Studies On The Construction Of CDNA Library In Drought-Induced Popμlus Simonii And Preparation Of Library-Screening Probe With The MRNA Differential Display

It's an important branch of plant research on stress resistance to enhance or improve the capability of drought and water deficit-tolerance in plant.With the advance of molecular biological technique,bioengineering becomes a rapid efficient methond of improving plant resistance to drought,water deficit,on the viewpoint of many researchers.Gene cloning,as an upstream technique of bioengineering,was being developed after 1990s In this thesis,the author elaborated the situation,characteristics,the advantages and disadvantages of new techniques in the field of plant gene cloning.Especially discussed the differential display reverse transformation (DDRT)PCR.To construct the cDNA library of drought-induced Popplus simonii and to prepare library-screening probe.The results were showed that:the drought-treated library comprised with approximately 2 × 10⁵ recombinant clones and the control library comprised with approximately 0.8 × 10⁵ recombinant clones.After amplification via culture,both of them increased their library capacity to 2.0 × 10¹⁰pfu/nil and 1.2 × 10⁽80pfu/ml responsively.By means of silver staining,DDRT-PCR products were displayed.There were about 5,000 bands observed in both treated sample and control sample,35 differential bands were acquired.Through the recovery of interesting bands and reamplication of products,sequencing,compared with EST BLAST online.some cDNA fragment with relative to the tolerance to drought,water-deficit,were found.as well as,to improve the recovery of interesting band from polyacrylamide gel.These results were essential to library screening following them.