Enhanced development of porcine embryos cloned from bone marrow mesenchymal stem cells

Jin, Hai-feng,Kumar, B Mohana,Kim, Jung-Gon,Song, Hye-Jin,Jeong, Yeon-Ji,Cho, Seong-Keun,Balasubramanian, Sivasankaran,Choe, Sang-Yong,Rho, Gyu-Jin UPV/EHU Press 2007 The international journal of developmental biology Vol.51 No.1

<P>In the present study, we have characterized an isolated population of porcine bone marrow mesenchymal stem cells (MSCs) for multilineage commitment and compared the developmental potential of cloned embryos with porcine MSCs and fetal fibroblasts (FFs). MSCs exhibited robust alkaline phosphatase activity and later transformed into mineralized nodules following osteoinduction. Furthermore, MSCs underwent adipogenic and chondrogenic differentiation by producing lipid droplets and proteoglycans, respectively. Primary cultures of FFs from a female fetus at ~30 day of gestation were established. Donor cells at 3-4 passage were employed for nuclear transfer (NT). Cell cycle analysis showed that the majority of MSCs in confluence were in the G0/G1 stage. Cumulus-oocyte complexes were matured and fertilized in vitro (IVF) as control. The cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs and FFs (84.54.6% vs. 52.25.4% and 50.85.2%, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs (20.62.5% and 18.43.0%) did not differ, but were significantly (P<0.05) higher than NT derived from FFs (9.52.1%). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs (34.45.2 and 0.380.08, respectively) were significantly (P<0.05) higher than those from FFs (22.65.5 and 0.180.12, respectively). Proportions of TUNEL positive cells in NT embryos from FFs (7.31.8%) were significantly (P<0.05) higher than in MSCs (4.61.3%) and IVF (2.50.9%). The results clearly demonstrate that multipotent bone marrow MSCs have a greater potential as donor cells than FFs in achieving enhanced production of cloned porcine embryos.</P>

Comparison of Developmental Competency of Porcine Embryos Cloned with Mesenchymal Stem Cells and Somatic Cells

Jin Hai-Feng,Kumar B. Mohana,Cho Sung-Keun,Ock Sun-A,Jeon Byeong-Gyun,Balasubramanian S.,Choe Sang-Yong,Rho Gyu-Jin The Korean Society of Animal Reproduction 2006 Reproductive & developmental biology Vol.30 No.2

The present study compared the developmental potential of cloned porcine embryos with mesenchymal stem cells (MSCs), fetal fibroblasts (FFs) and cumulus cells (CCs) by assessing the cleavage and blastocyst rate, total cell number, inner cell mass (ICM) ratio and apoptosis. MSCs were isolated by ficoll gradients from femur of -6 month old female pig, and maintained for primary cultures. FFs from a female fetus at ${\sim}30$ day of gestation were established, and CCs were obtained from cumulus oocyte complexes (COCs) aspirated from $3{\sim}6$ mm follicles in diameter. Donor cells at $3{\sim}4$ passage were employed for nuclear transfer (NT). COCs were matured and fertilized in vitro(IVF) as control. Cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs, FFs and CCs ($82.7{\pm}8.9%\;vs\;70.6{\pm}5.4,\;68.7{\pm}5.1\;and\;63.4{\pm}5.6%$, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs ($24.5{\pm}2.8\;and\;20.4{\pm}8.3%$) did not differ, but were significantly (P<0.05) higher than NT derived from FFs and CCs ($10.6{\pm}2.7\;and\;9.8{\pm}2.1%$). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs ($35.4{\pm}5.2\;and\;0.40{\pm}0.09%$, respectively) were significantly (P<0.05) higher than those from FFs and CCs ($24.9{\pm}6.2%\;vs\;0.19{\pm}0.16,\;23.6{\pm}5.5\;and\;0.17{\pm}0.16%$, respectively). Proportions of TUNEL positive cells in NT embryos from FFs and CCs ($6.9{\pm}1.5\;and\;7.4{\pm}1.7%$, respectively) were significantly (P<0.05) higher than in MSCs ($4.8{\pm}1.4%$) and IVF ($2.3{\pm}0.9%$). The results demonstrate that MSCs have a greater potential as donor cells than FFs and CCs in achieving enhanced production of cloned porcine embryos.

Blast diseases influenced on agronomic and eating quality traits of rice in Korea

Feng-peng Li,Won-Hee Ra,Yong-Jin Park,Il-Pyung Ahn 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07

Rice (Oryza sativa L.) is one of the most important staple foods that feed more than 50% of the world’s population. With the improving of people’s living standard, eating quality of rice become the most important aims in current breeding programs. Amylose content (AC) and gelatinization temperature (GT) are the two main measures to estimate the rice grain quality. In rice, a total of 27 genes directly involved the rice starch biosynthesis effecting on the rice eating quality. It clearly identified chromosome 6 to be rich in the genes related to AC and GT properties (GBSS I, SSIIa and SBE I) along with other genomic regions scattered in rice genome. Rice blast, caused by the fungal pathogen M. oryzae, is the most devastating disease of rice and severely affects crop stability and sustainability worldwide. Many fungal genes involved in pathogenicity and rice genes involved in effector recognition and defense responses have been identified over the past decade. A total of 99 and 22 blast resistance genes have been identified and cloned; in which 45% were found in japonica cultivars, 51% in indica cultivars, and the rest 4% in wild rice species. Among them, three major resistance gene clusters have been characeterized: the Pik locus on Chromosome 11, and the Pita locus on Chromosome 12, the Piz locus on Chromosome 6 closely to the starch synthesis-related genes. These results could be important clues for studying the relationship between resistance / susceptible materials and eating quality.

Cat fertilization by mouse sperm injection

Jin, Yong-Xun,Cui, Xiang-Shun,Yu, Xian-Feng,Lee, Sung-Hyun,Wang, Qing-Ling,Gao, Wei-Wei,Xu, Yong-Nan,Sun, Shao-Chen,Kong, IL-Keun,Kim, Nam-Hyung Cambridge University Press 2012 Zygote Vol.20 No.4

<B>Summary</B><P>Interspecies intracytoplasmic sperm injection has been carried out to understand species-specific differences in oocyte environments and sperm components during fertilization. While sperm aster organization during cat fertilization requires a paternally derived centriole, mouse and hamster fertilization occur within the maternal centrosomal components. To address the questions of where sperm aster assembly occurs and whether complete fertilization is achieved in cat oocytes by interspecies sperm, we studied the fertilization processes of cat oocytes following the injection of cat, mouse, or hamster sperm. Male and female pronuclear formations were not different in the cat oocytes at 6 h following cat, mouse or hamster sperm injection. Microtubule asters were seen in all oocytes following intracytoplasmic injection of cat, mouse or hamster sperm. Immunocytochemical staining with a histone H3-m2K9 antibody revealed that mouse sperm chromatin is incorporated normally with cat egg chromatin, and that the cat eggs fertilized with mouse sperm enter metaphase and become normal 2-cell stage embryos. These results suggest that sperm aster formation is maternally dependent, and that fertilization processes and cleavage occur in a non-species specific manner in cat oocytes.</P>

Transcriptome profiling of Shindongjin and Sugary mutant at grain-filling stages using RNA-Seq

Feng-Peng Li,Min-Young Yoon,Gang Li,Yong-Jin Park 한국육종학회 2013 한국육종학회 심포지엄 Vol.2013 No.07

Rice (Oryza sativa) is an excellent model monocot with a known genome sequence for studying developmental seeds. In the study, the seeds of 10th day after flowering (DAF) were conducted RNA-Seq of the variety Shindongjin and Sugary mutant using RNA-seq technique. Approximately 202 and 214 million high-quality paired-end reads (101-bp in size) were generated in Shindongjin and Sugary mutant, respectively. Comprehensive analysis on the transcript levels of genes which encode starch-synthesis enzymes is fundamental for the assessment of the function of each enzyme and the regulatory mechanism of starch biosynthesis in seeds. Quantitative real-time PCR was also used to validate the expression profiles of 28 rice genes encoding six classes of enzymes, viz., ADPglucose pyrophosphorylase (AGPase), starch synthase, starch branching enzyme, starch debranching enzyme, starch phosphorylase, and disproportionating enzyme at different developmental grain- filling stages (DAF 1-14) between Shindongjin and Sugary mutant. The results showed that the expression of most of starch synthesis genes were up-regulated except the cytosolic AGPase small subunit2b (AGPS2b), which sharply decreased at grain-filling stages in Sugary mutant. These results will expand our understanding of the complex molecular and cellular events in rice grain-filling stages and provide a fundamental understanding of future studies on developmental endosperm in rice and other cereal crops.

Comparison of Developmental Competency of Porcine Embryos Cloned with Mesenchymal Stem Cells and Somatic Cells

Hai-Feng Jin,B. Mohana Kumar,Sung-Keun Cho,Sun-A Ock,Byeong-Gyun Jeon,S. Balasubramanian,Sang-Yong Choe,Gyu-Jin Rho 한국동물생명공학회(구 한국동물번식학회) 2006 Reproductive & developmental biology Vol.30 No.2

Outbound Open Innovation for Enhanced Innovation Performance : An Empirical Study based on BTB Enterprises in China

Huan-Yong Ji,Jing Li,Chong-Feng Lan,Dan-Feng, Wang,Yong-Sheng Jin 보안공학연구지원센터 2016 International Journal of u- and e- Service, Scienc Vol.9 No.5

Open Innovation gains a wide spread attention in technology management research in recent years. Because existing research about the relationship between outbound open innovation and innovation performance is limited and context factors are ignored in existing research and based on Chesbrough’s open innovation model and theory of enterprise competence, this study introduces enterprise competence and knowledge spillovers into the field of outbound open innovation, explores their moderated roles between open innovation and innovation performance. We use high-tech firms as research sample to test the hypothesis that we proposed and find that: (1) outbound open innovation has a positive effect on innovation performance; (2) both enterprise competence and knowledge spillovers moderate the relationship between outbound open innovation and innovation performance. These conclusions have important implications for the development and application of open innovation theory.

Genome Wide Association Study (GWAS) approach for the blast disease resistance in rice

Won-Hee Ra,Feng-peng Li,Yong-Jin Park,Il-Pyung Ahn 한국육종학회 2014 한국육종학회 심포지엄 Vol.2014 No.07

One of the biotic stresses in rice production is rice blast disease caused by Magnaporthe oryzae, which is one of the most destructive fungal diseases in rice. We outlined an approach towards genome wide association study for the blast disease resistance in rice. In total, 295 rice accessions including 137 Heuristic Set accessions (HS) and 158 Korean Bred varieties (KB) were screened for the rice blast disease resistance. Firstly, Magnaporthe oryzae were inoculated to the rice seedlings of two weeks after germinations. Then, evaluation of the disease symptoms and checking the crossing point (CP) value were conducted one week after inoculation. To quantify the CP value, real-time polymerase chain reaction (PCR) was employed in combination with the primer pair and Taqman probe specific to Magnaporthe oryzae HYDROPHOBIN class 1 (MHP1) which is an indispensable unigene encoding HYDROPHOBIN for normal virulence expression. Based on these CP values from the PCR reactions containing a series of increasing concentration of cloned amplicon or fungal genomic DNA, correlation among the template’s copy number or its amount and amplification pattern was calculated. Reliability of this equation was further confirmed using the DNA samples from the rice leaves infected with compatible or incompatible strains of M. oryzae. These steps are still being undertaken, and after the complete process of disease resistance phenotyping for the whole population containing 295 accessions, GWAS will be performed to examine the associated genes involving in blast resistance mechanism using the whole genome resequencing data of 295 accessions. This approach would be a useful technique for identifying genetic loci responsible for natural variation in rice blast disease resistance and ultimately, new R genes which can improve the blast resistance in rice.

AtCPL5, a novel Ser‐2‐specific RNA polymerase II C‐terminal domain phosphatase, positively regulates ABA and drought responses in Arabidopsis

Jin, Yong‐,Mei,Jung, Jinwook,Jeon, Hyesung,Won, So Youn,Feng, Yue,Kang, Jae‐,Sook,Lee, Sang Yeol,Cheong, Jong‐,Joo,Koiwa, Hisashi,Kim, Minkyun Blackwell Publishing Ltd 2011 The New phytologist Vol.190 No.1

<P><B>Summary</B></P><P><P>Arabidopsis RNA polymerase II (RNAPII) C‐terminal domain (CTD) phosphatases regulate stress‐responsive gene expression and plant development via the dephosphorylation of serine (Ser) residues of the CTD. Some of these phosphatases (CTD phosphatase‐like 1 (CPL1) to CPL3) negatively regulate ABA and stress responses. Here, we isolated <I>AtCPL5</I>, a cDNA encoding a protein containing two CTD phosphatase domains (CPDs).</P><P>To characterize AtCPL5, we analyzed the gene expression patterns and subcellular protein localization, investigated various phenotypes of <I>AtCPL5</I>‐overexpressors and knockout mutants involved in ABA and drought responses, performed microarray and RNA hybridization analyses using <I>AtCPL5</I>‐overexpressors, and assessed the CTD phosphatase activities of the purified AtCPL5 and each CPD of the protein.</P><P>Transcripts of the nucleus‐localized AtCPL5 were induced by ABA and drought. <I>AtCPL5</I>‐overexpressors exhibited ABA‐hypersensitive phenotypes (increased inhibition of seed germination, seedling growth, and stomatal aperture), lower transpiration rates upon dehydration, and enhanced drought tolerance, while the knockout mutants showed weak ABA hyposensitivity. <I>AtCPL5</I> overexpression changed the expression of numerous genes, including those involved in ABA‐mediated responses. In contrast to Ser‐5‐specific phosphatase activity of the negative stress response regulators, purified AtCPL5 and each CPD of the protein specifically dephosphorylated Ser‐2 in RNAPII CTD.</P><P>We conclude that AtCPL5 is a unique CPL family protein that positively regulates ABA‐mediated development and drought responses in Arabidopsis.</P></P>

A study on genotypic variations of Pb accumulation and distribution among organs of rice plant

Yong-Feng Yan,Doug-Hwan Choi,Jin-Ern Lee,Byun-Woo Lee 한국작물학회 2008 한국작물학회 학술발표대회 논문집 Vol.2008 No.10

In order to investigate the genotypic differences in Pb accumulation of rice, thirty-five rice cultivars were irrigated by irrigation water containing 10 ppm Pb throughout all growth season under field condition. At harvest, thirty-five rice cultivars showed significant difference in Pb concentrations in straw, grain, chaff, brown rice, rice bran, and polished rice, but not in root. Pb concentration in polished rice ranged from 0.02 to 0.22 mg/kg. The total Pb accumulation in shoot varied from 0.77 to 2.74 mg/m2. There were no significant difference of Pb concentration in all of rice parts among four rice cultivar groups. Indica and tongil cultivar groups presented the higher Pb accumulation in shoot. The large genotypic variation suggested the possibility of breeding low Pb accumulating rice cultivar.