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首页> 外文期刊>African Journal of Biotechnology >Stable Agrobacterium-mediated transformation of the halophytic Leymus chinensis (Trin.)
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Stable Agrobacterium-mediated transformation of the halophytic Leymus chinensis (Trin.)

机译：稳定的农杆菌介导的嗜睡（Trin）的转化

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In this study, an efficient procedure for stable?Agrobacterium-mediated transformation of?Leymus chinensis?(Trin.) was established.?Agrobacterium tumefaciens?strain EHA105, harboring a binary vector pCAMBIA2300, was used for transformation, along with a sweet potato 2-cysteine peroxiredoxin (2-Cys Prx) gene under the control of the stress-inducible sweet potato anionic peroxidase 2 (SWPA2) promoter and the neomycin phosphotransferase (npt?= 2 * ROMAN II) gene under the control of the cauliflower mosaic virus (CaMV) 35 S promoter. We found that a one-month-old callus derived from mature seeds could be efficiently transformed. Seven-day preculture followed by inoculation with the addition of 100 μmolL-1?acetosyringone (AS) and then a 3 day co-cultivation were performed before selection. Selection of transgenic shoots was done in the presence of 150 mgL-1?kanamycin (KM). An optical density at a wavelength of 600 nm (OD600) of approximately 0.4 for?A. tumefaciensinfection solution and 20 min of infection time gave the highest transformation efficiency. Polymerase chain reaction (PCR) analysis of KM-resistant plants and newly regenerated rhizomes revealed stable transformation of the?2-Cys Prx?gene and the?npt?= 2 * ROMAN II?gene, with the highest transformation frequency of 4.93%. RT-RCR analysis was conducted using salt stressed transgenic plants, and the results suggested that?2-Cys Prx?had low transcription levels under non-stressed conditions, and increased transcription after 6 h of 200 mM NaCl stress. This gene continued to demonstrate high levels of transcription until 6 h after withdrawal of stress, with a slow recovery. The method reported herein provides a direct opportunity for improvement of the quality traits of?L. chinensis?via genetic transformation.
机译：在这项研究中，稳定的稳定性介导的α（Trin。）的有效方法？（Trin。）。Tumefaciensαagrobacteriumtumefaciensα菌株Eha105，含有二元载体pcambia2300，用于转化，以及甘薯2在Cauliflower马赛克病毒的控制下对应力诱导的甘薯阴离子过氧化物酶2（SWPA2）启动子和NPTα= 2 *罗马II）基因进行胁迫诱导的甘薯阴离子过氧化物酶2（SWPA2）启动子（NPTα= 2 *罗马II）基因的-Cysteine （CAMV）35 S启动子。我们发现，可以有效地改变来自成熟种子的一个月大的愈伤组织。七天的预培养，然后用100μmoll-1的加入接种，然后在选择前进行3天共培养。转基因芽的选择在150mgl-1的存在下进行，Kanamycin（KM）。波长为600nm（OD600）的光密度约为0.4的Δa。甘露啶素素溶液和20分钟的感染时间得到了最高的转化效率。 KM抗性植物和新再生根茎的聚合酶链式反应（PCR）分析显示出α2-Cys PRX的稳定转化α基因和α= 2 *罗马II？基因，其变性率最高为4.93％ 。使用盐胁迫转基因植物进行RT-RCR分析，结果表明？2-Cys PRX？在非应激条件下具有低的转录水平，并且在200mM NaCl应激6小时后随后转录。该基因继续证明高水平的转录直至戒断应激后6小时，恢复缓慢。本文报告的方法提供了改进ΔL的质量特征的直接机会。 Chinensis？通过遗传转化。

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