Fingerprinting Analysis Of Ogura-CMS Restorer Lines And Establishment Of Rapid Analysis Of Hybrid Seeds Purity

Canola is one of the main oil crops in China,and it is also the main source of edible oil.Because of an uncompleted supervision system,there are many unqualified varieties present in the market with a wrong brand or even a fake brand,which seriously damages the interest of famers,companies and breeders.Scientists are increasingly aware of the importance of establishing a special identity information for each varieties.With high stability,polymorphism and co-dominant,SSR markers are widely used in the identification and fingerprinting of varieties.This study used SSR markers to perform fingerprinting analysis of 87 breeding lines.Breeders and farmers are very concerned about seed purity.Hybrids must meet the official standard of seed purity before entering the markets.The method of molecular marker detection for seed purity has become unsatisfied with high-throughput demands,due to the constraints of its feature.Recently,q-PCR technology was widely used in biological research because of its sensitivity,high efficiency,accuracy,and high throughput.In this study,a new seed purity identification model has been constructed using q-PCR technology,and the purity of hybrids can be rapidly identified.The main results are as follows:1.We selected 36 markers from the 57 SSR markers and performed analysis of fingerprinting for 87 breeding lines in Brassica napus(including 2 maintainers,15 sterile lines,64 restorer lines of Ogura-CMS,and 6 lines of conventional lines).In the analysis,a total of 111 polymorphic bands were amplified,with an average of 3.08 bands per marker,and the average PIC value of markers was 0.47.UPGMA cluster analysis was performed using these markers,classifing 87 breeding lines into 7 categories,namely A to G.Among them,the largest class was A group,including 54 breeding lines totally and can be divided into two sub-categories.The first sub-category included 12 male sterile lines,and the second sub-category included 29 restorer lines.2.We selected 15 core markers with high polymorphism,which can distinguish the lines,and constructed 64 digital ID cards and QR codes for Ogura-CMS restorer lines.3.This study used P74-17 and Huayouza 62 as experimental breeding lines.Known concentration gradients of recovered genes were used to construct an identification model for the purity of hybrids,using q-PCR techniques.The equation was y=-3.3396x+0.3617,with a correlation coefficient as 0.9635 and the variance analysis F-value was 449.3.Analysing the purity of the 187 samples in the field and calculating the purity of the samples with the standard curve were used for regression analysis.The linear regression coefficient was 0.8604 and the variance analysis F value was 1140.2,indicating there were highly correlated.