首页|Development of an Agrobacterium-mediated CRISPR/Cas9 gene editing system in jute(Corchorus capsularis)
Development of an Agrobacterium-mediated CRISPR/Cas9 gene editing system in jute(Corchorus capsularis)
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Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy root system mediated by Agrobacterium rhizogenes strain K599 was developed for Meifeng 4,an elite jute variety widely cultivated in China.The transgenic hairy root system for jute was verified by subcellular localization and bimolecular fluorescence complementation(BiFC)assays.The CHLOROPLASTOS ALTERADOS 1(CcCLA1)gene,which is involved in the development of chloroplasts,was targeted for editing at two sites in Meifeng 4.Based on this hairy root transformation,the gRNA scaf-fold was placed under the control of cotton ubiquitin GhU6.7 and-GhU6.9 promoters,respectively,to assess the efficiency of gene editing.Results indicated the 50.0％(GhU6.7)and 38.5％(GhU6.9)editing events in the target 2 alleles(gRNA2),but no mutation was detected in the target 1 allele(gRNA1)in transgenic-positive hairy roots.CcCLA1 gene editing at gRNA2 under the control of GhU6.7 in Meifeng 4 was also carried out by Agrobacterium tumefaciens-mediated transformation.Two CcCLA1 mutants were albinic,with a gene editing efficiency of 5.3％.These findings confirm that the CRISPR/Cas9 system,incor-porating promoter GhU6.7,can be used as a gene editing tool for jute.
JuteAgrobacterium-mediated transformationGenome editingHairy root system
Key Laboratory of Ministry of Education for Genetic Breeding and Multiple Utilization of Crops,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China
Experiment Station of Ministry of Agriculture and Rural Affairs for Jute and Kenaf in Southeast China/Fujian Public Platform for Germplasm Resources of Bast Fiber Crops/Fujian Provincial Key Laboratory of Crop Breeding by Design/Key Laboratory of Ministry of Agriculture and Rural Affairs for Biological Breeding of Fujian and Taiwan Crops,Fujian
National Key Lab of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070,Hubei,China
国家自然科学基金福建省自然科学基金China Agriculture Research System of the Ministry of Agriculture and MARA
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