Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. var. durum cv Stewart) with improved efficiency

doi:10.1093/jxb/erq035

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	Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. var. durum cv Stewart) with improved efficiency
	He, Y.2; Jones, H. D.1; Chen, S.2; Chen, X. M.2; Wang, D. W.3; Li, K. X.1; Wang, D. S.2; Xia, L. Q.2
刊名	JOURNAL OF EXPERIMENTAL BOTANY
	2010
卷号	61 期号:6 页码:1567-1581
关键词	Acetosyringone Agrobacterium tumefaciens durum wheat cv Stewart picloram transformation
ISSN号	0022-0957
DOI	10.1093/jxb/erq035
通讯作者	He, Y.
英文摘要	An efficient Agrobacterium-mediated durum wheat transformation system has been developed for the production of 121 independent transgenic lines. This improved system used Agrobacterium strain AGL1 containing the superbinary pGreen/pSoup vector system and durum wheat cv Stewart as the recipient plant. Acetosyringone at 400 mu M was added to both the inoculation and cultivation medium, and picloram at 10 mg l(-1) and 2 mg l(-1) was used in the cultivation and induction medium, respectively. Compared with 200 mu M in the inoculation and cultivation media, the increased acetosyringone concentration led to significantly higher GUS (beta-glucuronidase) transient expression and T-DNA delivery efficiency. However, no evident effects of acetosyringone concentration on regeneration frequency were observed. The higher acetosyringone concentration led to an improvement in average final transformation efficiency from 4.7% to 6.3%. Furthermore, the concentration of picloram in the co-cultivation medium had significant effects on callus induction and regeneration. Compared with 2 mg l(-1) picloram in the co-cultivation medium, increasing the concentration to 10 mg l(-1) picloram resulted in improved final transformation frequency from 2.8% to 6.3%, with the highest frequency of 12.3% reached in one particular experiment, although statistical analysis showed that this difference in final transformation efficiency had a low level of significance. Stable integration of foreign genes, their expression, and inheritance were confirmed by Southern blot analyses, GUS assay, and genetic analysis. Analysis of T(1) progeny showed that, of the 31 transgenic lines randomly selected, nearly one-third had a segregation ratio of 3:1, while the remainder had ratios typical of two or three independently segregating loci.
学科主题	Plant Sciences ; PLANT SCIENCES
语种	英语
出版者	OXFORD UNIV PRESS
WOS记录号	WOS:000276735300002
内容类型	期刊论文
源URL	[http://111.203.20.206/handle/2HMLN22E/5433]
专题	作物科学研究所
作者单位	1.Ctr Crop Genet Improvement, Harpenden AL5 2JQ, Herts, England 2.Chinese Acad Agr Sci, Inst Crop Sci, Natl Key Facil Crop Gene Resources & Genet Improv, Beijing 100081, Peoples R China 3.Chinese Acad Sci, Inst Genet & Dev Biol, State Key Lab Plant Cell & Chromosome Engn, Beijing 100101, Peoples R China
推荐引用方式 GB/T 7714	He, Y.,Jones, H. D.,Chen, S.,et al. Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. var. durum cv Stewart) with improved efficiency[J]. JOURNAL OF EXPERIMENTAL BOTANY,2010,61(6):1567-1581.
APA	He, Y..,Jones, H. D..,Chen, S..,Chen, X. M..,Wang, D. W..,...&Xia, L. Q..(2010).Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. var. durum cv Stewart) with improved efficiency.JOURNAL OF EXPERIMENTAL BOTANY,61(6),1567-1581.
MLA	He, Y.,et al."Agrobacterium-mediated transformation of durum wheat (Triticum turgidum L. var. durum cv Stewart) with improved efficiency".JOURNAL OF EXPERIMENTAL BOTANY 61.6(2010):1567-1581.