차세대염기서열분석법을 통한 돌연변이감귤 아라온주, 제대온주의 유전체분석

Abstract

Recently, the development of next generation sequencing method (NGS) and cost reduction, researches using genome analysis have been actively conducted, and whole genome re-sequencing that aligns the leads of the sample to the previously coded reference gene. The cost is very low, unlike De novo assembly, which uses the overlap information of leads that sequence new samples without standard genome. In this study, we used the reference genes of the already registered C. unshiu Marc. Cv. Miyagawa-wase(Shimizu T, etal. 2017) and used the control groups Citrus unshiu and mutant Ara-unshiu, Jedae-unshiu overall genom analysis was conducted. In this experiment, the reason of Citrus unshiu already registered on NCBI was analyzed is that the nucleotide sequence may have naturally mutated depending on factors such as environmental effects. As a result, the SNPs were changed by 2,355,501 compared to the reference sequence registered with NCBI, and the insertions had 98,321 deletions with a difference of about 87,696, showing a considerable difference from the reference gene registered with NCBI. Through this, we are performing the registration as a new reference gene. Through these results, this study analyzed SNP and Indel of Ara-unshiu and Jedae-unshiu compared with Citrus unshiu. Sequence of filtered fragment after quality adjustment of FASTQ file reads obtained using Whole genome re-seq instrument using FastQC, PICARD tools after alignment using BWA Samtools Remove duplicates that occur during the sequencing process using, extract SNPs and InDels using Samtools, and analyze and summarize the occurrence region, frequency, and frequency of each DNA polymorphism using the SNPeff tool. And annotation is performed to classify the functions of each SNPs and Indels so that the skins are not smooth, and the valleys are vertically sloping from the nipple and have an uneven appearance. And the content of citrus flavonoids is increased compared to the fruits of C grown under the same conditions, and the contents of each of the components of Hesperidin and Narirutin are also increased compared to the contents of each of the fruits of Gungcheonsaeng grown under the same conditions. J gene related to (Inoue T1, Yoshinaga A., et al. 2015, Susoma Jannat, Md Yousof Ali., Et al. 2016), flowering time is the same as C, but fruit matures later than C, We have developed a molecular marker using ANP and Indel to identify genes for A, which is characterized by a 1 to 3 month delay in pigmentation time compared to C (Fujii H, Ohta S., et. al. (2016).