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Molecular cytogenetic map visualizes the heterozygotic genome and identifies translocation chromosomes in Citrus sinensis

查看全文 作　　者：Shipeng [1,2]Song;Hui [1,2]Liu;Luke [2]Miao;Li [3]He;Wenzhao [2,4]Xie;Hong [1,5]Lan;Changxiu [1,2]Yu;Wenkai [6]Yan;Yufeng [6]Wu;Xiao-peng [7]Wen;Qiang [1]Xu;Xiuxin [1]Deng;Chunli [1,2]Chen 高影响力作者 机构地区：[1]National Key Laboratory for Germplasm Innovation and Utilization for Fruit and Vegetable Horticultural Crops,Hubei Hongshan Laboratory,Wuhan,Hubei 430070,China;[2]College of Life Science and Technology,Huazhong Agricultural University,Wuhan,Hubei 430070,China;[3]National-local Joint Engineering Laboratory of Citrus Breeding and Cultivation/Horticulture Institute,Sichuan Academy of Agricultural Sciences,Chengdu,Sichuan 610066,China;[4]National Key Laboratory of Crop Genetics and Improvement,Huazhong Agricultural University,Wuhan,Hubei 430070,China;[5]Hubei Province Engineering Research Center of Legume Plants,College of Life Science,Jianghan University,Wuhan,Hubei 430056,China;[6]State Key Laboratory for Crop Genetics and Germplasm Enhancement,Bioinformatics Center,Academy for Advanced Interdisciplinary Studies,Nanjing Agricultural University,Nanjing,Jiangsu 210095,China;[7]Key Laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education),Institute of Agrobioengineering,College of Life Science,Guizhou University,Guiyang,Guizhou 550025,China高影响力机构 出　　处：《Journal of Genetics and Genomics》索引2023年第50卷第6期,共12页高影响力期刊 基　　金：supported by the National Natural Science Foundation of China(31970525,31672112);the National Key Research and Development Project of China(2019YFD1001401-GJ03);Advanced Foreign Experts Project(G2021157012L,G20200017071);Cultivating Fund Project of Hubei Hongshan Laboratory(2022hspy002);Fundamental Research Funds for the Central Universities(2662018PY099)from Chinese government;Breeding of breakthrough new citrus varieties and creation of new germplasms,Science and Technology Planning Project of Sichuan Province(2021YFYZ0023-02);Comprehensive experimental station of middle and late maturing citrus in Chengdu,China Agriculture Research System(CARS-26)。 摘　　要：Citrus sinensis is the most cultivated and economically valuable Citrus species in the world,whose genome has been assembled by three generation sequencings.However,chromosome recognition remains a problem due to the small size of chromosomes,and difficulty in differentiating between pseudo and real chromosomes because of a highly heterozygous genome.Here,we employ fluorescence in situ hybridization(FISH)with 9 chromosome painting probes,30 oligo pools,and 8 repetitive sequences to visualize 18 chromosomes.Then,we develop an approach to identify each chromosome in one cell through single experiment of oligo-FISH and Chromoycin A3(CMA)staining.By this approach,we construct a high-resolution molecular cytogenetic map containing the physical positions of CMA banding and 38 sequences of FISH including centromere regions,which enables us to visualize significant differences between homologous chromosomes.Based on the map,we locate several highly repetitive sequences on chromosomes and estimate sizes and copy numbers of each site.In particular,we discover the translocation regions of chromosomes 4 and 9 in C.sinensis“Valencia.”The high-resolution molecular cytogenetic map will help improve understanding of sweet orange genome assembly and also provide a fundamental reference for investigating chromosome evolution and chromosome engineering for genetic improvement in Citrus. 关 键 词：Chromosomalmarker FISH CMA banding Genomic heterozygosity Chromosomal translocation Sweet orange
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