Establishment and application of cDNA-AFLP technical system for xylem tissue of Betula luminifera.

To investigate genes related to wood property in Betula luminifera, a suitable cDNA-AFLP reaction system for xylem tissues of B. luminifera was found in this study based on the comparative analysis of main procedures, including RNA extraction, reverse transcription and purification, and optimizing of selective amplification system. The results showed that CTAB method in combination with the RNAiso reagent was appropriate for RNA isolation of xylem, and cDNA purifying was an essential step after its synthesis. The optimal conditions for selective amplification were obtained as follows: the amount of 4.0 μL of 40 times-diluted preamplification products, 1.6 μL of primer(10.0 μmol/ L), 0.2 μL of dNTP (2.5 mmol/ L), 0.4 μL of Mg 2 + (25.0 mmol/ L), and 0.2 μL of Taq polymerase (5 U/ μL). In addition, 31 out of 64 primer pairs were screened by application of different lignification tissues in the selective amplification. Differentially expressed transcripts during the lignification were identified, and their function was analysed.