Development Of Genetic Linkage Map And Screening Of Molecular Markers Linked With Flower Color In Anthurium Andraeanum

Flower color is an important goal in ornamental plants breeding. It is of great importance to construct molecular linkage map and establish molecular marker-assisted selection technical system of flower color for molecular breeding, enhancing the efficiency and shortening the cycle of cross breeding in Anthurium andraeanum. In this stduy, we built mapping population by crossing A.‘Mystral' with A.‘Alabama', developed SSR markers by analyzing transcriptome sequencing information of A.‘Mystral' and A.‘Alabama', constructed genetic linkage map by the use of SSR markers, and also screened the molecular markers linked with flower color in Anthurium andraeanum. The main results are as follows:1. Analysis of hybrid morphological characters and construction of mapping population.(1) Character separation occurred in the combination of A.‘Mystral' and A.‘Alabama', and the degree of separation varied with traits, the coefficiency of variation ranged from 7.6% to 23.1%, with that of flower traits, especially in the shape of spathe, being higher, and followed by leaf traits;(2) The ratio of the hybrid with and without aroma was 2:1 in 278 F1 offspring;(3) 94 individuals were selected from the F1 population to construct mapping population.2. Transcriptome sequencing of A.‘Mystral' and A.‘Alabama'and development of SSR markers.(1) 79735 unigenes were obtained through transcriptome sequencing and analysis by using of vegetative and reproductive organs of A.‘Mystral' and A.‘Alabama'as materials;(2) 10625 SSR locis were identified from 7655 genes by MISA software searching on unigenes with the size being more than 1 KB, which included six types of SSR, with the majority being one, two and three nucleotide repeat unit, accounted for 98.38% of the total number of SSR;(3) 299 pairs of primer were obtained from 10625 SSR markers by Primer Premier 5.0 software. 24 pairs of SSR primers randomly sampled from 299 pairs of SSR primers for polymorphic analysis, 14 polymorphic pairs of primer were found, with the polymorphism rate of 58.3%.3. Construction of genetic linkage map by the use of SSR marker.(1) 309 pairs of SSR primers were employed for polymorphic analysis. 160 primer pairs in parents and 134 in mapping population stably and repeatedly produced clear bands;(2) 134 pairs of primers were subjected for polymorphic analysis in mapping population, 183 polymorphic loci were identified;(3) The genetic map of Anthurium andraeanum was constructed by utilizing Join Map 4.0 software's CP model based on the linkage analysis of 183 polymorphic marker loci data tested from mapping population, which included 25 linkage groups covered 1428.1c M of the genome with a mean marker interval of 16.8c M.4. Development of SSR markers linked with flower color in Anthurium andraeanum.(1) 58 pairs of primers were developed by using of Primer Premier 5.0 software according to the sequences of 60 flower color genes obtained from the NCBI databases and the Web of Science database.(2) 10 gene pools of different flower color were prepared from 60 cultivars and employed to perform PCR amplification by the use of 58 pairs of SSR primers, six markers linked to flower color were eventurally obtained in Anthurium andraeanum.Based on the above researches, the genetic diversity of the hybrid population were known, mapping population was constructed, 309 SSR markers were developed, the genetic linkage map was preliminarily constructed,and SSR markers linked with flower color were sceened in Anthurium andraeanum. These results laid a foundation for further mapping of main breeding target gene and molecular breeding in Anthurium andraeanum.