LI Xueyan,ZHOU Lihong,WANG Weidong,et al.Establishment of in vitro Regeneration System of Lilium regale Wilson From Flower Organs[J].Northern Horticulture,2022,(11):59-66.[doi:10.11937/bfyy.20215129]
岷江百合花器官离体再生体系的建立
- Title:
- Establishment of in vitro Regeneration System of Lilium regale Wilson From Flower Organs
- 文献标志码:
- A
- 摘要:
- 以岷江百合（Lilium regale Wilson）为试材，采用花丝、花柱、子房为外植体进行组织培养，研究不定芽或愈伤组织诱导、愈伤组织继代、小鳞茎膨大及生根以及试管苗的炼苗移栽，以期为建立岷江百合花器官的组培快繁技术提供参考依据。结果表明：3种花器官外植体诱导能力存在差异，由易到难依次为花丝＞花柱＞子房，发生褐化程度也存在差异，其中花丝最易发生褐化，而子房则极不易发生褐化；不同外植体的最适诱导培养基不同，花丝最适诱导培养基为MS+BA 1.0 mg?L-1+NAA 0.5 mg?L-1，总诱导率高达91%；花柱诱导最佳培养基为MS+2,4-D 1.0 mg?L-1+KT 0.1 mg?L-1，总诱导率为63%左右；子房诱导最佳培养基为MS+2,4-D 2.0 mg?L-1+KT 0.05 mg?L-1，总诱导率为52%；花器官诱导过程中主要产生2种愈伤组织，BA与NAA诱导主要获得绿色、致密、坚硬的愈伤组织，经石蜡切片鉴定为非胚性愈伤组织，而2,4-D与KT诱导主要获得黄色、疏松、颗粒状的愈伤组织，石蜡切片鉴定为胚性愈伤组织；对胚性愈伤组织进行继代培养，愈伤组织最适继代培养基为MS+PIC 1.0 mg?L-1+NAA 0.2 mg?L-1；小鳞茎膨大及生根最适培养基为MS+90 g?L-1蔗糖；鳞茎越大（直径>10 mm），移栽成活率越高（90%），植株长势越好。
- Abstract:
- Lilium regale Wilson was used as the test material,the filaments,styles and ovules were used as explants for tissue culture.Adventitious bud or callus induction,callus subculture,bulb expansion and rooting,and seedling training and transplanting of test tube plantlets were studied in order to provide reference for the establishment on the tissue culture and rapid propagation technology of flower organs of Lilium regale Wilson.The results showed that the induction ability of explants of three flower organs was different,the order of induction ability from easy to difficult was filament>style>ovary.Filaments were the most prone to browning,while ovaries were the least prone to browning.The optimal medium of different explants was different.The most suitable medium for filament explants was MS+BA 1.0 mg?L-1+NAA 0.5 mg?L-1 with the highest induction rate reaching 91%,the best medium for style induction was MS+2,4-D 1.0 mg?L-1+KT 0.1 mg?L-1,showing 63% induction rate.Two kinds of callus were produced in the process of flower organ induction,and the best explants for ovary induction was MS+2,4-D 2.0 mg?L-1+KT 0.05 mg?L-1,with 52% induction rate.BA and NAA were mainly induced to produce green,dense and hard callus,which were identified as non-embryonic callus by paraffin section;2,4-D and KT were mainly induced to produce yellow,loose and granular callus,which were identified as embryonic by paraffin section.The embryo callus was subcultured,the most suitable subculture medium was MS+PIC 1.0 mg?L-1+NAA 0.2 mg?L-1;while the most suitable medium for small scale stem enlargement and rooting was MS+90 g?L-1 sucrose;the larger the bulb (>10 mm),the higher the transplant survival rate (90%),the better the plant growth.
参考文献/References:
[1]潘会堂,张启翔.花卉种质资源与遗传育种研究进展[J].北京林业大学学报,2000,22(1):81-86.[2]夏宜平,高晓辰,郑慧俊.百合种球酝酿国产化[J].中国花卉园艺,2003(22):11-13.[3]高洁,王元忠,黄衡宇.绿花百合胚性愈伤组织诱导与植株再生研究[J].植物研究,2016,36(1):52-57.[4]BAE K,YOON E.Plant regeneration through the callus culture induced from bulb scales of an endangered species Lilium cernum Komarvo.[J].Journal of Plant Biotechnology,2013,40(2):65-71.[5]张梅,张杰,王强,等.南川百合种子萌发及组织培养快繁技术研究[J].农业与技术,2020,40(18):140-143.[6]曾洲德.泸定百合四种不同外植体组织培养技术研究[D].成都:四川农业大学,2014.[7]刘芳,王晓丽,张彦妮,等.垂花百合花器官离体培养[J].草业科学,2012,29(12):1894-1898.[8]顾地周,赵淑玲,郭伟,等.条叶百合和垂花百合高效快繁体系的建立[J].福建农林大学学报(自然科学版),2009,38(3):243-247.[9]孙道阳,牛立新,张延龙,等.野生百合花器官愈伤组织诱导及生根研究[M]//张启翔.中国观赏园艺研究进展.北京：中国林业出版社，2016.[10]姚绍娥,杨美纯,凌征柱.亚洲百合花器官组织培养再生植株研究[J].安徽农业科学,2010,38(7):3459-3460.[11]张彦妮,李兆婷,张艳波,等.毛百合试管鳞茎形成和膨大的培养优化[J].江苏农业科学,2016,44(4):74-78．[12]赵静,帅明蓉,赵玉飞,等.南川百合组织培养与快繁技术体系研究[J].西南师范大学学报(自然科学版),2012,37(6):109-115.[13]徐茜,余鸿燕,徐燕,等.重庆野生百合花蕾诱导再生植株技术研究[J].中国农学通报,2014,30(7):121-125.[14]陈丽静,葛菱,张丽,等.朝鲜百合鳞茎诱导及再生体系建立[J].中国农学通报,2011,27(19):121-124.[15]SAHOO M R,DEVI M P,DASGUPTA M,et al.An efficient protocol for in vitro regeneration and conservation of Shirui lily (Lilium mackliniae Sealy):A lab-to-land approach to save the rare endangered Asiatic lily species[J].In Vitro Cellular & Developmental Biology-Plant,2018,54:701-710.[16]赵祥云,王树栋,陈新露,等.百合[M].北京:中国农业出版社,2000.[17]NHUT D T,van LE B,TANAKA M,et al.Shoot induction and plant regeneration from receptacle tissues of Lilium longiflorum[J].Scientia Horticulturae,2001,87(1):131-138.[18]ARZATEFERNANDEZ A M,NAKAZAKI T,OKUMOTO Y,et al.Efficient callus induction and plant regeneration from filaments with anther in lily (Lilium longiflorum Thunb.)[J].Plant Cell Reports,1997,16(12):836-840.[19]马怡迪.野生百合高效再生体系建立及愈伤组织增殖调控研究[D].杭州:浙江大学农业与生物技术学院,2015.[20]吴青青,王维泽,崔嵬,等.百合茎尖培养材料的筛选及其组培配方的优化[J].贵州农业科学,2019,47(9):69-73.[21]杨薇红,张延龙,童斌,等.亚洲百合花器官的组培快繁技术研究[J].中国农学通报,2004,20(5):193-195.[22]孙红梅,宋胜利,申屠玥,等.亚洲百合Strawberry and Cream花器官组培快繁技术研究[J].沈阳农业大学学报,2015,46(1):7-12.[23]徐洁婷,王月,唐克轩,等.麝香百合花部组织离体培养与植株再生[J].海交通大学学报(农业科学版),2008,26(1):13-16.[24]崔祺,贾桂霞.“OT”杂种系百合‘Bay watch’离体再生体系的建立[J].东北林业大学学报,2014,42(5):34-38.[25]李莺,李星,李生玲,等.‘黄天霸’百合花器官愈伤组织诱导及植株再生[J].热带作物学报,2013,34(8):1507-1512.[26]翟彦,张宗勤,贾敏,等.百合体细胞胚胎发生和植株再生[J].西北植物学报,2011,31(4):834-841.[27]TAKAYAMA S,MISAWA M.Differentiation in Lilium bulbscales grown in vitro:Effect of various cultural conditions[J].Physiol Plant,1979,46:184-190.[28]周玲云,高素萍,陈锋.蔗糖和光周期在泸定百合试管鳞茎膨大中的作用机制[J].浙江大学学报(农业与生命科学版),2016,42(4):435-441.[29]吕翠竹,王有国,王立.绿花百合组培芽增殖与试管鳞茎膨大的影响因素研究[J].西南林业大学学报(自然科学版),2020,40(1):38-45.[30]张洁,蔡宣梅,林真,等.百合试管鳞茎诱导及膨大技术的研究[J].福建农业学报,2010,25(3):328-331.
相似文献/References:
[1]白杜娟,杨宝生,白岗栓.秦巴山区野生百合资源及保护措施[J].北方园艺,2017,41(05):71.[doi:10.11937/bfyy.201705017]
BAI Dujuan,YANG Baosheng,BAI Gangshuan.Wild Lily Resources and Protection in Qin-ba Mountains[J].Northern Horticulture,2017,41(11):71.[doi:10.11937/bfyy.201705017]
[2]蒋瑶,陈文波,陈其兵.野生湖北百合[STBX]HSFA2[STBZ]耐热基因的克隆及表达分析[J].北方园艺,2022,(22):67.[doi:10.11937/bfyy.20220715]
JIANG Yao,CHEN Wenbo,CHEN Qibing.Cloning and Expression Characterization of Heat-resistant Gene [STBX]HSFA2[STBZ] in Wild Lily[J].Northern Horticulture,2022,(11):67.[doi:10.11937/bfyy.20220715]
备注/Memo
第一作者简介：李雪艳（1986-），女，博士，助理研究员，现主要从事百合栽培与育种等研究工作。E-mail：lixueyan321@163.com.责任作者：杨迎东（1973-），男，硕士，研究员，现主要从事百合栽培与育种及种球繁育等研究工作。E-mail：yangyingdong2011@163.com.基金项目：中国博士后科学基金资助项目（2021M693845）；辽宁省重点研发资助项目（2020JH2/10200016）；辽宁省科技特派资助项目(2020020015-JH5/101)；辽宁省民生科技计划资助项目（2021JH2/10200026）。收稿日期：2022-12-16