越橘原花青素合成相关基因VcLAR和VcANR的克隆和功能鉴定

Molecular cloning and functional identification of proanthocyanidin synthesis related genes VcLAR and VcANR of blueberry

本研究以越橘(Vaccinium corymbosum Duke)为试验材料,克隆原花青素合成基因VcLAR和VcANR,分析其表达模式及其对水杨酸和茉莉酸甲酯的响应,鉴定其在原花青素合成过程中的作用。结果表明,成功获得了越橘VcLAR(GenBank登录号为MH321470)和VcANR(GenBank登录号为MH321471)基因。VcLAR和VcANR在根、茎、幼叶、花以及不同发育阶段果实中均可表达,但相对表达量存在差异,在绿果中的相对表达量最高。在不同发育阶段果实中,原花青素含量在绿果中最高,在蓝果中最低。水杨酸处理可促进VcLAR和VcANR基因的表达,茉莉酸甲酯处理可抑制VcLAR和VcANR基因的表达。在转VcANR基因拟南芥株系中,原花青素的含量显著高于野生型。在转VcLAR基因拟南芥株系中,原花青素含量与野生型相比无明显变化。由此推测,VcLAR和VcANR在越橘果实原花青素积累过程中发挥重要作用,并受水杨酸和茉莉酸甲酯的调控。

In this study,using Vaccinium corymbosum Duke as experimental material,proanthocyanidin synthesis genes VcLAR and VcANR were cloned,the expression patterns and their responses to salicylic acid(SA)and methyl jasmonate(MeJA)were analyzed,and their roles in proanthocyanidin synthesis were identified.The results showed that VcLAR(GenBank accession No.MH321470)and VcANR(MH321471)were cloned from blueberry.The quantitative real time PCR(qRT-PCR)analysis results indicated that VcLAR and VcANR expressed in roots,stems,young leaves,flowers and different development stages of fruits.However,the expression levels were different,and the highest expression level was in green fruits.The content of proanthocyanidin was highest in green fruits and lowest in blue fruits during different development stages of fruits.The expression of VcLAR and VcANR was induced by SA treatment and suppressed by MeJA treatment.The content of proanthocyanidins in transgenic VcANR Arabidopsis was significantly higher than that in wild type.There was no significant change in proanthocyanidin content between wild type and transgenic VcLAR Arabidopsis lines.It is speculated that VcLAR and VcANR play an important role in the accumulation of proanthocyanidins in blueberry fruits and are regulated by SA and MeJA.