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Huang L.,Sichuan Agricultural University | Yan H.,Sichuan Agricultural University | Jiang X.,Sichuan Agricultural University | Yin G.,University of Arizona | And 6 more authors.
PLoS ONE | Year: 2014

Background: Quantitative real-time reverse-transcriptase PCR (qRT-PCR) is an important technique for analyzing differences in gene expression due to its sensitivity, accuracy and specificity. However, the stability of the expression of reference genes is necessary to ensure accurate qRT-PCR assessment of expression in genes of interest. Perennial ryegrass (Lolium perenne L.) is important forage and turf grass species in temperate regions, but the expression stability of its reference genes under various stresses has not been well-studied. Methodology/Principal Findings: In this study, 11 candidate reference genes were evaluated for use as controls in qRT-PCR to quantify gene expression in perennial ryegrass under drought, high salinity, heat, waterlogging, and ABA (abscisic acid) treatments. Four approaches - Delta C T, geNorm, BestKeeper and Normfinder were used to determine the stability of expression in these reference genes. The results are consistent with the idea that the best reference genes depend on the stress treatment under investigation. Eukaryotic initiation factor 4 alpha (eIF4A), Transcription elongation factor 1 (TEF1) and Tat binding protein-1 (TBP-1) were the three most stably expressed genes under drought stress and were also the three best genes for studying salt stress. eIF4A, TBP-1, and Ubiquitin-conjugating enzyme (E2) were the most suitable reference genes to study heat stress, while eIF4A, TEF1, and E2 were the three best reference genes for studying the effects of ABA. Finally, Ubiquitin (UBQ), TEF1, and eIF4A were the three best reference genes for waterlogging treatments. Conclusions/Significance: These results will be helpful in choosing the best reference genes for use in studies related to various abiotic stresses in perennial ryegrass. The stability of expression in these reference genes will enable better normalization and quantification of the transcript levels for studies of gene expression in such studies. © 2014 Huang et al.


Li M.,Sichuan Agricultural University | Li M.,Tsinghua University | Tian S.,Novogene Bioinformatics Institute | Jin L.,Sichuan Agricultural University | And 49 more authors.
Nature Genetics | Year: 2013

We report the sequencing at 131× coverage, de novo assembly and analyses of the genome of a female Tibetan wild boar. We also resequenced the whole genomes of 30 Tibetan wild boars from six major distributed locations and 18 geographically related pigs in China. We characterized genetic diversity, population structure and patterns of evolution. We searched for genomic regions under selection, which includes genes that are involved in hypoxia, olfaction, energy metabolism and drug response. Comparing the genome of Tibetan wild boar with those of neighboring Chinese domestic pigs further showed the impact of thousands of years of artificial selection and different signatures of selection in wild boar and domestic pig. We also report genetic adaptations in Tibetan wild boar that are associated with high altitudes and characterize the genetic basis of increased salivation in domestic pig.


PubMed | National Animal Husbandry Service, CAS Institute of Genetics and Developmental Biology, Chinese Academy of Agricultural Sciences and China Agricultural University
Type: | Journal: Scientific reports | Year: 2017

P-HYDROXYPHENYLPYRUVATE DIOXYGENASE (HPPD) is the first committed enzyme involved in the biosynthesis of vitamin E, and is characterized by catalyzing the conversion of p-hydroxyphenyl pyruvate (HPP) to homogentisic acid (HGA). Here, an HPPD gene was cloned from Medicago sativa L. and designated MsHPPD, which was expressed at high levels in alfalfa leaves. PEG 6000 (polyethylene glycol), NaCl, abscisic acid and salicylic acid were shown to significantly induce MsHPPD expression, especially in the cotyledons and root tissues. Overexpression of MsHPPD was found to significantly increase the level of -tocotrienol and the total vitamin E content in Arabidopsis seeds. Furthermore, these transgenic Arabidopsis seeds exhibited an accelerated germination time, compared with wild-type seeds under normal conditions, as well as under NaCl and ABA treatments. Meanwhile, the expression level of several genes associated with ABA biosynthesis (NCED3, NCED5 and NCED9) and the ABA signaling pathway (RAB18, ABI3 and ABI5) were significantly down-regulated in MsHPPD-overexpressing transgenic lines, as well as the total free ABA content. Taken together, these results demonstrate that MsHPPD functions not only in the vitamin E biosynthetic pathway, but also plays a critical role in seed germination via affecting ABA biosynthesis and signaling.


Zhao N.,Northeast Agricultural University | Xu X.-X.,Northeast Agricultural University | Zhang Y.-J.,Shanghaichenguan Dairy Co. | Zhang S.-Y.,National Animal Husbandry Service
Modern Food Science and Technology | Year: 2015

The effects of human lactoferricin (LfcinH) and bovine lactoferricin (LfcinB) on the proliferation and cytokine secretion of spleen lymphocytes in mice were studied, and the differences and correlation between LfcinH and LfcinB were compared and analyzed, respectively. The cell counting kit-8 (CCK-8) assay was used to measure the effects of different concentrations of LfcinH or LfcinB alone on the proliferation of mouse spleen lymphocytes and of different concentrations of LfcinH or LfcinB combined with a mitogen on the proliferation of mouse T and B lymphocytes. The IL-2, IL-4, TNF-α content was determined by enzyme-linked immunosorbent assay (ELISA). The results showed that LfcinH and LfcinB could significantly promote the proliferation of spleen lymphocytes and enhance the T lymphocyte proliferation induced by concanavalin A (ConA) and B lymphocyte proliferation induced by lipopolysaccharide (LPS). Moreover, Lfcin could significantly increase the secretions of IL-2, IL-4, TNF-α in spleen lymphocytes. The results also showed that Lfcin has a synergistic effect on the proliferation of T and B spleen lymphocytes with Con A or LPS; the mechanism may be related to the immune enhancement achieved by promoting cytokine secretion. © 2015, South China University of Technology. All right reserved.


Zhi-Hong C.,Chinese Academy of Agricultural Sciences | Zan W.,Chinese Academy of Agricultural Sciences | Xu-Jiang Y.,National Animal Husbandry Service | Xiang-Fang L.,National Animal Husbandry Service | Hong-Wen G.,Chinese Academy of Agricultural Sciences
African Journal of Biotechnology | Year: 2011

The silica gel drying method was utilized to reduce the moisture content of Elymus dahuricus seeds from an original content of 9.03 to 7.69, 6.10, 4.97, 4.47, 3.83, 2.58 and 1.26%. After sealing the seeds in aluminum foil bags, they were placed at -4°C, 4°C, room temperature and 45°C to store for 12 months and to identify their physiological and biochemical indicators. Results indicate that ultra-dry storage can: 1) Improve the storability and membrane permeability of E. dahuricus seeds, 2) increase the activity of E. dahuricus SOD, POD and CAT, and 3) lower MDA content. Moderate ultra-dry storage at room temperature can reach the conserved effect of low-temperature without intense ultra-drying. Therefore, the moderate ultra-dry can be use as effective measure for the conservation of E. dahuricus germplasm resources.


Lin F.,Tianjin Institute of Animal science and Veterinary Medicine | Lin F.,Henan Agricultural University | Huang C.-J.,Henan Agricultural University | Liu C.-S.,National Animal Husbandry Service | And 3 more authors.
Reproduction in Domestic Animals | Year: 2016

This study detected the distribution of laminin during embryonic formation by immunofluorescence. To determine the possible function of laminin on developmental ability of in vitro fertilized embryos, the presumptive zygotes were divided and transferred to CR1aa medium supplemented with different concentrations (0 μg/ml, 5 μg/ml, 10 μg/ml and 20 μg/ml) of laminin. To explore the association with sperm–oocyte fusion, oocytes and/or sperm were pre-incubated with laminin or anti-β1 antibody before insemination. Laminin was absent in mature oocytes and could be detected first at the 8-cell stage and then displayed an increasing tendency. Adding 10 μg/ml laminin to the culture medium improved embryonic development including cleavage rate, blastocyst rate, total cell numbers in the blastocyst and cell numbers in the inner cell mass. Laminin inhibited sperm–oocyte fusion when incubated with oocytes and/or sperm before in vitro fertilization, and only integrin-β1 of sperm was involved in sperm–oocyte binding. Inhibition may be caused by blocking β1, but why laminin inhibits fertilization is still unknown. The results suggest that laminin plays an important role during embryonic formation and has a negative function in sperm–oocyte fusion, but improves embryonic development. However, only integrin-β1 is involved in sperm–oocyte binding. © 2016 Blackwell Verlag GmbH


Qiang H.,Chinese Academy of Sciences | Chen Z.,National Animal Husbandry Service | Zhang Z.,Chinese Academy of Sciences | Wang X.,Chinese Academy of Sciences | And 2 more authors.
PLoS ONE | Year: 2015

Information on genetic diversity and population structure of a tetraploid alfalfa collection might be valuable in effective use of the genetic resources. A set of 336 worldwide genotypes of tetraploid alfalfa (Medicago sativa subsp. sativa L.) was genotyped using 85 genome-wide distributed SSR markers to reveal the genetic diversity and population structure in the alfalfa. Genetic diversity analysis identified a total of 1056 alleles across 85 marker loci. The average expected heterozygosity and polymorphism information content values were 0.677 and 0.638, respectively, showing high levels of genetic diversity in the cultivated tetraploid alfalfa germplasm. Comparison of genetic characteristics across chromosomes indicated regions of chromosomes 2 and 3 had the highest genetic diversity. A higher genetic diversity was detected in alfalfa landraces than that of wild materials and cultivars. Two populations were identified by the model-based population structure, principal coordinate and neighbor-joining analyses, corresponding to China and other parts of the world. However, lack of strictly correlation between clustering and geographic origins suggested extensive germplasm exchanges of alfalfa germplasm across diverse geographic regions. The quantitative analysis of the genetic diversity and population structure in this study could be useful for genetic and genomic analysis and utilization of the genetic variation in alfalfa breeding. © 2015 Qiang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


PubMed | National Animal Husbandry Service, Henan Agricultural University and Tianjin Institute of Animal science and Veterinary Medicine
Type: Journal Article | Journal: Reproduction in domestic animals = Zuchthygiene | Year: 2016

This study detected the distribution of laminin during embryonic formation by immunofluorescence. To determine the possible function of laminin on developmental ability of invitro fertilized embryos, the presumptive zygotes were divided and transferred to CR1aa medium supplemented with different concentrations (0g/ml, 5g/ml, 10g/ml and 20g/ml) of laminin. To explore the association with sperm-oocyte fusion, oocytes and/or sperm were pre-incubated with laminin or anti-1 antibody before insemination. Laminin was absent in mature oocytes and could be detected first at the 8-cell stage and then displayed an increasing tendency. Adding 10g/ml laminin to the culture medium improved embryonic development including cleavage rate, blastocyst rate, total cell numbers in the blastocyst and cell numbers in the inner cell mass. Laminin inhibited sperm-oocyte fusion when incubated with oocytes and/or sperm before invitro fertilization, and only integrin-1 of sperm was involved in sperm-oocyte binding. Inhibition may be caused by blocking 1, but why laminin inhibits fertilization is still unknown. The results suggest that laminin plays an important role during embryonic formation and has a negative function in sperm-oocyte fusion, but improves embryonic development. However, only integrin-1 is involved in sperm-oocyte binding.


PubMed | Beijing Forestry University and National Animal Husbandry Service
Type: | Journal: Scientific reports | Year: 2015

Published data have shown inconsistent results about the pharmacogenetics of XRCC1 gene on clinical outcomes of advanced lung cancer patients treated with platinum-based chemotherapy. This meta-analysis aimed to summarize published findings and provide more reliable association. A total of 53 eligible studies including 7433 patients were included. Patients bearing the favorable TrpTrp and TrpArg genotypes of Arg194Trp were more likely to better response rates to platinum-based chemotherapy compared to those with the unfavorable ArgArg genotype (TrpTrp+TrpArg vs. ArgArg: odds ratio (OR)=2.02, 95% CI, 1.66-2.45). The GlnGln and GlnArg genotypes of Arg399Gln were significantly associated with the poorer response rates compared to those with the ArgArg genotype (GlnGln +GlnArg vs. ArgArg: OR=0.68, 95% CI, 0.54-0.86). The GlnGln genotype might be more closely associated with shorter survival time and higher risks of death for patients (GlnGln vs. ArgArg: hazard ratio (HR)=1.14, 95% CI, 0.75-1.75). Our cumulative meta-analyses indicated a distinct apparent trend toward a better response rate for Arg194Trp, but a poorer response rate in Arg399Gln. These findings indicate a predictive role of XRCC1 polymorphisms in clinical outcomes. The use of XRCC1 polymorphisms as predictive factor of clinical outcomes in personalized chemotherapy treatment requires further verification from large well-designed pharmacogenetics studies.


PubMed | Chinese Academy of Sciences and National Animal Husbandry Service
Type: Journal Article | Journal: PloS one | Year: 2015

Information on genetic diversity and population structure of a tetraploid alfalfa collection might be valuable in effective use of the genetic resources. A set of 336 worldwide genotypes of tetraploid alfalfa (Medicago sativa subsp. sativa L.) was genotyped using 85 genome-wide distributed SSR markers to reveal the genetic diversity and population structure in the alfalfa. Genetic diversity analysis identified a total of 1056 alleles across 85 marker loci. The average expected heterozygosity and polymorphism information content values were 0.677 and 0.638, respectively, showing high levels of genetic diversity in the cultivated tetraploid alfalfa germplasm. Comparison of genetic characteristics across chromosomes indicated regions of chromosomes 2 and 3 had the highest genetic diversity. A higher genetic diversity was detected in alfalfa landraces than that of wild materials and cultivars. Two populations were identified by the model-based population structure, principal coordinate and neighbor-joining analyses, corresponding to China and other parts of the world. However, lack of strictly correlation between clustering and geographic origins suggested extensive germplasm exchanges of alfalfa germplasm across diverse geographic regions. The quantitative analysis of the genetic diversity and population structure in this study could be useful for genetic and genomic analysis and utilization of the genetic variation in alfalfa breeding.

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