干旱区生态环境知识资源中心(Arid): Optimization of Tissue Culturing and Genetic Transformation Protocol for Casuarina equisetifolia

Arid

DOI	10.3389/fpls.2021.784566
	Optimization of Tissue Culturing and Genetic Transformation Protocol for Casuarina equisetifolia
	Ren, Huimin; Xu, Yan; Zhao, Xiaohong; Zhang, Yan; Hussain, Jamshaid; Cui, Fuqiang; Qi, Guoning; Liu, Shenkui
通讯作者	Qi, GN ; Liu, SK (corresponding author)，Zhejiang A & F Univ, Sch Forestry & Biotechnol, State Key Lab Subtrop Silviculture, Hangzhou, Peoples R China.
来源期刊	FRONTIERS IN PLANT SCIENCE
ISSN	1664-462X
出版年	2022
卷号	12
英文摘要	Casuarina equisetifolia is widely used in agroforestry plantations for soil stabilization, ecosystem rehabilitation, reclamation, and coastal protection. Moreover, C. equisetifolia has remarkable resistance to typhoons, desert, low soil fertility, drought, and salinity, but not cold. Therefore, it is significant to breed high-quality Casuarina varieties to improve the tolerance and adaptability to cold weather by molecular techniques. The establishment of a rapid and efficient callus induction and regeneration system via tissue culture is pre-requisite for the genetic transformation of C. equisetifolia, which is so far lacking. In this study, we reported an efficient and rapid regeneration system using stem segment explants, in which callus induction was found to be optimal in a basal medium supplemented with 0.1 mg.L-1 TDZ and 0.1 mg.L-1 NAA, and proliferation in a basal medium containing 0.1 mg.L-1 TDZ and 0.5 mg.L-1 6-BA. For bud regeneration and rooting, the preferred plant growth regulator (PGR) in basal medium was 0.5 mg.L-1 6-BA, and a combination of 0.02 mg.L-1 IBA and 0.4 mg.L-1 IAA, respectively. We also optimized genetic a transformation protocol using Agrobacterium tumefaciens harboring the binary vector pCAMBIA1301 with beta-glucuronidase (GUS) as a reporter gene. Consequently, 5 mg L-1 hygromycin, 20 mg L-1 acetosyringone (As), and 2 days of co-cultivation duration were optimized to improve the transformation efficiency. With these optimized parameters, transgenic plants were obtained in about 4 months. Besides that, Agrobacterium rhizogenes-mediated transformation involving adventitious root induction was also optimized. Our findings will not only increase the transformation efficiency but also shorten the time for developing transgenic C. equisetifolia plants. Taken together, this pioneer study on tissue culturing and genetic transformation of C. equisetifolia will pave the way for further genetic manipulation and functional genomics of C. equisetifolia.
英文关键词	Casuarina equisetifolia regeneration system transformation system Agrobacterium tumefaciens Agrobacterium rhizogenes
类型	Article
语种	英语
开放获取类型	Green Published, gold
收录类别	SCI-E
WOS记录号	WOS:000751842500001
WOS关键词	AGROBACTERIUM-RHIZOGENES ; PLANT TRANSFORMATION ; EXPRESSION ; FRANKIA ; MYCORRHIZAL ; BIOLOGY ; FUNGI
WOS类目	Plant Sciences
WOS研究方向	Plant Sciences
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/376527
作者单位	[Ren, Huimin; Xu, Yan; Zhao, Xiaohong; Zhang, Yan; Cui, Fuqiang; Qi, Guoning; Liu, Shenkui] Zhejiang A & F Univ, Sch Forestry & Biotechnol, State Key Lab Subtrop Silviculture, Hangzhou, Peoples R China; [Hussain, Jamshaid] COMSATS Univ Islamabad, Dept Biotechnol, Abbottabad, Pakistan
推荐引用方式 GB/T 7714	Ren, Huimin,Xu, Yan,Zhao, Xiaohong,et al. Optimization of Tissue Culturing and Genetic Transformation Protocol for Casuarina equisetifolia[J],2022,12.
APA	Ren, Huimin.,Xu, Yan.,Zhao, Xiaohong.,Zhang, Yan.,Hussain, Jamshaid.,...&Liu, Shenkui.(2022).Optimization of Tissue Culturing and Genetic Transformation Protocol for Casuarina equisetifolia.FRONTIERS IN PLANT SCIENCE,12.
MLA	Ren, Huimin,et al."Optimization of Tissue Culturing and Genetic Transformation Protocol for Casuarina equisetifolia".FRONTIERS IN PLANT SCIENCE 12(2022).