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ACTA HORTICULTURAE SINICA ›› 2019, Vol. 46 ›› Issue (2): 345-355.doi: 10.16420/j.issn.0513-353x.2018-0330

• Research Notes • Previous Articles     Next Articles

Establishment and Application of YL-1 High-efficiency Genetic Transformation System in Cabbage(Brassica oleracea L. var. capitata)

CUI Huilin1,2,LI Zhiyuan2,FANG Zhiyuan2,YANG Limei2,ZHUANG Mu2,Lü Honghao2,LIU Yumei2,SONG Jianghua1,*,and ZHANG Yangyong2,*   

  1. 1College of Horticulture,Anhui Agriculture University,Hefei 230036,China;2Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China
  • Online:2019-02-25 Published:2019-02-25

Abstract: In order to select the cabbage genotypes with high-efficiency transformation,cotyledons with petiole and hypocotyl of four inbred lines were used to compare the regeneration frequency. The results showed that YL-1 was the optimal transformation donor and its regeneration frequency was significantly higher than the other three materials. The regeneration frequency of YL-1 cotyledons with petiole and hypocotyl had no significant difference and both of them could be used for genetic transformation. The transformation factors including light intensity,concentration of herbicide Basta and Agrobacterium concentration of infection were further optimized. The results showed that the optimum culture condition for YL-1 adventitious bud regeneration was light intensity 4 000 lx + Basta 8 mg · L-1. When the explants were inoculated by Agrobacterium with OD600 = 0.3 and 8 min,the regeneration frequency of herbicide-resistant buds was the highest. Eventually,a total of 47 herbicide-resistant transformants were obtained and 12 of them were PCR-positive,with the transformation frequency up to 2.18%,which indicated that bar gene had been transformed into YL-1. Gene expression analysis of two PCR-positive transformants were performed by transcriptome sequencing,and bar gene was transcribed in both of them. The establishment of high-efficiency genetic transformation system for YL-1 mediated by Agrobacterium tumefaciens would provide a basis for gene function identification,the editing and improvement of important agronomic traits in the future.

Key words: cabbage, Agrobacterium-mediated, genetic transformation, system optimization, herbicide resistant

CLC Number: