Optimization of transient expression by Agrobacterium in almond

Since permanent transfer of genes and transgenic plant production, particularly in perennial plants such as almonds are time and cost consuming, optimization of various factors affecting the transgenic plants is critical. Transient genetic expression using Agrobacterium (Agroinfiltration) in plant is a useful tool to study gene function indifferent gene expression systems. Since optimization of factors that could affect the transgenic methods is necessary for gene transfer programs, Because This study was aimed at developing an efficient system for transient expression via Agrobacterium in almond leaves for rapid prediction of systems performance. Here the lichenasereporter gene was usedto assessthe parametersinfluencing thelevel oftransient expression. The main advantagesof using thisgene includeeasy operation, high sensitivity and biosafety. Three different constructs carrying lichenase gene and differentregulatoryelementswereinfiltrated intoleaves ofmamaei and shahrood 12cultivars,by two Agrobacteriumstrain, LBA4404 andGV3501,using syringewithoutneedle.After threedays,the amount of active proteinwas measured. Statistical analysis showedthat the highestamount ofprotein was obtainedin theleavesof mamaei cultivar transformedby theexpression systemcausing accumulation ofproteins in theendoplasmic reticulum by LBA4404 strain. So forrapidproduction ofrecombinant protein in Almond, theagroinfiltration method by using the related expression system, LBA4404 strain and mamaei cultivar, isrecommended.