Eureka lemon zinc finger protein ClDOF3.4 interacts with citrus yellow vein clearing virus coat protein to inhibit viral infection

Citrus yellow vein clearing virus(CYVCV)is a new citrus virus that has become an important factor restricting the development of China’s citrus industry,and the CYVCV coat protein(CP)is associated with viral pathogenicity.In this study,the Eureka lemon zinc finger protein(ZFP)ClDOF3.4 was shown to interact with CYVCV CP in vivo and in vitro.Transient expression of ClDOF3.4 in Eureka lemon induced the expression of salicylic acid(SA)-related and hypersensitive response marker genes,and triggered a reactive oxygen species burst,ion leakage necrosis,and the accumulation of free SA.Furthermore,the CYVCV titer in ClDOF3.4 transgenic Eureka lemon plants was approximately 69.4%that in control plants 6 mon after inoculation,with only mild leaf chlorotic spots observed in those transgenic plants.Taken together,the results indicate that ClDOF3.4 not only interacts with CP but also induces an immune response in Eureka lemon by inducing the SA pathways.This is the first report that ZFP is involved in the immune response of a citrus viral disease,which provides a basis for further study of the molecular mechanism of CYVCV infection.

Effects of different dietary energy and protein levels and sex on growth performance,carcass characteristics and meat quality of F1 Angus x Chinese Xiangxi yellow cattle

Background：The experiment evaluated the effect of nutrition levels and sex on the growth performance,carcass characteristics and meat quality of F1 Angus × Chinese Xiangxi yellow cattle.Methods：During the background period of 184 d,23 steers and 24 heifers were fed the same ration,then put into a2×2×2 factorial arrangement under two levels of- dietary energy（TON：70/80%DM）,protein（CP：11.9/14.3%DM）and sex（S：male/female） during the finishing phase of 146 d.The treatments were-（1） high energy/low protein（HELP）,（2） high energy/high protein（HEHP）,（3） low energy/low protein（LELP） and（4） low energy/high protein（LEHP）.Each treatment used 6 steers and 6 heifers,except for HELP- 5 steers and 6 heifers.Results：Growth rate and final carcass weight were unaffected by dietary energy and protein levels or by sex.Compared with the LE diet group,the HE group had significantly lower dry matter intake（DMI,6.76 vs.7.48 kg DM/d）,greater chest girth increments（46.1 vs.36.8 cm）,higher carcass fat（19.9 vs.16.3%） and intramuscular fat content（29.9 vs.22.8%DM）.The HE group also had improved yields of top and medium top grade commercial meat cuts（39.9 vs.36.5%）.The dressing percentage was higher for the HP group than the LP group（53.4 vs.54.9%）.Steers had a greater length increment（9.0 vs.8.3 cm）,but lower carcass fat content（16.8 vs.19.4%） than heifers.The meat quality traits（shear force value,drip loss,cooking loss and water holding capacity） were not affected by treatments or sex,averaging 3.14 kg,2.5,31.5 and 52.9%,respectively.The nutritive profiles（both fatty and amino acid composition） were not influenced by the energy or protein levels or by sex.Conclusions：The dietary energy and protein levels and sex significantly influenced the carcass characteristics and chemical composition of meat but not thegrowth performance,meat quality traits and nutritive profiles.

Effects of Different Dietary Energy and Protein Levels on Meat Performance and Meat Quality of Jinghai Yellow Chickens

To investigate the effect of different dietary energy and protein levels on meat performance and meat quality of Jinghai yellow chickens, 480 43-day old Jinghai yellow chickens with similar weight were randomly divided into four experimental groups： experimental group 1 （protein 15%, metabolic energy 9.95 MJ/kg）, experimental group 2 （protein 16%, metabolic energy 10.95 MJ/kg）, experimental group 3 （protein 17%, metabolic energy 12.65 MJ/kg） and experimental group 4 （ protein 18%, metabolic energy 13.95 MJ/kg）, respectively. All chickens were slaughtered at 112-day old. The breast and leg muscles of Jinghai yellow chickens were collected, to determine the slaughter performance, conventional meat quality and muscle chemical indicators. The results indicated that dressing-out percentage and eviscerated yield percentage in four experimental groups were above 87.27% and 67.00%, respectively; other slaughter performance indicators exhibited no significant differences among various groups （P 〉 0.05 ） ; breast muscle color of hens in experimental group 4 varied significantly from that in other three groups （ P 〈 0.05 ） ; leg muscle color of hens in experimental group 2 varied extremely significantly from that in other three groups （ P 〈 0.01 ） ; water-holding capacity of breast muscles of hens in experimental group 3 was significantly higher than that in experimental group 4 （P 〈 0.05 ） ; thiamine content of breast muscles of cocks in experimental group 3 was significandy higher than that in experimental group 2 （ P 〈 0.05 ） ; however, other properties exhibited no significant differenees among various groups （P 〉 0.05 ）.

Effects of fish protein hydrolysate on growth performance and humoral immune response in large yellow croaker (Pseudosciaena crocea R.)

We investigated the effects of fish protein hydrolysate (FPH) on growth performance and humoral immune response of the large yellow croaker (Pseudosciaena crocea R.). One thousand and two hundred large yellow croakers [initial average weight: (162.75±23.85) g] were divided into four groups and reared in floating sea cages (3 m×3 m×3 m). The animals were fed with 4 diets: basal diet only (control) or diets supplemented with 5%, 10% and 15% (w/w) FPH. The results show that dietary FPH levels significantly influenced the growth and immunity of the large yellow croaker. Compared with the control group, total weight gain (TWG) in all treatment groups, relative weight gain (RWG) and specific growth rate (SGR) in fish fed with diets supplemented with 10% and 15% FPH were significantly increased (P<0.05). Similar results were observed in immune parameters [lysozyme activity, serum complements, immunoglobulin M (IgM)]. Lysozyme activity, complement C4 and IgM were also significantly increased (P<0.05) in fish fed with diets supplemented with 10% and 15% FPH, while complement C3 level was significantly increased (P<0.05) in all treatment groups. In general, with the supplementation of FPH, particularly at dose of 10%, the growth performance and immunity of the large yellow croaker can be improved effectively.

Improving the nutritional values of yellow mealworm Tenebrio molitor(Coleoptera: Tenebrionidae) larvae as an animal feed ingredient: a review

Yellow mealworm larvae(YML;Tenebrio molitor) are considered as a valuable insect species for animal feed due to their high nutritional values and ability to grow under different substrates and rearing conditions. Advances in the understanding of entomophagy and animal nutrition over the past decades have propelled research areas toward testing multiple aspects of YML to exploit them better as animal feed sources. This review aims to summarize various approaches that could be exploited to maximize the nutritional values of YML as an animal feed ingredient. In addition, YML has the potential to be used as an antimicrobial or bioactive agent to improve animal health and immune function in production animals. The dynamics of the nutritional profile of YML can be influenced by multiple factors and should be taken into account when attempting to optimize the nutrient contents of YML as an animal feed ingredient. Specifically, the use of novel land-based and aquatic feeding resources, probiotics, and the exploitation of larval gut microbiomes as novel strategies can assist to maximize the nutritional potential of YML. Selection of relevant feed supplies, optimization of ambient conditions, the introduction of novel genetic selection procedures, and implementation of effective post-harvest processing may be required in the future to commercialize mealworm production. Furthermore, the use of appropriate agricultural practices and technological improvements within the mealworm production sector should be aimed at achieving both economic and environmental sustainability. The issues highlighted in this review could pave the way for future approaches to improve the nutritional value of YML.

Influence of Amino Acid Supplementations in Juvenile Yellow Perch Fed Plant Protein Combinations

Distillers dried grains with solubles (DDGS) in combination with soy protein concentrate (SPC) with and without an essential amino acid (EAA) complex were assessed as protein alternatives in juvenile Yellow Perch Perca flavescens diets. Diets contained 5% FM, 40% SPC, and 20% or 40% DDGS each with or without EAA. No mortalities or health assessment differences were observed during the trial and all fish readily accepted the experimental diets. Diets supplemented with EAA produced greater weight gain, improved feed conversion, and apparent protein digestibility. Performance was consistently improved for fish fed diets containing amino acid supplements. Based on these results, Yellow Perch are able to utilize high levels of the plant proteins, accompanied with EAAs, as a FM replacer.

Effects of Crude Protein Level of Concentrate Supplement on Production Performance and Serum Biochemical Parameters in Post-fattening Hainan Yellow Cattle

[Objective]The paper was to investigate the effects of crude protein level of concentrate supplement on production performance and serum biochemical parameters of post-fattening Hainan Yellow Cattle.[Method]Twelve Hainan Yellow Cattles with the same genetic backgrounds and similar initial weight of(168.13±0.55)kg were randomly divided into three treatments.Cattle were fed with concentrate supplement containing different levels of crude protein.The trial lasted 60 d.[Result]The daily gain of 17.56%crude protein group was significantly higher than those of 15.55%and 13.56%crude protein groups(P<0.05),and the gross profit of 17.56%crude protein group was markedly higher than that of 13.56%crude protein group(P<0.05),but the feed gain ratio of 17.56%crude protein group was significantly lower than that of 13.56%crude protein group(P<0.05).Compared with 15.55%and 13.56%crude protein groups,the serum total protein content markedly increased(P<0.05),but the urea nitrogen level significantly decreased(P<0.05).[Conclusion]When crude protein level of concentrate supplement was 17.56%,post-fattening Hainan Yellow Cattle could obtain better production performance.

Construction of the Enhanced Yellow Fluorescent Protein Expression Vector Carrying IFN-γ Gene

Purpose: To construct the enhanced yellow fluorescent protein (EYFP) vector carryinginterferon-y gene (ifn-γ) in order to provide an ideal reporter in the expression of ifn-γand location of protein in vitro and in vivo.Method: According to the nucleotide sequence of ifn-y gene, a pair of oligonucleotideswas designed as primer whose two end contained nucleotide sequence of EcoR V and NotⅠ restriction endonuclease respectively. The gene encoding for inf-y was amplified usingPCR technique. After the PCR product was retrieved and purified, it was digested withEcoR V and Not Ⅰ restriction endonuclease, and then cloned into the plasmidpIRES-EYFP. The recombinant plasmid plRES-EYFPIFN-γwas identified by restrictionendonuclease enzyme analysis and DNA sequence analysis.Results: The ifn-γ was successfully amplified and verified by partial DNA sequenceanalysis. The recombinant plasmid was correctly screened.Conclusion: The EYFP expression vector carrying ifn-γgene was successfully established.This research work has formed a base for monitoring the ifn-y gene expression andprotein position in living cells.

Influence of sodium fluoride on alkaline phosphatase activity and bone gla protein synthesis in human yellow ligament cells in vitro

Objective To investigate the influence of sodium fluoride(NaF)on alkaline phosphatase(ALP)activity and bone gla protein(BGP)synthesis in yellow ligament cells from different surgical simples in vitro.Methods The human ligament cells

Comparative Study on Muscle Fiber Characteristics and Muscle Nutritional Quality between Two Groups of Yellow River Carps

The aim was to protect the Henan Yellow River carp germplasm resources and provide a scientific basis for the meat quality improvement of Yellow River carps. With artificially farmed and wild Henan Yellow River carps as the research objects, comparative study on muscle fiber diameter and density, routine nutritional composition, calcium and phosphorus contents and amino acids composition was conducted between the two groups of carps. The results showed that the moisture content was significantly higher （P 〈0.05）, the crude fat and crude protein contents were significantly lower （P〈0.05）, the essential amino acids and total amino acids contents were lower （P〉0.05）, the phosphorus content was higher （P 〉0.05）, and the calcium content was lower （P〉0.05） in the wild group compared with those in the farming group. The analysis of muscle fiber characteristics showed that there were significant differences in the average muscle fiber diameter （P〈0.05） and muscle fiber density （P〈0.01） between the two groups of Henan Yellow River carps.

Bright Yellow 2烤烟热激蛋白90生物信息学分析

【目的】分析Bright Yellow 2烤烟热激蛋白90(HSP90)的生物学信息,为揭示烘烤过程中烟叶的烘烤特性提供理论依据。【方法】从NCBI中获取Bright Yellow 2烤烟HSP90蛋白序列Nt HSP-1、Nt HSP-2和Nt HSP-3(Gen Bank登录号分别为BAL42333、BAL42332和BAM24708),利用生物信息学软件对各序列进行理化特性、跨膜区、信号肽、亚细胞定位、二级结构和三级结构预测分析。【结果】Bright Yellow 2烤烟Nt HSP-1、Nt HSP-2和Nt HSP-3的氨基酸数量分别为812、811和699个,均以酸性氨基酸含量较多,其中谷氨酸(Glu)含量最高,理论等电点均小于5.00,二级结构均以α-螺旋(H)和无规则卷曲(C)为主。Nt HSP-1和Nt HSP-2的亲水性较高,具有较高的稳定性,属于分泌蛋白,具有锚定结构,定位于内质网,其三级结构呈现"r"形,N端内部具有Mg2+结合位点、HATPase_c(ATP酶)和top6b(DNA拓扑异构酶VI)功能域;Nt HSP-3为非分泌型蛋白,定位于叶绿体,三级结构呈现"i"形,N端含有Mg2+结合位点和HATPase_c功能域。【结论】Nt HSP-1和Nt HSP-2可能与烟叶耐烤性有关,而Nt HSP-3可能与烟叶易烤性有关。

Monoclonal Antibodies Against the Whitefly-Transmitted Tomato Yellow Leaf Curl Virus and Their Application in Virus Detection

Tomato yellow leaf curl virus（TYLCV）is a species of the family Geminiviridae,causing serious yield losses in tomato production.The coat protein（CP）gene of TYLCV isolate SH2 was expressed in Escherichia coli BL21（DE3）using pET-32a as the expression vector.The recombinant protein was purified through Ni＋-NTA affinity column and used to immunize BALB/c mice.Three hybridoma cell lines（2B2,2E3 and 3E10）secreting monoclonal antibodies（MAbs）against TYLCV CP were obtained by fusing mouse myeloma cells（Sp 2/0）with spleen cells from the immunized BALB/c mouse.The titers of ascitic fluids of three MAbs ranged from 10-6 to 10-7 in indirect-ELISA.Isotypes and subclasses of all the MAbs belonged to IgG1,κ light chain.Triple antibody sandwich enzyme-linked immunosorbent assay（TAS-ELISA）showed that the MAb 3E10 could react with five begomoviruses infecting tomato,while the other two（2B2 and 2E3）mainly reacted with TYLCV.TAS-ELISA was set up using the MAb 3E10,and the established method could successfully detect virus in plant sap at 1：2 560（w/v,g mL-1）.Detection of field samples showed that begomoviruses were common in tomato crops in Zhejiang Province,China.

Establishment of a Spleen Cell Line from Large Yellow Croaker Pseudosciaena crocea and its Primitive Application in Foreign Gene Transfection

A large yellow croaker,Pseudosciaena crocea,spleen(LYCS)cell line was established and the feasibility of using it for foreign gene transfection was evaluaed in this study.Primary culture of LYCS cells was initiated from spleen tissue pieces,which were cultured at 25℃ in Dulbecco's modiced Eagle medium/F12 medium(DMEM/F12,1:1)(pH7.2),supplemented with 20% fetal bovine serum,carboxymethyl chitosan,chondroitin sulfate,basic fibroblast growth factor(bFGF)and insulin-like growth factor-I(IGF-I).The cultured LYCS cells,in fibroblast shape,proliferated to 100% confluency 20 days later.Chromosome analyses indicated that the LYCS cells exhibited chromosomal aneuploidy with a modal chromosome number of 48 which displayed the normal diploid karyotype of P.crocea(6m+6sm+36t,NF=60).A LYCS cell line,with a population doubling time of 48.7 h at passage 60,has been established and subcultured to passage 70.Transgenic feasibility test demonstrated that positive green fluorescence protein(GFP)expression was observed in LYCS cells after pcDNA3.1-GFP plasmid transfection.In conclusion,a continuous foreign gene trans-fection feasible LYCS cell line has been established successfully.The cell line might serve as a valuable tool for studies of transgenic breeding and has potential applications for different kinds of cytotechnological studies.

Effects of undigested protein-rich ingredients on polarised small intestinal organoid monolayers

Here, we describe the use of monolayers of intestinal epithelial cells derived from intestinal organoids and transcriptomics to investigate the direct effects of dietary protein sources on epithelial function. Mechanically dissociated 3 D organoids of mouse duodenum were used to generate a polarized epithelium containing all cell types found in the tissue of origin. The organoid-derived cell monolayers were exposed to 4%(w/v) of ‘undigested(non-hydrolysed)-soluble' fraction of protein sources used as feed ingredients [soybean meal(SBM) and casein], or alternative protein sources(spray dried plasma protein, and yellow meal worm), or controls for 6 h prior to RNA isolation and transcriptomics. All protein sources altered expression of unique biological processes in the epithelial cells. Exposure of intestinal organoids to SBM downregulated expression of retinol and retinoid metabolic processes as well as cholesterol and lipid biosynthetic pathways, consistent with the reported hypotriglyceridaemic effect of soy protein in vivo. These findings support the use of intestinal organoids as models to evaluate complex interactions between dietary ingredients and the intestinal epithelium and highlights some unique host effects of alternative protein sources in animal feed and potentially human food.

Molecular diversity of barley yellow dwarf virus-PAV from China and the Czech Republic

Wheat yellow dwarf disease(BYD),caused by different species of barley/cereal yellow dwarf viruses(B/CYDVs),is one of the most serious cereal diseases in China and the Czech Republic.Because genetic diversity of the virus directly influences disease epidemiology,the molecular diversity and population structure of 24 Chinese isolates and 16 the Czech Republic isolates of BYDV-PAV from different regions in two countries were analyzed by sequencing their coat protein(CP)and readthrough protein(RTP)domain(RTD)genes and comparing the sequences with six CP and 16 RTP sequences of BYDVPAV isolates from the NCBI database based on nucleotide identity position,phylogenetic analysis and nucleotide diversity.Nucleotide identities between the Chinese and the Czech Republic isolates for the CP were 76.6–99.4%,73.9–89.1%for RTD(ORF5),respectively.The Chinese and the other country isolates showed 74.7–99.2%nucleotide identity for RTP(ORF3+ORF5).Phylogenetic analysis of CP sequences showed that 20 Chinese isolates clustered in the same clade,but the other four Chinese isolates clustered in another clade with the isolates from the Czech Republic and other counties.The population of BYDV-PAV in China had greater nucleotide variability and was more divergent than that in the Czech Republic.Geographical and ecological factors but not hosts might contribute to the population differences in the two countries.

YGL9, encoding the putative chloroplast signal recognition particle 43 kDa protein in rice, is involved in chloroplast development

The nuclear-encoded light-harvesting chlorophyll a/b-binding proteins（LHCPs） are specifically translocated from the stroma into the thylakoid membrane through the chloroplast signal recognition particle（cp SRP） pathway. The cp SRP is composed of a cp SRP43 protein and a cp SRP54 protein, and it forms a soluble transit complex with LHCP in the chloroplast stroma. Here, we identified the YGL9 gene that is predicted to encode the probable rice cp SRP43 protein from a rice yellow-green leaf mutant. A phylogenetic tree showed that an important conserved protein family, cp SRP43, is present in almost all green photosynthetic organisms such as higher plants and green algae. Sequence analysis showed that YGL9 comprises a chloroplast transit peptide, three chromodomains and four ankyrin repeats, and the chromodomains and ankyrin repeats are probably involved in protein-protein interactions. Subcellular localization showed that YGL9 is localized in the chloroplast. Expression pattern analysis indicated that YGL9 is mainly expressed in green leaf sheaths and leaves. Quantitative real-time PCR analysis showed that the expression levels of genes associated with pigment metabolism, chloroplast development and photosynthesis were distinctly affected in the ygl9 mutant. These results indicated that YGL9 is possibly involved in pigment metabolism, chloroplast development and photosynthesis in rice.

Expression and characterization of Mac-1-FP fusion protein in CHO cells

Objective: To construct mammalian cell expression vectors for Mac-1 with CFP and YFP and apply FRET to study the dimerization and function of CD11 b( Mac-1 α subunit) and CD18(Mac-1 β subunit). Methods: The mammalian cell expression vector for CD11b fused with CFP at the carboxyl terminal was constructed to create recombinant plasmid of pCD11b-CFP. Then pCD11b-CFP was co-transfected with pYFP-CD18 into CHO cell, a fibroblast like cell line, as a target cell within which there are some signal pathways involved in inflammatory stimulation but without endogenous Mac-1. Then CHO cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were selected by Western blot and laser scanning confocal microscope. Results: The cyan and yellow fluorescence in co-transfected positive CHO cells were observed under a fluorescence microscope. CHO-Mac-1-FP cells stably expressing both CD11b-CFP and YFP-CD18 fusion proteins were obtained as demonstrated by Western blot successfully. The adhesive activity of CHO-Mac-1-FP cells with CHO-1CAM-1 cells was increased markedly by treatment with PMA, suggesting the translocation of GD11b-CFP and YFP-CD18 to the plasma membrane in CHO-Mac-1-FP cells and dimerization of CD11b-CFP and YFP-CD18 just as the function of the wild type Mac-1. Conclusion: CHO-Mac-1-FP cells with adhesive activity are established successfully, thus CHO-Mac-1-FP cells may be useful for the study of Mac-1 by FRET and for other purposes.

The Nutritional Value of Yellow Lupine (Lupinus luteus)for Growing Pigs

The aim of the study was a comparison of the composition and apparent total tract digestibility (ATTD) of nutrients in the seeds of two varieties of yellow lupine (YL). Moreover, an examination of the level of soybean meal (SBM) substitution by YL meal and in a combination with rapeseed meal (RSM) on the pigs’ performance was performed. In a digestibility trial, 30 male pigs were tested using a marker method. In two growth experiments, 60 pigs (in each) were fed diets where SBM was replaced by YL in 0%, 20%, 40%, 60%, 80% and 100% or by a mixture of RSM and YL or by both these components. The chemical composition of varieties differed among crude protein (CP), fiber, acid detergent fiber (ADF), neutral detergent fiber (NDF) and fat. The digestibility coefficients of protein, fat and gross energy were similar for both varieties. The substitution of SBM with YL did not negatively affect the pigs’ performance. A mixture of RSM with YL had no negative effect on growth parameters, except for the starter phase, when a level of above 15% RSM in the diet reduced the pigs’ gains. In conclusion, YL alone and with RSM may be a sustainable alternative to SBM in pig nutrition.

低蛋白日粮对黄羽肉鸡生长性能及胴体品质的影响

试验旨在研究低蛋白日粮对黄羽肉鸡生长性能、胴体品质及肉质的影响。试验选取1日龄慢速型母鸡360羽,设对照组、试验组,每组6个重复,每个重复30只鸡。试验组1~6周(w)、7~13 w、14~19 w阶段日粮粗蛋白浓度,分别设定为对照组的90%、95%、100%,配方中增加玉米副产物比例、降低豆粕与大豆油用量。结果显示,相比于对照组,试验组鸡各阶段体重明显下降(P<0.05),1~6 w增重明显降低(P<0.01)、料重比增高(P<0.01),全程料重比无显著变化(P>0.05);鸡腹脂率显著下降(P<0.05),胸肌a值、肉色L值优于对照组(P<0.05);单位增重所需日粮粗蛋白下降1.7%,饲料成本下降7.1%。综上所述,在本研究条件下,下调生长前、中期日粮的粗蛋白浓度会明显降低黄羽肉鸡全期生长速度,但能节约增重饲料成本,对胴体品质、肉质有一定的改善作用。

新型非粮蛋白源黄粉虫粉的营养特性及替代鱼粉对水产养殖动物的影响

鱼粉供应资源短缺,价格日益高涨,严重制约水产养殖业的可持续发展。我国新型非粮蛋白源资源丰富,种类繁多,具有可持续性强、营养价值高等特点,但在水产饲料中的应用并未得到充分挖掘。黄粉虫(Tenebrio molitor)具有蛋白质含量高、氨基酸组成均衡等优点,其虫粉是一种颇具潜力的优质非粮蛋白源。文章对黄粉虫粉的营养特性及其替代鱼粉对水产养殖动物的生长性能、肉品质、消化功能、免疫功能和抗氧化能力的影响进行概述,旨在为合理开发和应用新型非粮蛋白源提供参考。