Effects of EMS Mutagenesis on Seed Germination and Seedling Growth of Pomegranate

doi:10.15933/j.cnki.1004-3268.2025.08.013

Abstract

Abstract: This study investigated the mutagenesis effect of ethyl mesylate（EMS）on pomegranate seeds，and determined the appropriate mutagenesis dose for germplasm innovation of pomegranate seeds，to explore new techniques and methods for the breeding of new varieties of pomegranate. Pomegranate seeds were treated with EMS solutions of five different mass fractions（0，0.2%，0.4%，0.6%，0.8%），and the germination time，germination potential，germination rate，emergence rate，seedling formation rate，phenotypic variance，phenotypic variation rate and other indicators were measured，and the seedling height，ground diameter，root length，root number and other morphological indicators were measured.Physiological and biochemical indexes such as superoxide dismutase（SOD）activity，peroxidase（POD）activity，catalase（CAT）activity and ascorbate peroxidase（APX）activity were detected in the leaves of seedlings. The results showed that，after EMS treatment，the germination time of seeds was delayed，and the higher the concentration，the more the delay.The germination potential and germination rate of seeds were increased slightly after 0.2% treatment，and the other three treatments were significantly decreased，and the higher the concentration，the more the decrease.The seedling success rate gradually decreased with the increase of EMS mass fraction，reaching a minimum of 11.67% at 0.8%.The variation rate gradually increased with the increase of EMS mass fraction，reaching a maximum of 5.34% at 0.8%.The average plant height and average root length of seedlings were decreased，except for a slight increase after 0.2% treatment.The average ground diameter of seedlings was decreased，and the higher the concentration was，the greater the decrease was.The average root number of seedlings was decreased by the other three treatments except for 0.2% treatment，which had no change，and the higher the concentration was，the greater the decrease was.The activities of SOD，POD，CAT and APX in the leaves of seedlings were increased，and with the increase of the concentration，the increase amplitude showed a trend of first increasing and then decreasing，and gradually increased with the increase of concentration in 0.2%—0.6%，and the increase amplitude decreased significantly at 0.8%.EMS generally showed an inhibitory effect on the germination of pomegranate seeds and the growth of seedlings，and EMS with an appropriate mass fraction could enhance the stress resistance of plants.Considering the seedling formation rate，variation rate，physiological and biochemical indexes，the optimal EMS mutagenesis dose for pomegranate seeds ranges from half lethal dose（LD₅₀）to critical dose（LD₄₀），that is 0.54%—0.63%.

Key words: Pomegranate, Seed germination, Ethyl mesylate, Mutagenesis breeding, Seedling growth, Physiology and biochemistry

摘要： 探究甲磺酸乙酯（EMS）对石榴种子的诱变效应，确定EMS诱变石榴种子适宜的剂量，为石榴新品种选育探索新技术、新方法。用5种不同质量分数（0、0.2%、0.4%、0.6%、0.8%）的EMS溶液处理石榴种子，测定种子的发芽时间、发芽势、发芽率、出苗率、成苗率、表型变异数、表型变异率等指标，测量幼苗株高、地径、根长、根数等形态指标，检测幼苗叶片中超氧化物歧化酶（SOD）活性、过氧化物酶（POD）活性、过氧化氢酶（CAT）活性、抗坏血酸过氧化物酶（APX）活性等生理生化指标。结果表明，EMS处理后种子的发芽时间推迟，且质量分数越大推迟越多。种子的发芽势、发芽率除EMS 0.2%处理后略有上升外，其他3个处理均大幅度下降，且质量分数越大下降越多。成苗率随EMS质量分数的增大逐渐降低，在EMS 0.8%时最低，为11.67%，变异率随EMS质量分数的增大逐渐升高，在EMS 0.8%时最高，为5.34%。幼苗的平均株高、平均根长除EMS 0.2%处理略有增加外，其他3个处理均有所减少，且质量分数越大减少越多。幼苗的平均地径4个处理均有所减少，且质量分数越大减少越多。幼苗平均根数除EMS 0.2%处理没有变化外，其他3个处理均有减少，且质量分数越大减少越多。幼苗叶片SOD、POD、CAT、APX活性均有所升高，且随着质量分数的增加，升高幅度表现出先增大后减小的趋势，在EMS 0.2%~0.6%，随质量分数增加逐渐增大，EMS 0.8%处理时升高幅度出现了明显减小。EMS对石榴种子萌发及幼苗生长整体上表现为抑制作用，适宜质量分数的EMS能提高植物的抗逆性，综合考虑成苗率、变异率、生理生化指标，石榴种子的最适EMS诱变剂量为半致死剂量（LD₅₀）至临界剂量（LD₄₀）的范围，即0.54%~0.63%。

关键词: 石榴, 种子萌发, 甲磺酸乙酯, 诱变育种, 幼苗生长, 生理生化

CLC Number:

S665.4

GAO Xiaofeng, ZHANG Qiuyue, ZHOU Xiaojing, ZUO Weifang, HUANG Rantao, GUO Shuangshuang, YUAN Chaozheng, ZHENG Mingyan. Effects of EMS Mutagenesis on Seed Germination and Seedling Growth of Pomegranate[J]. Journal of Henan Agricultural Sciences, 2025, 54(8): 133-139.

高小峰, 张秋月, 周晓静, 左卫芳, 黄冉涛, 郭双双, 源朝政, 郑明燕. EMS 诱变对石榴种子萌发及幼苗生长的影响[J]. 河南农业科学, 2025, 54(8): 133-139.

Figures/Tables 4

References

［1］郝兆祥，李志强，张忠涛. 中国石榴盆景［M］.北京：中国林业出版社，2024.

HAO Z X，LI Z Q，ZHANG Z T. Chinese pomegranate bonsai［M］.Beijing：China Forestry Publishing House，2024.

［2］冯玉增. 中国石榴［M］. 北京：中国农业科学技术出版社，2023.

FENG Y Z.Chinese pomegranate［M］.Beijing：China Agricultural Science and Technology Press，2023.

［3］侯乐峰，罗华，毕润霞，等.我国石榴育种四十年回顾与展望［J］. 北方园艺，2022（24）：139‑147.

HOU L F，LUO H，BI R X，et al. Review and prospect of pomegranate breeding in China in the past 40 years［J］.Northern Horticulture，2022（24）：139‑147.

［4］侯乐峰，罗华，王艳芹，等. 耐贮软籽石榴新品种‘赤艳’的选育［J］. 中国果树，2021（11）：63‑64.

HOU L F，LUO H，WANG Y Q，et al. Breeding of a new storage resistance soft‑seed pomegranate cultivar ‘Chiyan’［J］. China Fruits，2021（11）：63‑64.

［5］秦改花，刘春燕，黎积誉，等. 石榴新品种‘妃红’［J］. 园艺学报，2022，49（S1）：37‑38.

QIN G H，LIU C Y，LI J Y，et al. A new pomegranate variety‘Feihong’［J］. Acta Horticulturae Sinica，2022，49（S1）：37‑38.

［6］秦改花，刘春燕，黎积誉，等. 石榴新品种‘酥籽’［J］. 园艺学报，2022，49（S1）：39‑40.

QIN G H，LIU C Y，LI J Y，et al. A new pomegranate variety‘Suzi’［J］. Acta Horticulturae Sinica，2022，49（S1）：39‑40.

［7］王庆军，朱薇，李雪，等. 鲜食中熟红皮石榴新品种江石榴的选育［J］. 中国果树，2023（2）：101‑102.

WANG Q J，ZHU W，LI X，et al. Breeding of a new fresh and mid‑ripening red‑skinned pomegranate cultivar ‘Jiangshiliu’［J］. China Fruits，2023（2）：101‑102.

［8］胡青霞，万然，史江莉，等. 石榴软籽新品种‘豫农早艳’的选育［J］. 果树学报，2021，38（1）：146‑149.

HU Q X，WAN R，SHI J L，et al. A new soft‑seeded pomegranate cultivar ‘Yunongzaoyan’［J］. Journal of Fruit Science，2021，38（1）：146‑149.

［9］李永锋，王先伟，范静，等. 石榴新品种露红宝石的选育［J］.中国果树，2022（10）：52‑54.

LI Y F，WANG X W，FAN J，et al. Breeding of a new pomegranate cultivar‘Luhongbaoshi’［J］.China Fruits，2022（10）：52‑54.

［10］李好先，曹尚银，骆翔，等. 超软籽石榴新品种‘中石榴5号’的选育［J］.果树学报，2020，37（12）：1987‑1990.

LI H X，CAO S Y，LUO X，et al. A new super soft‑seed pomegranate cultivar‘Zhongshiliu 5’［J］.Journal of Fruit Science，2020，37（12）：1987‑1990.

［11］李好先，陈利娜，姚方，等. 超软籽石榴新品种‘中石榴4号’的选育［J］.中国果树，2021（8）：76‑78.

LI H X，CHEN L N，YAO F，et al. Breeding of a new super soft‑seed pomegranate cultivar‘Zhongshiliu 4’［J］. China Fruits，2021（8）：76‑78.

［12］罗华，张晋，张聪宽，等. 软籽石榴新品种桔美人的选育［J］.果树学报，2024，41（4）：791‑795.

LUO H，ZHANG J，ZHANG C K，et al. A new soft‑seed pomegranate cultivar Jumeiren［J］. Journal of Fruit Science，2024，41（4）：791‑795.

［13］董颖苹，连勇，何庆才，等. 植物化学诱变技术在育种中的运用及其进展Ⅰ. 化学诱变技术及诱变效率［J］.种子，2005，24（7）：54‑58.

DONG Y P，LIAN Y，HE Q C，et al. Progress and application of plant chemical mutagenesis technology on breeding. Ⅰ. Chemical mutagenesis technology and mutagenic effect［J］. Seed，2005，24（7）：54‑58.

［14］张瑞成，李魏，潘素君，等. 化学诱变在种质资源改良上的应用［J］. 分子植物育种，2017，15（12）：5189‑5196.

ZHANG R C，LI W，PAN S J，et al. Application of chemical mutagenesis in improving germplasm resource［J］. Molecular Plant Breeding，2017，15（12）：5189‑5196.

［15］CHHABRA B，SINGH L，WALLACE S，et al. Screening of an ethyl methane sulfonate mutagenized population of a wheat cultivar susceptible to Fusarium head blight identifies resistant variants［J］. Plant Disease，2021，105（11）：3669‑3676.

［16］HOSSEN Q M M，BADIER RAHMAN S M，RAHMAN M N，et al. Development of early flowering，short life‑spanned jute（Corchorus spp.）mutant via ethyl methane sulfonate mutagenesis［J］. Journal of Crop Science and Biotechnology，2022，25（4）：489‑500.

［17］CHEN J G，WANG F，LEI B Q，et al.Modulation of growth duration，grain yield and nitrogen recovery efficiency by EMS mutagenesis under OsNRT2.3b overexpression background in rice［J］.Agriculture，2022，12（6）：799.

［18］杨子博，张勇，刘畅，等. 利用EMS诱变创制淮麦33抗赤霉病突变体［J］.南方农业学报，2023，54（4）：1010‑1017.

YANG Z B，ZHANG Y，LIU C，et al. Creation of Huaimai 33 Fusarium head blight resistant mutants by EMS mutation［J］. Journal of Southern Agriculture，2023，54（4）：1010‑1017.

［19］连晓荣，王晓娟，周玉乾，等. EMS诱变玉米自交系郑58后代变异分析及应用评价［J］.玉米科学，2024，32（1）：11‑19.

LIAN X R，WANG X J，ZHOU Y Q，et al. Variation analysis and application evaluation of progeny of maize inbred line Zheng 58 by EMS chemical mutation［J］.Journal of Maize Sciences，2024，32（1）：11‑19.

［20］曹亚萍，武银玉，刘博，等. 基于EMS诱变的济麦22抗旱节水突变体创制与鉴定［J］.麦类作物学报，2024，44（3）：315‑322.

CAO Y P，WU Y Y，LIU B，et al. Creation and identification of Jimai 22 mutants with drought‑resistant and water‑saving features induced by EMS［J］. Journal of Triticeae Crops，2024，44（3）：315‑322.

［21］吴茜，蒋丹，文欢，等. 水稻‘9311’突变体库构建及磷响应突变体筛选［J］.分子植物育种，2023，21（1）：168‑173.

WU Q，JIANG D，WEN H，et al. Construction of rice ‘9311’ mutant library and screening of mutants response to phosphorus［J］. Molecular Plant Breeding，2023，21（1）：168‑173.

［22］王黎明，陈君燚，姬敬敬，等. 小麦穗型突变体（库）的创制、鉴定及其遗传分析［J］.种子，2022，41（1）：117‑122.

WANG L M，CHEN J Y，JI J J，et al. Creation，identification and genetic analysis of library of spike‑type wheat mutants［J］. Seed，2022，41（1）：117‑122.

［23］康勋杰，刘士尧，冀占东. 利用EMS诱变技术创制弱感光水稻种质资源［J］.杂交水稻，2024，39（3）：48‑54.

KANG X J，LIU S Y，JI Z D. New rice germplasm with weak photosensitivity created by EMS mutagenesis［J］.Hybrid Rice，2024，39（3）：48‑54.

［24］周玉彩，肖栋，李英，等. 不同浓度EMS诱变不结球白菜的效果分析［J］.中国蔬菜，2024（7）：42‑49.

ZHOU Y C，XIAO D，LI Y，et al. Effects of different concentrations of EMS on mutagenic non‑heading Chinese cabbage［J］. China Vegetables，2024（7）：42‑49.

［25］朱泯珏，陈橙，史文甯，等. 达乌里胡枝子愈伤组织及种子EMS诱变研究［J］.种子，2024，43（3）：1‑10.

ZHU M J，CHEN C，SHI W N，et al. Study on EMS mutagenesis of callus and seeds of Lespedeza davurica［J］.Seed，2024，43（3）：1‑10.

［26］卢苗苗，谢明芳，白邦琴，等. 两种冷季型草坪草的EMS诱变及突变体筛选［J］.草业科学，2024，41（4）：810‑821.

LU M M，XIE M F，BAI B Q，et al.Ethyl methanesulfonate mutagenesis and mutant screening of two cold‑season turfgrasses［J］. Pratacultural Science，2024，41（4）：810‑821.

［27］孙明洋，王萍. 籽用美洲南瓜EMS诱变突变体库的构建及表型分析［J］.西北植物学报，2024，44（8）：1283‑1294.

SUN M Y，WANG P. Construction and phenotypic analysis of EMS mutant library of the seed‑used pumpkin［J］.Acta Botanica Boreali‑Occidentalia Sinica，2024，44（8）：1283‑1294.

［28］SURAKSHITHA N C，SOORIANATHASUNDARAM K，GANESAN N M. Determination of mutagenic sensitivity of hardwood cuttings of grapes ‘Red Globe’ and ‘Muscat’（Vitis vinifera L.）to gamma rays［J］.Scientia Horticulturae，2017，226：152‑156.

［29］张治安，陈展宇. 植物生理学实验技术［M］. 长春：吉林大学出版社，2008.

ZHANG Z A，CHEN Z Y. Experimental techniques of plant physiology［M］.Changchun：Jilin University Press，2008.

［30］王长泉，刘峰，李雅志. 果树诱变育种的研究进展［J］.核农学报，2000，14（1）：61‑64.

WANG C Q，LIU F，LI Y Z. Research progress on mutation breeding of fruit trees［J］.Journal of Nuclear Agricultural Sciences，2000，14（1）：61‑64.

［31］金鸽，张铭芳，魏蕾，等. EMS诱变对岷江百合种子萌发的影响［J］.种子，2023，42（3）：81‑87.

JIN G，ZHANG M F，WEI L，et al. Effect of EMS mutagenesis on Lilium regale Wilson seed germination［J］.Seed，2023，42（3）：81‑87.

［32］黎诗艳，阮景军，范昱，等. EMS诱变剂处理对苦荞种子萌发及主要农艺性状的影响［J］. 分子植物育种，2021，19（3）：914‑922.

LI S Y，RUAN J J，FAN Y，et al. Effects of EMS mutagenesis inducing treatment on seed germination and main agronomic traits of Tartary buckwheat［J］.Molecular Plant Breeding，2021，19（3）：914‑922.

［33］李颜方，杜艳伟，王高鸿，等. EMS诱变对晋谷21号种子萌发的影响［J］. 种子，2020，39（11）：83‑87.

LI Y F，DU Y W，WANG G H，et al. Effects of EMS mutagenesis on seed germination of rice variety Jingu 21［J］.Seed，2020，39（11）：83‑87.

［34］谢玲玲，周火强，弭宝彬，等. EMS诱变技术研究概况及应用进展［J］.湖南农业科学，2020（6）：92‑95.

XIE L L，ZHOU H Q，MI B B，et al. Research progress and application of EMS mutagenesis technology［J］.Hunan Agricultural Sciences，2020（6）：92‑95.

［35］沙国飞，鲁敏，安华明. EMS诱变对刺梨种子出苗及早期生长性状的影响［J］. 北方园艺，2022（2）：33‑39.

SHA G F，LU M，AN H M. Effects of EMS mutagenesis on Rosa roxburghii Tratt. seed emergence and its early growth traits［J］. Northern Horticulture，2022（2）：33‑39.

［36］伊风艳，孙海莲，王晓娟，等. EMS诱变对蒙古国黄花苜蓿种子萌发和幼苗生长的影响［J］.种子，2018，37（10）：41‑44.

YI F Y，SUN H L，WANG X J，et al. Effect of EMS mutagenesis on seed germination and seedling growth of Medicago falcata L.in Mongolia［J］. Seed，2018，37（10）：41‑44.

［37］ATTIA H，KARRAY N，LACHAÂL M. Light interacts with salt stress in regulating superoxide dismutase gene expression in Arabidopsis［J］.Plant Science，2009，177（3）：161‑167.

［38］SHAFI A，GILL T，SREENIVASULU Y，et a1.Improved callus induction，shoot regeneration，and salt stress tolerance in Arabidopsis overexpressing superoxide dismutase from Potentilla atrosanguinea［J］.Protoplasma，2015，252（1）：41‑51.

［39］GONDIM F A，GOMES‑FILHO E，COSTA J H，et al.Catalase plays a key role in salt stress acclimation induced by hydrogen peroxide pretreatment in maize ［J］. Plant Physiology and Biochemistry，2012，56：62‑71.

［40］温日宇，刘建霞，肖东东，等. EMS诱变对藜麦种子萌发及幼苗生理特性的影响［J］.分子植物育种，2024，22（8）：2656‑2662.

WEN R Y，LIU J X，XIAO D D，et al. Effects of EMS mutagenesis on seed germination and physiological characteristics of quinoa seedlings［J］.Molecular Plant Breeding，2024，22（8）：2656‑2662.

［41］王炜，陈军，陈琛，等. EMS诱变创制胡麻新种质的研究［J］.核农学报，2023，37（10）：1931‑1937.

WANG W，CHEN J，CHEN C，et al. Creating new germplasm of oil flax by EMS mutation［J］. Journal of Nuclear Agricultural Sciences， 2023， 37 （10）：1931‑1937.

［42］范昕琦，王海燕，陈静，等. EMS诱变对高粱成苗及M1主要农艺性状的影响［J］. 生物技术通报，2023，39（7）：173‑184.

FAN X Q，WANG H Y，CHEN J，et al. Effects of EMS mutagenesis on the seeding survival and major agronomic traits of sorghum in M1 generation［J］.Biotechnology Bulletin，2023，39（7）：173‑184.

［43］翟永琪，贾利，马绍鋆，等. EMS诱变对西瓜种子萌发及农艺性状的影响［J］.中国蔬菜，2023（1）：52‑59.

ZHAI Y Q，JIA L，MA S J，et al. Effects of EMS mutagenesis on seed germination and agronomic traits of watermelon［J］. China Vegetables，2023（1）：52‑59.

[1]	TIAN Ye, YANG Peihua, YANG Tingqian, LIU Xiaocen, LIU Yiqing, WANG Jiao, HU Haijun, JIANG Xinchen, ZHANG Wanshun, ZHU Yongxing. Effect of Exogenous Nano‐Silicon on Seed Germination of Pepper under Salt Stress [J]. Journal of Henan Agricultural Sciences, 2025, 54(6): 110-120.
[2]	ZHANG Zhen, ZHOU Lei, CHEN Yongzhong, ZHANG Ying, WANG Rui, MIAO Ziqi, LI Zhigang, LONG Ling, XU Yanming. Effects of Slow Release Fertilizer Dosage on the Growth and Nutrient Content of Camellia oleifera Container Seedlings [J]. Journal of Henan Agricultural Sciences, 2025, 54(6): 63-71.
[3]	DENG Cong, MA Lu, WANG Qingsong, FU Jian, WANG Yufeng, YANG Kejun. Effect of Bacillus on Seed Germination and Physiological and Biochemical Characteristics of Maize under Salt⁃Alkali Stress [J]. Journal of Henan Agricultural Sciences, 2025, 54(3): 20-30.
[4]	QI Xianke, LI Miao, LI Caihong, QU Chenling. Effects of PAW and Its Combined Action with LTP on Seed Germination and Seedling Growth of Wheat [J]. Journal of Henan Agricultural Sciences, 2024, 53(4): 20-29.
[5]	PANG Xiaowen, XIE Qinming. Effect of Two Seaweed Fermentation Solutions on Seed Germination and Seedling Growth of Cabbage [J]. Journal of Henan Agricultural Sciences, 2024, 53(12): 131-138.
[6]	TANG Qian, WANG Jiali, LI Wenqiang. Effects of Extracellular Polysaccharides on Seed Germination，Seedling Growth and Physiological Characteristics of Schizonepeta tenuifolia under Salt Stress [J]. Journal of Henan Agricultural Sciences, 2024, 53(10): 96-105.
[7]	GUO Menglu, LI Jianqi, LU Yu. Effects of Phytohormones and Chelating Agents on the Development and Remediation Effect of Portulaca oleracea L.under Cadmium Stress [J]. Journal of Henan Agricultural Sciences, 2023, 52(9): 102-109.
[8]	LEI Mengyao, GAO Xiaofeng, BAI Qingmin, DENG Ke, ZUO Weifang, LI Yuying. Evaluation of Cold Resistance of Pomegranate Branches from Different Varieties [J]. Journal of Henan Agricultural Sciences, 2023, 52(6): 120-130.
[9]	YUE Hewei, LI Lianzhen, WANG Yuqing, WANG Rongrong, LIU Yang, WEI Junjie, CHANG Jianfeng. Effect of IAA on Seed Germination and Physiological Characteristics of Polygonatum sibiricum Red. [J]. Journal of Henan Agricultural Sciences, 2023, 52(3): 64-72.
[10]	GUO Li, SUO Sumin, YANG Hongli, XU Minghui, CHANG Jietian. Seed Germination Characteristics and Salt Resistance of Wild Leptodermis oblonga Bunge [J]. Journal of Henan Agricultural Sciences, 2022, 51(8): 128-133.
[11]	YIN Juan, ZHU Qingsong, ZHAO Jingjing, CAO Yongyi, MA Guihua. Effects of Ultrasound on Seed Germination and Seedling Physiological Characteristics of Echinacea purpurea under Salt Stress [J]. Journal of Henan Agricultural Sciences, 2022, 51(8): 117-127.
[12]	LI Qian, JIA Yan, LI Xinyi, LI Qinmei, LIU Song, WANG Mengyu, HUANG Haiquan, HUANG Meijuan. Effects of Plant Growth‑Promoting Bacteria Containing ACC Deaminase on Seed Germination of Psammosilene tunicoides [J]. Journal of Henan Agricultural Sciences, 2022, 51(5): 45-52.
[13]	ZHANG Huan, GAO Xiaofeng, LEI Mengyao, TAO Aili, LI Yuying, YAO Gang. Multivariate Analysis of the Relationship between Soil Nutrients and Fruit Quality and Optimization of Soil Nutrients in Soft‑seed Pomegranate Orchards [J]. Journal of Henan Agricultural Sciences, 2022, 51(4): 111-119.
[14]	LI Jiewei, CHI Yongqing, WU Tianfang, ZHU Yangchun, DING Fenghua. Effects of Cadmium and Lead Combined Stress on Seed Germination and Seedling Growth of Herbaceous Flowers [J]. Journal of Henan Agricultural Sciences, 2022, 51(12): 122-130.
[15]	LIU Jiayao, CHENG Yan, WEI Zunmiao, LONG Wei, WANG Liang, MU Zhongsheng. Effects of Exogenous Hormones on Growth，Yield and Physiological Characteristics of Tigernut（Cyperus esculentus L.）under Drought Stress [J]. Journal of Henan Agricultural Sciences, 2022, 51(11): 42-49.