• Journal of Practical Agriculture & Fisheries Research
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• v.25 no.4
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• pp.90-102
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• 2024

Haploid/double haploid plants developed from isolated microspores can significantly accelerate plant breeding. Haploid plants can naturally double their chromosomes to create a pure homozygous line of diploid plants. We present a method for producing embryos from isolated microspores of hot peppers (Capsicum annuumL.). We analyzed the polyploidization levels of the regenerated plants. The donor plants produced the optimal stage of microspores following short-term growth under low-intensity light, which resulted in high rates of embryogenesis and cotyledonary embryogenesis. To find an efficient culture method, liquid, doubled-layer, and 2-step cultures were tested. Liquid culture yielded the highest number of embryos, whereas the highest efficiency for cotyledonary embryogenesis was afforded by the doubled-layer culture. When normal cotyledonary embryos were transplanted onto a regeneration medium, they developed into complete plants. From these, 208 plants were tested via flow cytometric analysis, and 35.6% and 72.7% of the chromosomes from the Milyang-jare and LV2319 genotypes, respectively, were found to be spontaneous double haploids. These results are the same as those obtained on analyzing horticultural characteristics, including the size of leaves and the size and shape of fruits. The present study provides information on the practical application of isolated microspore culture of hot peppers, factors that affect embryogenesis, and methods for polyploidy testing.

• Korean Journal of Breeding Science
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• v.42 no.5
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• pp.431-438
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• 2010

Anthers of sweet pepper $F_1$ cultivar 'Special' were cultured on Dumas De Vaulx (C medium), supplemented with $0.1mgL^{-1}$ 2, 4-D and $0.1mg{\cdot}L^{-1}$ kinetin with 3% sucrose, and 0.32% phytagel. The calluses obtained were further sub-cultured on Murashige and Skoog (MS) medium without growth regulators for regeneration. Regenerated plantlets were grown in plastic pots under plastic house and characterized their cytological and morphological characters in spring, 2008. Twenty percent plantlets were identified as haploid plants after chromosome and ploidy analysis. Haploid plants contained 12 chromosomes, high stomatal density with small stomatal length as compared to diploid plants. Stomatal length in haploids was 23.3% smaller than diploids. Haploid plants were characterized as small leaf and petiole size, poor vigor, thin stem and short plant height, short internodes and small flower buds, fruit size and fruit weight as compared to diploid plants and most of the haploid fruits were seedless. SP55, SP62, SP68, SP72 and SP77 are found high yielding double haploids with high total soluble content (8.6, 8.7, 9.2, 9.1 and $9.8^{\circ}Brix$, respectively) and desirable fruit shape, and recommended them to exploit as inbred lines for heterosis breeding.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.04a
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• pp.130-130
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• 2022

• Journal of Plant Biotechnology
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• v.44 no.1
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• pp.35-41
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• 2017

One of the most important species, Brassica rapa encompasses a variety of commercial vegetables, such as the Chinese cabbage, pak choi and oilseed crops. The LP08 of yellow sarson (Brassica rapa ssp, tricolaris) have a distinct morphology, with yellow seed color and a unique tetralocular ovary. LP21 of pak choi (Brassica rapa ssp, chinensis) have a dark brown seed color and bilocular ovary. In this study, we generated double haploid plants by crossing the LP08 (maternal variety) and LP21 (paternal variety), using microspore culture. A total of 66 accessions with various morphological characteristics were used for content analysis of flavonoids. The three flavonoids, quercetin, naringenin and kaempferol, showed differing contents in the two crossing parents. The Chinese cabbage type 'Chiifu' was used as the control. The highest accumulation of total flavonoids was observed in LP08. The lowest mean total flavonoids were found in 'Chiifu'. Among the 66 DH accessions, the quercetin contents of 18 accessions showed higher content than LP08. Kaempferol content was also high, and was found to be 79.7% of the total flavonoid content. Naringenin content was low at 2.8%, and was not detected in 22 accessions. Interestingly, the quercetin content positively correlated with the kaempferol content. These results can be used to identify genetic locus and genes related to useful traits. Phenotypic analysis of 66 DH accessions can further be used for natural selection of good breeding materials in B. rapa.

• Journal of Plant Biotechnology
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• v.38 no.1
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• pp.30-41
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• 2011

The influences of the agitation as well as the addition of medium during culture on the production of embryos were invested in isolated microspore culture of hot pepper (Capsicum annuum L.). When the culture medium was added during initial liquid culture step of liquid-double layer culture, the embryo yield and quality greatly increased. The most effective time point for medium addition was 5 days after the culture commenced. On the other hand, the effect of medium addition at later double layer culture step in liquid-double layer culture on the embryo production was less compared to that of medium addition during the initial liquid culture step. Agitating the culture for 1 week during later double layer culture step in liquid-double layer culture effectively increased the production of normal cotyledonary embryos. In the case of liquid culture, agitating the culture for 1 week from 7 days after the culture commenced was also effective for embryo development. However, when the total agitation time was longer (2 to 3 weeks) during liquid-double layer culture or liquid culture, the embryos developed abnormally in both cases. The normal cotyledonary embryos obtained in this study successfully developed to plants when transferred to regeneration media. These regenerated plants were either diploid or haploid, and there was a difference in the number of chloroplasts between guard cells of diploid and haploid. These results can be used as an important data for developing an efficient microspore culture system with high quality embryo production in hot pepper.

• Proceedings of the Korean Society of Crop Science Conference
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• 2023.04a
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• pp.167-167
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• 2023

• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.38-38
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• 2015

Throughout the world, clubroot disease is one of the most damaging diseases affecting Brassica oleracea. In order to perform QTL analysis of CR (clubroot resistance) loci in B. oleracea, we constructed a map, and analyzed CR-QTLs using the mean phenotypes of F3 progenies from the cross of a resistant double-haploid cabbage line (Anju) with a susceptible double-haploid broccoli line (GC). We identified one major QTL, pb-Bo(Anju)1 in C2 from Anju and four minor QTLs; pb-Bo(GC)1 in O5 from GC, pb-Bo(Anju)2, -3, -4 in C2, C3, and C7 from Anju, respectively. Additionally, we found that the accumulation of Pb-Bo(Anju)1 allele and the minor CR-QTLs is essential for resistance against various six isolates. Our finding markers closely linked to the CR-QTLs will help marker-assisted selection for CR. At present, we are undergoing toward map-based cloning for Pb-Bo(Anju)1 gene. The preliminary experiment delimited Pb-Bo(Anju)1 locus, encompassing among 450kB.

• Korean Journal of Microbiology
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• v.20 no.3
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• pp.134-146
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• 1982

This experiment was designed to elucidate the life cycle of 7 species (Rh.nigricans, Rh. delemar, Rh.oryzae, Rh.acidus, Rh.tritici, Rh. formosaensis and Rh. japonicus) in genus Rhizopus isolated from Korean soil, so as to seize the appropriate stage for detecting their chromosomal number and nuclear size under the method of thin layer slide culture using modified Rogers(1965a) medium. The results are summarized as the folowings ; 1. The haploid chromosome number are found 16 in Rh. japonicus are 8, respectively. 2. Comparing the 7 species of Rhizopus with each other, it may be concluded that the basic haploid chromosome number of genus Rhizopus distributed in Korean soil are 8 and that Rh. nigricans is double of the basic hapolid chromosome number (n = 16). Besides them, the other two species (Rh. tritici and Rh. formosaensis) are believed aneuploids.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.2
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• pp.165-171
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• 2009

The objective of this study was to characterize the main-effect QTLs, epistatic QTLs and QTL-by-environment interactions (QE), which are involved in the control of protein content. A population of 120 doubled haploid (DH) lines derived from a cross between 'Samgang' and 'Nagdong', was planted and determined for protein content over three years. Based on the population and a genetic linkage map of 172 markers, QTL analysis was conducted by WinQTLcart 2.5 and QTLMAPPER. Three main-effect QTLs affecting protein content of brown rice were detected from 2004 to 2006 on chromosomes 1 and 11. The qPC11.2 was repeatedly detected across two years. Seven pairs of epistatic loci were identified on eight chromosomes for protein content and collectively explained 39.15% of phenotype variation. These results suggest that epistatic effects might be an even more important component of the genetic basis for protein content and that the segregation of the DH lines for protein content could be largely explained by a few main-effect QTLs and many epistatic loci.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.282-282
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• 2022

Arsenic (As) is a toxic heavy metal that affects the major rice-growing regions of the world and can cause cancer in humans. Rice paddy fields in South Asia are mostly dependent on arsenic-contaminated water sources due to which rice takes up the arsenic from the soil through roots and accumulates it in plant different parts. Here, we present a quantitative trait locus (QTL) mapping study to find out candidate genes conferring As toxicity tolerance in rice (Oryza sativa L.) at the seedling stage. Three weeks old, 120 double haploid CNDH lines derived from a cross between the Indica variety Cheongcheong and the Japonica variety Nagdong and their parental lines were used by treating with 25 μM As. After 2 weeks ofAs stress, 5 traits such as; shoot length (SL), root length (RL), shoot fresh weight (SFW), root fresh weight (RFW), and chlorophyll contents (CHC) were measured. A linkage map of 12 rice chromosomes was constructed from genotypic data DH lines using 778 SSR markers. The linkage map covered a total genetic distance of 2121.7 cM of the rice genome with an average interval of 10.6 cM between markers. A total of seventeen QTLs (LOD>2) were mapped on chromosomes 1, 2, 3, 6, 7, 8, 9, 11, and 12 using composite interval mapping with trait-increasing alleles coming from both parents. Five QTLs for SL, Two QTLs for RL, Five QTLs for SHL, Three QTLs for RFW, and Two QTLs for CHC were detected. The QTLs related to CHC were selected for forther study.