甜瓜果肉硬度KASP标记的开发与应用  

Development and Utilization of KASP Marker for Identification of Pulp Firmness in Melon

作　　者：李肯 张伟 武云鹏 彭冬秀[2] 张若纬 LI Ken;ZHANG Wei;WU Yunpeng;PENG Dongxiu;ZHANG Ruowei(Tianjin Key Laboratory of Vegetable Genetics and Breeding,Tianjin Kernel Vegetable Research Institute,Tianjin 300381,China;State Key Laboratory of Vegetable Biobreeding,Tianjin Academy of Agricultural Sciences,Tianjin 300392,China)

机构地区：[1]天津科润农业科技股份有限公司蔬菜研究所,天津市蔬菜遗传育种企业重点实验室,天津300381 [2]蔬菜生物育种全国重点实验室,天津市农业科学院,天津300392

出　　处：《园艺学报》2024年第4期773-786,共14页Acta Horticulturae Sinica

基　　金：天津市重点研发计划科技支撑重点项目(20YFZCSN00410);天津市农业科学院青年创新研究与实验项目(2020021)。

摘　　要：为提高不同果肉硬度类型甜瓜材料的选择效率,以脆肉甜瓜自交系20S11和软肉甜瓜自交系20S75为亲本,构建F_(2)与BC_(1)F_(1)群体,分析了果肉硬度性状的遗传规律为单基因遗传,脆肉相对于软肉为显性。根据F_(2)分离群体构建果肉硬度极端性状混池,利用全基因组重测序结合图位克隆技术,将果肉硬度基因CmPf1定位在10号染色体上约54.71 kb的区域内。通过重测序分析发现,软肉甜瓜20S75中编码GATL3(galacturonosyltransferase-like 3)的基因MELO3C012216(CmGATL3)存在1处终止密码突变,位于编码区第567位碱基处(C–G),导致蛋白翻译提前终止,造成转移酶蛋白结构域完全缺失。利用qRT-PCR对CmGATL3进行表达模式分析,结果显示其在脆肉甜瓜20S11中的表达量显著高于软肉甜瓜20S75。基于上述变异位点,开发PF-KASP分子标记并对56份甜瓜自交系材料进行基因型检测,其中脆肉材料均表现为C︰C型,软肉材料均表现为G︰G型,标记呈共显性。进一步利用PF-KASP对F_(2)群体进行基因型鉴定,与果肉硬度表型鉴定结果相比较,准确率达到100%。In order to improve the selection efficiency of melon materials with different pulp firmness types.The crispy melon inbred line 20S11 and the soft melon inbred line 20S75 were used as parents to construct F_(2) and BC_(1)F_(1) populations,analyzed the fruit pulp firmness trait is inherited by a single gene,and crispy is dominant relative to soft.A mixed pool of extreme fruit flesh hardness traits was constructed based on the F_(2) segregation population.Using whole genome sequencing combined with map based cloning technology,the pulp firmness gene CmPf1 was located in a region of approximately 54.71 kb on chromosome 10.Through resequencing analysis,it was found that the gene MELO3C012216(CmGATL3)encoding GATL3(galacturonosyltransferase-like 3)in 20S75 had a termination codon mutation located at the 567th base(C–G)of the coding region,leading to early termination of protein translation and resulting in complete deletion of the transferase protein domain.The expression pattern analysis of CmGATL3 using qRT-PCR showed that the expression level in crispy melon 20S11 was significantly higher than that in soft melon 20S75.Based on the above mutation sites,PF-KASP molecular marker was developed and performed genotype detection on 56 melon inbred line materials.Among them,the crisp materials were C︰C type,and the soft materials were G︰G type,and the markers were codominance.Further utilizing PF-KASP for genotype identification of F_(2) population,compared with the results of pulp firmness phenotype identification,the accuracy reached 100%.

关 键 词：甜瓜 果肉硬度 基因定位 KASP标记 辅助选择育种 

分 类 号：S652[农业科学—果树学]