Ploidy Identification System Of Camellia Sinensis

In this article,we explored anther culture of Camellia Sinensis,established the way of ploidy identification basing on FISH and estimated DNA variation among Tea species.Main conclusions were as follows:1.Anther culture of Camellia Sinensis. In this study, ’shuchazao’ and ’longjing-43’ were used to investigate influence of different factors with N6 basal medium, including different species, different medium.2.Microscopic examination of the root. This study established chromosomes technique:taking roots at10.am, pretreating in 8--hydroxide quinolino under 4 for 4 ℃hours,fixing in Carnoy’s Fluid for 4 hours,dissociating in 1mol/L hydrochloric acid for 5min,dyeing with phenol magenta for 10 min,then observing under the microscope.3.FISH.We had analyzed ’shuchazao’ with the probes of 45 S r DNA. six signals were detected on chromosomes. The results indicated that ’shuchazao’ was diploid4.Estimation of nuclear DNA content. We selected internal standardization among chicken red blood,maize and soybean.the optimum was soybean.We use soybean to estimate DNA content of different tea species.including ’shuchazao’、’longjing-43’、 ’baihaozao’、’fuyun-6’ and so onChoosing ’longjing-43’ as standardization to analysis diploid cultivar, triploid cu ltivar and Callus of ’Shangfuzhou.’5.Determination of DNA content of different tissues. Young leaves, flowers and filament were used to estimate DNA content.The relative DNA content of young leaves was 197.13, flowers was 204.26, filament was 193.31. Compare with Young leaves, flowers and filament were more suitable for FCM, followed by its fewer secondary metabolites.6.The influence of cold stress on FCM analysis. ’Shuchazao ’leaves were treated in the refrigerator at 4 ℃for 2 hours,4 hours,8 hours,12 hours and 24 hours.the result shows that the relative DNA content become lower with the time goes by.cold stress have influence on estimating of relative DNA content.Then put the leaves with 4 ℃treatment in 25 ℃incubator for 2 hours,4 hours,8 hours,12 hours and 24 hours.the result indicated that the relative DNA content turn to normal for 12 hours...