• Title/Summary/Keyword: Breeding resource

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High-Throughput Development of Polymorphic Simple Sequence Repeat Markers Using Two Whole Genome Sequence Data in Peucedanum japonicum

  • Lee, Junki;Joh, Ho Jun;Kim, Nam-Hoon;Lee, Sang-Choon;Jang, Woojong;Choi, Beom Soon;Yu, Yeisoo;Yang, Tae-Jin
    • Plant Breeding and Biotechnology
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    • v.5 no.2
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    • pp.134-142
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    • 2017
  • Resource plants are important and have strong potential for a variety of utilities as crops or pharmaceutical materials. However, most resource plants remain wild and thus their utility for breeding and biotechnology is limited. Molecular markers are useful to initiate genetic study and molecular breeding for these understudied resource plants. We collected various wild collections of Peucedanum japonicum which is indigenous resource plants utilized as oriental medicine and leafy vegetables in Korea. In this study, we produced two independent whole genome sequences (WGSs) from two collections and identified large scale polymorphic simple sequence repeat (pSSR) based on our pipeline to develop SSR markers based on comparison of two WGSs. We identified a total of 452 candidate pSSR contigs. To confirm the accuracy and utility of pSSR, we designed ten SSR primer pairs and successfully applied those to seven collections of P. japonicum. The WGS and pSSR candidates identified in this study will be useful resource for genetic research and breeding purpose for the valuable resource plant, P. japonicum.

Relationship among porcine lncRNA TCONS_00010987, miR-323, and leptin receptor based on dual luciferase reporter gene assays and expression patterns

  • Ding, Yueyun;Qian, Li;Wang, Li;Wu, Chaodong;Li, DengTao;Zhang, Xiaodong;Yin, Zongjun;Wang, Yuanlang;Zhang, Wei;Wu, Xudong;Ding, Jian;Yang, Min;Zhang, Liang;Shang, Jinnan;Wang, Chonglong;Gao, Yafei
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.219-229
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    • 2020
  • Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORTTM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

Relationship between porcine miR-20a and its putative target low-density lipoprotein receptor based on dual luciferase reporter gene assays

  • Ding, Yueyun;Zhu, Shujiao;Wu, Chaodong;Qian, Li;Li, DengTao;Wang, Li;Wan, Yuanlang;Zhang, Wei;Yang, Min;Ding, Jian;Wu, Xudong;Zhang, Xiaodong;Gao, Yafei;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.922-929
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    • 2019
  • Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

Breeding Resource Materials of Silkworm Bombyx mori L., Adaptive to Tropical Climates

  • P. Sudhakara Rao;R. K. Datta;K. M, Vijaya-Kumari;M. Ramesg Babu
    • International Journal of Industrial Entomology and Biomaterials
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    • v.4 no.2
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    • pp.109-115
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    • 2002
  • With the objective of selecting suitable breeding resource material, 10 polyvoltine and 10 bivoltine breeds were drawn from the germplasm collection of Central Sericultural Research and Training Institute, Mysore, and evaluated for 3 seasons comprising one year (6 trials). Data were collected on seven traits of economic importance such as fecundity, pupation rate, cocoon yield, cocoon weight, cocoon shell weight, cocoon shell ratio and filament length, and statistically analysed with two-way classification, Joint scoring method and evaluation index. Significant seasonal variations (P < 0.01) were observed in both polyvoltine and bivoltine breeds. Polyvoltines BL27, BL36 and BL54 and bivoltines CSR2, CSR4 and Daizo scored highest ranking values in all the three methods. Hybridization was initiated based on larval markings and cocoon shapes. Seasonal variations were discussed.

Reference Gene Screening for Analyzing Gene Expression Across Goat Tissue

  • Zhanga, Yu;Zhang, Xiao-Dong;Liu, Xing;Li, Yun-Sheng;Ding, Jian-Ping;Zhang, Xiao-Rong;Zhang, Yun-Hai
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.12
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    • pp.1665-1671
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    • 2013
  • Real-time quantitative PCR (qRT-PCR) is one of the important methods for investigating the changes in mRNA expression levels in cells and tissues. Selection of the proper reference genes is very important when calibrating the results of real-time quantitative PCR. Studies on the selection of reference genes in goat tissues are limited, despite the economic importance of their meat and dairy products. We used real-time quantitative PCR to detect the expression levels of eight reference gene candidates (18S, TBP, HMBS, YWHAZ, ACTB, HPRT1, GAPDH and EEF1A2) in ten tissues types sourced from Boer goats. The optimal reference gene combination was selected according to the results determined by geNorm, NormFinder and Bestkeeper software packages. The analyses showed that tissue is an important variability factor in genes expression stability. When all tissues were considered, 18S, TBP and HMBS is the optimal reference combination for calibrating quantitative PCR analysis of gene expression from goat tissues. Dividing data set by tissues, ACTB was the most stable in stomach, small intestine and ovary, 18S in heart and spleen, HMBS in uterus and lung, TBP in liver, HPRT1 in kidney and GAPDH in muscle. Overall, this study provided valuable information about the goat reference genes that can be used in order to perform a proper normalisation when relative quantification by qRT-PCR studies is undertaken.

Investigation of the Proper Density for Rearing Protaetia brevitarsis Larvae (흰점박이꽃무지(Protaetia brevitarsis)의 유충사육 시 적정밀도 규명)

  • Choi, Sung-Up;Choi, In-Hag;Son, Jin-Sung;Chung, Tae-Ho
    • Journal of Environmental Science International
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    • v.30 no.2
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    • pp.191-194
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    • 2021
  • This study aimed to provide information and data for the management of insect breeding farms by identifying the appropriate density when rearing Protaetia brevitarsis larvae. The breeding box of the insects was filled with 2 L of fermented sawdust on a 50 × 35 × 15 cm sheet of plastic, and the density of the treatments was 200, 300 and 400 P. brevitarsis in the first, second, and third larval stages. Each treatment was repeated five times, and the sawdust medium was replaced three times (10 intervals). The experiments were conducted for a total of 30 days from March 1, 2020. Overall, 200 P.brevitarsis in the first, second, and third larval stages reared in the breeding box had a higher average survival rate and lower average mortality rate, thereby contributing to efficient production. In conclusion, this result suggests a way to increase production efficiency through the environmental management of insect breeding farms.

Chicken Breeding with Local Breeds in China - A Review

  • Jiang, X.;Groen, A.F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.10
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    • pp.1482-1498
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    • 2000
  • This paper reviews the meat-type chicken breeding with local breeds in China. The quality chickens are defined as purebred final products of local breeds, and semi-quality chickens as crossbreds of local breeds with specialized broiler (sire or dam) lines from western breeding organizations. The present status of the chicken production and the market in China, in comparison with the western countries, is reviewed, indicating that there is large market demand for (semi-) quality chickens in the present and future China. Breeding for (semi-) quality chickens emphasizes the sensory quality of chicken meat. The present status of breeding for (semi-) quality chickens with the local breeds is illustrated, including breeding goals and the existing breeding programs. The potential role of local breeds in breeding programs in China is discussed in relation to both providing higher quality (than commercial hybrid broilers) of chicken meat for the local market and meeting the objectives of genetic resource conservation. Besides, further research topics on breeding for (semi-) quality chickens are suggested.