Abstract: Castor is one of the most important oil crops with high economic value.In this work, the cDNA-AFLP system of castor seed was established and optimized to lay a foundation of research fatty acid gene expression in different castor materials or different developmental stages.The main results were as follows:total RNA was extracted and then reversed transcription into cDNA.cDNA digested with EcoR Ⅰ and Mse Ⅰ was joined with EcoR Ⅰ and Mse Ⅰ joint.The jointing products diluted by ddH₂O according to the ratio of 1∶ 40 were used in pre-amplification.And the optimized selective amplification system was performed with 10×PCR Buffer 2.0 μL、dNTPs(10 mmol/L) 1.4 μL、Mg²⁺ (25 mmol/L) 1.4 μL, jointing product 1.0 μL, E_x primer(50 pmol/μL) 1.2 μL, M_y primer(50 pmol/μL) 1.2 μL, LA Taq(5 U/μL) 0.2 μL、ddH₂O 11.6 μL.With the optimized PCR reaction system, we estimated 256 primer combinations with SDS-PAGE and gained 10 combinations which were suitable for cDNA-AFLP analysis of castor seed.

Key words: Castor, Fatty acid, cDNA-AFLP

CLC Number: 

• S565.6