[1]穆京妹,吕兴娜,聂玲玲,等.万寿菊花药培养双单倍体诱导技术初探[J].北方园艺,2026,(17):28-36.[doi:10.11937/bfyy.20260501]
　MU Jingmei,LYU Xingna,NIE Lingling,et al.Preliminary Exploration of Doubled Haploid Induction Through Anther Culture in Marigold[J].Northern Horticulture,2026,(17):28-36.[doi:10.11937/bfyy.20260501]
点击复制

万寿菊花药培养双单倍体诱导技术初探

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 期数: 2026年17 页码: 28-36 栏目: 出版日期: 2026-09-15
Title:
Preliminary Exploration of Doubled Haploid Induction Through Anther Culture in Marigold
文章编号:
1001-0009(2026)17-0028-09
作者:
穆京妹1; 吕兴娜2; 聂玲玲2; 曹世哲2; 高伟2; 3
(1.河北工程大学 生命科学与食品工程学院,河北 邯郸 056038;2.晨光生物科技集团股份有限公司,河北 邯郸 056250;3.河北省植物资源综合利用重点实验室,河北 邯郸 056107)
Author(s):
MU Jingmei1; LYU Xingna2; NIE Lingling2; CAO Shizhe2; GAO Wei2; 3
(1.School of Life Science and Food Engineering,Hebei University of Engineering,Handan，Hebei 056038;2.Chenguang Biotech Group Limited Co.，Ltd.，Handan，Hebei 056250;3.Key Laboratory of Comprehensive Utilization of Plant Resources of Hebei Province，Handan，Hebei 056107)
关键词:
万寿菊; 花药培养; 双单倍体; 染色体鉴定; 秋水仙素加倍
Keywords:
marigold; anther culture; doubled haploid; chromosome identification; colchicine-induced doubling
分类号:
S681.9
DOI:
10.11937/bfyy.20260501
文献标志码:
A
摘要:
以国外万寿菊主栽品种‘XH-1’、国内主栽品种‘CD-F1’和自主培育品种‘1-26-5’的花药为试材，采用花药离体培养方法，研究了花蕾预处理方法与花药取材方法、花药愈伤诱导培养基、分化培养基及生根培养基配方对万寿菊花药培养的影响，以期获得万寿菊新种质，缩短选育年限。结果表明：当花蕾长度为17.77~21.31 mm、小花长度为3.00~4.00 mm时，花药愈伤诱导率和花药培养苗获得率最高。通过单因素试验确定最佳愈伤诱导培养基为MS固体培养基+2 mg·L-1 6-BA+0.5 mg·L-1 NAA+15 g·L-1蔗糖+50 mL·L-1椰子汁+0.25 g·L-1酪蛋白氨基酸+0.05 g·L-1抗坏血酸，诱导率达85.8%；最佳不定芽诱导培养基为MS固体培养基+2 mg·L-16-BA+0.7 mg·L-1 NAA+2.5 g·L-1AgNO3，不定芽诱导率达90%；MS培养基与1/2MS培养基均可诱导不定芽生根，生根率为95%。利用该方法获得32株万寿菊花药培养苗，经染色体鉴定得到1株单倍体苗，通过秋水仙素加倍成功获得双单倍体苗。该研究成功获得了万寿菊新种质，丰富了万寿菊种质资源。
Abstract:
Anthers of the foreign main cultivar ‘XH-1’,the domestic main cultivar ‘CD-F1’,and the self-bred cultivar ‘1-26-5’ were used as explants.Anther culture in vitro was employed to investigate the effects of bud pretreatment methods,anther sampling methods,and the compositions of callus induction medium,differentiation medium,and rooting medium on marigold anther culture,with the aim of obtaining new marigold germplasms and shortening the breeding period.The results showed that the optimal bud length (17.77-21.31 mm) and floret length (3.00-4.00 mm) gave the highest callus induction rate and plantlet regeneration rate.Through single-factor experiments,the optimal callus induction medium was determined as MS solid medium supplemented with 2 mg·L-1 6-BA,0.5 mg·L-1 NAA,15 g·L-1 sucrose,50 mL·L-1 coconut milk,0.25 g·L-1 casein hydrolysate,and 0.05 g·L-1 ascorbic acid,achieving a callus induction rate of 85.8%.The optimal adventitious bud induction medium was MS solid medium supplemented with 2 mg·L-1 6-BA,0.7 mg·L-1 NAA,and 2.5 g·L-1 AgNO3,giving an adventitious bud induction rate of 90%.Both MS and 1/2MS media induced rooting of adventitious buds,with a rooting rate of 95%.Using this method,32 marigold plantlets derived from anther culture were obtained,among which one haploid plantlet was identified by chromosome analysis,and a doubled haploid plantlet was successfully obtained after colchicine treatment.This study successfully obtained new marigold germplasms,enriching the genetic resources of marigold.

参考文献/References:

［1］RAUT THANESHWARI S.Commercial application of lutein:Marigold flower pigment［J］.Ecology,Environment and Conservation,2022:S311-S316.［2］张世远,高伟,吕兴娜,等.喷施腐殖酸肥对万寿菊现蕾期农艺性状与生理指标的影响［J］.北方园艺,2024(16):41-48.［3］胡相伟,郑杰,汪斌强,等.Na2SO4胁迫对万寿菊幼苗生长及其生理特性的影响［J］.北方园艺,2025(14):56-65.［4］张华丽,宋利娜,孙丽萍,等.万寿菊杂交结实条件及促进其结实技术［J］.北方园艺,2025(17):19-26.［5］王炜,陈琛,叶春雷,等.花药培养与麦谷蛋白亚基分子标记结合选育小麦新品种的研究［J］.麦类作物学报,2019,39(3):277-282.［6］沈胜,尹延旭,李宁,等.辣椒花药培养研究进展［J/OL］.分子植物育种,(2023-12-20)［2026-05-06］.https://link.cnki.net/urlid/46.1068.S.20231219.1718.037.［7］贾利,马绍鋆,江海坤,等.茄子细胞工程技术应用研究进展［J］.安徽农业大学学报,2022,49(4):565-573.［8］李艳敏,蒋卉,符真珠,等.芍药花药愈伤组织诱导及体细胞胚发生［J］.植物学报,2021,56(4):443-450.［9］李甫,唐道城,梁顺祥.生长调节剂对万寿菊花药培养的影响［J］.青海大学学报(自然科学版),2007,25(1):51-53.［10］KUMAR K R,SINGH K P,BHATIA R,et al.Optimising protocol for successful development of haploids in marigold (Tagetes spp.) through in vitro androgenesis［J］.Plant Cell,Tissue and Organ Culture (PCTOC),2019,138(1):11-28.［11］宋雪娇,陶承光,王平.色素万寿菊小孢子发育时期与花器形态的相关性［J］.湖北农业科学,2010,49(9):2154-2157.［12］李甫.万寿菊小孢子时期鉴定及花药培养研究［D］.西宁:青海大学,2010.［13］GERMAN M A.Anther culture for haploid and doubled haploid production［J］.Plant Cell,Tissue and Organ Culture (PCTOC),2011,104(3):283-300.［14］李慧,赵林姝,古佳玉,等.小麦花药培养体系优化及高再生力基因型的筛选［J］.植物遗传资源学报,2022,23(3):738-745.［15］牛文霞.观赏植物花药培养及其影响因素研究进展［J］.种子科技,2022,40(19):27-29.［16］齐迎春,叶要妹,刘国锋,等.不同基因型孔雀草高效植株再生体系的建立［J］.中国农业科学,2005,38(7):1414-1417.［17］ARISANDI D P,PARADISA F V,SUGIHARTO B,et al.Effect of ethylene inhibitor,type of auxin,and type of sugar on anther culture of local East Java aromatic rice varieties［J］.Journal of Crop Science and Biotechnology,2020,23(4):367-373.［18］ISLAM M T,ARIF M R,HASAN M T,et al.Anther culture in crop plants:Progress and perspectives［J］.Plant Breeding and Biotechnology,2023,11(2):69-96.［19］YONG J W H,GE L,NG Y F,et al.The chemical composition and biological properties of coconut (Cocos nucifera L.) water［J］.Molecules,2009,14(12):5144-5164.［20］沈跃鹏.不同生长调节剂对小麦成熟胚愈伤组织诱导效果研究［J］.农业科技通讯,2023(7):54-58.［21］王玉书,沈静怡,耿伟,等.小型西瓜花药愈伤组织诱导及防褐化研究［J］.北方园艺,2018(10):41-44.［22］孟凡娟,刘沛公,刘天娇,等.蓖麻花药愈伤组织诱导的研究［J］.作物杂志,2009(6):59-63.［23］李颜方,杜艳伟,张正,等.农杆菌介导谷子成熟胚遗传转化体系的建立与优化［J］.作物杂志,2019(3):73-79.［24］李惠.谷子不同外植体再生体系建立及优化的研究［D］.太谷:山西农业大学,2015.［25］LIU Z Y,JI J J,JIANG F,et al.Establishment of a genetic transformation system for Codonopsis pilosula callus［J］.Plant Biotechnology,2022,39(3):251-257.

相似文献/References:

[1]缑艳霞,张明方.西瓜花药离体培养影响因子研究[J].北方园艺,2013,37(10):117.
　GOU Yan-xia,ZHANG Ming-fang.Study on Factors Affecting Anther Culture in virto of Citrullus lanatus[J].Northern Horticulture,2013,37(17):117.
[2]伍亚平,唐道城.万寿菊雄性不育系离体保存及快繁体系的建立[J].北方园艺,2013,37(01):116.
　WU Ya-ping,TANG Dao-cheng.Conservation in vitro and Establishment of the Propagation System on Male Sterile Lines of Marigold[J].Northern Horticulture,2013,37(17):116.
[3]周春娥,朱凤荣,周延清.怀地黄花药培养影响因素的初步研究[J].北方园艺,2012,36(04):128.
　ZHOU Chun-e,ZHU Feng-rong,ZHOU Yan-qing.Study on Influential Factors on Anther Culture of Rehmannia glutinosa Libosch[J].Northern Horticulture,2012,36(17):128.
[4]胡雨彤,时连辉,刘登民,等.牛粪代替泥炭作栽培基质对万寿菊生长的影响[J].北方园艺,2013,37(20):60.
　HU Yu-tong,SHI Lian-hui,LIU Deng-min,et al.Effects of Different Ratios of Decomposed Cattle Dung as Peat Substitute on Substrate Properties and Growth of Tagetes erecta [J].Northern Horticulture,2013,37(17):60.
[5]赵 庆,唐道城.万寿菊根尖染色体观测方法的优化[J].北方园艺,2013,37(20):98.
　ZHAO Qing,TANG Dao-cheng.Optimization of Microscope Observation Method on Root-tip Chromosomes of Marigold[J].Northern Horticulture,2013,37(17):98.
[6]郑杰,李素琴,黄红梅.万寿菊组织培养体系的建立[J].北方园艺,2012,36(06):111.
　ZHENG Jie,LI Su-qin,HUANG Hong-mei.Study on the Tissue Culture System of Tagetes erecta L.[J].Northern Horticulture,2012,36(17):111.
[7]王立凤,姜 明,姜述君,等.施钾对万寿菊生物学性状及产量的影响[J].北方园艺,2012,36(22):69.
　WANG Li-feng,JIANG Ming,JIANG Shu-jun,et al.Effect of Potassium Fertilizer Application on Biology Character and Yield of Tagetese recta[J].Northern Horticulture,2012,36(17):69.
[8]汪殿蓓,张胜红,李建华.响应曲面法优化万寿菊叶黄素的提取工艺[J].北方园艺,2012,36(22):111.
　WANG Dian-pei,ZHANG Sheng-hong,LI Jian-hua.Optimization of Extraction Conditions of Xanthophyll from Tagetes erecta L. by Response Surface Methodology[J].Northern Horticulture,2012,36(17):111.
[9]刘敏,厉悦,梁艳,等.不同浓度氯化钠胁迫对万寿菊幼苗生长及生理特性的影响[J].北方园艺,2013,37(24):63.
　LIU Min,LI Yue,LIANG Yan,et al.Effect of Different Concentration of NaCl Stress on the Growth and Some Physiological Characteristics of Tagetes erecta Seedlings[J].Northern Horticulture,2013,37(17):63.
[10]冯倩倩,陈东亮,程曦,等.万寿菊黑斑病的研究进展[J].北方园艺,2015,39(20):181.[doi:10.11937/bfyy.201520045]
　FENG Qianqian,CHEN Dongliang,CHENG Xi,et al.Research Progress of Black Spot on Marigold[J].Northern Horticulture,2015,39(17):181.[doi:10.11937/bfyy.201520045]

备注/Memo

第一作者简介：穆京妹（1987-），女，博士，讲师，现主要从事菊科植物育种和分子标记开发等研究工作。E-mail:1029468136@qq.com.责任作者：高伟（1980-），男，博士，高级工程师，现主要从事天然产物提取及分析等研究工作。E-mail:gw@ccgb.cn.基金项目：邯郸市科学技术研究与发展计划资助项目（2211201008）；河北工程大学横向课题资助项目（HK2023000543）；河北工程大学博士专项基金资助项目(SJ2101003086)。收稿日期：2026-02-06

更新日期/Last Update: 2026-09-17