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根据森林草莓中独角金内酯合成过程中的关键基因D27的序列信息设计引物,获得了D27基因的完整开放阅读框架。采用MEGA5.0软件,对森林草莓与其他物种D27基因编码的氨基酸序列进行聚类分析,发现森林草莓D27基因与其他物种D27基因具有较高的同源性。设置正常供磷、缺磷和在缺磷条件下接种丛枝状菌根真菌处理,利用电感耦合等离子发射光谱仪测定草莓植株的磷含量,结果表明缺磷条件下接种丛枝状菌根真菌,草莓植株中磷含量极显著提高,而缺磷处理的磷含量极显著降低。实时荧光定量PCR结果表明,与正常供磷处理相比,缺磷和接种丛枝状菌根真菌处理条件下,D27基因的表达量分别上调7倍和11倍,这说明D27基因的表达既受到磷水平调控,又受到菌根形成的调控。
According to the sequence information of the key gene D27 in the synthesis of furil in forest strawberry, primers were designed and the complete open reading frame of D27 gene was obtained. Using MEGA5.0 software to cluster the amino acid sequence of D27 gene of forest strawberry and other species, we found that D27 gene of forest strawberry has higher homology with D27 gene of other species. Set the normal phosphorus, phosphorus deficiency and inoculation under the condition of phosphorus-deficient mycorrhizal fungi treatment, the use of inductively coupled plasma-atomic emission spectrometry determination of phosphorus content of strawberry plants, the results show that under phosphorus-deficient conditions inoculated with arbuscular mycorrhizal fungi, strawberry Phosphorus content in plants increased significantly, while the phosphorus content of phosphorus deficiency significantly reduced. Real-time PCR results showed that the expression of D27 gene was up-regulated 7-fold and 11-fold, respectively, under phosphorus deficiency and inoculation with arbuscular mycorrhizal fungi, which indicated that the expression of D27 gene was affected by Phosphorus level regulation, but also by the formation of mycorrhizal regulation.