|
Tung tree (Vernicia fordii (Hemsl.)) is endemic to China economic trees , has a long history of cultivation , tung oil is an important industrial oils . The flower bud differentiation plant reproductive process is directly related to the yield and quality of fruit . In this paper, the different stages of the tung tree during flower bud differentiation into flower the key genes Tunglfy expression characteristics of the hormone signal response and different hormone stimulation of photosynthesis , tung tree flower bud differentiation mechanisms and exogenous hormone regulation provides a theoretical basis . The main findings are as follows : 1 , by comparing the different concentrations of CTAB different extraction methods , different precipitation method , the amount of different samples of the tung tree total RNA extraction quality impact proposed modified CTAB method . Detected by agarose gel electrophoresis , 18S , 28S , 5.8S visible , 28S : 18S close to 2:1 ; of OD260/OD280 in between 1.7 to 1.9 . Higher purity and integrity of the total RNA improved CTAB method . 2 , analysis of plant actin (β-Actin) genes conserved sequences , primers were designed . The tung tree flower bud total RNA was reverse transcribed first-strand cDNA as a template , using RT-PCR amplified length about 400bp cDNA fragment was cloned into pMD18-T vector . Sequencing and sequence analysis showed that the size of 396bp cDNA fragment , the fragment with castor have a higher nucleotide homology to reach the 97% . 3, to tung trees actin as reference gene , the tung tree flower bud differentiation at different stages of the critical period , the different hormone treatment tung buds RNA was reverse transcribed cDNA as a template , with Tunglfy gene primers and actin gene primers PCR amplification , agarose gel electrophoresis of PCR products , with a brightness value ratio of the target gene and reference gene electrophoresis as Tunglfy relative expression levels , to find 100mg / L abscisic acid expression of Tunglfy has a significant role in promoting , three different concentrations of 6 - BA the tung tree Tunglfy expression inhibited . 4 , select GA 200mg / L , 100mg / L ; the ABA 200 mg / L , 50mg / L ; the 6- BA100mg / L and 25 mg / L treatment tung production materials , Li-cor Li-6400 portable photosynthesis measurement system measurement the tung tree net photosynthetic rate and other indicators , the analysis found that gibberellin has a significant role in promoting the role of the high concentration of net photosynthetic rate stronger than the low concentration of the tung tree photosynthesis .
|