甘蓝BoFLCs基因标记开发及优异单倍型聚合  

Development of BoFLCs Gene Markers and Pyramiding of Elite Haplotypes in Cabbage(Brassica oleracea L.var.capitata)

作　　者：王娇 张斌 吴元康 杨丽梅[2] 庄木[2] 吕红豪[2] 王勇 季家磊 闫超 张扬勇[2] WANG Jiao;ZHANG Bin;WU Yuankang;YANG Limei;ZHUANG Mu;LYU Honghao;WANG Yong;JI Jialei;YAN Chao;ZHANG Yangyong(College of Horticulture,Qingdao Agricultural University,Shandong Province Engineering Laboratory of Genetic Improvement of Horticultural Crops,Qingdao 266109,Shandong,China;Institute of Vegetables and Flowers,Chinese Academy of Agriculture Sciences,State Key Laboratory of Vegetable Biobreeding,Key Laboratory of Horticultural Crop Biology and Germplasm Creation,Ministry of Agriculture and Rural Affairs,Beijing 100081,China)

机构地区：[1]青岛农业大学园艺学院,山东省园艺作物基因改良工程实验室,山东青岛266109 [2]中国农业科学院蔬菜花卉研究所,蔬菜生物育种全国重点实验室,农业农村部园艺作物生物学与种质创制重点实验室,北京100081

出　　处：《中国蔬菜》2024年第5期40-50,共11页China Vegetables

基　　金：湖北省乡村振兴科技支撑项目(2022BBA121);国家大宗蔬菜产业技术体系项目(CARS-23)。

摘　　要：开花抑制因子FLOWERING LOCUS C(FLC)编码MADS-box家族蛋白,在拟南芥的春化途径中起关键作用。为了探究甘蓝BoFLCs在春化过程中的作用,利用CRISPR/Cas9技术对甘蓝自交系GL的BoFLCs基因进行编辑,显著提早了其开花时间,明确了BoFLCs的基因功能。以晚花甘蓝自交系20-2221(P_(1),BoFLCs-w)和中早花甘蓝自交系20-2229(P_(2),BoFLCs-z)为亲本,构建BC_(1-2)P_(2)和BC_(2)F_(1)分离群体。通过序列比对分析,发现BoFLC_(1)、BoFLC3和BoFLC5在双亲间均存在序列差异,并根据这些差异序列开发了其特异性分子标记。利用BoFLCs特异性分子标记对BC_(2)F_(1)群体进行筛选,最终获得1个包含BoFLC_(1)-w/w、BoFLC5-w/w纯合单倍型以及BoFLC3-w/z杂合单倍型的植株(BC_(2)F_(1)-29),其抽薹时间和开花时间分别比回交亲本20-2229晚8.00 d和13.00 d;4个包含BoFLC_(1)-w/w、BoFLC3-w/w纯合单倍型以及BoFLC5-w/z杂合单倍型的植株(BC_(2)F_(1)-55、BC_(2)F_(1)-119、BC_(2)F_(1)-121和BC_(2)F_(1)-163),平均抽薹时间和开花时间比20-2229分别晚9.75 d和(13.50 d;3个包含BoFLC_(1)-w/w、BoFLC3-w/w)和BoFLC5-w/w纯合单倍型的植株(BC_(2)F_(1)-82、BC_(2)F_(1)-106和BC_(2)F_(1)-192),平均抽薹时间和开花时间分别比20-2229晚11.33 d和15.67 d。本研究开发了BoFLCs特异性分子标记,并实现了BoFLCs优异基因型的高效聚合,改良了亲本20-2229的抽薹性。FLOWERING LOCUS C(FLC)encodes MADS-box family protein and plays a key role in the vernalization pathway of Arabidopsis.To explore the role of BoFLCs in the vernalization pathway of cabbage,we knocked out BoFLCs in cabbage inbred line‘GL’by CRISPR/Cas9.All the knock-out lines had significantly earlier flowering time,providing insight into the function of BoFLCs gene.The BC_(1-2)P_(2) and BC_(2)F_(1) segregating populations were constructed using late-flowering cabbage inbred line‘20-2221’(P_(1),BoFLCs-w)and mid-early-flowering cabbage inbred line‘20-2229’(P_(2),BoFLCs-z).Sequence analysis showed that BoFLC_(1),BoFLC3 and BoFLC5 had sequence differences between parents,and specific molecular markers were developed based on these sequence differences.The BC_(2)F_(1) population was screened by the BoFLCs specific molecular markers.One plant containing BoFLC_(1)-w/w and BoFLC5-w/w homozygous haplotype,and BoFLC3-w/z heterozygous haplotype(BC_(2)F_(1)-29)was obtained.The bolting time and flowering time of BC_(2)F_(1)-29 were 8.00 and 13.00 days later than those of backcross parent‘20-2229’,respectively.Four plants containing BoFLC_(1)-w/w and BoFLC3-w/w homozygous haplotype,and BoFLC5-w/z heterozygous haplotype(BC_(2)F_(1)-55,BC_(2)F_(1)-119,BC_(2)F_(1)-121,BC_(2)F_(1)-163)were obtained.The average bolting time and flowering time of these four plants were 9.75 and 13.50 days later than those of‘20-2229’,respectively.In addition,three plants containing BoFLC_(1)-w/w,BoFLC3-w/w and BoFLC5-w/w homozygous haplotype(BC_(2)F_(1)-82,BC_(2)F_(1)-106,BC_(2)F_(1)-192)were obtained.The average bolting time and flowering time of these three plants were 11.33 and 15.67 days later than those of‘20-2229’,respectively.In this study,BoFLCs specific molecular markers were developed and the excellent BoFLCs genotypes were efficiently pyramided and the bolting properties of parent‘20-2229’was improved.

关 键 词：甘蓝 FLC 晚抽薹 标记开发 单倍型聚合 

分 类 号：S635[农业科学—蔬菜学]