摘要
利用含有过表达抑制子载体的根癌农杆菌分别在番茄源叶和果实中进行瞬时表达,快速验证转化酶抑制子(inhibitors of invertases,INH)转基因的表达情况及对果实发育过程中转化酶活性的影响。Micro-Tom在源叶注菌处理后,Lin6的表达含量增加,表明转化酶在植株中响应病菌侵染;但转INH在源叶几乎没有明显表达变化。果实注射后,注射空菌(EHA105)菌液和空载体(pBI121-2A11)菌液后,转化酶(invertases,Inv)基因Lin5、Lin6表达量没有明显改变,但Inv的活性明显升高1倍左右,这表明转录数量并不能反映翻译后的蛋白质量;在注射转基因(p1300-2A11-INH)菌液后3d,INH的表达明显增加,是其它处理的数倍,这是由于在果实特异性启动子2A11的作用下,INH在果实中特异地表达。而且注射p1300-2A11-INH菌液3～5d后,Inv的活性显著下降,尤其胶质胎座中最为明显。这一结果表明INH主要在翻译后水平调控Inv的活性。
The purpose of this study was to use an Agrobacterium-mediated transient expression system containing the expression carrier inhibitor to prove the effect of INH gene expression and the invertase activities in the process of tomato fruit development. Micro-Tom treatment after Agroinfiltration in source leaves, the expression of Lin6 was increased, indicating that cell wall invertase should response to pathogen infection in plants. Micro-Tom were injected with EHA 105 or injected that contains pBI121-2A 11 plasmid, expression oft he invertase gene （Lin5, Lin6） did not change obviously. Whereas tomato was injected strain containing p1300--2A ll-INH plasmid after 3 d, expression of INH was significantly increased several times. This would be contributed by the function of the fruit specific promoter 2A11, resulting in INH specific expression in the fruit, while activity of Inv decreased significantly after 3 d to 5 d infection which injected strain containing pl3OO-2All-INH plasmid, especially expression in pectinic placenta was most obvious. The results in this research presented that the activity ofinverase was regulated by inhibitor in the post-translational level.
出处
《分子植物育种》
CAS
CSCD
北大核心
2013年第2期217-224,共8页
Molecular Plant Breeding
基金
国家自然科学基金项目(30971999)
辽宁省重点实验室项目(2009S092)共同资助
关键词
番茄
转化酶抑制子
瞬时表达
实时荧光定量PCR
Tomato, Invertase inhibitor, Transient expression, Real-time fluorescence quantitative, PCR
