• Journal of the Korean Society of Tobacco Science
• /
• v.7 no.1
• /
• pp.93-96
• /
• 1985

This study was conducted to establish the maternal haploid method for the practical breeding of M tabacum using the interspeciflc hybridization between M tabacum and N. aflicana. The frequency of surviving seedling per seed capsule of interspecific hybridization was 4.15. Among them, the frequencies of maternal haploid and hybrid were 1.20 and 2.95, respectively. The chromosome numbers of n=24 for maternal haploid and 2n=47 for hybrid were identified in surviving seedling from interspecific hybridiztion. Except the chromosome number, distinguishable morphological differences of material haploid from hybrid were observed at seedling stage.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.40 no.5
• /
• pp.649-654
• /
• 1995

This study was conducted to investigate the utility and agronomic characteristics and use of cytoplasmic. male-sterile (CMS) haploid plants derived from interspecific cross between (male-sterile NC82$\times$burley21) F$_1$ plant (Nicotiana tabacum L.) and Nicotiana africana. Abundant seeds of high germinability were obtained when Nicotiana tabacum (cytoplasmic male-sterile F$_1$ plants) is pollinated by Nicotiana africana. Most of seedlings died at the cotyledonary stage. The remaining seedlings are viable F$_1$ hybrids or maternal haploids that can be easily distinguished. Number of interspecific Fl hybrids and matermal haploids per capsule obtained from the interspecific cross between cytoplasmic male-sterile tobacco F$_1$ plants and N.africana yielded 2.2 and 0.5 plants, respectively. Out of 149 CMS haploid plants obtained from interspecific cross, 102 plants showed green type while the others were yellow type for leaf and stem. This results agreed with the genetic ratios expected among haploid plants from the F$_1$ hybrids heterozygous for two recessive genes of yellow color of burley tobacco plant. Out of 83 CMS haploid plants inoculated with TMV, 48 plants showed resistant, while the others was susceptible. It agreed with the expected genetic ratios for a single dominant gene of TMV resistance. CMS haploid plant will be useful as a source material for breeding of CMS doubled haploid lines

• Molecules and Cells
• /
• v.20 no.2
• /
• pp.201-209
• /
• 2005

We have constructed an AFLP-based linkage map of Japanese red pine (Pinus densiflora Siebold et Zucc.) using haploid DNA samples of 96 megagametophytes from a single maternal tree, selection clone Kyungbuk 4. Twenty-eight primer pairs generated a total of 5,780 AFLP fragments. Five hundreds and thirteen fragments were verified as genetic markers with two alleles by their Mendelian segregation. At the linkage criteria LOD 4.0 and maximum recombination fraction 0.25(${\theta}$), a total of 152 markers constituted 25 framework maps for 19 major linkage groups. The maps spanned a total length of 2,341 cM with an average framework marker spacing of 18.4 cM. The estimated genome size was 2,662 cM. With an assumption of equal marker density, 82.2% of the estimated genome would be within 10 cM of one of the 230 linked markers, and 68.1% would be within 10 cM of one of the 152 framework markers. We evaluated map completeness in terms of LOD value, marker density, genome length, and map coverage. The resulting map will provide crucial information for future genomic studies of the Japanese red pine, in particular for QTL mapping of economically important breeding target traits.

• Journal of Aquaculture
• /
• v.19 no.3
• /
• pp.166-172
• /
• 2006

Induced androgenesis, a form of artificial parthenogenesis is an important tool for the generation and use of genetically isogenic or clonal fish strain. An optimized protocol for the genetic inactivation of mud loach (Misgurnus mizolepis) oocytes (i.e. production of androgenetic haploid) was developed using UV-irradiation. Various dose levels of UV significantly affected the fertilizing capacity of the eggs, hatchability of embryos and incidence of haploidy. Based on the extensive examinations of treatment conditions on embryo viability and haploid incidence, the optimum dose level of UV irradiation was turned out to be $10,800ergs/mm^2$ with 56.9% of hatching success and 94.6% of haploidy. The overall yield of putative androgen under optimized treatment condition was more than 50% out of total eggs inseminated. The success of androgenetic reproduction of haploid genome was verified by flow cytometry and PCR amplification of transgene that is exclusive to either one of parental sexes. However, a small portion $(8\sim11%)$ of presumed androgenetic haploid larvae was proven to contain residual DNA fragment(s) from maternal parent.

• Journal of Plant Biotechnology
• /
• v.44 no.1
• /
• pp.35-41
• /
• 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 Aquaculture
• /
• v.20 no.3
• /
• pp.184-189
• /
• 2007

Treatment conditions for the induced diploid gynogenesis, a maternal-exclusive form of artificial parthenogenetic reproduction, were optimized in bagrid catfish (Leiocassis ussuriensis, Siluriformes). Optimal amounts of ultraviolet (UV) irradiation for the genetic inactivation of spermatozoa in bagrid catfish and Pseudobagrus fulvidraco were proven to be ranged from 3,600 to 4,800 $ergs/mm^2$ based on the examination of viability of embryos and haploid incidence. Haploid embryos were restored to diploidy by preventing the extrusion of the second polar body using cold shock treatment. Thermal treatments (4 or $6^{\circ}C$ for 30, 40 or 50 min) were carried out 3, 5 or 7 min post insemination. Best scores for embryo viability (38.6% of total eggs taken) and incidence of normal diploidy (87.9% of hatched larvae) were observed at the embryo group treated at $4^{\circ}C$ for 40 min, 5 min after insemination. Restoration of gynogenetic diploidy was confirmed based on the absence of haploid syndrome, cell size and/or nucleolar organizing region (NOR) counts.

• Journal of Aquaculture
• /
• v.12 no.3
• /
• pp.167-173
• /
• 1999

Blocking the 1st mitotic cleavage was performed in mud loach (Misgurmus mizolepis) using UV-irradiated cyprinid loach (M. anguillicaudatus) sperm and ternal shocks Optimum UV range for inactivation of cyprinid loach sperm and thermal shocks. Optimum UV range for inactivation of cyprinid loach sperm was between 3,150 to 4,050 ergs/m$m^2$. Heat shock treatment ($41^{\circ}C$ for 3mins) with various treatment initiation times ranged from 22 to 50 min post insemination resulted wide range of success for induced gynogenesis. Best result was obtained when haploid egges were shocked at 28 min after insemination (corresponding to metaphase division of the 1st cleavage); 26% of total eggs inseminated were viable diploid gynogens. The hatching success and early survival of the both meiotic and mitotic gynogenetic groups were significantly lower than those of control crosses (P<0.05). Maternal origin of induced gynogenetic mud loach was verified by multi-locus DNA fingerprinting.

• Journal of the Korean Society of Tobacco Science
• /
• v.14 no.2
• /
• pp.104-115
• /
• 1992

The present study was conducted to compare the relative efficiency of four different breeding methods in tobacco varietal development. A single Fl hybrid plant from cross of two flue-cured cultivars of Nicotiana tabacum L Bright Yellow 4(BY4) and NC95, was selfed. F2population above cross was screened for resistance against bacterial wilt caused by Pseudommonas solanacearum E.F.Smith under the naturally infested field conditions, and the 30 lines were developed from F2 individual plant by anther culture (ADH), maternal method utilizing Nicotiana afpicana (MDH), single- seed descent(SSD) and Bulk breeding method, respectively. All characters except content of total alkaloids of ADH and MDH which wore bred by haploid methods reduced more than that of lines bred by conventional methods(SSD & Bulk) : however, the yields were 8% lower than other lines. The total alkaloid content of ADH was higher than that of MDH, and yield was reduced about 4n even though the number of leaf was identical with the MDH. All other characters of ADH were also reduced significantly. In the lines bred by conventional methods, population developed by SSD showed significantly wider leaf width, shorter plant height, later days to flower, and lower in percent reducing sugar than those by Bulk. The populations derived from haploid method showed greater phenotypic variance and wider range of variation than conventionally developed ones. The results obtained indicate that selection will provide a significantly greater genetic gain for leaf number and leaf length in the ADU and MDH populations, and for plant height and days to flower in the SSD and Bulk populations.

• Journal of Aquaculture
• /
• v.10 no.1
• /
• pp.39-47
• /
• 1997

Induction of hybrid and allotriploid between female rainbow trout, oncorhynchus mykiss and male cherry salmon, O. masou were performed. Triploid rate in allotriploid was 92% and the survival rates of allotriploid exceeded that of their hybrid counterparts. Karyological studies revealed that the hybrid had exactly the same intermediate diploid karyotypers as that of two parental species, while allotriploid had two sets of the maternal haploid complement and one set of the paternal haploid complement. Hybrid revealed an intermediate size in celluar and nuclear size of erythrocyte, while allotriploid revealed larger cellular and nuclear size of erythrocytes than hybrid. The number of nucleolar organizer regions of hybrid were 1 or 2, while those of allotriploids were 1, 2 or 3. These results proved that allotriploidization can lead to increase cytogenetical characters compared to counterpart hybridization.

• Korean Journal of Veterinary Research
• /
• v.30 no.2
• /
• pp.123-127
• /
• 1990

By nuclear transplantation technology twenty eight mice have been produced after transfer of heterozygous biparental eggs. Also heterozygous gynogenetic eggs with two female pronuclei and heterozygous androgenetic eggs with two male pronuclei have been obtained by injecting a male or female pronucleus with Sendai virus into the perivitelline space of enucleated haploid zygotes at pronuclear stage. The success rate of enucleation, karyoplast injection and fusion of both the pronuclei was 80.3, 83.4 and 81.8%, respectively. The overall pronuclei fusion rates by this technique were 56, 50 and 56% in biparental, gynogenetic and androgenetic eggs, respectively. The evidence was ascertained that the gynogenetic and androgenetic eggs were also able to develop in vitro up to blastocyst stage, even though their developmental potential was greatly diminished beyond 2-cell stage. The gynogenetic eggs were able to develop in vivo up to day 10 of pregnancy, while the androgenetic eggs failed to develop in vivo during the same period.