甜柠檬实生后代四倍体创制及其分子细胞遗传组成鉴定  

Development of tetraploid sweet lemon seedling progeny and identification of its molecular and cytogenetic composition

作　　者：何礼[1] 张国艳 李文婷 关斌[1] 陈克玲[1] 刘建军[1] 何建[1] HE Li;ZHANG Guo-yan;LI Wen-ting;GUAN Bin;CHEN Ke-ling;LIU Jian-jun;HE Jian(National-Local Joint Engineering Laboratory of Citrus Breeding and Cultivation/Horticulture Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066,China)

机构地区：[1]柑橘育种与栽培国家地方联合工程实验室/四川省农业科学院园艺研究所,成都610066

出　　处：《西南农业学报》2025年第1期41-47,共7页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(32272671);四川省自然科学基金项目(23NSFSC1003);四川省“十四五”果树育种攻关项目(2021YFYZ0023-02);四川省农业科学院1+9“揭榜挂帅”重点学科领域科技攻关任务(1+9KJGG001、1+9KJGG003);国家现代农业(柑橘)产业技术体系(CARS-26);四川水果创新团队(SCCXTD-2024-4)。

摘　　要：【目的】创制甜柠檬(Citrus lumia Risso)多倍体种质,创建其分子和细胞遗传组成鉴定新体系,为种质资源创新和鉴定利用提供新方法体系。【方法】通过实生苗根尖染色体计数筛选甜柠檬多倍体材料,与二倍体对比分析多倍体的叶形、气孔等植物学性状。利用47对分布于柑橘9条染色体上的SSR标记,分析多倍体甜柠檬的亲本来源。利用柑橘全部9条染色体特异的寡核苷酸荧光原位杂交(Oligo-FISH)涂染探针,精准鉴定多倍体甜柠檬的染色体组成。【结果】染色体计数表明获得四倍体甜柠檬材料1份(2n=4x=36)。与二倍体对照相比,其表现出叶形指数变小、叶片增厚、气孔指数变大、气孔密度降低。47对SSR标记扩增结果显示,四倍体和二倍体甜柠檬拥有完全一致的遗传条带。Oligo-FISH涂染显示,在四倍体中每个染色体特异探针均识别4条同源染色体,无染色体间重排。分子标记和细胞遗传学鉴定结果共同表明,该四倍体甜柠檬系二倍体自然加倍得到的同源四倍体。【结论】创新了柑橘类多倍体材料创制及分子细胞遗传组成鉴定体系,丰富了甜柠檬特色种质资源。【Objective】The paper aimed to create polyploid germplasm of sweet lemon(Citrus lumia Risso),and establish a new system for identifying its molecular and cytogenetic composition,which provided a novel method system for the innovation,identification and utilization of germplasm resources.【Method】Polyploid material of sweet lemon was screened by chromosome counting of root tips from seedlings.Leaf shape,stomata and other botanical characters between polyploid and the diploid control plants were compared and analyzed,respectively.The parental origin of polyploid sweet lemon was analyzed using 47 pairs of SSR markers distributed across 9 chromosomes of Citrus.The chromosomal composition of the polyploid sweet lemon was identified by fluorescence in situ hybridization with oligonucleotide probes(Oligo-FISH),each of which was specificed to one of the 9 chromosomes of Citrus,respectively.【Result】A tetraploid sweet lemon material was obtained and confirmed by chromosome counting(2n=4x=36).Compared with its diploid control,the tetraploid sweet lemon showed a smaller leaf shape index,thicker leaves,a larger stomatal index and a lower stomatal density.The results of 47 pairs of SSR markers distributed across 9 chromosomes showed that the tetraploid and diploid sweet lemons had identical genetic bands.In the tetraploid,FISH painting with oligonucleotide probes revealed that each chromosome specific probe recognized 4 homologous chromosomes,without any chromosomal rearrangements were detected.The results of molecular markers and cytogenetic identification together indicated that the tetraploid sweet lemon was an autotetraploid derived from natural chromosomal doubling of the diploid.【Conclusion】The development of Citrus polyploid material and its genetic composition identification system through molecular and cytogenetic methods are innovated.Additionally,the study enriches the characterized germplasm resources of sweet lemon.

关 键 词：柑橘 甜柠檬 多倍体 寡核苷酸荧光原位杂交 SSR分子标记 

分 类 号：S666.5[农业科学—果树学]