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首页> 外文期刊>In Vitro Cellular and Development Biology. Plant: Journal of the Tissue Culture Association >Extracellular enzymes of Erwinia carotovora eliminate the need for azacytidine treatment for high frequency transformation of Arabidopsis thaliana.
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Extracellular enzymes of Erwinia carotovora eliminate the need for azacytidine treatment for high frequency transformation of Arabidopsis thaliana.

机译：胡萝卜欧文氏菌的细胞外酶消除了氮杂胞苷处理对拟南芥高频转化的需要。

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摘要

To enhance the chance of tagging specific stress-responsive genes of A. thaliana by monitoring the preferential insertion of the T-DNA into the actively transcribed loci, root explants were exposed to low temperature (LT), abscisic acid (ABA) and extracellular enzymes (EXE) of the plant pathogen Erwinia carotovora prior to transformation by Agrobacterium tumefaciens. Both LT and ABA reduced the frequency of transformation; with these treatments, the average transformation frequencies were 8.1% and2.6%, respectively. However, in explants pretreated with EXE the transformation frequency was 89.0%, similar to that obtained in control materials (92.6%). Transgenic calluses developed from these explants did not require any treatment with azacytidine for efficient shoot regeneration. Furthermore, this treatment enhanced multiple insertion of the T-DNA into the plant genome; within a population of EXE-treated transgenic plants, the number of lines harbouring at least three copies of the integrated T-DNA was much higher (61%) than that observed in an untreated population (34%).
机译：为了通过监视将T-DNA优先插入主动转录的基因座来增加标记拟南芥特定胁迫反应基因的机会，将根外植体暴露于低温（LT），脱落酸（ABA）和细胞外酶中根癌土壤杆菌转化之前的植物病原体胡萝卜欧文氏菌（EXE）的基因（EXE）。 LT和ABA均降低了转化频率;通过这些处理，平均转化频率分别为8.1％和2.6％。但是，用EXE预处理的外植体的转化频率为89.0％，与对照材料的转化频率相似（92.6％）。从这些外植体产生的转基因愈伤组织不需要氮杂胞苷的任何处理即可有效地再生芽。此外，这种处理增强了T-DNA向植物基因组中的多次插入。在经过EXE处理的转基因植物种群中，具有至少三拷贝的整合T-DNA的品系数量（61％）比未处理种群（34％）高得多。

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