Tomato spotted wilt virus (TSWV) causes one of the most destructive viral diseases that threaten tomato (Solanum lycopersicum) worldwide. So far, eight TSWV resistance genes, Sw1a, Sw1b, sw2, sw3, sw4, Sw-5b, Sw-6, and Sw-7 have been identified and Sw-5b has been incorporated into tomato for prevention of TSWV. The objectives of this research are first to discover single nucleotide polymorphisms (SNPs) in Sw-5 alleles and then to develop SNP markers to distinguish resistant genotypes against TSWV for marker-assisted breeding in tomato. First, DNA sequences of Sw-5b alleles from both resistant and susceptible cultivars amplified using known Sw-5 gene-based marker was analyzed. The single functional SNP (G→A) was detected as non-synonymous substitution because this SNP causes change of arginine (Arg599) to glutamine (Gln599). Next, the primer pair for high resolution melting analysis (HRM) was designed around this SNP. To determine accuracy of this SNP marker to distinguish resistant Sw-5b genotypes against TSWV, genotypes of 32 commercial tomato cultivars were checked. The newly developed SNP marker could select six cultivars carrying resistant Sw-5b genotype, which was 100% correlated with genotypes based on the gene-based marker. These results indicate that the SNP maker developed in this study could be useful for better tracking resistance to TSWV in tomato breeding.

We report the C language implementation of a program that merges SNP data from all samples and estimates various statistical quantities related to SNP. The software combines the SNP information from different samples according to the SNP position in the nucleotide sequence. The combined SNP information is converted into HapMap format that can be used as an input for genome-wide association analysis for quantitative traits. The software additionally provides estimates of the minor allele frequency, the heterozygosity ratio, and the In/Del frequency. The software is prepared as a stand-alone program and is downloadable from http://info.kongju.ac.kr/snpmerge/.

The goal of marker-assisted backcrossing is to reduce the number of generations significantly by using genome-based molecular markers. Among other types of molecular markers, SNP (single nucleotide polymorphism) is mostly used in genetic diversity analysis due to its abundance. To develop high-throughput SNP marker for MAB system, we selected 20 Chinese cabbage lines each represent traits as inner leaf color, disease resistance, head type and maturity etc. Then, we sequenced the transcriptomes of 20 lines by using Illumina Hiseq2000. The average transcriptome size was 1.37 Gbase, and the average of short reads mapping rate was about 62.15% (30xcoverage). We identified 13,976 SSR markers and 380,198 SNPs by aligning contigs of 20 Chinese cabbage lines. To develop SNP marker set, we chose 409 SNPs that covers the whole Brassica rapa transcriptome. The filtering criteria were depth, polymorphism, segregation ratio, lack of adjacent SNP and copy number. We positioned the selected SNP markers to the Chinese cabbage linkage map. Clustering dendrogram was produced using SNP marker and three different clusters were generated. The result showed that the genotyping data is partially linked to the phenotyping data. We assume that the developed SNP marker set can be applied in the Chinese cabbage MAB system soon.

Magnoliae Flos (Sini in Korean) is one of the most important oriental medicinal plants. In the Korean Herbal Pharmacopeia, the bud of the all species in Manolia denudate and Manolia genus were regarded as the botanical sources for ‘Sini’. Most the dried bud of Manolia denudata, Manolia biondii and Manolia sprengeri were used as ‘Xin-yi’ in China. Therefore, the purpose of this study was to determine and compare the ‘Magnolia’ species, four species including Manolia denudata, M. biondii, M. liliiflora and M. Kobus were analysis of sequencing data revealed DNA polymorphisms. The based on tRNA coding leucine/phenylalanine (trnL-F) and NADH-plastoquinone oxidoreductase subunit 5 (ndhF) sequences in chloroplast DNA. For the identification of ‘Magnolia’ species, polymerase chain reaction (PCR) analysis of chloroplast DNA regions such as ndhF have proven an appropriate method. A single nucleotide polymorphism (SNP) has been identified between genuine “Sini” and their fraudulent and misuse. Specific PCR primers were designed from this polymorphic site within the sequence data, and were used to detect true plants via multiplex PCR.

To successful molecular breeding, identification and functional characterization of breeding related genes and development of molecular breeding techniques using DNA markers are essential. Although the development of a useful marker is difficult in the aspect of time, cost and effort, many markers are being developed to be used in molecular breeding and developed markers have been used in many fields. Single nucleotide polymorphisms (SNPs) markers were widely used for genomic research and breeding, but has hardly been validated for screening functional genes in olive flounder. We identified single nucleotide polymorphisms (SNPs) from expressed sequence tag (EST) database in olive flounder; out of a total 4,327 ESTs, 693 contigs and 514 SNPs were detected in total EST, and these substitutions include 297 transitions and 217 transversions. As a result, 144 SNP markers were developed on the basis of 514 SNP to selection of useful gene region, and then applied to each of eight wild and culture olive flounder (total 16 samples). In our experimental result, only 32 markers had detected polymorphism in sample, also identified 21 transitions and 11 transversions, whereas indel was not detected in polymorphic SNPs. Heterozygosity of wild and cultured olive flounder using the 32 SNP markers is 0.34 and 0.29, respectively. In conclusion, we identified SNP and polymorphism in olive flounder using newly designed marker, it supports that developed markers are suitable for SNP detection and diversity analysis in olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Molecular markers are useful for selecting to include superior character genetic like as strong immune system and rapid growth in fish. The marker is also very important part of breeding technology in Olive flounder (Paralichthys olivaceus). Single nucleotide polymorphisms (SNPs) marker is already in use widely for genomic research and breeding. But this SNPs marker hardly has been validated for screening functional genes in Olive flounder. We study identify single nucleotide polymorphisms (SNPs) on Expressed sequence tag (EST) database, develop usable SNP marker and apply to wild sample and cultured of olive flounder. As a result, Out of total 4.327 ESTs, 693contigs and 514 SNP from total contigs were detected while these substitutions include 297 transitions and 217 transversions. 144 developed markers were applied in 16 samples (wild 8, culture 8), Out of total marker, only 32 markers had detected polymorphic in sample. Polymorphism of 32 markers was observed in the variety genes region involved in immunity and protein synthesis. And the 32 marker were identified 21 transitions, 11 transversions, and indel was not detected in polymorphic SNPs. The analysis on heterozygosity by sample showed 0.34 in wild sample and 0.29 in cultured sample. In conclusion, we was identified SNP and Polymorphism by designed new marker, it supports that development marker is suitable for SNP detection and diversity analysis in Olive flounder. The outcome of this study can be basic data for researches for immunity gene and characteristic with SNP.

Soybean is an important worldwide crop of dietary protein and oil resources for human foods and animal feeds. However, soybean breeding and improvement has been experienced challenges by a narrow germplasms. SNP genotyping array is regarded as a promising tool for dissecting wild and cultivated germplasms to find important genes by high-density genetic mapping and genome-wide association studies (GWAS). Here, we present the establishment of a large soyaSNP array and its use for diversity analysis and high density linkage mapping. More than 4 million high-quality SNPs identified from 16 high-depth and 31 low-depth soybean genome resequencing data were used to select 180,961 SNPs for the Axiom® SoyaSNP array. Our validation analysis for a set of 222 diverse soybean lines showed that a total of 171,161 markers were good quality for genotyping. Of the converted SNPs, 82.6% SNPs had a marker spacing of less than 9 kb and 17.4% SNPs greater than 9 kb, thereby suggesting that our array is likely suitable for GWAS of soybean germplasms. In the GWAS for seed protein content in the wild soybean germplasms with the size of 1,135 accessions, 22 loci on 12 chromosomes showed significant association (-logP>4). The highest associated peaks were shown at the 28 Mbp region on Gm05 (-logP=5.89), at 45 Mbp on Gm03 (-logP=5.32), and at 2.8 Mbp on Gm17 (-logP=5.00). Of the 22 associations, 8 corresponded with the location of previously reported seed protein QTLs and 14 regions is thought to be new QTLs for seed protein content in wild soybean. This array is being used to construct high-density genetic maps in two recombinant inbred lines and nested-association mapping populations with 30 combinations used Daepung cultivar as hub-parents, with an objective to confirm large structural variations of chromosomes using the ultra-high-density maps.

With the development of next generation sequencing (NGS) technology, the variation of sequences represented as SNP between cultivars becomes available at genome level. The major domestic cultivars with high yield have been developed by breeding of indica and japonica, it is important to localize the region of origin according to the genotype for further characterization of unique features of cultivars. For the localization of SNP at genome level, the paired end sequences of 6 major domestic rice cultivars, Ilmi, Ilpoom, Sulgaeng, Bakjinju, Hwayoung and Woonkwang were compared against Japonica and Indica Rice Genomes as reference genomes. The genomic DNAs were prepared from callus tissues and paired-end of the fragments were sequenced with NGS Sequencer, Illumina HISeq. About 50x coverage of paired-end sequences were trimmed according to the quality of the sequences, and errors were corrected with statistical analysis of kmers of 15. The trim-corrected sequences were mapped and variants were analyzed against reference genomes. The overall change rate of Ilmi against Nipponbare IRGSP 1.0 and Indica BGI 93-11 reference genomes were 0.92 base/1kb (1/1,079 base) and 8.09 base/1kb (1 base/123 bases), respectively. Among 6 cultivars, overall rate of Bakjinju showed the lowest overall change rate of 0,53 base/1kb, and Hwayoung showed highest frequency of 0.92 base/1kb. Compared to high level in the range of change rate of 7.0-9.3 base/1kb against indica, domestic cultivars showed lower range of change rate 0.2-3.3 base/1kb with unique local high peak against japonica genome depend on the chromosomes. Compared to assembly of genome sequences, the variation of nucleotides compared to reference sequences is much faster and simple to characterize the genotype. The types of variation and the effect on functional categories will be presented.

Capsicum diversity is getting lower in modern crops because of the genetic erosion. In Capsicum, breeders have been mainly focused on agriculturally important traits such as disease resistances, high yield and pungency. This narrow breeding pool hampered to develop improved cultivars. It has become a hot issue to conservation of genetic diversity and exploitation of wild germplasm in Capsicum. However, although a large number of accessions are maintained in Capsicum germplasm collections, their use for crop improvement is limited by the scarcity of information on genetic diversity, population structure and proper phenotypic assessment. The identification of representative and manageable subset of accessions would facilitate access to the diversity available in large collections. A genome wide germplasm characterization using molecular markers can offer reliable tools for adjusting the quality and representativeness of core samples. We investigated patterns of molecular diversity at 48 single nucleotide polymorphisms (SNPs) in 4056 accessions from 11 Capsicum species from 89 different countries. Using these genetic variations and 32 different morphological traits, 250 core set was selected in whole Capsicum germplasm. The core collection could be a primary source for distributing germplasm to pepper breeders and other national programs as well as for evaluation

Genotyping-by-sequencing (GBS) is a robust and rapid tool to develop SNP markers. Reduced sequencing complexity and multiplex sequencing of GBS has reduced genotyping cost for complex genome. However multiplex sequencing brings low sequencing depth which can lead to reduced number of markers. Therefore to find the appropriate condition for GBS is needed. In this research we demonstrated the use of ApeKI and selective primers for GBS of pepper (Capsicum spp.). Selective primers which amplify the GBS library with one or two flanking sequences to ApeKI site were used to increase sequencing depth. By in silico digestion, we developed six different selective primers amplify 4,000-400,000 regions. We made the GBS library with eight pepper accessions in four species using six selective primers and sequenced. Proper selective primers and pooling rate for each species will be determined. This approach will be useful for genotyping Capsicum breeding lines or populations by developing high quality SNP markers.

An important worldwide plant source of dietary protein and oil, modern breeding and improvement of soybean is suffered from a narrow cultivated germplasm relative to other crop species likely because of underuse of wild soybeans as breeding resources. SNP genotyping array is regarded as a promising tool for dissecting wild and cultivated germplasms to find important adaptive genes by high-density genetic mapping and genome-wide association studies (GWAS). Here, we present the establishment of a large soybean SNP array and its use for diversity analysis and high density linkage mapping. More than 4 million high-quality SNPs identified from 16 high-depth and 31 low-depth soybean genome resequencing data were used to select 180,961 SNPs for the AxiomÒ SoyaSNP array. Our validation analysis for a set of 222 diverse soybean lines showed that a total of 171,161markers were of good quality for genotyping. Of the converted SNPs, 82.6% 82.6% SNPs had a marker spacing of less than 9 kb and 17.4% SNPs greater than 9 kb with the 297 inter-SNP spacings of >100 kb and with 812 kb of the largest spacing, thereby suggesting that our array is likely suitable for GWAS of soybean germplasm. This array is being used to construct high-density genetic map in populations generated from intermatings of two cultivated and two wild soybeans, with an objective to confirm large structural variations of chromosomes using the ultra-high-density maps

토마토(Solamum lycopersicum L.)는 전 세계적으로 생산량이 연간 146백만 톤이며 재배면적은 4,339천ha에 달하는 채소작물로 경제적 가치가 매우 높다. 최근 국내 및 미국, 남미, 유럽, 동남아시아, 일본, 중국 등지로 토마토황화잎 마름바이러스 (Tomato yellow leaf curl virus; TYLCV)가 급격히 확산되고 있어 토마토 생산량과 농가소득 감소 등 의 심각한 피해를 입히고 있다. 토마토의 TYLCV 저항성 유전자는 야생종에서 Ty-1, Ty-2, Ty-3, Ty-4, Ty-5 등이 보고 되고 있으며 이들 유전자원을 활용한 계통 및 품종 육성이 활발하게 이루어지고 있다. 자사에서도 TYLCV의 피해 를 최소화할 수 있는 저항성 품종 육성을 주요 목표로 하여 품종 육성을 활발하게 진행하고 있다. 최근에는 내병계 품종 육성에 분자마커를 활용함으로써 신품종 개발기간을 단축시키고자 하는 노력이 많이 이루어지고 있다. Ty-1, Ty-2, Ty-3에 대한 CAPS 또는 SCAR 마커 개발이 보고되고 있으나(Hoogstraten et al., 2005; Garcia et al., 2007; Ji et al., 2007), gel-based 마커로 검정에 많은 시간과 노력이 필요하다. 본 연구에서 TYLCV 내병계 육성을 위해 보다 신 속하고 효율적인 분자마커 검정을 실시하고자 Ty-1과 Ty-3의 SNP마커를 개발하였다. 기 개발 마커를 활용하여 내 병계 계통 및 도입품종 20개의 Ty-1과 Ty-3의 염기서열을 분석하여 20개의 프라이머 셋트를 제작하였다. Ty-1 SNP2와 Ty-3 hrm3 마커를 선발하고 계통 및 품종으로 HRM (high resolution melting)분석을 통해 실효성을 판단하였 다. 또한 자사의 시판 이병계와 내병계 종자를 대상으로 검정을 실시하여 Ty-1 SNP2와 Ty-3 hrm3 마커의 실효성을 확인하였다. 개발된 마커를 활용하여 2014년 봄작기 파종 F2 분리세대 50 계통과 도입품종 21 품종에 대한 검정을 실시하여 품종육성을 위한 자료로 활용하였다.

콩에서 유전자지도 작성은 마커간의 상대적인 위치를 설정하여 금후 genome 상에 목표 유전자 위치를 확보하고 이를 표지화하는데 매우 요긴한 수단으로 활용된다. 최근 많은 연구자들이 콩에서 고밀도의 유전자지도 작성을 위해 노력하였고, 현재 soybean consensus map이 soybase(www.soybase.org) 에 공유되고 있다. 본 연구에서는 고밀 도의 콩 유전자 지도 작성을 위해 genotyping은 한국과 중국의 콩 genome resequencing결과 얻어진 약 4백만 개의 SNP중에서 선정된 약 180만개의 SNP로 만들어진 180k Axiom SoyaSNP array를 이용하였고, 실험집단은 큰올콩/신 팔달콩 및 큰올콩/익산10호의 교배후대로 작성된 F12 RIL집단을 활용하였다. 유전자지도 작성결과 큰올콩/신팔달 콩 집단에서 사용된 166,279개의 SNP 중 27,308개의 SNP가 다형성을 보였고 이중 6,535개의 SNP가 유전자지도상 에 표기되어 지도의 총 거리는(total coverage)는 약 3,313cM을 나타내었다. 한편 큰올콩/익산10호 집단에서는 사용 된 166,279개의 SNP 중 23,581개의 SNP가 다형성을 보였고 이중 6,597개의 SNP가 유전자지도상에 표기되어 지도 의 총 거리는(total coverage)는 약 5,017cM을 나타내었다. 본 연구를 통해 작성된 유전자지도에서 마커간 평균거리 는 큰올콩/신팔달콩 0.51cM, 큰올콩/익산10호 0.76cM으로 나타나 매우 고밀도의 유전자 지도가 작성되었음을 나 타내었다.

Single nucleotide polymorphisms (SNPs) are the most frequent type among variations found in genomic regions and are valuable markers for genetic mapping, genetic diversity studies and association mapping in plants. There are three basic species known as Korean native which are Pyrus ussuriensis, P. pyrifolia, and P. fauriei. Genetic relationship among Korean pear cultivars compared with their parents was identified that they are closely related P. pyrifolia, P. ussuriensis and/or hybrids between two species. Lack of genetic resources, including molecular markers to study pears are very severe. Recently developed next generation sequencing (NGS) platforms offer opportunities for high-throughput and inexpensive genome sequencing and rapid marker development. The objective of this study was to develop polymorphic SNP markers in ‘Whangkeumbae’ and ‘Minibae’, which were chosen as the representative cultivars of P. pyrifolia and P. ussuriensis × pyrifolia in each among Korean pears, using genomic sequences generated by NGS technology. In this study, more than 18.6 Gbp and 15.8 Gbp sequences were obtained from NGS of ‘Whangkeumbae’ and ‘Minibae’, respectively. ‘Whangkeumbae’ and ‘Minibae’ contained 2,712,288 and 2,747,224 SNPs, respectively. In SNPs validations between ‘Whangkeumbae’ and ‘Minibae’, the number of polymorphic SNPs were 2,516,438 and non-polymorphic SNPs were 1,179,391. For HRM primer design, 2,125,479 HRM candidate primers were obtained from polymorphic SNPs and 343,731 SNP primers were developed. This study shows that the utility of NGS technology to discover efficiently a large number of SNPs and SNP primers can provide valuable information in the genome study of Pyrus spp.

Single nucleotide polymorphisms (SNPs) are the most abundant variation in plant genomes. As DNA markers, SNPs are rapidly replacing simple sequence repeats (SSRs) and sequence tagged sites (STSs) markers, because SNPs are more abundant, stable, easy to automation, efficient, and increasingly cost-effective. We developed a 96-plex indica/japonica SNP genotyping set for genetic analysis and molecular breeding in rice using Fluidigm platform. Informative SNPs for indica/japonica populations were selected from 1536 Illumina SNPs and 44K Affymetrix SNP chip data of Rice Diversity and our resequencing data sets. Selected SNPs were evenly distributed across 12 chromosomes and average physical distance between adjacent SNP markers was 4.38Mb. We conducted genetic diversity analysis of 49 Bangladesh germplasm and check varieties to test a 96-plex indica/japonica SNP genotyping set we developed. High-throughput Fluidigm SNP genotyping system will serve a more efficient and valuable tool for genetic diversity analysis, DNA fingerprinting, quantitative trait locus (QTL) mapping and background selection for crosses between indica and japonica in rice. This work was supported by a grant from the Next-Generation BioGreen 21 Program (Plant Molecular Breeding Center No. PJ008125), Rural Development Administration, Republic of Korea.

The fruit shape is an important character in tomato. OVATE is one of genes controlling fruit elongation in tomato. Two loci suppress the ovate mutation, sov1 and sov2, on chromosome 10 and chromosome 11 respectively. sov1 appears to control neck constriction in the fruits (Rodriguez et al, 2013). We sequenced the genomes of Gold Ball Livingston and Yellow Pear using the Illumina Hiseq 2000 generating 101 PE reads and developed molecular markers tightly linked to sov1. The locus was confirmed by fruit shape index analysis, marker genotyping and progeny testing of recombinants. We find mapped sov1 to a 145 kb interval corresponding to a region comprising two candidate genes. One of the candidate genes for sov1 is SlOFP20 another member of the Ovate Family Protein class. Although there is no difference expression of SlOFP20 in the parents at anthesis, when the gene is expressed very high, the mutation appears to be a 34 kb promoter deletion of SlOFP20 in Yellow Pear, conferring a pear shaped and neck-constricted fruit.

With the development of next generation sequencing (NGS) technology, the variation of sequences represented as SNP between cultivars becomes available at genome level. The major domestic cultivars with high yield have been developed by breeding of indica and japonica, it is important to localize the region of origin according to the genotype for further characterization of unique features of cultivars. For the localization of SNP at genome level, the paired end sequences of 6 major domestic rice cultivars, Ilmi, Ilpoom, Sulgaeng, Baekjinju1ho, Hwayoung and Woongwang were compared against Japonica and Indica Rice Genomes as reference genomes. The genomic DNAs were prepared from callus tissues and paired-end of the fragments were sequenced with NGS Sequencer, Illumina HISeq2000. About 50x coverage of paired-end sequences were trimmed according to the quality of the sequences, and errors were corrected with statistical analysis of kmers of 15. The trim-corrected sequences were mapped and variants were analyzed against reference genomes. The overall change rate of Ilmi against Nipponbare IRGSP 1.0 and Indica BGI 93-11 reference genomes were 0.92 base/1kb (1/1,079 base) and 8.09 base/1kb (1 base/123 bases), respectively. Among 6 cultivars, overall rate of Baekjinju1ho showed the lowest overall change rate of 0,53 base/1kb, and Hwayoung showed highest frequency of 0.92 base/1kb. Compared to high level in the range of change rate of 7.0-9.3 base/1kb against indica, domestic cultivars showed lower range of change rate 0.2-3.3 base/1kb with unique local high peak against japonica genome depend on the chromosomes. Compared to assembly of genome sequences, the variation of nucleotides compared to reference sequences is much faster and simple to characterize the genotype. The types of variation and the effect on functional categories will be presented.

We are currently developing a high-throughput single nucleotide polymorphism (SNP) genotyping service at IRRI to accelerate progress in rice breeding by providing rapid and cost-effective marker services. SNP marker development and validation is being performed based on cloned genes and QTLs, GWAS hits, and whole genome sequence data to identify predictive SNP markers at important genes for key traits for the breeding programs. Trait-based and targeted SNPs are being deployed in sets of 24 and 96 SNPs on a Fluidigm EP1 system. At the same time, 384 SNP sets and a 6K SNP chip developed by Susan McCouch at Cornell University are being used for higher density genome scans on an Illumina system. Genotyping by sequencing (GBS) approaches with 96 and 384 barcoded samples per sequence lane are also being evaluated in comparison to SNP array technology based on the number of loci, call rates, turnaround times, and cost per sample. An efficient sample processing workflow with an integrated LIMS is also being optimized to enable high throughput genotyping with sample tracking to minimize errors. Moreover, web-based SNP data analysis tools have been deployed through the IRRI Galaxy workbench to speed up SNP data analysis. Future efforts will focus on large-scale deployment of GBS across breeding materials to enable QC genotyping, tracking of donor introgressions, and integration of genome-wide prediction into the variety development pipelines. The large-scale application of high-density markers will help transform IRRI’s rice breeding programs and increase the rate of genetic gain towards developing high-yielding, stress-tolerant varieties for target environments and market segments

본 연구에서는 토마토 MAB에 활용하고자 토마토 7 품종의 genome-wide SNPs 데이터베이스를 구축하고, MAB를 위한 분자마커 선발 프로그램을 개발하였다. 토마토 전사체 데이터를 NCBI-SRA에서 다운로드 하여 in silico 분석으로 SNP를 추출하였다. 전사체 데이터에서 추출된 SNP를 재료로 7 품종의 토마토 계통을 이용해 총 21개 교배조합별 SNP 분자마커를 선발하였고, primer가 이용 가능한 마커를 이용하여 데이터베이스를 구축하였다. 마커를 선발하기에 앞서 염색체의 분획으로 두 가지 방법을 사용하였는데, 물리적 거리에 따른 분획과 유전거리에 따른 분획 방법이다. 물리적 거리를 이용한 분획은 각 염색체를 동일한 크기의 5개의 구획으로 나누고, 한 구획 당 교배조합별 차이를 보이는 3개의 SNP를 선발하였다. 교배조합이 바뀔 때마다 이용 가능한 SNP가 자동으로 primer 정보와 함께 제공되도록 하였다. 유전거리를 반영한 분획 방법은 각 염색체의 유전적 거리를 측정하여 물리적 거리에 차등을 두어 염색체 구획을 설정하였다. 즉 재조합이 자주 일어나는 염색체 양끝 말단 부분은 구획을 조밀하게 나누어 MAB 마커 또한 많이 할당하여 자세히 조사하도록 구성하였다. 유전거리에 따른 마커 선발에는 1,924개의 tomato- EXPEN 2000 map 분자마커와 SNP 마커를 이용하였다. 교배조합별로 이용할 수 있는 마커를 12개 염색체 상에 그래픽적으로 제공함으로써 사용자가 쉽게 이해하고 이용할 수 있는 MAB 위한 마커 선발 프로그램을 개발하였다. 이러한 토마토 MAB용 분자마커를 제공하는 프로그램은 실제적인 여교잡 선발 육종에 적용하여 분자마커의 활용을 높이고, 육종효율을 증진시킬 것이다.