以东试早柚为母本创制柑橘三倍体种质资源  

Creation of triploid seedling plants of Citrus by crossing Dongshizao pummelo female with tetraploid male parents

作　　者：刘承浪 冯迪 曹宗洪 陶亚文 徐祥增 高世德[2] 岳建强[3] 谢宗周[1] 叶俊丽[1] 柴利军[1] 郭文武[1] 邓秀新[1] LIU Chenglang;FENG Di;CAO Zonghong;TAO Yawen;XU Xiangzeng;GAO Shide;YUE Jianqiang;XIE Zongzhou;YE Junli;CHAI Lijun;GUOWenwu;DENG Xiuxin(National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops/College of Horticulture and Forestry Sciences,Huazhong Agricultural University,Wuhan 430070,Hubei,China;Yunnan Institute of Tropical Crops,Jinghong 666100,Yunnan,China;Institute of Tropcal and Subtropical Cash Crops,Yunnan Academy of Agricultural Sciences,Baoshan 678000,Yunnan,China)

机构地区：[1]果蔬园艺作物种质创新与利用全国重点实验室·华中农业大学园艺林学学院,武汉430070 [2]云南省热带作物科学研究所,云南景洪666100 [3]云南省农业科学院热带亚热带经济作物研究所,云南保山678000

出　　处：《果树学报》2023年第10期2041-2049,共9页Journal of Fruit Science

基　　金：国家现代农业(柑橘)产业技术体系(CARS-26);云南省科技厅对外科技合作专项(202003AD150014)。

摘　　要：【目的】三倍体植物由于减数分离紊乱,难以形成可育的雌雄配子,属于天然的不育类型,配置以2x×4x多个杂交组合,旨在创制柑橘三倍体种质,丰富柑橘无核材料。【方法】以单胚性二倍体品种东试早柚为母本与四倍体柑橘材料ZP(纸皮,四倍体甜橙)、PT(四倍体葡萄柚)、NH(四倍体,诺瓦橘柚+HB柚体细胞杂种)为父本进行倍性杂交,授粉后85 d和100 d采摘幼果并对未成熟种子实施幼胚离体挽救培养,获得再生植株后用流式细胞仪和InDel标记对其倍性及遗传来源进行鉴定。【结果】从3个倍性杂交组合的168株再生植株中,通过倍性检测获得三倍体幼苗128株且均为双亲杂交后代,其中东试早柚×ZP共计60株、东试早柚×PT共计60株、东试早柚×NH共计8株。【结论】通过倍性杂交高效创制三倍体柑橘新种质,为柑橘早熟无核育种及相关基础研究提供了珍贵的育种材料。【Objective】Citrus is mainly cultivated for fresh consumption in China,so seedless citrus has become the dominant position in market consumption and in breeding programs.In order to enhance the competitiveness of the citrus industry and meet the market demands,it has been the goal of breeders to cultivate seedless varieties of citrus.Triploids are naturally seedless material whose chromosomes are disrupted during meiosis,making it difficult to form normal fertile gametes,resulting in seedless fruits.Triploid citrus typically has larger fruits and possesses stronger resistance and adaptation to the environmental conditions due to chromosome doubling.Ploidy crosses is the most effective strategy to obtain seedless citrus varieties,using monoembryonic diploid and tetraploid ploidy as parents.Therefore,we performed several 2x×4x crosses to create triploid citrus.【Methods】In this study,we selected the Dongshizao pummelo[C.grandis(L.)Osbeck Dongshizao pummelo]as the female parent,which is a local specialty cultivar from Yunnan province,characterized by early-maturing,seedlessness,high sugar and low acidity.Then,we used a late-maturing and productive autotetraploid ZP[C.sinensis(L.)Osbeck Paperrind orange],PT[C.paradisi(L.)Osbeck grapefruit]and an allotetraploid somatic hybrids NH[(C.reticulata Blanco×C.paradisi Macf.)+C.grandis(L.)Osbeck Hirado Buntan pummelo],as the male parent for the artificial crosses.In the earlier step,the pollinated styles were stained with aniline blue to observe the cross-compatibility between the parents,and in the subsequent steps,the immature seeds obtained from young fruits at 85 and 100 days after pollination(DAP)were cultured in vitro.When seeds grew into seedlings,the ploidy levels were analyzed by flow cytometry.In addition,we also resequenced the maternal Dongshizao pummelo,then the data was mapped to the HWB[Citrus grandis(L.)Osbeck Wanbai pummelo]reference genome to obtain InDel(insertion/deletion)variant loci,which contained 50-200 bp differences.According to these loci,pr

关 键 词：柑橘 东试早柚 胚挽救 倍性育种 INDEL 

分 类 号：S666[农业科学—果树学]