不同量化RT-PCR的方法在棉花纤维GhCAD基因表达中的研究

以陆地棉（Gossypium hirsutum L.）高代品系CJ01在开花后5、10、15、20、25d（DPA）共5个不同发育时期的棉花纤维为材料，分别利用常规半定量RT-PCR法、QuantumRNA?18S内标相对定量法以及实时荧光定量 RT-PCR法，对棉花肉桂醇脱氢酶（GhCAD）基因在棉花纤维不同发育时期中的表达进行相对定量分析，并测定了内标表达的稳定性。结果表明3种方法获得相似的结果，GhCAD基因在5～15DPA时表达量较低，20、25 DPA时表达量升高。3种方法虽各有利弊，但均为研究基因量化表达的有力工具。

Comparative Study on the Different Methods of RT-PCR for Ghcad Gene in Developing Cotton Fiber

In this study,cotton fibers from 5,10,15,20,25 days post-anthesis(DPA)of strain CJ01 in upland cotton (Gossypium hirsutum L.)were used as materials.The relative quantitative expression of GhCAD gene at different developmental stages of cotton fiber were analyzed by conventional semi-quantitative RT-PCR,QuantumRNA~(TM) 18S internal standard relative quantitative RT-PCR and real-time RT-PCR respectively.The stability of internal standard expression was also tested.Results showed that difierent methods obtained similar results:the relative quantitative expression of GhCAD Was lower during 5～15 DPA and increased during 20～25 DPA in cotton fiber.Though three methods have their advantages and disadvantages,all of them are useful tools for testing of gene relative quantitative expression.