质, 利用EMS对冬小麦品种宁麦36进行诱变, 通过乙酰乳酸合成酶(acetolactate synthetase, ALS)抑制剂类除草剂甲氧咪草烟对M2代诱变群体进行抗除草剂单株筛选, 结合株高测定, 最终获得2株稳定遗传的矮秆突变体WK120和WK121。2个突变体较野生型宁麦36, 具有较高的甲氧咪草烟抗性, 且株高分别下降37%和29%。ALS基因的基因型鉴定结果表明, WK120和WK121的D亚基因组上的ALS均发生G-A的单点突变, 导致第627位的丝氨酸转变为天冬酰胺。农艺性状及产量相关性状分析表明, 较野生型宁麦36, WK120、WK121的千粒重分别提高4.29%和5.17%, 每穗粒数分别提高1.9%和3.1%, 穗长分别提高1.8%和2.7%。赤霉素(GA3)处理结果表明, WK120和WK121均属于赤霉素反应敏感型种质。分子标记检测和Sanger测序结果表明, 野生型和突变体中均无已知赤霉素敏感矮秆基因(Rht4、Rht8、Rht9、Rht11和Rht22), 暗示突变体中可能存在新的矮化基因位点。综上, 抗除草剂矮秆型宁麦36不仅对ALS抑制剂类除草剂具有较高的耐受性, 且农艺性状获得改善, 本研究为后续的抗除草剂矮秆小麦品种的培育提供了遗传资源和理论参考。

关键词: 小麦, 矮秆突变体, 甲氧咪草烟, 赤霉素, 农艺性状

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

表1

赤霉素敏感型小麦部分矮秆基因"

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

图1

抗性小麦基因组ALS基因序列 A: WK120突变位点; B: WK121突变位点。红框处代表由G-A的单点突变。"

图2

小麦野生型宁麦36和矮秆突变体WK120、WK121对甲氧咪草烟的抗性水平 图下方数字表示浓度, 单位g a.i. hm-2。标尺为10 cm。"

表2

小麦野生型宁麦36和矮秆突变体WK120、WK121主要农艺性状的比较"

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

表3

小麦野生型宁麦36和矮秆突变体WK120、WK121粒型和千粒重的比较"

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

图3

小麦野生型宁麦36和突变体WK120、WK121的主要表型特征 A: 穗长; B: 粒长; C: 茎秆横切面; D: 粒宽; E: 株高。图A~E标尺分别为5 cm, 1 cm, 10 mm, 1 cm和5 cm。"

图4

外源赤霉素(GA3)处理下野生型宁麦36与突变体WK120、WK121的苗期形态响应 A、B分别为赤霉素对宁麦36和突变体WK120、WK121苗期第一叶长度和胚芽鞘伸长影响的显著性分析。C为赤霉素(GA3)与清水(MOCK)处理下各株系的表型对比。GA: 赤霉素。不同小写字母和大写字母分别代表在0.05和0.01水平差异显著。图C标尺为1 cm。"

图5

小麦矮秆相关基因分子标记检测及Sanger测序 A、B分别为分子标记检测及Sanger测序。M: marker, 100~5000 bp。泳道1~3依次表示野生型宁麦36、突变体WK120、突变体WK121; 图中已标注出各基因目标片段大小。B图中红框处代表野生型(WT)、WK120和WK121在目标区间碱基均为CG, 突变体不含T类型拷贝。"

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