Studies On Tomato Transformation With BFL Gene By Agrobacterium Tumefaciens

Adverse environment such as weather and soil conditions that affect the tomato yield and quality,in particular the deteriorating environmental conditions in recent years,so that the living organisms on earth, including tomato growing suffered serious harm,which improved tomato resistance is even more urgent and important. With the rapid development of modern molecular biology,some resistance functional gene is gradually separated and cloned,and applied modified tomato resistance.The gene of Bountiful(BFL) used in this paper is isolated form Arabidopsis thaliana, an AT-hook DNA binding transcription co-factor,identified via a gain-of function Arabidopsis mutant bfl-D. Both at home and abroad there has been no use of this mutant applied into tomato. This study used leaf disc and agrobacterium mediated transformation system into tomato. The results during the experiment are as follows:1. Development of an efficient tomato transformation systemWe used leaf disc and agrobacterium mediated transformation system into tomato 'M82'. We selected proper medium and determined 50mg/L kanamycine being the effective selection antibiotic concentration in transforming tomato.Maintaining 50mg/L kanamycine in the whole process of transformation reduce significantly the escapes and increase the efficiency to detect positive transformants. In the present case,70% of all the rooted plants were detected positive.2. Transformants analysis of BFL in tomatowe have introduced Arabidopsis BFL gene into 'M82' Tomato by Agrobacterium tumefaciens and then obtain 53 transgenic plants, most of which show large wrinkled leaves,vigorous growth habit and leaves of increased size. Initial PCR and RT-PCR analysis shows that there are 38 plants integrated exogenous BFL gene. And histochemical stanining method on leaf tissues of these plants revealed that the expression of GUS reporter gene in plant leaves. In addition, analysis of the leaf structure suggested that the epidermal cell size of transgenic paints is 1.5 times larger than that of WT.