Development Of Genomic SSR Markers And Application In Constructing Genetic Linkage Map Of Salvia Miltiorrhiza Bge.

Salvia miltiorrhiza Bge is a traditional Chinese herb plant,its root has been used for centuries to treat diseases such as angina pectoris,cerebral vascular diseases,degenerative diseases,acute or chronic liver injuries,and has extensive and important curative effects in practice.The construction of molecular genetic linkage map and analysis of quantitative traits loci not only provide information for the early selection of important traits but also produce the important significance on map-based cloning and molecular breeding.The genetic linkage map of S.was constructed in 206 F1 individuals from an intraspecific cross by using SSR markers,and identified the QTLs for the main agronomic traits of S..The results obtained in this study were as the follows:1.Genomic SSR marker development of S.Restriction site associated with DNA sequencing(RAD-seq)was conducted on S.ZH23 by using Illumina-HiSeq 2000 sequencer.The gene sequence was then assembled,and SSR was detected by bioinformatics method.Totally,there was 2.40 Gb in Salvia miltiorrhiza Bge.The average read length was 323 bp.There are 6865 microsatelites(SSRs)ranged from 2 to 6 bp.Among them,6058 microsatelites(SSRs)could be used to design primers,accounting for 88.24%.There were more dinucleotide and trinucletide than others,the number was 2745 and 2544,accounting for 45.31% and 41.99%,respectively.Through the analysis,AT/TA?AG/TC and CT/GA were the most frequent motifs in dinucleotide.However,in trinucletide,they were AAT/TTA?AAG/TTC?ATT/TAA?CTT/GAA?AGA/TCT and ATA/TAT.6058 SSR primer pairs were designed using Primer Software.2.Construction of genetic linkage mapUsing 206 F1 individuals derived from a hybrid between ZH23 and BH18,we constructed a genetic linkage map consisting of 8 linkage groups covering 397.5 cM and including 111 simple sequence repeat(SSR)markers based on a double pseudo-testcross mapping strategy.The average distance per marker was 3.6 cM.The length of linkage groups varied from 24.8 cM~102.0 cM and each of them included 6~24 markers respectively.It was estimated that the Salvia miltiorrhiza genome genetic distance was 428.8 cM representing 92.7% genome coverage3.Primary quantitative trait locations(QTLs)for agronomic traits of S.11 main agronomic traits in the period of duration of F1 mapping population such as acervate petiole length,acervate leaf length and cauline petiole length showed continuous and normal distribution.There are significant correlation between different traits,in which 10 groups showed extremely significant correlation(P<0.01),and 4 groups showed significant correlation(P<0.05).QTL results were shown as follows: 74 QTLs were detected for these 11 traits using interval mapping,and phenotypic variance explained ranged from 6.2% to 13.1%.There were 15 QTLs for flower number per ring.Only 2 QTLs were detected for footstalk length.In these 11 phenotypic traits,10 QTL were greater than 10% was ten.The largest one,QTL-CPL-5,was 13.1%,which was located on the LG3 controlling cauline leaf length.LOD value was 5.56.