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ACTA HORTICULTURAE SINICA ›› 2019, Vol. 46 ›› Issue (4): 623-634.doi: 10.16420/j.issn.0513-353x.2018-0626

• Research Papers • Previous Articles     Next Articles

Knock-out Analysis of VviPDS1 Gene Using CRISPR/Cas9 in Grapevine

GUO Ye,WAN Dongyan,CHAI Zhuangzhuang,WANG Yuejin,and WEN Yingqiang*   

  1. College of Horticulture,Northwest A & F University,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China,Ministry of Agriculture,State Key Laboratory of Crop Stress Biology for Arid Areas, Yangling,Shaanxi 712100,China
  • Online:2019-04-25 Published:2019-04-25

Abstract: VviPDS1,phytoene desaturase in grapevine,was selected as the target gene. A gene knockout vector,using the CRISPR/Cas9 system,was constructed. Different types of mutations were detected in grape protoplasts by transient transformation of protoplasts from grape leaves. In addition,the vector was constructed and transformed to Vitis vinifera L.‘Thompson Seedless’through Agrobacterium- mediated transformation. Seventy-one kanamycin resistant lines were obtained through resistance screening. Fifty-three transgenic lines,74.64%,were confirmed by T-DNA specific PCR. Sequencing revealed that twenty transgenic lines had different types of mutations at the target site and the editing efficiency was 37.74%. Among them,9 lines with biallelic mutations were shown the albino phenotype. The prediction of amino acids of biallelic mutations showed them have different degrees of mutation behind the 202th amino acids. These biallelic mutations were shown dwarf and different albino phenotypes. These indicated that the CRISPR/Cas9 system allows genome editing in the transient of grape cells or stable expression,which can produce homozygous knockout in transgenic plants of grape.

Key words: grapevine, CRISPR/Cas9, PDS1, gene editing

CLC Number: