• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.7
• /
• pp.925-937
• /
• 2016

In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.36 no.8
• /
• pp.960-964
• /
• 2007

This study was performed to examine the toxicity of Psoralea corylifolia L. by the single-dose oral toxicity tests in rat and bacterial reverse mutation assay. In single-dose oral toxicity tests, 5 mL ethanol extract of P. corylifolia L. were directly injected into 10 rats (5 males and 5 females) at a dosage of 2 g/kg. Death practice was not detected during breeding periods (14 days), and $LD_{50}$ was calculated over 2 g/kg. No difference were observed with control group in the growth rate and histological observations. In bacterial reverse mutation assay, his(-) Salmonella Typhimurium TA98, TA100, TA1535, TA1537 and trp(-) Escherichia coli WP2uvrA (pKM101) were used for assessing the toxicity of ethanol extracts of P. corylifolia L.. No significant difference in formation of the colonies and no dose-dependent increase was observed regardless of the addition of S9 mix. The results showed that ethanol extracts of P. corylifolia L. did not have single-dose oral toxicity and mutagenic toxicity.

• Asian-Australasian Journal of Animal Sciences
• /
• v.27 no.9
• /
• pp.1254-1262
• /
• 2014

Lymphoid enhancer binding factor 1 (LEF-1) is a member of the T-cell specific factor (TCF) family, which plays a key role in the development of breast endothelial cells. Moreover, LEF-1 gene has been identified as a candidate gene for teat number trait. In the present study, we detected two novel mutations (NC_010450.3:g. 99514A>G, 119846C>T) by DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism in exon 4 and intron 9 of LEF-1 in Guanzhong Black, Hanjiang Black, Bamei and Large White pigs. Furthermore, we analyzed the association between the genetic variations with teat number trait in these breeds. The 99514A>G mutation showed an extremely significant statistical relevance between different genotypes and teat number trait in Guanzhong (p<0.001) and Large White (p = 0.002), and significant relevance in Hanjiang (p = 0.017); the 119846C>T mutation suggested significant association in Guanzhong Black pigs (p = 0.042) and Large White pigs (p = 0.003). The individuals with "AG" or "GG" genotype displayed more teat numbers than those with "AA"; the individuals with "TC" or "CC" genotype showed more teat numbers than those with "TT". Our findings suggested that the 99514A>G and 119846C>T mutations of LEF-1 affected porcine teat number trait and could be used in breeding strategies to accelerate porcine teat number trait improvement of indigenous pigs breeds through molecular marker assisted selection.

• Journal of Sericultural and Entomological Science
• /
• v.5
• /
• pp.25-38
• /
• 1965

I. Breeding of tussah silkworm(preliminary report). The preliminary examination for bleeding has been carried out since 1963 in tussah silkworms. 1) The strain(l-MG-B)of the heaviest silk quantity was the green silkworm and brown cocoon in univoltine, and the strains(2-G-B, 2-MG-B) of the heaviest silk quantity were also the green silkwom and brown cocoon in bivoltine in both spring and fall in 1965. 2) It looks like the voltinism, the body color and the cocoon color have reached to pure line up to 1965. II. Best place for the winter of tussah pupa. This work was aimed to find out good ways for the winter of tussah pupa. 1) The hatch of bivoltine was better than that of univoltine. 2) The cocoons covered with the leaves were good in the emergence of moth. 3) The cocoons which were kept at natural temperature till the first emergence of moths would show bad in both hatch and emergence. 4) If some of the pupae kept under natural condition were controled at proper temperature for a few days, hatch and laying eggs were best. 5) The best places for the winter were the egg storage and the rearing room. III. Relation between incubation temperature and voltinism. 1) When the tussah pupa are kept at natural temperature during winter, the moths do not come out of the pupa. 2) There is no difference between about 18$^{\circ}C$ and about 25$^{\circ}C$ during incubation in hatching ratio. 3) The tussah silkworms of univoltine in mortality are stronger than that of bivoltine. 4) There is not any relation between voltinism and high or low temperature for pupa and eggs. IV. Induced mutation by gamma-ray and neutron in tussah silkworm. This work was carried out in order to induce the mutation by treating the pupa or the eggs of tussah silkworm with gamma my and neutron. The results obtained are as follows. 1. Though the whole pupa treated with neutron become moths, the moths have no ability to copulate each other. The only moths emerged from pupa treated with neutron, 4000${\gamma}$ are able to lay all un-fertilized eggs, some of which have a hole on the surface and nothing of contents. 2. The non-diapause eggs are treated with neutron in spring, but the hatching ratio is 50∼60 percent, but the whole eggs treated with gamma ray are never hatched. 3. The sensitivity of the pupa to neutron is weaker than that of the eggs. 4. The hatching ratio is in direct proportion to the gamma ray dose. 5. Author find out a new mutant which is excellent in the cocoon quality, so he will do the progeny test next hear.

• Horticultural Science & Technology
• /
• v.28 no.5
• /
• pp.796-801
• /
• 2010

This study was carried out to establish a system for mutation breeding by irradiation of gamma-ray in $Rosa$ $hybrida$ Hort. The rooted cuttings of two roses, 'Spidella' and 'Cabernet' were irradiated with different gamma-ray doses (0, 30, 50, 70, 90, 110, 130, 150 and 170 Gy) from a $^{60}Co$ source to reveal an optimal dose for induction of mutants. The irradiated plants were planted in a greenhouse, and investigated on the appearance of petal color mutants and shoot growth by gamma ray dose. The 50% lethal doses ($LD_{50}$) of plant were 110 Gy for 'Spidella' and 150 Gy for 'Cabernet', respectively. The 50% decrease dose of shoot length was observed at 70-90 Gy dose for 'Spidella', and 110 Gy dose for 'Cabernet'. Solid, chimeric and mosaic petal mutants with various colors were induced from pink petal of 'Spidella' and red petal of 'Cabernet' when 30-170 Gy dose was irradiated. The mutants obtained from 'Spidella' had white, ivory, pinky ivory, light pink and deep pink petal colors. The mutants obtained from 'Cabernet' had pink, deep pink, purple red (magenta), orange red and purple petal colors. It was suitable to irradiate 70-90 Gy dose for 'Spidella' and 90-110 Gy dose for 'Cabernet' for the induction of various mutants considering plant survival rate, shoot growth and mutant occurrence rate.

• Korean Journal of Plant Resources
• /
• v.29 no.1
• /
• pp.128-135
• /
• 2016

This study examined radiation damage and the optimal gamma-ray dose for mutation breeding in oat (Avena sativa L. cv. Samhan). The seed germination rate decreased as the dose increased over 500 Gy. The median lethal dose (LD₅₀) was approximately 392 Gy. The median reduction dose (RD₅₀) for plant height, tiller number, root length, and flash weight was 411, 403, 394, and 411 Gy, respectively. The optimal dose of gamma irradiation for inducing oat mutation appears to be in the range 300-400 Gy. We performed the comet assay to observe nuclear DNA damage induced by gamma-ray irradiation. This assay showed a clear difference with gamma-ray treatments. DNA damage increased temporarily 7 days after treatment depending on the dose, while no significant difference was identified in response to 300 Gy 30 days after the gamma-ray treatments. The growth characteristics of the M₂ generation decreased as the dose increased over 400 Gy.

• Journal of Korean Society of Forest Science
• /
• v.25 no.1
• /
• pp.80-84
• /
• 1975

The seeds, scions and plants of chestnut tree (Castanea. crenata) and Chinese chestnut tree (C. bungeana) were irradiated by gamma ray in order to know their biological effects on germination, plant growth and mutation at several accumulative doses and dose rates. The results of this study could be summarized as follows: 1. In general, the radio-sensitivity of scions was more sensitive to irradiation, showing 50% reduction dose of the control for grafting percentage at 3.1 kR as compared with 4.6 kR for the above reduction dose in germination rates of seeds. 2. The seeds treated by 5 kR dose resulted in some albino mutants at a rate of 0.84%. There was a general tendency that the seedling height reduced significantly as the irradiation dose increased. 3. The scions treated by an acute irradiation showed their 50% reduction dose in grafting at 3.2 kR for Chinese chestnut and at 3.1 kR for chestnut, respectively, while their irradiation doses increased three times, having 10.2 kR for the 50% reduction dose in the case of semi-acute irradiation. 4. When Chinese chestnut trees were irradiated during their dormant period with a total dose of 7.5 kR to 4.9 kR at a dose rate of 150 R to 98 R per day, there were induced giant leaf bud-sports at a frequency of 16.6%. The averag leaf area of the giant leaf bud-sports were measured at 96.36 square centimeters, while the area of normal leaf was only 26.28 square centimeters.

• Journal of Radiation Industry
• /
• v.8 no.2
• /
• pp.89-95
• /
• 2014

This study was conducted to determine the optimal dose of gamma-ray on the growth and nucleus DNA damage for mutation breeding in licorice. Gamma-rays irradiated to dry seeds with various doses (0 to 1000 Gy). Significant decreases in germination rate (%), survival rate (%) and growth characteristics (plant height, number of leaves, root length and fresh weight) were observed by dose of increased. $LD_{50}$ (lethal dose) was approximately 400 Gy to 500 Gy. Also, reduction doses ($RD_{50}$) of plant height, number of leaves, root length and flash weight were 428 Gy, 760 Gy, 363 Gy and 334 Gy, respectively. It is supplest that the optimal dose of gamma irradiation for licorice mutation induction might be about 400 Gy in this study. We also conducted comet assay to observe nucleus DNA damage due to gamma irradiation. In comet assay, a clear difference was identified over 300 Gy treatments. With increasing doses of gamma-ray in the range of 100 to 1000 Gy, the rate of head DNA was decreased significantly from 92.88% to 73.09%. Tail length(${\mu}m$) was increased as the dose of increased over 300 Gy. Growth characteristics (Germination rate, Survival rate, plant height, number of leaves, root length and fresh weight) were highly negatively ($P{\leq}0.01$) correlated with dose. While the tail length was highly positively ($P{\leq}0.01$) correlated with dose.

• Molecules and Cells
• /
• v.41 no.11
• /
• pp.979-992
• /
• 2018

Potato (Solanum tuberosum L.) is the third most important food crop, and breeding drought-tolerant varieties is vital research goal. However, detailed molecular mechanisms in response to drought stress in potatoes are not well known. In this study, we developed EMS-mutagenized potatoes that showed significant tolerance to drought stress compared to the wild-type (WT) 'Desiree' cultivar. In addition, changes to transcripts as a result of drought stress in WT and drought-tolerant (DR) plants were investigated by de novo assembly using the Illumina platform. One-week-old WT and DR plants were treated with -1.8 Mpa polyethylene glycol-8000, and total RNA was prepared from plants harvested at 0, 6, 12, 24, and 48 h for subsequent RNA sequencing. In total, 61,100 transcripts and 5,118 differentially expressed genes (DEGs) displaying up- or down-regulation were identified in pairwise comparisons of WT and DR plants following drought conditions. Transcriptome profiling showed the number of DEGs with up-regulation and down-regulation at 909, 977, 1181, 1225 and 826 between WT and DR plants at 0, 6, 12, 24, and 48 h, respectively. Results of KEGG enrichment showed that the drought tolerance mechanism of the DR plant can mainly be explained by two aspects, the 'photosynthetic-antenna protein' and 'protein processing of the endoplasmic reticulum'. We also divided eight expression patterns in four pairwise comparisons of DR plants (DR0 vs DR6, DR12, DR24, DR48) under PEG treatment. Our comprehensive transcriptome data will further enhance our understanding of the mechanisms regulating drought tolerance in tetraploid potato cultivars.

• Journal of Plant Biotechnology
• /
• v.45 no.1
• /
• pp.17-29
• /
• 2018

This study was conducted to investigate a morphological trait in 294 rice accessions including Korean breeding lines. We also carried out a genome-wide association study (GWAS) to detect significant single nucleotide polymorphism markers and candidate genes affecting major agronomic traits. A Manhattan plot analysis of GWAS using morphological traits showed that phenotypic and statistical significance was associated with a chromosome in each group. The significance of SNPs that were detected in this study was investigated by comparing them with those found previously studied QTL regions related to agronomic traits. As a result, SNP (S8-19815442), which is significant with regard to leaf angle, was located in the known QTL regions. To observe gene mutations related to leaf angle in a candidate gene, Os08g31950, its sequences were compared with sequences in previously selected rice varieties. In Os08g31950, a single nucleotide mutation occurred in one region. To compare relative RNA expression levels of candidate gene Os08g31950, obtained from GWAS analysis of 294 rice accessions and related to lateral leaf angle, we investigated relative levels by selecting 10 erect leaf angle varieties and 10 horizontal leaf angle varieties and examining real-time PCR. In Os08g31950, a high level of expression and various expression patterns were observed in all tissues. Also, Os08g31950 showed higher expression levels in the erect leaf angle variety group and higher expression rates in the leaf than in the root. The candidate gene detected through GWAS would be useful in developing new rice varieties with improved yield potential through future molecular breeding.