利用横向薄层培养技术建立番木瓜植株再生体系  

Establishment of Plant Regeneration System of Papaya by Transverse Thin Cell Layer Culture

作　　者：魏岳荣[1] 幸洁华 陈爱君 戴沛美 周陈平 杨敏 邝瑞彬[1] 黄炳雄[1] 杨护[1] Wei Yuerong;Xing Jiehua;Chen Aijun;Dai Peimei;Zhou Chenping;Yang Min;Kuang Ruibin;Huang Binxiong;Yang Hu(Guangdong Province Key Laboratary ofTropical and Subtropical Fruit Tree Rescarch,Key Laboratory of South Subtropical Fruit Biology and Genetic Resource Utilization(MOA),Institute of Fruit Tree Rescarch,Guangdong Academy of Agricultural Sciences,Guangzhou,510640;College of Biology and Food Engineering,Guangdong University of Education,Guangzhou,510303)

机构地区：[1]广东省农业科学院果树研究所农业部南亚热带果树生物学及遗传资源利用重点实验室广东省热带亚热带果树研究重点实验室,广州510640 [2]广东第二师范学院生物与食品工程学院,广州510303

出　　处：《分子植物育种》2021年第6期1962-1968,共7页Molecular Plant Breeding

基　　金：广东省重点领域研发计划项目(2019B020214005);广东省科技计划公益研究与能力建设项目(2017A03030-3037);广东省现代农业产业技术体系创新团队建设项目(优稀水果产业)(2019KJ116);2019年“攀登计划”广东大学生科技创新专项资金项目(pdjh2019a0352)共同资助。

摘　　要：建立一个高效稳定的植株再生系统是番木瓜生物技术育种和品种改良的关键。本研究通过横向薄层培养技术,以‘一尺瓜’番木瓜品种为材料,通过对田间成年结果树截干后萌生的侧芽进行消毒处理,经过初代培养和增殖培养后获得大量丛生分化芽。以健壮分化芽为材料横向切取1~2.5 mm厚度的薄层6~10片,经不定芽诱导和增殖培养后进行生根培养,成功获得植株再生。研究结果表明,采用300 mg/L利福平溶液浸泡并在摇床上140 r/min振荡处理1 h、75%酒精处理30 s、0.1%的氯化汞溶液(W/V)处理7~8 min的组合递进式消毒处理方法,可使侧芽接种培养成功率达到85%以上。分化芽薄层在MS培养基+30 mg/L蔗糖+0.1 mg/L6-BA+0.2 mg/L NAA+0.3 mg/L KT+0.1 mg/L IBA的条件下诱导培养20 d,单个分化芽切取的薄层平均可获得7.04个不定芽,其中第3号薄片出芽能力最强,平均出芽数可达到3.4个,最高5.0个。采用将叶尖代替分化芽茎基部插入培养基的茎段悬空植叶促根方法进行分化芽的促根培养,生根培养基组成为1/2MS培养基+3%蔗糖(W/V)+2 mg/L IBA,20~25 d后平均出根率可达到87.5%,根系无结构疏松和愈伤化现象,假植成活率可高达95%以上,有效改善了常规生根技术存在的根系质量差和假植成活率低的问题。本研究结果可提高番木瓜组培快繁效率,并为番木瓜诱变育种和分子育种提供技术支撑。Establishing an efficient and stable plant regeneration system is the key to biotechnology breeding and variety improvement of Carica papaya L..In this experiment,’Yi Chigua’variety(Carica papaya L.)was used as experimental material for transverse thin cell layer(t TCL)culture.The lateral buds germinated from the adult plants in the field after cut off for 20~30 days were taken as explants.A large number of cluster-like differentiated shoots were obtained from the lateral buds through disinfection,initial culture and proliferation.A combined progressive disinfection method was established,which is as followings:explants with 300 mg/L rifampicin solution shaked in a speed of 140 r/min for 1 hour,then immersed into 70%ethanol for 30 seconds and surface disinfected with 0.1%(W/V)mercuric chloride for 7~8 minutes.The survival rate of lateral buds inoculation and culture was over 85%.Healthy and stout shoots were transversely sliced into 6~10 TCL sections approximately 1~2.5 mm thick and inoculated onto MS medium with 3%sucrose(W/V)supplemented with 0.1 mg/L 6-BA+0.2 mg/L NAA+0.3 mg/L KT+0.1 mg/L IBA.The average number of adventitious shoot buds induced from TCL sections was 7.04 per explant in 20 d culture period.No.3 section exhibited a high ability of adventitious shoot bud induction with an average number of 3.4,and the highest number of 5.0.Rooting of shoots was achieved to 87.5%average rooting rate with good root quality in 20~25 days when the leaves instead of the shoot base were inserted into half-strength MS medium with 3%sucrose(W/V)supplemented with 2 mg/L IBA.These plantlets were transplanted into loose medium and grew into healthy seedlings with over 95%survival rate.This technique can effectively improve the problem of poor root quality and low survival rate in conventional rooting techniques.The results of this study are helpful to improve the propagation efficiency and provide technical support for mutation breeding and molecular breeding of Carica papaya L..

关 键 词：番木瓜 薄层培养 分化芽 生根 植株再生 

分 类 号：S667.9[农业科学—果树学]