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首页> 外文期刊>African Journal of Biotechnology >Increasing the amylose content of maize through silencing of sbe2a genes
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Increasing the amylose content of maize through silencing of sbe2a genes

机译：通过沉默sbe2a基因提高玉米的直链淀粉含量

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摘要

Improved amylose?content in maize has been achieved by reducing the starchbranching enzyme (SBE)?activity via transgenic?maize inbred line?transformed by a high-efficient RNAi expression vector, which?may provide the foundation for maize quality improvement. The sense and anti-sense fragments of maize SBE gene?sbe2awere cloned by reverse transcript PCR and high efficient RNAi expression vector was constructed based on plant expression plasmid?pCAMBIA1301. Then the reconstruct was introduced into?maize inbred line Tie7922 by pollen tube pathway transformation. Four transgenic plants were obtained. The integration of interest gene?sbe2a?into maize genome has been confirmed by PCR amplification and Southern hybridization. The?sbe2a?transcription was suppressed obviously by the analyses of RT-PCR, SBE activity and amylase content on the four transgenic plants. The SBE activity was significantly less than that of wild type maize, and was at most reduced by 77.9%; the amylose content was at most increased by 87.8%. In conclusion, RNAi expression vector pRSBE2a containing?sbe2a?gene was successfully constructed. Through genetic transformation, RNAi technique efficiently silences endogenous?sbe2a?gene to reduce the SBE activity, and?high-amylose maize lines?are?obtained.
机译：通过使用高效RNAi表达载体转化的转基因玉米自交系降低淀粉分支酶（SBE）的活性，可以提高玉米中的直链淀粉含量，这可以为提高玉米品质提供基础。利用植物表达质粒pCAMBIA1301构建了玉米SBE基因sbe2awere的正义和反义片段，通过逆转录PCR和高效RNAi表达载体进行了克隆。然后通过花粉管途径转化将重组体导入玉米自交系Tie7922。获得了四株转基因植物。通过PCR扩增和Southern杂交已证实将目的基因“ sbe2a”整合到玉米基因组中。通过对四种转基因植物的RT-PCR，SBE活性和淀粉酶含量的分析，明显抑制了sbe2a转录。 SBE活性显着低于野生型玉米，最多降低了77.9％。直链淀粉含量最多增加了87.8％。综上所述，成功构建了含有？sbe2a？基因的RNAi表达载体pRSBE2a。通过遗传转化，RNAi技术有效地沉默了内源性的“ sbe2a”基因以降低SBE活性，并获得了“高直链淀粉”玉米品系。

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