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为了筛选在柑橘韧皮部中特异表达的启动子,构建GRP、Rolc、RSSl异源韧皮部特异启动子控制GUS报告基因的植物表达载体,转化枳[Poncirus trifoliata(L.)Raf.]上胚轴。GUS组织化学染色分析表明,3个启动子均显示了维管束组织特异性。其中,GRP启动子活性强,且具有严格的韧皮部组织特异性;Rolc启动子活性最强,在根和茎中呈组成型表达;RSSl启动子活性最弱。Real-time RT-PCR和GUS酶活性分析进一步证明了GRP启动子具有较强的韧皮部组织特异性,该启动子有望应用于柑橘抗黄龙病基因工程育种。
In order to screen the promoter specifically expressed in phloem of citrus, a plant expression vector containing GUS reporter gene with GRP, Rolc and RSSl heterologous phloem-specific promoters was constructed and transformed into the hypocotyl of Poncirus trifoliata (L.) Raf. GUS histochemical staining analysis showed that all three promoters showed vascular bundle tissue specificity. Among them, GRP promoter activity is strong, and has strict phloem tissue specificity; Rolc promoter activity strongest, root and stem were constitutively expressed; RSSl promoter activity weakest. Real-time RT-PCR and GUS enzyme activity analysis further proved that the GRP promoter has strong phloem tissue specificity, and the promoter is expected to be applied to genetic engineering of citrus anti-Huanglong disease.