摘要
将大麦β-1,3-葡聚糖酶同功酶基因(GIII)启动子(PGIII)与其报告基因gus(β-葡聚糖酸醛苷酶基因)耦联,构建植物表达载体,通过农杆菌介导法转化水稻。PCR、DNA印迹法结果显示,构建的pGIII-gus表达载体已整合到水稻基因组DNA中。GUS组织化学染色、RNA印迹法及荧光法结果显示,该启动子驱动的gus在水稻叶片中为低水平表达;而用水扬酸(SA)与稻瘟菌来源的激发子处理,可诱导gus的高水平表达。T1代种子的GUS组织化学染色结果也表明,SA与激发子可以诱导高水平的PGIII活性。这些结果表明PGIII是一种强诱导型启动子,并可能是一种病原菌诱导型的启动子。
A promoter of the gene encoding β-1,3-glucanase isoenzyme GIII was amplified frombarley genomic DNA using PCR. The GIII genepromoter, designated PGIII, was ligated upstream ofthe gus report gene and pGIII-gus fusion fragmentwas then cloned into a binary vectorpCAMBIA1300 for Agrobacterium-mediatedtransformation of rice (Oryza sativa L. cv. Taipei309). PCR analyses indicated that the fusion genewas present in all T0 transgenic plants (Fig.2). Theintegration of the gene into rice genomic DNA wasfurther confirmed by Southern blot (Fig.3). His-tochemical staining and spectrofluorophotometricanalyses of transgenic rice leaves showed that theGUS activity was increased after treatment with SAand fungal elicitor (Fig.4A, C). Northern blot analysisalso showed expression of such induction (Fig.4B). Histochemical staining of T1 rice seeds displayedmarked GUS activity after induction with SA andelicitor, while no GUS activity was detected in un-treated T1 rice seeds (Fig.5). The results presentedhere strongly suggested that PGIII could be patho-gen-inducible promoter.
出处
《植物生理与分子生物学学报》
CAS
CSCD
北大核心
2005年第2期143-148,共6页
Journal Of Plant Physiology and Molecular Biology
基金
国家863高技术研究发展计划(No.2001AA212191)
中国博士后科学基金(No.2003034446)
国家自然科学基金(No.30470913)
广东省自然科学基金重点项目(No.04105513)资助~~
