中国农业科学 ›› 2020, Vol. 53 ›› Issue (23): 4961-4968.doi: 10.3864/j.issn.0578-1752.2020.23.020

• 研究简报 • 上一篇    

以本地早橘和槾橘为母本倍性杂交创制柑橘三倍体

解凯东1(),彭珺1,袁东亚1,强瑞瑞1,谢善鹏1,周锐1,夏强明1,伍小萌1,柯甫志2,刘高平3,GROSSER Jude W4,郭文武1()   

  1. 1华中农业大学园艺林学学院/园艺植物生物学教育部重点实验室,中国武汉 430070
    2浙江省农业科学院柑桔研究所,中国浙江台州 318020
    3浙江省台州市黄岩区果树技术推广总站,中国浙江台州 318020
    4美国佛罗里达大学柑橘研究与教育中心,美国佛罗里达 33850
  • 收稿日期:2020-09-01 接受日期:2020-10-14 出版日期:2020-12-01 发布日期:2020-12-09
  • 通讯作者: 郭文武
  • 作者简介:解凯东,Tel:027-87287393;E-mail: xiekaidong@mail.hzau.edu.cn
  • 基金资助:
    国家自然科学基金国际合作重点项目(31820103011);湖北省科技支撑计划(2020BBA036);广东省科技计划(2018B020202009);中央高校基本科研业务费专项资金(2662019QD048);中央高校基本科研业务费专项资金(2662018PY007)

Production of Citrus Triploids Based on Interploidy Crossing with Bendizao and Man Tangerines as Female Parents

XIE KaiDong1(),PENG Jun1,YUAN DongYa1,QIANG RuiRui1,XIE ShanPeng1,ZHOU Rui1,XIA QiangMing1,WU XiaoMeng1,KE FuZhi2,LIU GaoPing3,GROSSER Jude W4,GUO WenWu1()   

  1. 1College of Horticulture & Forestry Sciences, Huazhong Agricultural University/Key Laboratory of Horticultural Plant Biology (Ministry of Education), Wuhan 430070, China
    2Citrus Research Institute, Zhejiang Academy of Agricultural Sciences, Taizhou 318020, Zhejiang, China
    3Huangyan Fruit Tree Technology Promotion General Station, Taizhou 318020, Zhejiang, China
    4Citrus Research and Education Center, IFAS, University of Florida, Lake Alfred FL 33850, USA
  • Received:2020-09-01 Accepted:2020-10-14 Online:2020-12-01 Published:2020-12-09
  • Contact: WenWu GUO

摘要:

【目的】基于二倍体与四倍体倍性杂交策略创制柑橘三倍体新种质。【方法】以二倍体为母本,四倍体为父本进行人工授粉,授粉后85 d采摘幼果并对未成熟种子实施幼胚离体挽救培养;再生植株后,用流式细胞仪和根尖染色体计数法对再生植株进行倍性鉴定;并用SSR标记对三倍体后代进行分子鉴定。【结果】以本地早橘和槾橘为母本,8个异源四倍体体细胞杂种和1个双二倍体为父本,配置9个倍性杂交组合,共授粉2 749朵花,坐果489个,平均坐果率17.8%;培养幼嫩种子2 239粒,经幼胚离体挽救培养,共再生植株260株;用流式细胞仪和根尖染色体计数检测再生植株倍性,获得三倍体141株;SSR分子鉴定表明从槾橘×NS组合随机选取的50株三倍体后代全部为双亲的有性后代;三倍体后代在温室生长一年后,采用嫁接(枳砧)将这些三倍体定植于田间。【结论】这些三倍体新种质为本地早橘和槾橘无核新品种培育奠定了宝贵的材料基础。

关键词: 柑橘, 无核育种, 三倍体, 幼胚离体挽救培养, 流式细胞仪

Abstract:

【Objective】The aim of this study was to create the citrus triploids based on diploid and tetraploid ploidy cross strategy. 【Method】The artificial pollination was conducted with diploid as female parent and tetraploid as male parent. Young fruits were sampled at 85 d after pollination (DAP) and immature seeds were extracted and subjected to in vitro embryo rescue. Following plantlet regeneration from the embryos, their ploidy level was determined by flow cytometry and root-tip chromosome counting, as well as the genetic origin determined using simple sequence repeat (SSR) markers. 【Result】A total of nine interploidy crosses were carried out by using Bendizao tangerine and Man tangerine as female parents and eight allotetraploid somatic hybrids and one doubled diploids as male parents. From all crosses, 2 749 flowers were pollinated and 489 fruits were set, with an average fruit setting rate of 17.8%. By conducting in vitro immature embryo rescue, totally 260 plants were regenerated from 2 239 seeds cultured. By determining their ploidy level using flow cytometry and root-tip chromosome counting, 141 seedlings were proven to be triploids. SSR analysis showed that all of 50 randomly selected triploid plants from Man tangerine × NS were the hybrids of their both parents. After one year growing in greenhouse, all triploids were grafted onto Poncirus trifoliata in the field to accelerate flowering. 【Conclusion】These triploid citrus plants obtained herein provided elite materials for potential seedless variety selection.

Key words: Citrus, seedless breeding, polyploid, embryo rescue, flow cytometry