Hybrid Momordica Charantia.L Varieties Purity Identification Research Based On Ultrathin-layer Iso-electric Focusing Electrophoresis

Extracted bitter gourd seed storage proteins and conducted electrophoresis,screened out a kind of storage proteins which was appropriate for ultrathin-layer iso-electric focusing electrophoresis.Establish a electrophoresis optimizing system that would be used for several hybrid bitter gourd varieties purity identification.Explore a way of simple、 fast、 accurate and costeffective melon seeds purity identification that based on ultrathin-layer iso-electric focusing electrophoresis.1、 Extracted4kinds of bitter gourd storage proteins,include of Glubulin、 Glutelin、 Prolamin and Water soluble protein.Took advantage of ultrathin-layer iso-electric focusing electrophoresis to sift proteins.Electrophoresis atlas results showed that Glubulin and Glutelin samples salinity overtop,contained impurities,and the protein samples engendered some degree of denaturation,which resulted in electrophoresis bands skew、 fuzzy、 trailing and electrophoresis lane out-of-straight. Prolamin content was0.93mg/ml,but electrophoresis atlas bands showed to be few,lightcolored and fuzzy.Water soluble protein electrophoresis bands soigne,quantity,lane straight and limbe background,which meet the requirements of ultrathin-layer iso-electric focusing electrophoresis to identify seeds purity.2、 Screened out the optimal UTLIEF condition was:ultrathin-layer polyacrylamide of0.15mm thickness;cold acetone degrease3times in-29℃refrigerator;add to0.3ml extractives for0.1g sample;extracting1.5hour under4℃;add sample volume10μl.3、 Five bitter gourd F1hybrids were subjected to seed water soluble protein ultrathin-layer iso-electric focusing electrophoresis and field grow-out test. Protein marker bands were found in all parent lines of the five hybrids, and the electrophoresis of these five F1hybrids contained both male marker bands and female marker bands. The results of UTLIEF tests were identical to that of the grow-out tests, suggesting that UTLIEF could be employed as a rapid and practical method for varietal purity testing in door for hybrid bitter gourd seed.However,it was not easy to find the marker bands in varieties of resembled genetic constitution.4、 Both experimental facilities and chemicals were domestic,while part of mountings were insteaded of ordinary materials,which reduced the cost.Meanwhile,it’s found that domestic facilities were existed deficiencies in aspects of quality、 precision and consumable material mating.Which was not able to meet high precision and high flux seed inspection.