Background : Atractylodes japonica koidz (AJ) is a perennial herb that belongs to Atractylodes genus. The dried rhizome of AJ is known as ‘Baek-chul’. The ‘Baek-chul’ is used as important traditional medicine in north-east Asia. It is considered to be effective for the treatment of stomach disorder, virus, diuresis, inflammation, arthritis. AJ is heavily depend on import from china and only few studies have been carried out. In this study, we develop SSR marker to build a foundation of breeding, to analyze genetic diversity and to construct core collection.
Methods and Results : AJ resources was collected from each different place. To find simple sequence repeat (SSR) marker, we sequenced genomic DNA of AJ resources using Illumina HiSeq 2000 System. As a result of next generation sequencing (NGS), we obtained putative SSR loci. From these SSR primers, 553 SSR primer sets were designed successfully and confirmed polymorphism by in silico analysis. Nucleotide motifs ranged from tri- to penta-. Among these, 48 primer were tested in 4 individuals by capillary electrophoresis. Finally, selected 28 SSR marker were showed clear band and polymorphism by Electrophoresis.
Conclusion : In this study, we developed 28 polymorphic SSR marker using NGS, and it could be used for analyzing genetic diversity of A. japonica. These marker would be useful for breeding of new cultivar in the future.
Background: Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth with radical leaves during the first year and shows reproductive growth with cauline leaves and bolting during the second year. In addition, the shape of the plant varies within the same species. For this reason, there are limitations to classifying the species by visual examination. However, there is not sufficient genetic information or molecular tools to analyze the genetic diversity of the plant. Methods and Results: Approximately 34.59 Gbp of raw data containing 342,487,502 reads was obtained from next generation sequencing (NGS) and these reads were assembled into 357,211 scaffolds. A total of 84,106 simple sequence repeat (SSR) regions were identified and 14,133 primer sets were designed. From the designed primer sets, 95 were randomly selected and were applied to the genomic DNA which was extracted from five plants and pooled. Thirty-nine primer sets showing more than two bands were finally selected as SSR markers, and were used for the genetic relationship analysis. Conclusions: The 39 novel SSR markers developed in this study could be used for the genetic diversity analysis, variety identification, new variety development and molecular breeding of A. triphylla.
Background : Adenophora triphylla var. japonica (Regel) H. Hara shows vegetative growth by radical leaf until 1 year after sowing and shows reproductive growth during the second year and there is a characteristic of bolting by turning into cauline leaf. In addition, the phenotypes of plants varies even though they are belonging to the same species. For this reason, there is a limit for the classification of the species by the method of visual examination. Methods and Results : Simple sequence repeat (SSR) markers were developed based on the genomic sequence of A. triphylla using next generation sequencing to prepare the basis of molecular breeding and analyze the genetic diversity. Ninety-five primer sets including tri-, tetra- and penta-nucleotide motif types were randomly selected and they were applied to mixed genomic DNA and finally 39 primer sets showing from two to four bands were selected and used for genetic relationship analysis. Conclusions : Using the next generation sequencing, 39 polymorphic SSR markers were developed.