目的利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系,为进一步研究map3k15在肾脏疾病方面的作用提供动物模型。方法1)分析map3k15在斑马鱼中的表达模式;2)利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系;3)观察...目的利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系,为进一步研究map3k15在肾脏疾病方面的作用提供动物模型。方法1)分析map3k15在斑马鱼中的表达模式;2)利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系;3)观察map3k15缺失对斑马鱼表型的影响。结果1)map3k15在斑马鱼前肾中表达,且MAP3K15/Map3k15蛋白在多物种间具有高度保守性;2)成功构建了斑马鱼map3k15-/-突变体,并保留+2 bp及+1 bp 2种突变品系;3)map3k15突变斑马鱼在胚胎发育过程中出现卵黄囊、心包及头部的水肿,且随发育时间延长,症状逐渐加重。结论成功构建了map3k15敲除的纯合斑马鱼品系,为未来研究map3k15在肾脏发育及疾病中的作用提供了重要的模型。展开更多
Wheat grain morphology,particularly grain length(GL)and width(GW),is a key determinant of yield.To improve the suboptimal grain dimensions of the local anthocyanin-rich variety Guizi 1(GZ1),we crossed it with Zhongyan...Wheat grain morphology,particularly grain length(GL)and width(GW),is a key determinant of yield.To improve the suboptimal grain dimensions of the local anthocyanin-rich variety Guizi 1(GZ1),we crossed it with Zhongyan 96-3(ZY96-3),an elite germplasm known for faster grain filling and superior grain size.A genotyping-by-sequencing(GBS)approach was applied to an F_(2)population of 110 individuals derived from GZ1×ZY96-3,resulting in the identification of 23,134 high-quality SNPs.Most of the SNPs associated with GL and GW were clustered on chromosomes 2B,3A,and 3B.QTL mapping for GL revealed two major loci,GL1 on chromosome 2B and GL2 on chromosome 3B,and eight candidate genes were identified within their corresponding intervals(2B:63.6–70.4 Mb;3B:631.5–633.3 Mb).These genes encode proteins potentially involved in grain size regulation,including a TOR2 regulation-associated protein,erect spike 2(EP2),fibroblast growth factor 6(FGF6),cellulose synthase-like(CSLD),RelA/pot homologue three family protein,and three GDSL esterase/lipase(GLIP)proteins.Additionally,we detected a QTL associated with GW on chromosome 3A and identified two candidate genes,TOR2 regulation and starch synthase within the 61.4–68.5 Mb interval.Overall,this study provides a strong theoretical and technical basis for wheat genetic improvement and offers valuable resources for precise QTL mapping and candidate gene discovery.展开更多
文摘目的利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系,为进一步研究map3k15在肾脏疾病方面的作用提供动物模型。方法1)分析map3k15在斑马鱼中的表达模式;2)利用CRISPR/Cas9基因编辑技术构建斑马鱼map3k15敲除纯合品系;3)观察map3k15缺失对斑马鱼表型的影响。结果1)map3k15在斑马鱼前肾中表达,且MAP3K15/Map3k15蛋白在多物种间具有高度保守性;2)成功构建了斑马鱼map3k15-/-突变体,并保留+2 bp及+1 bp 2种突变品系;3)map3k15突变斑马鱼在胚胎发育过程中出现卵黄囊、心包及头部的水肿,且随发育时间延长,症状逐渐加重。结论成功构建了map3k15敲除的纯合斑马鱼品系,为未来研究map3k15在肾脏发育及疾病中的作用提供了重要的模型。
基金Funding for this project was provided by the National Natural Science Foundation of China(Grants No.32160456,32360474,32360486,32260496)the Key Laboratory of Functional Agriculture of Guizhou Provincial Higher Education Institutions(Grant No.Qianjiaoji(2023)007).
文摘Wheat grain morphology,particularly grain length(GL)and width(GW),is a key determinant of yield.To improve the suboptimal grain dimensions of the local anthocyanin-rich variety Guizi 1(GZ1),we crossed it with Zhongyan 96-3(ZY96-3),an elite germplasm known for faster grain filling and superior grain size.A genotyping-by-sequencing(GBS)approach was applied to an F_(2)population of 110 individuals derived from GZ1×ZY96-3,resulting in the identification of 23,134 high-quality SNPs.Most of the SNPs associated with GL and GW were clustered on chromosomes 2B,3A,and 3B.QTL mapping for GL revealed two major loci,GL1 on chromosome 2B and GL2 on chromosome 3B,and eight candidate genes were identified within their corresponding intervals(2B:63.6–70.4 Mb;3B:631.5–633.3 Mb).These genes encode proteins potentially involved in grain size regulation,including a TOR2 regulation-associated protein,erect spike 2(EP2),fibroblast growth factor 6(FGF6),cellulose synthase-like(CSLD),RelA/pot homologue three family protein,and three GDSL esterase/lipase(GLIP)proteins.Additionally,we detected a QTL associated with GW on chromosome 3A and identified two candidate genes,TOR2 regulation and starch synthase within the 61.4–68.5 Mb interval.Overall,this study provides a strong theoretical and technical basis for wheat genetic improvement and offers valuable resources for precise QTL mapping and candidate gene discovery.