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Knotweed (Polygonum Cuspidatum Sieb et zucc) is a perennial herb Polygonum Ke Luo, is commonly used in traditional Chinese medicine, the main origin for the East, Gansu, Sichuan, Yunnan and other places. Polygonum cuspidatum extract with a variety of functional roles, there are anti-inflammatory, prevention of cardiovascular disease and other pharmacological activity, while its main active ingredient is resveratrol (resveratrol) and emodin (emodin). As a phytoalexin resveratrol, not only can significantly improve the UV plant, fungi and other injuries resilience, but also plays a protective effect on the cardiovascular system. In this study, knotweed tender stems as explants, with its induced callus of materials, through the dark culture after ultraviolet irradiation treatment, plus H202 and H202 quencher studied callus growth and white resveratrol content changes, and through semi-quantitative RT-PCR amplification of gene expression levels of the RS discussed the situation. The main results are as follows: 1. Different UV irradiation and the different sampling time Polygonum cuspidatum callus H2O2 content in different time after UV irradiation, the dark culture callus culture time 1-5h callus H202 overall content trend is increased and then decreased, H202 levels were in the first 2h, 4h a peak, all treatments were higher than H202 CK, which deal with four callus H202 content is higher. In the first 2h, the processing 4 peak 2.68nmol · g-1, increased by 54% compared with CK. In the first 4h, the processing 4 peak 2.84nmol · g-1, increased by 37% compared with CK. Dark incubation time 4-20d callus H202 modest overall change in the content of the sampling time on the H202 content was not influenced. Two different times and different sampling times ultraviolet radiation on callus morphology and resveratrol (Res) Content of ultraviolet irradiation at different times after treatment cuspidatum callus, 1-5 hours, callus no significant change in color still green, fluffy shape. Fourth day onwards, with the passage of time, callus tissue aging, slow growth, the aging than CK about 8 days in advance. The content of each treatment Res trend is short (1-5h) erratic ups and downs, to reach the first four days, Res were significantly increased, and peaked in the first 12 days to deal with three treatment Res content than CK differences are significant; treatment after the first 12d, the content of each treatment decreased Res. Throughout Res content change trend in the treatment 3 Res content has been at a high level. 3 medium plus H2O2 and H202 quencher Res callus morphology and H202 according to the relative content of the highest content of callus content ratio of the H202, H202 and H202 were added in the medium quencher, following generation of callus on the medium after inoculation, 1 to 5 hours, no significant change in the callus, and the fourth day of H2O2 callus has significant deterioration, and the medium was added to quench H2O2 agent after callus subculture trends and CK similar morphological changes in the first four days no obvious aging, 12d after arrival, callus appeared aging. H202 and H202 plus calli quencher still in accordance with a short sampling time (1-5h) and long (4-20d) to be sampled, after removing callus Res HPLC determination, the results show more plus H202 Res wound tissue content 1-5 hours and 4 days were slightly higher than CK, and after 4 days, Res were significantly lower than CK, and significant differences in 12d. Join H202 quencher callus ,1-5h and CK within the Res showed no big difference when ,4-20d Res content increased slightly, but significantly lower than CK. Detection of gene expression levels 4.RS containing resveratrol highest callus, plus H202 callus and control callus were extracted and subjected to electrophoresis of RNA integrity test results show that the three samples 28S , 18S, 5.8S showed clearly that good quality total RNA extracted and relatively complete, consistent follow-RT-PCR amplification experiments. The resulting RNA was extracted semi-quantitative RT-PCR amplification, after the reaction, the product take 1.2μ1 to add a scavenger TAE of Rnase buffer, 90V/cm 20min after electrophoresis in ethidium pictures. Three samples were amplified 600bp about internal reference gene actin, and the brightness and width no difference, indicating stable internal reference gene expression, using specific primers, Res highest group in the above 1000bp 1200bp amplified to the desired specificity about fragment A, while the control group and the group did not amplify plus H202 to this fragment shows that, after UV irradiation, RS gene expression level has been improved.
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