Recent advances in next-generation sequencing (NGS) and single nucleotide polymorphism (SNP) genotyping promise to greatly accelerate crop improvement if properly deployed. High-throughput SNP genotyping offers a number of advantages over previous marker systems, including an abundance of markers, rapid processing of large populations, a variety of genotyping systems to meet different needs, and straightforward allele calling and database storage due to the bi-allelic nature of SNP markers. NGS technologies have enabled rapid whole genome sequencing, providing extensive SNP discovery pools to select informative markers for different sets of germplasm. Highly multiplexed fixed array platforms have enabled powerful approaches such as genome-wide association studies. On the other hand, routine deployment of trait-specific SNP markers requires flexible, low-cost systems for genotyping smaller numbers of SNPs across large breeding populations, using platforms such as Fluidigm’s Dynamic Arrays™, Douglas Scientific’s Array Tape™, and LGC’s automated systems for running KASP™ markers. At the same time, genotyping by sequencing (GBS) is rapidly becoming popular for low-cost high-density genome-wide scans through multiplexed sequencing. This review will discuss the range of options available to modern breeders for integrating SNP markers into their programs, whether by outsourcing to service providers or setting up in-house genotyping facilities, and will provide an example of SNP deployment for rice research and breeding as demonstrated by the Genotyping Services Lab at the International Rice Research Institute.
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Amplified fragment length polymorphism (AFLP) is a molecular marker technique based on DNA and is extremely useful in detection of high polymorphism between closely related genotypes like Korean wheat cultivars. Six sequence characterized amplified regions (SCARs) have been developed from inter simple sequence repeat (ISSR) analysis which enabled the identification and differentiation of 13 Korean wheat cultivars from the other cultivars. We used six combinations of primer sets in our AFLP analysis for developing additional cultivar-specific markers in Korean wheat. Fifty-eight of the AFLP bands were isolated from EA-ACG/MA-CAC, EA-AGC/MA-CTG and EA-AGG/MA-CTA primer combinations. Of which 40 bands were selected to design SCAR primer pairs for Korean wheat cultivar identification. Three of 58 amplified primer pairs, KWSM006, KWSM007 and JkSP, enabled wheat cultivar identification. Consequently, 23 of 32 Korean wheat cultivars were classified by eight SCAR marker sets.
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Hybrid sterility is the major obstacle that can act as a barrier to genetic recombination and limits favorable gene transfer during inter-subspecific crosses in rice. However, hybrid sterility can be overcome by utilization of wide-compatible varieties (WCV). In the present study, two F2 populations generated from crosses between a wide-compatibility line (HWC-line) and two Korean varieties, Dasan and Hwacheong were evaluated for eight agronomic traits. A total of 157 molecular markers consisted of 116 STS, 40 SSR and 1 FNP were used to create genetic linkage maps spanning all 12 chromosomes. Twenty-one main-effect QTLs (M-QTLs) were identified in HWC-line/Dasan (HD) population. These loci are populated with two QTLs for culm length, one for spikelet per panicle, one for spikelet fertility, four for grain length, five for grain width, five for grain shape, and three for 100 grain-weight. In HWC-line/Hwacheong (HH) population, 17 QTLs were detected including two QTLs for culm length, one for panicle length, one for spikelet fertility, five for grain length, three for grain width, three for grain shape, and two for 100-grain weight. Of the total 34 QTLs found in the two F2 populations, 9 QTLs associated with culm length, grain length, grain width and grain shape were newly identified in this study. These QTLs will be useful for further understanding the genetic basis of agronomic traits and for developing markers for selection in rice breeding. In addition, HWC-line could be used as a bridge for inter-subspecies crosses overcoming hybrid sterility.
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Pre-harvest sprouting (PHS) of wheat reduces grain yield and has deleterious effects on wheat flour quality. PHS resistance is strongly associated with grain color and seed dormancy. Resistance to PHS is a priority of wheat breeding program. We have studied a population of 122 Doubled Haploid (DH) lines from the cross between two cultivars, ‘Keumkang’ and ‘Olgeuru’, in order to detect QTLs correlated with PHS resistance. PHS was evaluated using mist spray in the humid chamber (PHS-MS), and the germination index (GI) was determined for two years. The variety ‘Keumkang’ has white grains and higher percentage of PHS rate (30.2%), while ‘Olgeuru’ has red grains and lower PHS rate (2.8%). There was significant difference in grain color according to the GI. PHS-MS was positively correlated with GI (r = 0.732,
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Quantification of potato glycoalkaloids (PGA) exerting toxicity to humans has some limits if applied to potato breeding populations on a large scale due to its high cost and time consumption. The aim of this study was to investigate key genes involved in PGA biosynthesis and their tuber expression patterns in two potato cultivars, Atlantic with low PGA content (18.6 mg/100g FW) and Haryoung with high PGA content (40.1 mg/100g FW), and to test the utility of these PGA gene transcript levels as selectable markers in potato breeding program. Pot grown potato plants of both ‘Atlantic’ and ‘Haryoung’ were exposed to drought stress where the transcript accumulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase 1 (
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Confirmation of T-DNA inheritance and stability requires testing of several parameters such as phenotypic performance of transgenic lines, transgene integrity structure and expression stability for at least three consecutive generations, and Mendelian inheritance analysis, which are very important to transgenic breeding and commercialization of genetically modified crops (GMOs). In the present study, we measured nine agronomic and yield-related traits of Iksan526 and confirmed stable phenotypic performance comparing to the donor variety Dongjin over three years. Transgene integrity structure, flanking sequence, functional element copy numbers, absence of backbone sequence and transgene expression were all verified in Iksan526 over three consecutive generations, indicating a stable inheritance of the transgene. In addition, phenotypic and genotypic segregation were analyzed on F2 and F3 populations derived from a cross of Dongjin/Iksan526, and results complied with Mendel’s law of segregation based on single transgene locus. This study demonstrated detailed analyses and confirmation of transgene event Iksan526 which is crucial for commercialization of GMO.
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Drought is one of the most important abiotic factors affecting wheat production and development of tolerant genotypes is limited by the lack of effective selection criteria. A genetic analysis of drought tolerance indices at seedling stage (i.e. root length, shoot length, root/shoot ratio and seedling dry weight) was performed for a seven-parent half diallel cross of bread wheat (
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A tissue-specific and developmentally expressed gene was isolated from Chinese cabbage (
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An increasing preference for good eating quality of rice among consumers has become one of the important considerations in rice breeding. Amylose content is a leading factor affecting eating quality of rice. Amylose composition is determined by the relative activity of soluble starch synthase (SSS) and granule-bound starch synthase (GBSS). This study focused on modifying the expression of
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