Abstract:
Berberine bridge enzyme(BBE,EC1.5.3.9) is a key enzyme in berberine biosynthesis. In this study,
HbBBE1 gene was cloned in rubber tree by RT-PCR. The
HbBBE1 promoter contained light, gibberellin, salicylic acid, wounding, fungal elicitor, defense, and stress responsive elements. The predicted protein of HbBBE1 contains a signal peptide, a trans membrane region, an FAD binding domain, and a BBE domain which is found specifically in the berberine bridge enzymes and berberine bridge-like enzymes. It shared the highest similarity with BBE protein from
Manihot esculenta with 80.1% identical residues. Comparing expression among different tissues of rubber tree demonstrated that
HbBBE1 has a particularly high expression level in the latex.
HbBBE1 expression was significantly upregulated by biotic and abiotic stress, i.e. powdery mildew infection, drought stress, high intensity light irradiation, H
2O
2, and both tapping treatment in virgin trees and mechanical wounding in seedlings. Furthermore,
HbBBE1 transcript was remarkably induced by application of exogenous hormones, including gibberellic acid(GA
3), salicylic acid(SA), ethephon(ET), methyl jasmonate(MeJA), and abscisic acid(ABA). Among these treatments,
HbBBE1 transcript was quickly induced by GA
3 stimulation, and reached a maximum(6.3-fold) at 0.5 h. The increased expression of
HbBBE1 was highly induced by ABA, ET, and H
2O
2 stimuli, and it accumulated dozens of times comparing that at the untreated control.To summarize, our study suggested that
HbBBE1 played multiple roles in response to biotic and abiotic stress in rubber tree.