Study On Purity Identification Of Rape Seeds By Esterase Isozymes And SSR Markers

In recent years,the ways of the seed purity identification are more and more rich.This paper reviewed on the basis of predecessors’ morphological marker, cell markers, biochemical markers and molecular markers(RFLP, RAPD, AFLP, SSR, EST-SSR), hyper-spectral imaging in seed purity identification, and objective over-viewed the basic principle and progress of the advantages and disadvantages of each technology.In this paper,author used the esterase isozyme electrophoresis and the method of SSR marker to analysis hybrid rape Shanyou 2014 and its parents;Mechanized harvesting Shanyou 18 and its parents seed purity identification as well as the variety purity and authenticity were verified in the field.The main results were as follows:1 the esterase isozyme electrophoresis1) Formulation screening different esterase isoenzyme stain, because reagent is expensive, and every time when you use, a new preparation is initiated, so in the case, it is meaningful that appropriate screening reagent dosage to ensure that the enzyme spectrum dyeing dyeing agent formula is clear and tidy.2)The best dye formula was determined, material 1521 C germination experiments indoors, starting from the third day author randomly take samples for four consecutive days to determine the best materials.From the map found that fifth day enzyme expression of esterase isozyme quantity is more, dyeing with clear and tidy.3)Using the traditional method of esterase isozyme electrophoresis appraised Shanyou 18, Shanyou 2014 and 1521 C. Experiments show that: the enzyme spectrum can only distinguish between parent and hybrids, but unable to distinguish between maternal and hybrids.Therefore, with the esterase isozyme cannot effectively distinguish between Shanyou 18;Shanyou 2014 hybrid seed purity.4)With the improved esterase isozyme electrophoresis method,author directly took Shanyou 18 dry seeds, Shanyou 2014 dry seeds to do experiment.Results show that geting rid of concentrated glue, not only saved reagent, time, and its enzyme spectrum and compared with seedling experiment, the experiment not only easy to operate, easy to promote, but also enzymes were clear and neat,and the expression of enzyme spectrum amount were more than seedlings as materials.5)With the 1521 C gathered in a year,1521 C dry seeds validated experiment, found the map of are the same, no difference.This show that using dry seed rape hybrid carried out directly as purity identification experiment’ s material not only saved time operation, simple and accurate but also not was limited by time.2.SSR marker identificationUsing the method of SSR marker to identificate purity of Shanyou 2014; Shanyou 18 and its parents, primer CB10159 can amplify the father in their hybrid F1, maternal complementary specific band, the primer can be used to identify 362 A and 830 C;9 A, 1521 C and its corresponding generation F1;BRAS100 is a specific strong primer, only amplification complementary belt in 362 A and 830 C.Therefore, SSR markers can be used for both sets of materials for the purity identification.3.Use the field planting experiment to identifyPoor flowering on Shanyou 2014 hybrid sterility plant is 2, complex number is 0, the total number of 492;Shanyou 18 hybrid sterility is 6, complex number is 0, the total number of 372.The results are slightly higher than the results of SSR markers.