首页|Establishing VIGS and CRISPR/Cas9 techniques to verify RsPDS function in radish
Establishing VIGS and CRISPR/Cas9 techniques to verify RsPDS function in radish
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国家科技期刊平台
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Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification in modern plant biotechnology.However,the investigation of gene silencing and editing in radish remains limited.In this study,a bleaching phenotype was generated through the knockdown of RsPDS using tobacco rattle virus(TRV)-and turnip yellow mosaic virus(TYMV)-mediated gene silencing vectors.The TYMV-mediated gene silencing efficiency was higher than the TRV-based VIGS system in radish.The expression level of RsPDS was significantly inhibited using VIGS in'NAU-067'radish leaves.The rootless seedlings of'NAU-067'were infected with Agrobacterium rhizogenes using the 2300GN-Ubi-RsPDS-Cas9 vector with two target sequences.Nine adventitious roots were blue with GUS staining,and four of these adventitious roots were edited at target sequence 1 of the RsPDS gene as indicated by Sanger sequencing.Furthermore,albino lines were generated with A.tumefaciens-mediated transformation of radish cotyledons.Five base substitutions and three base deletions occurred at target sequence 2 in Line 1,and three base insertions and three base substitutions occurred at target sequence 1 in Line 2.This study shows that VIGS and CRISPR/Cas9 techniques can be employed to precisely verify the biological functions of genes in radish,which will facilitate the genetic improvement of vital horticultural traits in radish breeding programs.
National Key Laboratory of Crop Genetics & Germplasm Enhancement and Utilization/Key Laboratory of Horticultural Crop Biology and Genetic Improvement(East China)of Ministry of Agriculture and Rural Affairs/College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China
College of Horticulture and Landscape Architecture,Yangzhou University,Yangzhou 225009,China
College of Life Sciences,Nanjing Agricultural University,Nanjing 210095,China
Jiangsu Seed Industry Revitalization Project,China中央高校基本科研业务费专项国家自然科学基金earmarked fund for Jiangsu Agricultural Industry Technology System,ChinaJiangsu Postgraduate Scientific Research Innovation Plan,ChinaPriority Academic Program Development of Jiangsu Higher Education Institutions,China(PAPD)
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