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Global population will increase to over nine billion by 2050 with the doubling in demand for meat and milk. To overcome this challenge, it is necessary to breed highly efficient and productive livestock. Furthermore, livestock are also excellent models for human diseases and ideal bioreactors to produce pharmaceutical proteins. Thus, genetic engineering of domestic animals presents a critical and valuable tool to address these agricultural and biomedical applications. Overall, genetic engineering has evolved through three stages in history: transgenesis, gene targeting, and gene editing. Since the birth of the first transgenic pig, genetic engineering in livestock has been advancing slowly due to inherent technical limitations. A major breakthrough has been the advent of somatic cell nuclear transfer, which, for the first time, provided the technical ability to produce site-specific genome-modified domestic animals. However, the low efficiency of gene targeting events in somatic cells prohibits its wide use in agricultural and biomedical applications. Recently, rapid progress in tools and methods of genome engineering has been made, allowing genetic editing from mutation of a single base pair to the deletion of entire chromosomes. Here, we review the major advances of genetic engineering in domestic animals with emphasis placed on the introduction of latest designer nucleases.
The H9N2 and H5N1 avian influenza viruses (AIVs) have been circulating in poultry in China and become endemic since 1998 and 2004, respectively. Currently, they are prevalent in poultry throughout China. This endemicity makes them actively involved in the emergence of the novel lineages of other subtypes of influenza viruses, such as the well-known viruses of the highly pathogenic avian influenza (HPAI) H5N2 and the 2013 novel H7N7, H7N9 and H10N8 subtypes, thereby threatening both the poultry industry and public health. Here, we will review briefly the prevalence and evolution, pathogenicity, transmission, and disease control of these two subtypes and also discuss the possibility of emergence of potentially virulent and highly transmissible AIVs to humans.
Reducing irrigation water use by improving water use efficiency (WUE) in grain production is critical for the development of sustainable agriculture in the North China Plain (NCP). This article summarizes the research progresses in WUE improvement carried out at the Luancheng station located in the Northern part of NCP for the past three decades. Progresses in four aspects of yield and WUE improvement are presented, including yield and WUE improvement associated with cultivar selection, irrigation management for improving yield and WUE under limited water supply, managing root system for efficient soil water use and reducing soil evaporation by straw mulch. The results showed that annual average increase of 0.014 kg·m-3 for winter wheat and 0.02 kg·m-3 in WUE were observed for the past three decades, and this increase was largely associated with the improvement in harvest index related to cultivar renewal and an increase in chemical fertilizer use and soil fertility. The results also indicated that deficit irrigation for winter wheat could significantly reduce the irrigation water use, whereas the seasonal yield showed a smaller reduction rate and WUE was significantly improved. Straw mulching of summer maize using the straw from winter wheat could reduce seasonal soil evaporation by 30–40 mm. With new cultivars and improved management practices it was possible to further increase grain production without much increase in water use. Future strategies to further improve WUE are also discussed.
Crop planting structure optimization is a significant way to increase agricultural economic benefits and improve agricultural water management. The complexities of fluctuating stream conditions, varying economic profits, and uncertainties and errors in estimated modeling parameters, as well as the complexities among economic, social, natural resources and environmental aspects, have led to the necessity of developing optimization models for crop planting structure which consider uncertainty and multi-objectives elements. In this study, three single-objective programming models under uncertainty for crop planting structure optimization were developed, including an interval linear programming model, an inexact fuzzy chance-constrained programming (IFCCP) model and an inexact fuzzy linear programming (IFLP) model. Each of the three models takes grayness into account. Moreover, the IFCCP model considers fuzzy uncertainty of parameters/variables and stochastic characteristics of constraints, while the IFLP model takes into account the fuzzy uncertainty of both constraints and objective functions. To satisfy the sustainable development of crop planting structure planning, a fuzzy-optimization-theory-based fuzzy linear multi-objective programming model was developed, which is capable of reflecting both uncertainties and multi-objective. In addition, a multi-objective fractional programming model for crop structure optimization was also developed to quantitatively express the multi-objective in one optimization model with the numerator representing maximum economic benefits and the denominator representing minimum crop planting area allocation. These models better reflect actual situations, considering the uncertainties and multi-objectives of crop planting structure optimization systems. The five models developed were then applied to a real case study in Minqin County, north-west China. The advantages, the applicable conditions and the solution methods of each model are expounded. Detailed analysis of results of each model and their comparisons demonstrate the feasibility and applicability of the models developed, therefore decision makers can choose the appropriate model when making decisions.
Dehydrated castor oil was epoxidized using phosphoric acid as a catalyst and acetic acid peroxide as an oxidant to produce epoxidized castor oil (ECO). Ring-opening polymerization with stannic chloride was used to produce polymerized ECO (PECO), and sodium hydroxide used to give hydrolyzed PECO (HPECO). The HPECO was characterized by Fourier transform infrared, 1H and 13C nuclear magnetic resonance spectroscopies, gel permeation chromatography, and differential scanning calorimetry. The weight-average molecular weight of soluble PECO and HPECO were 5026 and 2274 g·mol-1, respectively. PECO and HPECO exhibited glass transition. Through neutralizing the carboxylic acid of HPECO with different counterions, castor oil-based polymeric surfactants (HPECO-M, where M= Na+, K+ or triethanolamine ion) exhibited high efficiency to reduce the surface tension of water. The critical micelle concentration (CMC) values of HPECO-M ranged from 0.042 to 0.098 g·L-1 and the minimum equilibrium surface tensions at CMC (gcmc) of HPECO-M ranged from 25.6 to 30.0 mN·m-1. The water-hexadecane interfacial energy was calculated from measured surface tension using harmonic and geometric mean methods. Measured values of water-hexadecane interfacial tension agreed well with those calculated using the harmonic and geometric mean methods.
Scientific irrigation and nitrogen management is important for agricultural production in arid areas. To quantify the effect of water and nitrogen management on yield components, biomass partitioning and harvest index (HI) of maize for seed production with plastic film-mulching, field experiments including different irrigation and N treatments were conducted in arid north-west China in 2013 and 2014. The results indicated that kernel number per plant (KN) was significantly affected by irrigation and N treatments. However, 100-kernel weight was relatively stable. Reducing irrigation quantity significantly increased stem partitioning index (PIstem) and leaf partitioning index (PIleaf), and decreased ear partitioning index (PIear) at harvest, but lowering N rate (from 500 to 100 kg N·hm-2) did not significantly reduce PIstem, PIleaf, and PIear at harvest. HI was significantly reduced by reducing irrigation quantity, but not by reducing N rate. Linear relationships were found between KN, PIstem, PIleaf, PIear at harvest and HI and evapotranspiration (ET).
Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failures in sows and respiratory diseases in pigs of all ages. PRRS virus (PRRSV) is its causative agent and has caused huge economic losses in the swine industry. Porcine sialoadhesin (pSn) is a putative receptor of PRRSV. Previous studies have shown that a pSn V-set Ig-like domain is significant in PRRSV infection. However, its structural details are not fully known, hindering our deep understanding of PRRSV infection. In this study, we successfully cloned, expressed and purified the pSn V-set Ig-like domain in Drosophila S2 cells. Then we tried to crystallize the target protein and predicted its structure. This will establish the foundation for the further structural study of pSn, deepen our understanding of the invasion mechanism of PRRSV, and support the structural information for the development of clinical drugs and vaccines against PRRSV.
MicroRNAs (miRNAs), small non-coding RNAs, are involved in many aspects of biological processes. Previous studies have indicated that miRNAs are important for hair follicle development and growth. In our study, we found by qRT-PCR that miR-148b was significantly upregulated in sheep wool follicle bulbs in anagen phase compared with the telogen phase of the hair follicle cycle. Overexpression of miR-148b promoted proliferation of both HHDPC and HHGMC. By using the TOPFlash system we demonstrated that miR-148b could activate Wnt/β-catenin pathway and b-catenin, cycD, c-jun and PPARD were consistently upregulated accordingly. Furthermore, transcript factor nuclear factor of activated T cells type 5 (NFAT5) and Wnt10b were predicted to be the target of miR-148b and this was substantiated using a Dual-Luciferase reporter system. Subsequently NFAT5 was further identified as the target of miR-148b using western blotting. These results were considered to indicate that miR-148b could activate the Wnt/β-catenin signal pathway by targeting NFAT5 to promote the proliferation of human hair follicle cells.
As virulence-determining genes, RR1 and RR2 encode the small subunit and large subunit of viral ribonucleotide reductase (RR) in pseudorabies virus which have been extensively studied in mice. However, their role in pigs has not been adequately investigated. In this study, we deleted RR1 and RR2 genes based on a TK/gE/gI triple gene-deleted pseudorabies virus and tested its efficacy in pigs as a vaccine candidate. The rescued virus showed similar growth properties and plaque size in vitro as its parent strain. In an animal study, the virus could elicit humoral immune responses shown by generation of gB-specific antibodies and virus neutralizing antibodies. However, vaccination could not provide protection against virulent pseudorabies virus challenge since vaccinated pigs showed clinical pseudorabies-specific syndromes. The deficiency in protection may due to the generation of late and low levels of gB antibodies and virus neutralizing antibodies.
In the present study, follistatin (FST) gene expression vectors with either a bicistronic gene transfer cassette alone, or a bicistron gene cassette carrying a matrix attachment region (MAR) were constructed and transfected to bovine fetal fibroblasts. Evaluations of both the integration and expression of exogenous FST indicated that the pMAR-CAG-FST-IRES-AcGFP1-polyA-MAR (pMAR-FST) vector had higher capacity to form monoclonal transgenic cells than the vector without MAR, though transient transfection and integration efficiency were similar with either construct. Remarkably, protein expression in transgenic cells with the pMAR-FST vector was significantly higher than that from the bicistronic vector. Exogenous FST was expressed in all of the pMAR-FST transgenic mice at F0, F1 and F2. Total muscle growth in F0 mice was significantly greater than in wild-type mice, with larger muscles in fore and hind limbs of transgenic mice. pMAR-FST transgenic mice were also found with more evenly distributed muscle bundles and thinner spaces between sarcolemma, which suggests a correlation between transgene expression-associated muscle development and the trend of muscle growth. In conclusion, a pMAR-FST vector, which excluded the resistant genes and frame structure, enhances and stabilizes FST gene expressions in both transfected cells and transgenic mice.