Even though klotho deficiency in mice exhibits multiple aging-like phenotypes, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. The objective of this study was to generate homozygous klotho knockout porcine cell lines and cloned embryos. A CRISPR sgRNA specific for the klotho gene was designed and sgRNA (targeting exon 3 of klotho) and Cas9 RNPs were transfected into porcine fibroblasts. The transfected fibroblasts were then used for single cell colony formation and 9 single cell–derived colonies were established. In a T7 endonuclease I mutation assay, 5 colonies (#3, #4, #5, #7 and #9) were confirmed as mutated. These 5 colonies were subsequently analyzed by deep sequencing for determination of homozygous mutated colonies and 4 (#3, #4, #5 and #9) from 5 colonies contained homozygous modifications. Somatic cell nuclear transfer was performed to generate homozygous klotho knockout cloned embryos by using one homozygous mutation colony (#9); the cleavage and blastocyst formation rates were 72.0% and 8.3%, respectively. Two cloned embryos derived from a homozygous klotho knockout cell line (#9) were subjected to deep sequencing and they showed the same mutation pattern as the donor cell line. In conclusion, we produced homozygous klotho knockout porcine embryos cloned from genome-edited porcine fibroblasts.

Minipigs are regarded as one of the most important laboratory animal in that anatomical and physiological properties are similar to human and their reproduction efficiency is relatively higher compared to other large animal species. Particularly, several diseases that cannot be mimicked in rodent models are successfully occurred or induced in pig models therefore it has been interested in a valuable model for human diseases. Pigs are also ‘standard’ species in xenotransplantation research. To maximize experimental outcome using minipigs, establishment and management of proper animal facility, right animal husbandry and control of pathogens are very important. In this review, we summarized several international guidelines related with minipigs published by several companies or governments and discuss optimal conditions for providing informative ideas to the researchers who want to use minipigs in their future studies.

Background : The effective components of Omija(Schisandra chinensis Bailllon) are lignans (schizandrins and gomisins), and this components were contented mostly in seed part on Omija, which have various physiological functionalities such as anti-cancer, anti-inflammatory, and antioxidant activities. Methods and Results : This study was carried out to determine effective condition(CO2, CO2+ethanol) on extraction using supercritical carbon dioxide extraction (SFE) system and to find interrelation on effective components and antioxidant activity of extracts and residues obtained after extraction. Effective components were analysed lignans and phenolic compounds and antioxidant activirty was determined for DPPH radical scavenging ability on methanol extracts of SFE-extract and SFE-residue. On SFE with ethanol, SFE extract was separated two phase, upper(water phase) and lower(oil phase). SFE-extract showed the highest total lignans content(61.36 mg/g, 72.14 mg/g on lower, 50.58 mg/g on upper) and the lowest total phenolic compounds(6.52 mg/100g) and SFE-residue showed the lowest total lignans content(1.45 mg/g) and the highest total phenolic compounds(16.23 mg/100g) by extracted on CO2+ethanol treatment. SFE-residue methanol extract showed the highest DPPH radical scavenging abilities and SFE-extract upper showed the lowest. Conclusion : Thus, this results showed SFE-extract showed the highest total lignans content, but SFE-residue showed the highest DPPH radical scavenging ability although the lowest total lignans content.

Background : This study for the stable production and supply of seeds of Angelica gigas Nakai(Man-chu Korean Angelica), when seeds harvested using nets, seed productivity was investigated. Methods and Results : Planting density is 50 × 25cm, Fertilizer per 10a was sprayed amount of N-P2O5-K2O = 16-17-10kg. And the amount of compost per 10a was sprayed 3000kg. Seed harvesting nets were used for a time formed the endosperm of the seeds (later in mid-August to late). And net for seed production was used for onion nets (at least 13 × 18cm). Shoot growth conditions were as follows. Bolting rate was 89.0% in the untreated, the treated group was 93.1%. The length and thickness of each stem was 129.3 ~ 130.8cm, 1.8cm. The number of nodes per plant was 6.7 ~ 7.5 pieces, and the number of petiole was 14.8 ~ 15.5 per plant. The number of umbel was 10.3 ~ 11.1 piece per plant, and number of deleted umbel was 7.1 ~ 7.2 piece. Seed weight per plant was 24.2g of the net treatment, but ripening seeds 19.6g, 1000 grain weight were all treated and untreated 2.8g. The total seed weight per plant, the net treated was 24.2g, was the weight of the ripening seeds 19.6g. The weight of the ripening seeds were heavier than those of the control. However, the weight of 1000 grain were both treated and untreated 2.8g. When treated nets, the total seed yield per 10a was 88.0kg production, increased by 60.9% compared to untreated. In addition, the ripening seed production per 10a was 71.5kg production, increased by 50.1% compared to untreated. Researching after germination Seed Production, germination rate was 50.8% in the control group and the treatment group was 54.9%. When applying the germination rate, high-quality seed production per 10a was able to produce 39.2kg, compared to control obtain the results increased by 65%. Conclusion : Through the above results, When producing angelica seed, use of net for seed production is thought to be used as a way to prevent early shattering and insect damage.(True bug, etc.).

Abscission is an important developmental process used to shed organs such as leaves, flowers and fruits. Despite the detailed characterization of growth dynamics and hormonal balance during the early steps of fruit development, the molecular aspects remain unclear. Abscission of young fruit occurs by separation of cells in anatomically distinct regions between the pedicel and junction. Differences of gene expression between central pedicel and lateral pedicel were investigated by NGS. Partial cDNAs from 15 clones from both the central pedicel and lateral pedicel were selected for nucleotide sequence determination and homology searches, and 12 clones were subsequently selected for further analysis. In preliminary series of Real Time PCR analysis, 9 genes were confirmed as showing a higher expression level in lateral pedicel than in central pedicel. Many of these genes are expressed in a central or lateral pedicel in specific manner, and the expression profiles of the representative genes were confirmed. To clarify the mechanism of MdIAA14 transcription factor gene underlying abscission zone development, we are investigating the expression patterns between central and lateral pedicels in different apple cultivar using real-time PCR and constructing the vector for transformation into apple.

The precise, fast, and cost-effective identification of important fruit crop cultivars is essential for practical breeding and plant breeder’s rights. Traditional methods for identification of persimmon cultivars are based on the evaluation of sets of morphological characteristics. However, the identification using only morphological traits is difficult to distinguish among genetically closely related cultivars. This study was conducted to develop more reliable DNA markers for identification of the 32 persimmon cultivars in Korea and Japan. In total, 309 random amplified polymorphic DNA (RAPD) markers were identified using 40 different random primers. The 4 (OPP-08) to 14 (UBD159) polymorphic bands were detected with an average of 7.7. The resulting 57 RAPD fragments were selected, and their sequences were determined for developing sequence-characterized amplified region (SCAR) markers. As a result, 15 of 57 RAPD fragments were successfully converted to SCAR markers. A single polymorphic band of the same size as the RAPD fragments or smaller DNA fragments were amplified depending on primer combinations in the 15 SCAR markers. Among these markers, a combination of eight SCAR markers (PS225_200, PSN05_420, PSF13_523, PSN11_540, PS372_567, PS485_569, PSP08_635, and PS631_735) provided sufficient polymorphisms to identify 32 persimmon cultivars depending on number and size of amplicons. These newly developed markers will be useful as a fast and reliable tool to identify persimmon cultivars.

Seeds and seedlings of Acacia bivenosa D.C. Prod, A. salicina Lindl., A. saligna (Labill.) H. Wendl. and A. tumida F. Muell. ex Benth were germinated and grown in NaCl solutions to investigate the relative salt tolerance among the different species, and t