| The nurse seed grafting is a new plant propagation of grafting technique. The time of study on this technique which only apply to some plants, for example Camellia oleifera, Ginkg o biloba, Castanea mollissima, Carya cathayensis, is not long in china, which is in its infancy. The nurse seed grafting technique is not on a large scale applied yet. On scientific research, it was carried out preliminary exploration on the grafting method, selection scion, grafting time, cultivated media. The paper adopted the nurse seed grafting union as the experiment material, studied the mechanism of healing process of the nurse seed grafting union.Conventional paraffin sectioning was adopted to conduct the anatomical examination of the nurse seed grafting union during the development. The results showed that at day 4 after grafting the nurse seed grafting union come to being the isolation layer; at day 8 after grafting, come forth the callus in the root stock; at day 16 after grafting, the callus was found in the scion; at day 22 after grafting, the root stock connected the scion; at day 29 after grafting, the nurse seed grafting union formed the vascular cambium; at day 35 after grafting, the vascular bundles was formed, which indicated that the nurse seed grafting union in Camellia oleifera survived.The death rate, soluble sugar content, the relative conductivity, SOD, CAT and POD activity in camellia oleifera grafted chimera through nurse seed grafting were studied for inoculating the mechanism of physiological affinity. The results show that the content and activities of these index increase gradually in early period after grafting and decrease gradually in later period after grafting. The times on the peak value of these index are different, but they appear centrally at 18 days approximately after grafting. The death rate is extreme- significantly and significantly positive correlation with the POD activity and the relative conductivity respectively, but has no significant correlation with the SOD activity.To discuss on the difference of development between the nurse seed grafting union in Camellia oleifera and the healing wound (the contrasts), the test materials were interval sampled during 2-55 days after treatment and some physiological phenylpropanoid metabolic indices were measured. The results showed that the phenylalanine ammonialyase (PAL) activities of the grafting union were highly significant lower than the healing wound; that during 14-35 days after treatment the peroxidase (POD) activities of the grafting union were highly significant higher; The weight of seedling, the content of cellulose, the content of chlorogenic acid and the polyphenoloxidase (PPO) activities of the grafting union had no significant difference; The number and the time of break point in the grafting union physiological indices differentiated from the healing wound. By correlation analysis, the correlations of the PPO activities and the weight of seedling of the grafting union, the content of chlorogenic acid and the content of cellulose, the content of chlorogenic acid and the weight of seedling were equal to the healing wound; The correlations of the content of chlorogenic acid and the PPO activities, the content of chlorogenic acid and the POD activities, the content of cellulose and the weight of seedling were obviously higher than that of the healing wound; But the correlations of the PPO activities and the POD activities, the PPO activities and the content of cellulose were lower than that of the healing wound.In order to find out variation rule in the development of the nurse seed grafting union and different from the nurse seed grafting union and the healing wound, the contents of endogenous hormones IAA, GA3, ZR and ABA from both of them were determined by ELISA. The results showed that for the contents of IAA, ZR and GA, the nurse seed grafting union and the healing wound conform to the stage of development of their, the change in the contents of ABA became more complicated. The endogenous hormone change in the nurse seed grafting union was greater than in the healing wound, which showed that the development of the nurse seed grafting union was more complicated than the healing wound. The contents of IAA and GA3 from the nurse seed grafting union as a whole was below the healing wound, the contents of ZR and ABA approximately was the same to the healing wound, which showed that endogenous hormones IAA and GA3 were very importance to the development of the nurse seed grafting union.The difference of development between the nurse seed grafting union in Camellia oleifera and the healing wound, the mother tree of the scion, the root stock and the germinate buds from the scion were examined by AFLP(Amplified Fragment Length Polymorphism) technology.2 primers pairs were selected from 64 primer pairs to amplify the genetic DNA. Eighty-two loci were obtained, which sixteen were exhibited polymorphism, the average percentage of polymorphic loci was from 13% to 23%. The results showed that the number of bands changed relatively large during the development of the nurse seed grafting union, especially at day 4,16,29 and 35 after grafting; the changes in the number of bands during the development of the healing wound were small; the number of bands in the germinate buds from the scion were more than the mother tree of the scion and the root stock; from E1Ml primer pair, there were three bands not in the mother tree of the scion but in the root stock and the germinate buds from the scion; there were three bands not in the mother tree of the scion and the root stock but in the germinate buds from the scion; there was one bands not in the germinate buds from the scion but in the mother tree of the scion and the root stock. According to the above analysis, the gene changes during the development of the nurse seed grafting union in Camellia oleifera would be left behind to further research.Proteomics method was used to compare and identify the differential proteins of the development of the nurse seed grafting union in Camellia oleifera. Over 1000 protein spots were detected on 2 DE gels, mage Master TM 2D Platinum software7.0 was applied on gel pattern analysis,40 differentially expressed proteins were detected.34 differential proteins were identified by MALDI-TOF-TOF/MS and protein database searching, the identification rate was 85%. Biological function analysis showed these identificated proteins related with photosynthesis, respiration, energy metabolism, transcription and translation, secondary metabolism, the cells growth and differentiation, regulation and stress tolerance. The differential expression of these proteins showed that the healing of the nurse seed grafting union in Camellia oleifera changed a series of physical and chemical metabolism. |
PDF Full Text Request