• Title/Summary/Keyword: Molecular Breeding

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Double Mutations in eIF4E and eIFiso4E Confer Recessive Resistance to Chilli Veinal Mottle Virus in Pepper

  • Hwang, JeeNa;Li, Jinjie;Liu, Wing-Yee;An, Song-Ji;Cho, Hwajin;Her, Nam Han;Yeam, Inhwa;Kim, Dosun;Kang, Byoung-Cheorl
    • Molecules and Cells
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    • v.27 no.3
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    • pp.329-336
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    • 2009
  • To evaluate the involvement of translation initiation factors eIF4E and eIFiso4E in Chilli veinal mottle virus (ChiVMV) infection in pepper, we conducted a genetic analysis using a segregating population derived from a cross between Capsicum annuum 'Dempsey' containing an elF4E mutation ($pvr1^2$) and C. annuum 'Perennial' containing an elFiso4E mutation (pvr6). C. annuum 'Dempsey' was susceptible and C. annuum 'Perennial' was resistant to ChiVMV. All $F_1$ plants showed resistance, and $F_2$ individuals segregated in a resistant-susceptible ratio of 166:21, indicating that many resistance loci were involved. Seventy-five $F_2$ and 329 $F_3$ plants of 17 families were genotyped with $pvr1^2$ and pvr6 allele-specific markers, and the genotype data were compared with observed resistance to viral infection. All plants containing homozygous genotypes of both $pvr1^2$ and pvr6 were resistant to ChiVMV, demonstrating that simultaneous mutations in elF4E and eIFiso4E confer resistance to ChiVMV in pepper. Genotype analysis of $F_2$ plants revealed that all plants containing homozygous genotypes of both $pvr1^2$ and pvr6 showed resistance to ChiVMV. In protein-protein interaction experiments, ChiVMV viral genome-linked protein (VPg) interacted with both eIF4E and eIFiso4E. Silencing of elF4E and eIFiso4E in the VIGS experiment showed reduction in ChiVMV accumulation. These results demonstrated that ChiVMV can use both eIF4E and eIFiso4E for replication, making simultaneous mutations in eIF4E and eIFiso4E necessary to prevent ChiVMV infection in pepper.

Regeneration and Agrobacterium - Mediated Transient Transformation of Button Daisy (Leucanthemum vulgare)

  • Franklin G.;Alaiwi W. Abou;Goldman S.L.
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.37-43
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    • 2005
  • Explants of button daisy were screened for their regeneration potential and transient GUS gene expression. Medium containing MS salts minerals and $B_5$ vitamins supplemented with $0.1\;\cal{mg/L}$ BA and $0.1\;\cal{mg/L}$ TDZ showed the best regeneration. Disc florets and receptacles were the most responsive explants in regeneration and transient gene expression respectively. Regenerated plants were successfully rooted and established in the green-house conditions. Infection and co-cultivation of explants with Agrobacterium tumefaciens containing pCAMBIA 1301 resulted in transient GUS foci. Among the different explants, receptacles showed the highest percentage of transient GUS gene expression. Enzymatic and molecular analyses of transformed calli confirmed the integration of GUS gene.

Possibility of breeding super rice cultivars using gene linkage

  • Tran, Dang Xuan;Tran, Dang Khanh;Truong, Thi Tu Anh
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.27-27
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    • 2017
  • The rapid growth of world population, reduction of cultivated areas for crop production, and detrimental effects of pests, diseases, and climate changes have required to breed new rice cultivars with high yield, accepted quality, but strong resistance to abiotic and biotic stresses. However, traditional breeding needs much time to breed a new cultivar, whereas the successful use of molecular breeding is still questionable. We have developed a novel mutation which allow to cross many rice cultivars together with low segregation, that allow to breed a new cultivar in only several cropping. The mechanism has been unknown, but we suggest that gene linkage may play a crucial role, of which the semi dwarf gene might be the center gene for gene linkage occurrence. The phenomenon of this possible gene linkage is contrary to Mendel rules, but it is promising to breed new rice cultivars, of which, the most elite genes in rice might be able to gather in a targeted rice variety.

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Animal Breeding: What Does the Future Hold?

  • Eisen, E.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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    • pp.453-460
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    • 2007
  • An overview of developments important in the future of animal breeding is discussed. Examples from the application of quantitative genetic principles to selection in chickens and mice are given. Lessons to be learned from these species are that selection for production traits in livestock must also consider selection for reproduction and other fitness-related traits and inbreeding should be minimized. Short-term selection benefits of best linear unbiased predictor methodology must be weighed against long-term risks of increased rate of inbreeding. Different options have been developed to minimize inbreeding rates while maximizing selection response. Development of molecular genetic methods to search for quantitative trait loci provides the opportunity for incorporating marker-assisted selection and introgression as new tools for increasing efficiency of genetic improvement. Theoretical and computer simulation studies indicate that these methods hold great promise once genotyping costs are reduced to make the technology economically feasible. Cloning and transgenesis are not likely to contribute significantly to genetic improvement of livestock production in the near future.

Biofuel: Current Status in Production and Research

  • Yu, Ju-Kyung;Park, Soon Ki
    • Korean Journal of Breeding Science
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    • v.42 no.2
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    • pp.121-128
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    • 2010
  • Finding alternative and renewable energy sources has become an important goal for plant scientists, especially with the demand for energy increasing worldwide and the supply of fossil fuel being depleted. The most important biofuel to date is bioethanol which is produced from sugars (sucrose and starch) found in corn and sugarcane. Second generation bioethanol is targeting studies that would allow the use of the cell wall (lignocellulose) as a source of carbon by non-food plants. Plant scientists, including breeders, agronomists, physiologists and molecular biologists, are working towards the development of new and improved energy crops especially, how to design crops for bioenergy production and increased biomass generation for biofuel purposes. This review focuses on: i) the current status of first generation bioenergy production, ii) the limitations of first and second generation bioenergy, and iii) ongoing research to overcome challenging issues in second generation bioenergy.

Development of an Apple F1 Segregating Population Genetic Linkage Map Using Genotyping-By-Sequencing

  • Ban, Seung Hyun;Choi, Cheol
    • Plant Breeding and Biotechnology
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    • v.6 no.4
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    • pp.434-443
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    • 2018
  • Genotyping-by-sequencing (GBS) has been used as a viable single nucleotide polymorphism (SNP) validation method that provides reduced representation sequencing by using restriction endonucleases. Although GBS makes it possible to perform marker discovery and genotyping simultaneously with reasonable costs and a simple molecular biology workflow, the standard TASSEL-GBS pipeline was designed for homozygous groups, and genotyping of heterozygous groups is more complicated. To addresses this problem, we developed a GBS pipeline for heterozygous groups that called KNU-GBS pipeline, specifically for apple (Malus domestica). Using KNU-GBS pipeline, we constructed a genetic linkage map consisting of 1,053 SNP markers distributed over 17 linkage groups encompassing a total of 1350.1 cM. The novel GBS pipeline for heterozygous groups will be useful for marker-assisted breeding programs, and diverse heterozygous genome analyses.

Genome-wide association study for frozen-thawed sperm motility in stallions across various horse breeds

  • Nikitkina, Elena V.;Dementieva, Natalia V.;Shcherbakov, Yuri S.;Atroshchenko, Mikhail M.;Kudinov, Andrei A.;Samoylov, Oleg I.;Pozovnikova, Marina V.;Dysin, Artem P.;Krutikova, Anna A.;Musidray, Artem A.;Mitrofanova, Olga V.;Plemyashov, Kirill V.;Griffin, Darren K.;Romanov, Michael N.
    • Animal Bioscience
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    • v.35 no.12
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    • pp.1827-1838
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    • 2022
  • Objective: The semen quality of stallions including sperm motility is an important target of selection as it has a high level of individual variability. However, effects of the molecular architecture of the genome on the mechanisms of sperm formation and their preservation after thawing have been poorly investigated. Here, we conducted a genome-wide association study (GWAS) for the sperm motility of cryopreserved semen in stallions of various breeds. Methods: Semen samples were collected from the stallions of 23 horse breeds. The following semen characteristics were examined: progressive motility (PM), progressive motility after freezing (FPM), and the difference between PM and FPM. The respective DNA samples from these stallions were genotyped using Axiom Equine Genotyping Array. Results: We performed a GWAS search for single nucleotide polymorphism (SNP) markers and potential genes related to motility properties of frozen-thawed semen in the stallions of various breeds. As a result of the GWAS analysis, two SNP markers, rs1141327473 and rs1149048772, were identified that were associated with preservation of the frozen-thawed stallion sperm motility, the relevant putative candidate genes being NME/NM23 family member 8 (NME8), olfactory receptor family 2 subfamily AP member 1 (OR2AP1), and olfactory receptor family 6 subfamily C member 4 (OR6C4). Potential implications of effects of these genes on sperm motility are herein discussed. Conclusion: The GWAS results enabled us to localize novel SNPs and candidate genes for sperm motility in stallions. Implications of the study for horse breeding and genetics are a better understanding of genomic regions and candidate genes underlying stallion sperm quality, and improvement in horse reproduction and breeding techniques. The identified markers and genes for sperm cryotolerance and the respective genomic regions are promising candidates for further studying the biological processes in the formation and function of the stallion reproductive system.

Stillbirth rates and their association with swine leucocyte antigen class II haplotypes in Microminipigs

  • Imaeda, Noriaki;Ando, Asako;Matsubara, Tatsuya;Takasu, Masaki;Nishii, Naohito;Miyamoto, Asuka;Ohshima, Shino;Kametani, Yoshie;Suzuki, Shingo;Shiina, Takashi;Ono, Tetsushi;Kulski, Jerzy K.;Kitagawa, Hitoshi
    • Animal Bioscience
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    • v.34 no.11
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    • pp.1749-1756
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    • 2021
  • Objective: Microminipig (MMP) is a miniature pig with an extra small body size for experimental use. In the present study, the occurrence of stillbirths and their genetic association with swine leukocyte antigen (SLA) class II haplotypes were evaluated in a population of MMPs. Methods: The occurrences of stillbirth and genetic association with SLA class II haplotypes using 483 stillborn and 2,246 live piglets, and their parents were compared among the three groups of newborn piglet litters; an all stillborn (AS) group consisting of only stillborn piglet litters, a partial stillborn (PS) group consisting of stillborn and live piglet litters, and an all alive (AA) group consisting of only live piglet litters. Results: The incidence of stillborn piglets was 483/2,729 (17.7%). Distributions of litter sizes, numbers of stillborn piglets in a litter, parities, and gestation periods were distinct among the three groups. The frequencies of low resolution haplotype (Lr)-0.7 or Lr-0.23 were higher in the AS group than in the PS or AA groups. In sires, the frequency of Lr-0.7 associated with the AS group was significantly higher in the AS group than with the AA group. In dams, the frequency of Lr-0.23 was significantly higher in the AS group than in the PS or AA groups, whereas the frequency of Lr-0.7 was not significantly different. Conclusion: The incidence of stillborn piglets in MMPs appears to be higher than those in other pig breeds. Several traits related with stillbirths such as the number of stillborn piglets and parities of the AS group were different from those of the PS and AA groups. Specific SLA class II haplotypes were associated significantly with a high incidence of stillbirths and could be used as genetic markers to adopt breeding strategies to lower the rate of stillbirth in MMPs.

Analysis of Genetic Diversity of the Thai Swamp Buffalo (Bubalus bubalis) Using Cattle Microsatellite DNA Markers

  • Triwitayakorn, K.;Moolmuang, B.;Sraphet, S.;Panyim, S.;Na-Chiangmai, A.;Smith, Duncan R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.5
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    • pp.617-621
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    • 2006
  • Recently the numbers of the Thai swamp buffalo (Bubalus bubalis), a native species of Thailand, have been rapidly declining, leading to a requirement for conservation programs for this breed. Such studies of the genetic diversity of this species are essential for conservation decisions and to assist the rational implementation of breeding programs. In this study, the genetic diversity of 80 Thai swamp buffalo, randomly selected from seven different research stations of the Thai Department of Livestock Development, were studied using ten cattle microsatellite markers. Polymorphic PCR products were observed at all microsatellite loci, with percentages of polymorphic loci ranging from 80.00 to 100.00%. The population from Payao showed the lowest level of polymorphism. The mean number of alleles per locus was 4.7 with the highest number of alleles being eight (ETH152) and the lowest being three (HAUT27 and ILSTS030). The average unbiased heterozygosity for all seven populations was 0.61 and varied between 0.5314 (Samui) and 0.6798 (Surin). The genetic distance according to NEI's (1972) ranged from 0.0722 to 0.4427. The populations from Surin and Burirum are the closest populations, while populations from Samui and Payao are the most divergent. The information generated by this study will greatly aid in the establishment of effective breeding and conservation programs for the Thai swamp buffalo.

Knockdown of endogenous SKIP gene enhanced insulin-induced glycogen synthesis signaling in differentiating C2C12 myoblasts

  • Xiong, Qi;Deng, Chang-Yan;Chai, Jin;Jiang, Si-Wen;Xiong, Yuan-Zhu;Li, Feng-E;Zheng, Rong
    • BMB Reports
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    • v.42 no.2
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    • pp.119-124
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    • 2009
  • PI(3,4,5)$P_3$ produced by the activated PI3-kinase is a key lipid second messenger in cell signaling downstream of insulin. Skeletal muscle and kidney-enriched inositol phosphatase (SKIP) identified as a 5'-inositol phosphatase that hydrolyzes PI(3,4,5) $P_3$ to PI(3,4)$P_2$, negatively regulates the insulin-induced glycogen synthesis in skeletal muscle. However the mechanism by which this occurs remains unclear. To elucidate the function of SKIP in glycogen synthesis, we employed RNAi techniques to knockdown the SKIP gene in differentiating C2C12 myoblasts. Insulininduced phosphorylation of Akt (protein kinase B) and GSK-3$\beta$ (Glycogen synthase kinase), subsequent dephosphorylation of glycogen synthase and glycogen synthesis were increased by inhibiting the expression of SKIP, whereas the insulin-induced glycogen synthesis was decreased by overexpression of WT-SKIP. Our results suggest that SKIP plays a negative regulatory role in Akt/ GSK-3$\beta$/GS (glycogen synthase) pathway leading to glycogen synthesis in myocytes.