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| Cloning of Squalene Epoxidase Genes (SQEs) and Correlation Analysis Between Their Expression and Ursolic Acid (UA) Content in Suspension Cells of Loquat (Eriobotrya japonica L.) Under Temperature Stress |
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Abstract Ursolic acid (UA) is an important active component in loquat (Eriobotrya japonica L.), and squalene epoxidase (SQE) is one of the key rate-limiting enzyme in pentacyclic triterpenoids biosynthesis, such as UA. To clone SQEs gene, find the suitable stress temperature in loquat suspension cultured cells, and further to find out the correlation between the different expression of SQEs and the content of UA under suitable stress temperature, cells of Eriobotrya japonica L. in the late logarithmic growth phase of cell suspension had been chose. Both the cell growth and the accumulation of UA were investigated at the low temperature of 15 ℃ and the high temperature of 35 ℃ treatments. At the same time, the homology-based cloning method was employed to determine the different transcription levels of two genes (ejSQE1 and ejSQE2) at 15 ℃ treatment. The amino acid sequence similarity between ejSQE1 (GenBank accession No. JQ294053.2) and ejSQE2 (GenBank accession No. JQ294054.2) was 69.89%, and the significant differences between sequences were in the heads and tails. The results indicated that the total UA production of the 48 h at 15 ℃ treatment was the highest of all. Moreover, the expression of ejSQE1 and ejSQE2 peaked in 48 h treatment, and expressions of both genes declined significantly in 24 h treatment at 15 ℃(P＜0.05). The results revealed that there was significantly positive correlation between expressions of ejSQE1 and ejSQE2, as well as expression of ejSQE2 and accumulation of UA with any treatment longer than 24 h at 15 ℃. This study reveals the gene sequences and expression characteristics of SQEs under stress temperature, and provides the foundations for the further researches on UA regulatory mechanisms in loquat cell suspension cultures.
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Received: 12 September 2014
Published: 01 March 2015
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