柑橘13个多胚品种同源四倍体高效发掘与分子鉴定  

Efficient exploration and SSR identification of autotetraploids from the seedlings of thirteen apomictic Citrus genotypes

作　　者：陈昊 谢善鹏 解凯东[1] 肖公傲 周锐 伍小萌 吴群 邓家锐 敖义俊 刘高平 郭文武[1] CHEN Hao;XIE Shanpeng;XIE Kaidong;XIAO Gong’ao;ZHOU Rui;WU Xiaomeng;WU Qun;DENG Jiarui;AO Yijun;LIU Gaoping;GUO Wenwu(National Key Laboratory for Germplasm Innovation&Utilization of Horticultural Crops,Huazhong Agricultural University,Wuhan 430070,Hubei,China;Chenzhou Institute of Agricultural Science,Chenzhou 423000,Hunan,China;Quzhou Academy of Agricultural and Forestry Sciences,Quzhou 324000,Zhejiang,China;Chenggu Fruit Technology Guidance Station,Chenggu 723200,Shaanxi,China;Huangyan Fruit Tree Technology Promotion General Station,Taizhou 318020,Zhejiang,China)

机构地区：[1]华中农业大学果蔬园艺作物种质创新与利用全国重点实验室,武汉430070 [2]郴州市农业科学研究所,湖南郴州423000 [3]衢州市农业林业科学研究院,浙江衢州324000 [4]城固县果业技术指导站,陕西城固723200 [5]浙江省台州市黄岩区果树技术推广总站,浙江台州318020

出　　处：《果树学报》2023年第11期2297-2306,共10页Journal of Fruit Science

基　　金：国家自然科学基金项目(32172525);南宁市科技重大专项(20222061);浙江省黄岩区农业揭榜挂帅项目(JBGS2022-HYNY02);华中农业大学深圳营养健康研究院项目(SZYJY2022009);国家柑橘产业技术体系项目(CARS-26)。

摘　　要：【目的】基于柑橘珠心细胞存在自然加倍的特点,实生播种发掘千山红蜜橘等13个品种的同源四倍体。【方法】种子催芽萌发后实生播种,待幼苗长出3片以上真叶后,依据形态特征初选法“观根辨叶看油胞”从实生苗中筛选疑似多倍体,再通过流式细胞仪倍性分析与根尖染色体压片计数对疑似多倍体进一步鉴定倍性,并通过SSR分子标记分析鉴定所获多倍体的遗传来源。【结果】基于形态初选,分别从9个地方特色品种千山红蜜橘、八月橘、衢橘、早橘、扁平橘、瓯柑、狮头柑、冰糖橙、锦蜜冰糖橙和4个砧木品种磨坪香橙、日本香橙、枳雀、油皮金柑的343、499、892、385、519、290、457、241、119、690、828、114、129株实生苗中发掘获得2、1、3、2、7、3、1、3、1、3、17、1、2株疑似多倍体;用流式细胞仪对以上46株疑似多倍体进行倍性鉴定,获得45株四倍体和1株衢橘六倍体,采用根尖染色体计数法验证了上述结果;SSR分子鉴定表明,13个品种的45株四倍体所扩增条带大小与其相应二倍体亲本完全一致,推测其可能是由二倍体亲本珠心细胞自然加倍形成的同源四倍体。【结论】发掘的四倍体资源不仅丰富了我国柑橘多倍体类型,而且为我国柑橘无核育种和矮化广适砧木选育奠定了珍贵的材料基础。【Objective】Citrus is a crucial part of Chinese fruit crops.There are abundant citrus germplasm resources in China,but many excellent local varieties are gradually eliminated by the market due to the problem of numerous seeds within the fruit.The fruits of triploid plants are generally seedless because of their sterile male and female gametes.Therefore,triploid production is a promising strategy to breed seedless cultivars in citrus.Triploids can be obtained by interploidy crossing between diploids and tetraploids.However,the tetraploid germplasm is rare,which limits the application of this strategy.Exploration of tetraploids is an important prerequisite for triploid production with the aim to improve the seedy local cultivars in our country.For the rootstock improvement,tetraploid plants are also valuable resources because of their higher metabolite content,and better resistance than their diploid parents.In this study,we planned to explore tetraploid plants from 13 local cultivars in our country by using the traits of spontaneous doubling of the nucellar cells in polyembryonic citrus varieties.The exploration of tetraploids from the above 13 local cultivars will not only provide excellent tetraploid parents for the production of triploid plants,but also lay the foundation for the basic research about the effect of genome duplication on some important trait change,such as dwarfing,extensive adaptivity and higher medicinal value in tetraploids.【Methods】After the mature fruits were harvested,the seeds were extracted and the seed coats were peeled off,and then they were placed in a thermostat to accelerate germination.When the seeds germinated,they were sown in pots and cultivated in a plant growth chamber.After the seedlings grew up with three or more leaves,putative polyploids were screened according to the morphological feature showing lower height,shorter taproots,less lateral roots,thicker and rounder leaves and declined oil gland density.The ploidy levels of these putative polyploids were further confi

关 键 词：柑橘 多倍体 流式细胞仪 SSR分子标记 无核育种 

分 类 号：S666[农业科学—果树学]