宁春4号小麦EMS突变体库的构建及表型变异分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0027

摘要/Abstract

摘要：

目的为加快盐碱地小麦优质种质资源的创制，丰富遗传多样性，构建宁春4号EMS突变体库。方法采用0.5%甲基磺酸乙酯（EMS）溶液处理10 000粒小麦品种宁春4号种子，构建M₁代突变体库，对其表型性状突变率进行统计分析。此外，对M₁代材料进行250 mmol/L的NaCl溶液定向筛选，得到耐盐突变体，并对突变体进行表型鉴定，以及形态指标、物质积累、生理指标的测定。结果 M₁代成活3 709株，成株率37.09%，其中，有表型突变性状的1 226株，突变率为33.05%。1 226株突变类型丰富，包含叶部性状突变151株，穗部性状突变212株，分蘖突变161株，株高突变133株，育性和生育期突变130株，其他性状突变75株，突变率分别为13.89%、5.72%、4.34%、3.59%、3.50%和2.02%。经过盐胁迫处理后，宁春4号与突变体相比表现为，叶片黄化萎蔫，苗高、最大根长、初生根数、地上部鲜重、根鲜重、地上部干重、根冠比均表现显著差异，而根干重无显著变化；在生理指标方面，突变体的丙二醛（MDA）含量、细胞膜透性显著低于野生型，脯氨酸（Pro）含量、过氧化物酶（POD）和过氧化氢酶（CAT）活性显著高于野生型，可溶性糖（SS）含量无显著差异。结论以宁春4号为野生型材料诱变产生的突变体类型丰富，为快速定位小麦优异性状基因并研究其功能提供丰富的种质资源。

关键词: 小麦, 宁春4号, EMS诱变, 突变体库构建, 表型变异统计分析

Abstract:

Objective To accelerate the creation of high-quality germplasm resources for saline wheat and to enrich genetic diversity, we constructed Ningchun No. 4 EMS mutant library. Method In this study, 10 000 seeds of the wheat variety Ningchun No. 4 were treated with 0.5% ethyl methanesulfonate (EMS) solution, and a library of M₁ generation mutants was constructed and statistically analyzed for the mutation rates of their phenotypic traits. In addition, the M₁ generation material was subjected to targeted screening with 250 mmol/L NaCl solution to obtain salt-tolerant mutants, and the mutants were phenotyped as well as measured for morphological indices, substance accumulation, and physiological indices. Result The 3 709 plants survived in M₁ generation with a survival rate of 37.09%, of which 1 226 had phenotypic mutant traits with a mutation rate of 33.05%. The 1 226 plants were rich in mutation types, containing 151 mutations for leaf traits, 212 mutations for spike traits, 161 mutations for tillers, 133 mutations for plant height, 130 mutations for fertility and fecundity, and 75 mutations for other traits, with mutation rates of 13.89%, 5.72%, 4.34%, 3.59%, 3.50%, and 2.02%, respectively. After salt stress treatment, Ningchun No. 4 showed yellowing and wilting of leaves, significant differences in seedling height, maximum root length, number of primordial roots, aboveground fresh weight, root fresh weight, aboveground dry weight, and root-crown ratio compared to the mutant, while root dry weight showed no significant change. In terms of physiological indices, malondialdehyde (MDA) content and cell membrane permeability of the mutants were significantly lower than those of the wild type, proline content, peroxidase (POD) and catalase (CAT) activities were significantly higher than those of the wild type, and there was no significant difference in the soluble sugar content. Conclusion The types of mutants produced by EMS mutagenesis from Ningchun No. 4 are abundant, providing rich germplasm resources for rapid localization of wheat genes for excellent traits and study of their functions.

Key words: wheat, Ningchun No. 4, EMS mutagenesis, mutant library construction, statistical analysis of phenotypic variation

豆飞飞, 任毓昭, 王石磊, 刘春颖, 王晓东, 王昭懿, 刘彩霞, 刘凤楼, 王掌军, 李清峰. 宁春4号小麦EMS突变体库的构建及表型变异分析[J]. 生物技术通报, 2025, 41(8): 92-101.

DOU Fei-fei, REN Yu-zhao, WANG Shi-lei, LIU Chun-ying, WANG Xiao-dong, WANG Zhao-yi, LIU Cai-xia, LIU Feng-lou, WANG Zhang-jun, LI Qing-feng. Construction and Phenotypic Variation Analysis of Ningchun No. 4 Wheat EMS Mutant Library[J]. Biotechnology Bulletin, 2025, 41(8): 92-101.

图/表 14

表1 不同浓度的EMS处理的种子发芽率

Table 1 Germination rates of seeds treated with different concentrations of EMS

EMS浓度 EMS concentration （%）	发芽率 Germination rate （%）	相对致死率 Relative lethality rate （%）
0 （CK）	92.0a	0
0.3	54.2b	41.1
0.5	45.7c	50.3
0.7	35.6d	61.3

表2 M1的突变类型及比率

Table 2 Mutation types and ratios in M1

性状 Trait	表型 Phenotype	突变数量 Number of mutations	突变比率 Mutation ratio （%）
叶部 Leaf	间绿 Mesophyllous green	36	0.97
	单边失绿带 Single-sided loss of green tap	98	2.64
	两侧失绿带 Loss of green belt on both sides	22	0.59
	半边失绿 Lose one half of one’s head	7	0.19
	中间失绿带 Lose the center green zone	54	1.46
	叶变黄 Lose color	206	5.55
	卷叶 Leafroll	92	2.48
穗 Spike	双头穗 Double-headed spikelet	42	1.13
	三头穗 Three-headed spikelet	16	0.43
	弯穗 Bent spikelet	47	1.27
	密穗 Dense spikelet	2	0.05
	不育穗 Sterile spikelet	3	0.08
	畸形穗 Distorted spike	30	0.81
	短穗 Short spikelet	72	1.94
分蘖 Tiller	独秆 Solitary culm	96	2.59
	寡蘖 Few tiller	49	1.32
	多蘖 Multiple tiller	16	0.43
株高 Plant height	高秆 Tall stalks	12	0.32
株高 Plant height	矮秆 Short stalks	121	3.26
育性和生育期 Fertility and maturity	早熟 Early-maturingt	27	0.73
	晚熟 Late-maturing	53	1.43
	不育 Sterile	50	1.35
其他 Others	叶环腐斑 Leaf ring rot	23	0.62
	叶黄斑 Leaf yellow spot	20	0.54
	叶白斑 Leaf white spot	21	0.57
	叶白线 White lines on the leaf	9	0.24
	叶黑线 Black lines on the leaf	2	0.05
合计 Total		1 226	33.05

图1 M1代叶部突变性状A：间绿；B：单边失绿带；C：两侧失绿带；D：半边失绿；E：中间失绿带；F：叶变黄；G：卷叶

Fig. 1 Leaf mutations in the M1 generationA: Mesophyllous green. B: Single-sided loss of green tap. C: Loss of green belt on both sides. D: Lose one half of one’s head.E: Lose the center green zone. F: Lose color. G: Leafroll

图2 M1代穗部突变性状A：双头穗；B：三头穗；C：弯穗；D：密穗；E：不育穗；F：畸形穗；G：短穗

Fig. 2 Panicle mutant traits in the M1 generationA: Double-headed spikelet. B: Three-headed spikelet. C: Bent spikelet. D: Dense spikelet. E: Sterile spikelet.F: Distorted spike. G: Short spikelet

图3 M1代分蘖突变性状A：寡蘖突变体；B：多蘖突变体

Fig. 3 Mutant traits in tillers in M1 generationA: Few tiller mutant. B: Multiple tiller mutant

图4 M1代株高突变性状A：高秆突变体；B：矮秆突变体

Fig. 4 Plant height mutation traits in M1 generationA: Tall-stalk mutant. B: Short-stalk mutant

图5 M1代生育期和育性突变性状1：晚熟突变体；2：不育突变体

Fig. 5 Growth period and fertility mutation traits of M1 generation1: Late-maturing mutant. 2: Sterile mutant

图6 M1代病状突变性状A：叶环腐斑；B：叶黄斑；C：叶白斑；D：叶白线；E：叶黑线

Fig. 6 Diseased mutation traits of the M1 generationA: Leaf ring rot. B: Leaf yellow spot. C: Leaf white spot. D: White lines on the leaf. E: Black lines on the leaf

图7 盐胁迫下突变体植株的表型鉴定A：正常条件下；B：250 mmol/LNaCl；每个处理3次生物学重复

Fig. 7 Phenotypic characterization of mutant plants under salt stressA: Under normal conditions; B: 250 mmol/L NaCl. Three biological replicates per treatment

表3 盐碱胁迫对突变体形态指标的影响

Table 3 Effects of saline-alkali stress on mutant morphological indexes

品种 Material	NaCl （mmol/L）	苗高 Seedling height （cm）	最大根长 Maximum root length （cm）	初生根数 Number of initial roots
宁春4号 Ningchun No. 4	0	14.12a	8.92a	5.47a
宁春4号 Ningchun No. 4	250	15.16b	9.96b	5.62b
突变体 Mutant	0	14.12a	9.77c	5.46a
突变体 Mutant	250	18.44c	12.34d	5.74c

表4 盐碱胁迫对突变体苗期物质积累的影响

Table 4 Effects of saline-alkali stress on the accumulation of substances at the seedling stage of mutant

品种 Material	NaCl （mmol/L）	地上部鲜重 Shoot fresh weight （g/plant）	根鲜重 Root fresh weight （g/plant）	地上部干重 Shoot dry weight （g/plant）	根干重 Root dry weight （g/plant）	根冠比 Root-shoot ratio （%）
宁春4号 Ningchun No. 4	0	0.181a	0.049a	0.024a	0.008a	27.07a
宁春4号 Ningchun No. 4	250	0.186b	0.061b	0.029b	0.009a	32.80b
突变体 Mutant	0	0.182a	0.052c	0.025a	0.008a	28.57c
突变体 Mutant	250	0.239c	0.089d	0.038c	0.010a	37.24d

图8 盐胁迫下突变体植株丙二醛含量（A）和细胞膜透性（B）的变化

Fig. 8 Changes in malondialdehyde content (A) and cell membrane permeability (B) in mutant plants under salt stress

图9 盐胁迫下突变体植株脯氨酸（A）和可溶性糖含量（B）的变化

Fig. 9 Changes in proline (A) and soluble sugar content (B) in mutant plants under salt stress

图10 盐胁迫下突变体植株抗氧化酶活性的变化

Fig. 10 Changes in antioxidant enzyme activities of mutant plants under salt stress

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