Germplasm Evaluation And Preliminary Study On Cultivated Physiological Of Lonicrea Japonica Thunb

In order to provide theoretical basis for Lonicrea japonica induction and standard cultivation, the Growth development and physiological characteristic was studied. And in order to provide basis for the related molecular research, DNA barcoding and RTq-PCR techniques were discussed for the identification and key enzyme genes on the secondary metabolism, The results were as follows:1. Phenological law and growth indexs of different germplasms of Lonicrea japonica were studied. The results showed that the date of sprouting and new shoots growing stage begins with mid February and early March, but had no significant differences between different gerplasms. Squaring period and blooming stage begins with late April and early May. Fruiting stage most begins with late November and early December, and then get into the winer period.2. DNA barcoding sequences(ITS2 and psbA-trnH) were selected for the identication of Lonicera japonica. The ITS2 and psbA-trnH amplification and sequencing efficiency were high. The rate of successful amplification kept 100%, The sequencing efficiency of ITS2+ psbA-trnH were 100%. Both of them have much variable sites, so as to identify different varieties of Lonicera japonica efficiently.3. Analysis of chlorogenic acid, luteolin glycosides, total flavone among varieties of Lonicera japonica. The results showed that significant differences exist between different varieties. All variety types showed higher content of chlorogenic acid than Chinese pharmacopoeia addiction 2010 except for the golden flower of "Juhua 1", "Damao","Yate". All variety types showed higher content of luteolin glycosides than Chinese pharmacopoeia addiction 2010.4. On the preliminary stage of branch, the Paelobutrazo and DPC with different concentration can increase the number of flower shoots and delaying branches'growth and improving the drying rate of flower, but had no significant impacts on leaf indexs. Different concentrations of Paelobutrazo and DPC can increased the contents of flavonoids and chlorogenic acid in flowers, The highest can be up to 34.03%. When spraying the plant growth regulator, the urea and borax can promote the absorption except the Paelobutrazo.5. To detect the influence of spraying Micronutrient and Amino Acids on Growth and Yield?Quality of Lonicera japonica. The results showed that foliage spraying the Phe, Lyr,Zn2+ and Cu2+ with different concentrations has no appreciable effect on leaf area. By spraying the Phe, Lyr with different concentrations can affect the C-metabolism remarkable, but made opposite effection on N-metabolism. There was no significant difference between the content of flavonoids in flower than control except Zn2+. All of the content of Total flavonoids was lower than control, but the Chlorogenic acid was opposite. The spraying time or concentration have more influence on the ion content of leaf. The Zn2+?Cu2+?Fe2+ content of flower was significantly lower than control except 30 mg·L-1 CuSO4 treatment.6. To observe the influence of Penicillin on Photosynthetic characteristic of Lonicera japonica. As the rise of concentration, the antioxidant activities in leaf were rised gradually, the treatment with high concentration (800 mg·L-1) can inhibit the POD activities; The Penicillin with low concentration(400 mg·L-1) can increase the ?PS ?, qP and ETR effectively.when spraied the Penicillin with 600 mg·L-1, the content of chlorophyl a/b were increased by 8.6%?58.8%. when the concentration were 800 mg·L-1, The stomatal conductance (Gs) were enhanced by 162.9%, but exert some negative effects on the ?PS ? and ETR, and the content of chlorophyll was also decreased.7. The contents of CGA of different tissues in Lonicera japonica were detected by HPLC, and the expression of aroB gene related to the CGA biosynthetic pathway were analyzed. The results showed that the ranks of CGA contents of different tissues were Flower>Young leaf>Old leaf>Young stem>Root>Callus, all the differences were significant. The realtime quantitative-PCR results showed that the rank of expression of aroB gene were Flower> Young leaf> Callus> Old leaf> Young stem> Root.