Study On The Dynamic Changes Of Nutrition Composition And The Differential Proteome During The Ripening Stages In Mulberry

The mulberry is abundant of sixteen kinds of amino acids,seven kinds of vitamins and some minerals including reducing sugars,sucrose,zinc,manganese,calcium,and pectin,fruit acid,carotene,flavonoids,turin,anthocyanin,tannin,cyanidin,volatile oil,it is a natural and pollution-free food,and so popular to consumers,has already been included in the health products which can be used as food and drug.there is a good prospect in comprehensive utilization of mulberry resources.But the mulberry has characteristics of a short ripening period and a short time on sale,so it is of intolerance storage.Therefore,it is an important research topic to keep mulberries fresh,and of a Theoretical and Practical Significance on its mechanism of physiological and molecular regulation during the ripening stages.In this study,we used two mulberry cultivars,Da 10 and SG01,as experimental materials, take the comparative analysis on the changes of nutritional components during the different ripening stages,then analysis on the differential expression proteins applying proteomics and molecular biology technology.Finally,particial sequence of cell wall invertase is cloned,the results are as follows:1.During the development of mulberry ripening,there are a decrease of contents in protein,ash and calcium,and an increase of contents in soluble sugar,sugar acid ratio and vitamin C,taking on the similar tendency between Da 10 and SG01,however,the contents of soluble sugar,sugar acid ratio and vitamin C in Da 10 have more than that in SG01.2.Using the leaf and fruit of mulberry as material,we have optimized two-dimensional gel electrophoresis sample preparation technology,and have proved that protein sample preparation optimized is more effective on the colour of protein sample,solubility and the effect of electrophoresis separation in the leaf and fruit of mulberry.3.Using two methods of SDS-PAGE and 2DE to compare on the differential protein expression in different tissues.We gain the 22 protein components which differential expression was more than two times,2 spots which specific expressed in the green-ripening stage and 3 spots in the half-ripening stage.Then using 2DE anlysis on the differential proteins during the different ripening stages,there are 32 spots which differential expression was more than two times,in which 6 spots were expressed most in the full ripening stage.4.We choose 8 spots,which differential expression was more than two times,to carry in-gel digestion and mass spectrometry,by retrieval database and protein function prediction,it shows that 3 spots are related to photosynthesis,3 spots related to low temperature stress,1 spots related to glucose metabolism.5.According to the results of the protein identification,based on former research,we cloned the cell wall invertase(CWI),analysised on the sequence of CWI and bioinformatics,it showed that CWI has more high homology with Populus alba×Populus grandidentata and Soianum lycopersicum,and low homology with Oryza sativa,Triticum aestivum,and this CWI seguence we get has high conservation.