黄烷酮3-羟化酶基因表达分析及转基因大豆新种质创制

黄烷酮3-羟化酶(F3H)是大豆异黄酮代谢途径重要酶类。研究F3H在不同大豆品种中的表达差异,并应用转基因途径创制新种质是提高大豆异黄酮含量的重要途径。以高异黄酮含量大豆品种中豆27和低异黄酮含量大豆品种楚秀为试材,采用q PCR技术,分析F3H在大豆不同发育时期、组织部位中的表达差异;并利用花粉管通道转化技术,获得转F3H阳性植株。结果表明:F3H在2个大豆品种R1~R7期叶片中的表达模式不同,中豆27的F3H表达量在R1期最高,而后开始下降,R2~R5期维持较低水平,R6期略有上升,R7期又出现下降;楚秀F3H表达量在R1~R4期较低,R5期出现表达高峰,R6期开始下降。F3H在2个大豆品种R5~R8期籽粒中的表达模式基本相同,表达量均从R5期开始下降,且R6~R8期维持较低水平。基于此,构建F3H RNAi反义载体,并利用花粉管通道技术转入不同大豆品种,获得了5个转基因新材料。

Expression Analysis of Flavanone 3-Hydroxylase Gene and Innovation of Transgenic Elite Germplasms in Soybean

Flavanone 3-hydroxylase (F3H) is an important enzyme involved in the metabolism pathway of soybean isoflavones. Analysis of the expression pattern of F3H in different soybean varieties and cultivation of the transgenic germplasms with F3H are important methods to enhance the isoflavone of soybean. In this study, the expression patterns of F3H at different developmental stages and organs in both high isoflavone content soybean variety Zhongdou 27 and low isoflavone content germplasm Chuxiu were ana-lyzed by qPCR and transgenic elite germplasms with RNA interference vector of F3H were obtained. The results indicated that F3H showed quite different expression patterns between the two soybean varieties in the leaf. The expression level of F3H in Zhongdou 27 began falling at the R1 stage,maintained lower ex-pression level at R2-R5 stage,increased a little bit at R6 stage and got down again at R7 stage. While in the leaf of Chuxiu, F3H showed lower expression level at R1-R4 stages, highest level at R5 stage and falling expression level at R6 stage.The expression patterns of F3H were almost the same in the seeds of these two soybean varieties, beginning falling at stage R5 and maintaining the lower level at stages R6-R8. Based on these results,the RNA interference vector of F3H was constructed and in-troduced into soybean varieties through the pollen-tube pathway transgenic technology,and five trans-genic elite germplasms were obtained by now.