Reliable and Efficient Agrobacterium tumefaciens-Mediated Genetic Transformation of Dianthus spiculifolius  

作　　者：Jingang Wang Shiyue Liu Hongping Ma Ye Tao Shuang Feng Shufang Gong Jinzhu Zhang Aimin Zhou 

机构地区：[1]College of Horticulture and Landscape Architecture,Northeast Agricultural University,Harbin 150030,China [2]Key Laboratory of Saline-Alkali Vegetation Ecology Restoration,Northeast Forestry University,Ministry of Education,Harbin 150040,China [3]College of Life Sciences,Northeast Forestry University,Harbin 150040,China

出　　处：《Horticultural Plant Journal》2020年第3期199-204,共6页园艺学报（英文版）

基　　金：supported by grants from the National Natural Science Foundation of China(Grant Nos.31902052 and 31972450);the National Key Research and Development Program of China(Grant No.2016YFC0500300);the Natural Science Foundation of Heilongjiang Province of China(Grant No.C2018021);the‘Academic backbone’Project of Northeast Agricultural University of China(Grant No.18XG08).

摘　　要：Dianthus spiculifolius is a perennial herbaceous flower with strong environmental adaptability and is an important ornamental ground cover plant.In this study,seeds of D.spiculifolius were used as explants for callus induction,adventitious bud differentiation,and rooting by adding different concentrations of 2,4-dichlorophenoxyacetic acid(2,4-D),6-benzyl aminopurine(6-BA),and naphthaleneacetic acid(NAA)to Murashige and Skoog medium.The calli generated were co-cultured with Agrobacterium tumefaciens EHA105 containing pBI121-GUS or pBI121-GFP plasmids for 30 min,and transgenic regenerated plants were obtained by kanamycin(30mg·L^−1)screening.RT-PCR confirmed the stable expression of the exogenous GUS and GFP genes in the D.spiculifolius.Theβ-glucuronidase(GUS)histochemical staining confirmed GUS gene expression in transgenic calli,adventitious buds,and regenerated plants of D.spiculifolius.The green fluorescent protein(GFP)visual analysis showed GFP gene expression in transgenic calli.Furthermore,subcellular localization analysis showed that the three organelle marker proteins were not only successfully expressed but also accurately localized to their corresponding organelles in D.spiculifolius callus cells.These results indicated a successful establishment of a reliable and efficient A.tumefaciens-mediated genetic transformation system,which will contribute to functional gene research and genetic improvement of D.spiculifolius.

关 键 词：Agrobacterium tumefaciens Dianthus spiculifolius CALLUS Tissue culture Plant transformation 

分 类 号：S682.19[农业科学—观赏园艺]