Identification Of Maize Authenticity And Purity Based On Perfect SSR And Indel Markers On Chloroplast

China is a big country for maize planting so the quality of maize is very important,the authenticity and purity of maize varieties can't be ignored besides good varieties in order to promise its quality.In recent years,with the extensive application of molecular markers,the authenticity and purity identification of maize become more simple and accurate.This study select new SSR marks based on nuclear genome and Insertion/Deletion(InDel)markers in the maize chloroplast genome by whole genome of maize's Re-sequencing data.Using SSR markers to establish multiplex PCR amplification system and a new DNA fingerprinting database so that it is convenient for numerous maize authenticity and purity identification.Using Chloroplast InDel markers to identified maize normal and reciprocal cross hybrids that can't be identified based on the maize nuclear genome.Part?: Construction of Perfect SSR multiplex PCR system and database.The study developed and designed a batch of SSR markers based on maize re-sequencing data and selected 40 pairs of primers suitable for multiplex PCR form 111 pairs of primers based on chromosomal location,amplification of the amplified fragment,fluorescence color and other information.Ensuring suitable concentration for multiplex PCR amplification and condition.24 representative varieties maize is used for selecting and optimizing 32 pairs of primers from 40 pairs of primers.Finally,four sets of 8-fold PCR amplification and 2 sets of 16-fold electrophoresis is set up to establish mini DNA standard fingerprinting database by 95 nationally validated maize hybrids.Part?: Identification of maize normal hybrid and reciprocal cross hybrid based on chloroplast InDel markers.The maize normal hybrid and reciprocal cross hybrid are different in some characters,and even affect the yield characters in some varieties.The identification way of reciprocal cross previously had limitation especially on material types and time.Primers were developed with maize leaf materials based on Insertion/Deletion(InDel)markers' flanking sequence in the maize chloroplast genome.Eight primers were successfully developed from 11 InDel loci.All of the 8 primers could stably amplify and the peak type is beautiful.Results indicated CPMIDP01 and CPMIDP03 were stable and could be combined into 2-plex PCR,it can save more time and materials,because of establishing a rapid identification method for maize normal and reciprocal cross hybrids.This makes the identification of normal and reciprocal cross hybrids in maize not limited to the seed material.In addition,the detection using leaves makes the identification time unlimited,it still can be carried out even after sowing.Maize elite inbred lines were divided into three groups(group A,group B and group C)by chloroplast InDel markers,of which hybrids were combined with different groups could identified by this method.At present,maize breeding is mainly based on domestic germplasm and abroad germplasm in China.The method can be used to identify of normal and reciprocal cross of maize varieties widely which supplement identification on nuclear genome,improving variety authenticity and traceability information.