干旱区生态环境知识资源中心(Arid): Assessment of biolistic and Agrobacterium-mediated genetic transformation methods in Cenchrus ciliaris

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DOI	10.1007/s13237-020-00332-1
	Assessment of biolistic and Agrobacterium-mediated genetic transformation methods in Cenchrus ciliaris
	Laishram, Sundari Devi; Goyal, Shipra; Shashi; Kulkarni, Vishvas M.; Kumar, Suresh; Bhat, Vishnu
通讯作者	Bhat, V
来源期刊	NUCLEUS-INDIA
ISSN	0029-568X
EISSN	0976-7975
出版年	2020
卷号	63 期号:3 页码:303-312
英文摘要	Cenchrus ciliaris, commonly known as buffelgrass, is an apomictic perennial range grass usually grown in arid/semi-arid regions. Because of the difficulties faced in conventional breeding of this polymorphic polyploid grass, the development of an efficient protocol for genetic transformation is warranted. Such a protocol would enable functional genomic studies needed to elucidate the mechanism of apomixis in this species. The embryogenic calli for genetic transformation experiments were obtained by in vitro culture of the immature inflorescences of buffelgrass cv. IGFRI-3108. Four developmental stages of immature inflorescences were compared for callus induction, and the most mature stage produced most callus. Embryogenic calli were bombarded at three distances, 6 cm (L1), 9 cm (L2), or 12 cm (L3) with a marker geneuid A present in pCAMBIA1301, under the same vacuum (85 kPa) and at constant pressure (900 psi). TheAgrobacterium-mediated genetic transformation was also performed using the same construct. Transient and stable expression as well as PCR amplification of theGUSgene was used for comparative analysis as well as for validation of the transformants. Transient GUS expression was present in a significantly higher percentage of bombarded calli (56.33%) than ofAgrobacteriumtreated calli (11.17%), but the number of GUS positive cells per callus was similar. Among the three different bombardment distances, transient GUS expression was highest at L2, but stable GUS expression was highest at L1. Shoot development from the bombarded calli could be accomplished, which failed from theAgrobacterium-mediated transformed calli. Thus, the results indicate thatC.ciliariscv. IGFRI-3108 can be successfully transformed through Biolistic particle bombardment, while Agrobacterium-mediated transformation requires further optimization of transformation protocols.
英文关键词	Buffelgrass Embryogenic calli Agrobacteriumtumefaciens Biolistic Genetic transformation GUS expression Shoot development
类型	Article
语种	英语
收录类别	ESCI
WOS记录号	WOS:000567636500001
WOS关键词	VITRO PLANT-REGENERATION ; SOMATIC EMBRYOGENESIS ; CALLUS ; PRODUCTIVITY ; EXPRESSION ; FREQUENCY ; INDUCTION ; CULTURES ; L.
WOS类目	Cell Biology
WOS研究方向	Cell Biology
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/334581
作者单位	[Laishram, Sundari Devi; Goyal, Shipra; Shashi; Bhat, Vishnu] Univ Delhi, Dept Bot, Plant Dev Biol, New Delhi 110007, India; [Kulkarni, Vishvas M.] Bhabha Atom Res Ctr, Nucl Agr & Biotechnol Div, Plant Cell Culture Technol Sect, Mumbai 400085, Maharashtra, India; [Kumar, Suresh] ICAR Indian Agr Res Inst, Div Biochem, New Delhi, India
推荐引用方式 GB/T 7714	Laishram, Sundari Devi,Goyal, Shipra,Shashi,et al. Assessment of biolistic and Agrobacterium-mediated genetic transformation methods in Cenchrus ciliaris[J],2020,63(3):303-312.
APA	Laishram, Sundari Devi,Goyal, Shipra,Shashi,Kulkarni, Vishvas M.,Kumar, Suresh,&Bhat, Vishnu.(2020).Assessment of biolistic and Agrobacterium-mediated genetic transformation methods in Cenchrus ciliaris.NUCLEUS-INDIA,63(3),303-312.
MLA	Laishram, Sundari Devi,et al."Assessment of biolistic and Agrobacterium-mediated genetic transformation methods in Cenchrus ciliaris".NUCLEUS-INDIA 63.3(2020):303-312.