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通过实时定量PCR方法分析了mi R395abcd与其靶基因ATRX在两个木薯品种的4种低温处理下的差异表达情况,靶基因的定量引物设计在与mi RNA结合区的上游、结合边缘、结合区3个位置。结果显示mi R395abcd与其靶基因在两个木薯品种、4种低温处理和3个靶基因引物位置的表达量均不同,且mi R395abcd在C4和SC124两个品种间4种低温处理的6组两两比较中的相对表达量呈现完美的反向关系。进一步比较发现mi R395abcd与靶基因的负相关性在C4中最好,尤其是位于mi RNA与m RNA结合区上游的a引物的消长关系在AH(驯化后寒害:25~14℃驯化5 d后再降至4℃)和NAH(非驯化寒害:25℃直接降至4℃)中表现得最典型。具体而言,mi R395abcd在C4的AH和NAH处理中上调而在SC124的AH和NAH处理中下调表达,前期表型分析中AH和NAH处理下C4的低温胁迫症状小于SC124,预示着mi R395abcd的上调表达有利于木薯减轻低温伤害,其表达差异能够表征C4和SC124在耐低温性能的表型差异,且mi R395abcd对其靶基因ATRX的调控在木薯低温响应过程中起着正向保护作用。
The differential expression of mi R395abcd and its target gene ATRX under the four low temperature treatments of two cassava varieties were analyzed by real-time quantitative PCR. The quantitative primers of the target gene were designed in the upstream of miRNA binding region, the binding edge, the binding region 3 Location. The results showed that the expression of mi R395abcd and its target gene in two cassava varieties, four kinds of low temperature treatment and three target gene primers were different, and mi R395abcd in four varieties of C4 and SC124 low temperature treatment of six groups of two two The relative amount of expression in the comparison presented a perfect reverse relationship. Further comparison showed that the negative correlation between mi R395abcd and target gene was the best in C4, especially the increase and decrease of a primer upstream of mi RNA and m RNA binding region in AH (domesticated chilling injury: after domesticated at 25 ~ 14 ℃ for 5 d Then down to 4 ℃) and NAH (non-domesticated cold: 25 ℃ directly down to 4 ℃) in the most typical. In particular, mi R395abcd was up-regulated in AH and NAH treatment of C4 and down-regulated in AH and NAH treatment of SC124. Symptoms of low temperature stress of C4 under AH and NAH treatment were lower than those of SC124 in pre-phenotypic analysis, indicating that mi R395abcd Upregulation of Camptotheca acuminata is beneficial to cassava in reducing cold damage, and its differential expression can characterize the phenotypic differences of C4 and SC124 in low temperature tolerance. The regulation of ATRX by mi R395abcd plays a positive role in the low temperature response of cassava.