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Cisgenic inhibition of the potato cold induced phosphorylase L gene expression and decrease in sugar contents

机译:马铃薯冷诱导磷酸化酶L基因表达的顺生抑制和糖含量的降低

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To decrease the accumulation of reducing and non-reducing sugar in potato tubers stored at low temperature, a single gene silencing vector pARTPhL-IR, harboring a part of starch phosphorylase L gene as inverted repeats with pdk intron within was constructed and transformed into potato (Solanum tuberosum?L.) cultivars Agria and Marfona. Polymerase chain reaction (PCR) of?nptII gene and pdk intron indicated that the RNA interference construct was transformed successfully into the genome. Real time RT-PCR analysis of starch phosphorylase L gene in stored microtubers for 90 days at 4°C showed that the expression level of this gene in transgenics ranged from 1.63 to 7.54% of that in the non-transgenic plants. Analysis of sugar content in these plants showed that the total sugar content in transgenic microtubers was significantly reduced compared to the control, up to 35% in line M4. The accumulation of reducing sugars in transgenic lines at 4°C was reduced from 9.13 (in Agria) to 5.57?mg/g?fresh weight (transgenic line A5) and from 9.56 (in Marfona) to 6.52?mg/g?fresh weight (transgenic line M4), implying that silencing of starch phosphorylase L gene reduced starch breakdown during cold storage conditions.
机译:为了减少低温保存的马铃薯块茎中还原糖和非还原糖的积累,构建了一个基因沉默载体pARTPhL-IR,该载体内含一部分淀粉磷酸化酶L基因作为pdk内含子的反向重复序列,并转化为马铃薯(马铃薯(Solanum tuberosum?L。)品种Agria和Marfona。 nptII基因和pdk内含子的聚合酶链反应(PCR)表明RNA干扰构建体已成功转化到基因组中。在4°C下储存90天的淀粉块茎中淀粉磷酸化酶L基因的实时RT-PCR分析表明,该基因在转基因植物中的表达水平是非转基因植物的1.63%至7.54%。对这些植物中糖含量的分析表明,与对照相比,转基因微块茎中的总糖含量显着降低,最高达到M4品系的35%。转基因品系中还原糖的积累在4°C时从9.13(在Agria中)降低到5.57?mg / g新鲜重量(转基因品系A5)和从9.56(在Marfona中)降至6.52?mg / g新鲜重量。 (转基因品系M4),这意味着淀粉磷酸化酶L基因的沉默可减少冷藏条件下的淀粉分解。

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