Kim, Joo-Yean;Min Jung;Kim, Hyo-Soon;Lee, Yun-Hee;Park, Soon-Ho;Lim, Yong-Pyo;Min, Byung-Whan;Yang, Seung-Gyun;Harn, Chee-Hark
Journal of Plant Biotechnology
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v.4
no.3
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pp.129-134
/
2002
We report the development of a new selection system for the transformation of pepper plants by mannose. In order to achieve this, we first tested several factors related to regeneration conditions. Among the 30 inbred lines examined, line P9l5 was able to generate shoots at the highest rate from both cotyledons and hyporotyls in MS media. A dosage curve for optimizing the selection conditions was established by mixing mannose (range 0-50 g/L) and sucrose (range 0-30 g/L). The least selection pressure on shoot formation was created by a mixture of sucrose and mannose at 20 g/L and 15 g/L, respectively, and the threshold for ultimate tissue death was 50 g/L of mannose irrespective of the sucrose concentration. However, we found that mannose itself was not the sole inhibitor of pepper shoot development. High concentrations of sucrose (30 g/L) contributed additively to the inhibition of shoot formation at higher mannose concentrations. Genotype preference is a major factor that enhances regeneration ability in mannose media, as was observed in MS media. P9l5 and P410 line had high regeneration rates under mannose selection conditions in the presence of Agrobacterium infection. Different virulence levels of Agrobacterium strains did change the regeneration rates, probably due to interaction with the specificities of the inbred lines. Taken together, P9l5 offers the best pepper inbred line for transformation and we recommend a selection condition of 20 g/L of sucrose and 15 g/L or more of mannose up to 50 g/L in media.
Park, Choul Ji;Nam, Won Shik;Lee, Myeong Seok;Kang, Ji-Yun;Kim, Kyung Kil
The Korean Journal of Malacology
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v.30
no.4
/
pp.415-419
/
2014
Inbreeding depression may be an avoidable phenomenon for abalone culture. However, only a few studies were carried out on inbreeding depression. In the present study, to demonstrate inbreeding depression in growth trait of Pacific abalone, H. discus hannai, inbreeding and outbreeding families were produced in 2010. Inbred and outbred families from each experiment were reared in same tank until 10 month for the same breed environment. The individual of inbred and outbred were distinguished by paternity test using microsatellite DNA. The shell length between inbred and outbred families was compared. At the results, significantly higher shell length was observed in the outbred families at 10 mon (P < 0.05). These results indicate that inbreeding depression is obviously observed in growth traits in the first generation of full-sib family of the H. discus hannai.
Yang Zhi Hong;Jin Hua;Plaha Prikshit;Woong Bae Tae;Jiang Guo Bin;Woo Jong Gyu;Yun Han Dae;Lim Yong Pyo;Lee Hyo Yeon
Journal of Plant Biotechnology
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v.6
no.4
/
pp.235-239
/
2004
We studied the effect of genotype, explant, age of explant, medium (plant growth regulators and gelling agents), and standardized an efficient regeneration protocol for inbred lines of Chinese cabbage (Brassica rapa ssp. pekinensis). Of the different concentrations of NAA and BA tested, the combination of $5\;\cal{mg/L}\;BA\;and\;0.5\;\cal{mg/L}$ NAA gave the highest frequency of shoot regeneration. The cotyledonary explants had more shoot regeneration frequency ($\ge40\%$) than the hypocotyl ex-plants. Besides, the cotyledonary explants, excised from the 4-day old seedlings, showed higher shoot regene-ration ($56.7\%$). Of the three gelling agents and their concentrations used, 16 g/L agar was found to be the best for shoot regeneration. Shoot regeneration frequency in-creased significantly by supplementing the medium with $4\;\cal{mg/L}\;of\;AgNO_3$ MS medium devoid of NAA showed higher frequency of rooting in the regenerated shoots than the ones supplemented with NAA. Our improved regeneration protocol will be especially useful for the genetic transformation of Chinese cabbage inbred lines to develop transgenic hybrids.
A set of rice recombinant inbred lines was developed from a cross between a Tongil type variety, Nonganbyeo, and an indica variety, BG276, by the single seed descent method. The number of the lines in the population was 272. All the agronomic characters studied except ADV (alkali-digestion value) showed continuous variation among the RILs, implying that their inheritance mode should be quantitative. The patterns of the variation in the RILs were either normal or skewed distribution. ADVs of RILs were segregated into two groups with 1:1 ratio, indicating that ADVs in this KIL population might be controlled by one major gene. Transgressive variations were also observed in all characters. Heritability values of the characters varied from 0.488 in brown/rough rice ratio to 0.895 in alkali-digestion value. In the analysis of genotypic and phenotypic correlations, the character of yield was positively correlated with 8 different agronomic characters. The number of panicles per hill was negatively correlated with culm length, panicle length, and number of spikelets per panicle. Grain length was positively correlated with grain width, grain thickness, grain length/width ratio, white belly, ADV, and amylose. However, grain length/width ratio was negatively correlated with grain width. White core was also negatively correlated with white belly and ADV.
Lee Jai-Heon;Kim Kee-Young;Chung Dae-Soo;Chung Won Bok;Kwon Oh-Chang
Proceedings of the Korean Society of Crop Science Conference
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1999.05a
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pp.162-163
/
1999
The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.
As a preliminary experiment to establish the process on the sexing of mouse embryos by chromosomal analysis, present studies were carried out with inbred (ICR, C57BL) and F1 hybrid [(ICR${\times}$C57BL) = F1 ${\times}$ ICR] mice to investigate the blastomere numbers and mitotic indices (M.I.) to the developmental stage of embryos recovered, the optimum periods of anti-mitotic agent administration, the successful rates of sexing and sex-ratio. The results obtained were summarized as follows: 1. The blastomere numbers (mean${\pm}$S.E.) of the morula and blastocyst were 18${\pm}$0.4 and 54${\pm}$0.7, respectively. 2. Whereas the M.I. of F1 hybrid (16${\pm}$0.2%) was higher than that fo inbred ICR (15${\pm}$0.1%) and C57BL (12${\pm}$0.6%) in the different strains, the morula (7${\pm}$0.6%) was higher than that of blastocyst (6${\pm}$0.4%) in the case of embryo stages. 3. Following to anti-mitotic agents treated, the M.I. of embryos cultured with Colcemid (17${\pm}$1.1%) was superior to that fo embryos cultured with Velban (12${\pm}$0.9%) and the Colcemid injection (7${\pm}$0.4%). 4. The successful rate of sexing in the blastocyst (38.7%; 124/320) was superior to the morula (35.9%; 52/145), and the F1 hybrid (48.1%) was higher than that of inbred ICR (42.4%) and C57 BL (28.2%). 5. In the successful rate of sexing to the methods of administration, the embryos cultured with Colcemid (46.0%) was superior to that of embryos cultured with Velban (39.0%) and the Colcemid injection (38.8%). 6. Of 98 embryos sexed after culture with Colcemid, 89(90.8%) were observed between 2 and 4 hrs. In the case of Velban treatment, 83.1% (74/89) was observed between 2$\frac{1}{2}$ and 4$\frac{1}{2}$ hrs. 7. Out of 761 prepared embryos it was possible to sex 311; 157 were male and 154 were female, i.e.a sex-ratio of 50% a, pp.oximately.
Brassica rapa is one of the most valuable vegetable crops worldwide. Cultivated varieties of B. rapa exhibit diverse developmental and morphological appearances, which includes important vegetables, oilseeds, and fodder crops. In this study, various phenotypes of recombinant inbred lines (RILs) of B. rapa were investigated, including their responses to five different pathogenic Botrytis cinerea isolates, responses to aphid and thrips during flowering stages, days to flowering, and plant heights. Responses of 113 RILs to five different B. cinerea isolates showed variations, suggesting that genetic factors controlling resistance or tolerance against each isolate were dependent on isolate/genotype pairs. Correlation analysis was performed to understand the nature of genetic factors and the relationship among these phenotypes. Although high levels of correlation were not detected between phenotypes assessed in this study, statistically significant correlation was detected for several combinations. Significant positive correlations were found for different B. cinerea isolates, supporting that certain levels of commonality could exist in genetic components controlling resistance against different B. cinerea isolates. Based on correlation analysis using numbers of insects counted on plants, it was speculated that genetic factors responsible for aphid tolerance or repellence might be also involved in the response against thrips. Relationship between vegetative growth and tolerance against B. cinereal or insects is rather more complicated. However, it was observed that shorter plants appeared to have a certain level of tolerance or repellence against both aphids and thrips. Data presented in this study could be used to assist further genetic studies and breeding efforts to obtain Botritis and insect resistance for B. rapa.
Amylogram properties such as peak viscosity, hot viscosity, cool viscosity, breakdown, pasting consistency and setback were investigated by the Rapid Visco Analyzer and interpreted by the relationship among amylogram properties according to the varietal groups classified by the rapidity of grain filling (RGF) which was calculated by the percentage of grain weight at 15 days after heading to 40 days after heading. The 164 rice recombinant inbred lines from the cross of Milyang23 and Gihobyeo were used to get the basic information regarding the amylogram properties. The used recombinant inbred lines could be grouped into 4 varietal groups such as slow maturing (less than 40% of RGF), mid-slow maturing (41-60% of RGF),mid-fast maturing (61-80% of RGF), and fast maturing (more than 81% of RGF) groups based on the RGF. The peak viscosity and setback showed regular tendency according to the varietal groups classified by the RGF. Positive significant correlations were found between pasting consistency and setback, however negative significant correlations were found between breakdown and setback in all varietal groups.
Objective: Testicular growth and development are strongly influenced by androgen. Although both testis weight and plasma testosterone level are inherited traits, the interrelationship between them is not fully established. Males of DDD/Sgn (DDD) mice are known to have extremely heavy testes and very high plasma testosterone level among inbred mouse strains. We dissected the genetic basis of testis weight and analyzed the potential influence of plasma testosterone level in DDD mice. Methods: Quantitative trait loci (QTL) mapping of testis weight was performed with or without considering the influence of plasma testosterone level in reciprocal $F_2$ intercross populations between DDD and C57BL/6J (B6) mice, thereby assessing the influence of testosterone on the effect of testis weight QTL. Candidate genes for testis weight QTL were investigated by next-generation sequencing analysis. Results: Four significant QTL were identified on chromosomes 1, 8, 14, and 17. The DDDderived allele was associated with increased testis weight. The $F_2$ mice were then divided into two groups according to the plasma testosterone level ($F_2$ mice with relatively "low" and "high" testosterone levels), and QTL scans were again performed. Although QTL on chromosome 1 was shared in both $F_2$ mice, QTL on chromosomes 8 and 17 were identified specifically in $F_2$ mice with relatively high testosterone levels. By whole-exome sequencing analysis, we identified one DDD-specific missense mutation Pro29Ser in alpha tubulin acetyltransferase 1 (Atat1). Conclusion: Most of the testis weight QTL expressed stronger phenotypic effect when they were placed on circumstance with high testosterone level. High testosterone influenced the QTL by enhancing the effect of DDD-derived allele and diminishing the effects of B6-derived allele. Since Pro29Ser was not identified in other inbred mouse strains, and since Pro29 in Atat1 has been strongly conserved among mammalian species, Atat1 is a plausible candidate for testis weight QTL on chromosome 17.
Wang A. G;Kim, S. U.;Y. H. Han;Kim, S. K.;D. Y. Yu
Korean Journal of Animal Reproduction
/
v.27
no.1
/
pp.69-75
/
2003
The objective of this study is to improve the efficiency of embryo transfer in generation of transgenic mice using inbred mouse (C57BL/6J) embryos. The embryos of C57BL/6J and BCF1 mice were superovulated by the standard protocol. One-cell stage of embryos were microinjected and the resulted one-or two-cell were transferred into one-or two-side oviducts of foster mother, respectively. When one-cell stage of embryos were transferred into one-side oviduct of 0.75 d.p.c. foster mother, the results were not ideal because of showing pregnancy ratios of 68.8$\pm$7.83% for C57BL/6J and 48.3$\pm$14.22% for BCF1, and development ratios of pups vs transferred embryos of 11.9$\pm$5.51% for C57BL/6J and 10.5$\pm$8.03% fur BCF1 . However, when two-cell stage of embryos were transferred into two-side oviducts of 0.5 d.p.c. foster mother, we got better results of 94.4$\pm$9.64% and 100$\pm$0% pregnancy ratio, and 22.1$\pm$0.4% and 21.8$\pm$0.38% development ratio for C57BL/6J and BCF1, respectively. Therefore, transferring two-cell stage of C57BL/6J embryos into two-side oviducts of 0.5 d.p.c. foster mother may be competitive to the result in BCF1 embryos. Even if there are a lot of other factors affecting these results, we conclude from these experiments that transfer of two-cell embryos into two-side oviducts of 0.5 d.p.c. foster mother is a more efficient and safe method than others in generating transgenic mice using inbred mice embryos.
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