Cloning, expression and single nucleotide polymorphism analysis of phenylalanine ammonia-lyase gene in Eucalyptus urophylla

Objective  Through obtaining phenylalanine ammonia-lyase (PAL) gene sequence of Eucalyptus urophylla, to analyze the expression patterns of PAL gene in nine different water-fertilizer treatments and its single nucleotide polymorphism (SNP) in E. urophylla populations, and provide a basis for studying the correlation between PAL gene and stress resistance of E. urophylla.

Method  Primers were designed according to PAL gene sequence of E. grandis, and PAL gene sequence of E. urophylla was obtained by gene cloning and sequencing. The expression patterns of PAL gene in different treatments were obtained by reverse transcription quantitative polymerase chain reaction (RT-qPCR).

Result  The total length of PAL gene sequence was 4 507 bp, including 166 bp 5′-untranslated region, 411 bp 3′-untranslated region, 1 759 bp intron region and 2 172 bp protein coding region (with two exons, 422 and 1 750 bp respectively). The expression of PAL gene was the most abundant in AT4 treatment, and lowest in AT5 treatment. The PAL gene of E. urophylla was highly similar to that of E. grandis. The SNP sites of PAL gene in E. urophylla were mainly concentrated in intron region, indicating that PAL gene was stably inherited in long-term evolution history.

Conclusion  The complete length of PAL gene sequence in E. urophylla is 4 507 bp. PAL gene is stably inherited and plays an important role in stress resistance of E. urophylla.