Construction and Screening of EMS Induced Mutant Library of Luffa

doi:10.13560/j.cnki.biotech.bull.1985.2025-0994

Abstract

Abstract:

Objective This study is aimed to screen for elite germplasm resources of luffa (Luffa spp.) by constructing an ethyl methanesulfonate (EMS) mutant library, and provide abundant foundational materials for genetic improvement, new variety breeding, and gene function research in luffa. Method Using seeds of the luffa advanced inbred line S199 as experimental material, EMS mutagenesis was conducted with different combinations of EMS concentration (1.4%, 1.8%, and 2.2%) and treatment duration (6, 10, and 14 h). The germination rate and seedling survival rate of the treated seeds were used as evaluation indicators to determine the optimal mutagenesis conditions and subsequently construct the mutant library. Phenotypic observation of the mutant progeny was carried out, focusing on the investigation, preservation, and statistical analysis of major agronomic traits-including leaf, fruit, plant architecture, and reproductive organ characteristics-in M₂ generation individuals. Result The optimal mutagenesis condition for luffa seeds was treated with 1.8% EMS for 10 h, resulting in germination and seedling survival rates of 51.67% and 46.33%, respectively, which was close to the semi-lethal dose. Seedlings under this condition generally had leaf wrinkling. A mutant library consisting of 526 M₂ families was constructed. A total of 78 mutant individuals were screened, including 29 with leaf variations, 18 with plant architecture variations, 26 with fruit shape variations, and 5 with variations in reproductive organs, encompassing 17 distinct phenotypes. The overall mutation rate was 4.60%. A mutant with glossy fruit peel was identified, its fruit surface was smooth and shiny, and it showed significantly improved post-harvest storage and transport tolerance. Genetic analysis indicated that this trait was controlled by a single recessive gene, making it a valuable material for further research. Conclusion A mutant library of luffa comprising 526 M₂ families is successfully constructed, with an overall phenotypic mutation rate of 4.60%.

Key words: luffa, EMS, mutant library, construction, phenotype analysis, new germplasm

MIN Zi-yang, LI Jia-jia, WU Qi, HU Xin-jun, HAN Xiao-xia, HAN Rong, LI Yong-qi. Construction and Screening of EMS Induced Mutant Library of Luffa[J]. Biotechnology Bulletin, 2026, 42(5): 272-279.

Figures/Tables 8

Table 1 Effects of different EMS concentration and treatment time on the germination rates and survival rates of luffa seeds

处理时间 Treatment time （h）	EMS浓度 EMS concentration （%）	发芽率 Germination rate （%）	成苗率 Survival rate （%）
6	0	83.67 a	77.33 a
	1.4	72.67 c	64.67 c
	1.8	54.67 e	42.33 f
	2.2	43.67 g	30.00 g
10	0	82.67 a	75.33 a
	1.4	61.00 d	52.00 d
	1.8	51.67 f	46.33 e
	2.2	44.00 g	27.33 h
14	0	77.00 b	70.00 b
	1.4	51.67 f	42.33 f
	1.8	34.33 k	27.33 h
	2.2	16.67 l	12.00 i

Fig. 1 Effects of EMS treatment on germinating and seedling phenotype of luffa seedsA-C: Germination potential, seedling rate and seedling growth status after PBS (phosphate buffer solution) treatment for 10 h. D-F: Germination potential, seedling rate and seedling growth after 1.8% EMS treatment for 10 h. White bar=1 cm, red bar=5 cm

Table 2 Statistics on mutation types of M2 generation luffa induced by EMS

突变器官 Mutant organ	突变表型 Mutant phenotype	突变株数 Number of mutated plants	突变频率 Mutation frequency （%）
叶片 Leaf	黄色嵌合 Yellow mosaic	11	0.65
	浅绿色 Light green	5	0.30
	深绿色 Dark green	5	0.30
	皱缩 Crimple	5	0.30
	圆叶 Round	3	0.18
株型 Plant type	矮化 Dwarf	3	0.18
	细弱 Weak	4	0.24
	花打顶 Blunt with blossom	2	0.12
	卷曲 Twisted	6	0.36
	徒长 Spindling	3	0.18
瓜型 Fruit shape	瓜长变短 Shorter	7	0.42
	瓜色变亮 Color brighter	1	0.06
	瓜粉增多 Powder increase	5	0.30
	瓜色嵌合 Color mosaic	7	0.42
	瓜长变长 Longer	6	0.36
生殖器官 Reproductiveorgans	雌花簇生 Clustered female	3	0.18
生殖器官 Reproductiveorgans	雄花败育 Yellowing male	2	0.12

Fig. 2 Mutant phenotypes of leaves in M2 populationA: Wild type. B: Yellow mosaic leaves. C: Light green leaves. D: Dark green leaves. E: Crimple leaves. F: Round leaves. Bar=1 cm

Fig. 3 Mutant phenotypes of plant type in M2 populationA: Wild type. B: Dwarf. C: Weak. D: Blunt with blossom. E: Twisted. F: Spindling. Bar =10 cm

Fig. 4 Mutant phenotypes of fruit shape in M2 populationA: Wild type. B: Fruit length shorter. C: Fruit color brighter. D: Fruit powder increase. E: Fruit color mosaic. F: Fruit length longer. Bar =10 cm

Fig. 5 Mutant phenotypes of reproductive organs in M2 populationA: Wild type female flowers. B: Wild type male flowers. C: Clustered female flowers. D: Yellowing male flowers. Bar =1 cm

Fig. 6 Mutant with glossy fruit peelA: Wild type fruit. B: Fruit with glossy fruit peel. Bar =5 cm

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