Low crude protein formulation with supplemental amino acids for its impacts on intestinal health and growth performance of growing-finishing pigs

Background Low crude protein(CP)formulations with supplemental amino acids(AA)are used to enhance intestinal health,reduce costs,minimize environmental impact,and maintain growth performance of pigs.However,extensive reduction of dietary CP can compromise growth performance due to limited synthesis of non-essential AA and limited availability of bioactive compounds from protein supplements even when AA requirements are met.Moreover,implementing a low CP formulation can increase the net energy(NE)content in feeds causing excessive fat deposition.Additional supplementation of functional AA,coupled with low CP formulation could further enhance intestinal health and glucose metabolism,improving nitrogen utilization,and growth performance.Three experiments were conducted to evaluate the effects of low CP formulations with supplemental AA on the intestinal health and growth performance of growing-finishing pigs.Methods In Exp.1,90 pigs(19.7±1.1 kg,45 barrows and 45 gilts)were assigned to 3 treatments:CON(18.0%CP,supplementing Lys,Met,and Thr),LCP(16.0%CP,supplementing Lys,Met,Thr,Trp,and Val),and LCPT(16.1%CP,LCP+0.05%SID Trp).In Exp.2,72 pigs(34.2±4.2 kg BW)were assigned to 3 treatments:CON(17.7%CP,meeting the requirements of Lys,Met,Thr,and Trp);LCP(15.0%CP,meeting Lys,Thr,Trp,Met,Val,Ile,and Phe);and VLCP(12.8%CP,meeting Lys,Thr,Trp,Met,Val,Ile,Phe,His,and Leu).In Exp.3,72 pigs(54.1±5.9 kg BW)were assigned to 3 treatments and fed experimental diets for 3 phases(grower 2,finishing 1,and finishing 2).Treatments were CON(18.0%,13.8%,12.7%CP for 3 phases;meeting Lys,Met,Thr,and Trp);LCP(13.5%,11.4%,10.4%CP for 3 phases;meeting Lys,Thr,Trp,Met,Val,Ile,and Phe);and LCPG(14.1%,12.8%,11.1%CP for 3 phases;LCP+Glu to match SID Glu with CON).All diets had 2.6 Mcal/kg NE.Results In Exp.1,overall,the growth performance did not differ among treatments.The LCPT increased(P<0.05)Claudin-1 expression in the duodenum and jejunum.The LCP and LCPT increased(P<0.05)CAT-1,4F2hc,and B0AT expressions in the jejunum.In Exp.2,overall,the VLCP reduced(P<0.05)G:F and BUN.The LCP and VLCP increased(P<0.05)the backfat thickness(BFT).In Exp.3,overall,growth performance and BFT did not differ among treatments.The LCPG reduced(P<0.05)BUN,whereas increased the insulin in plasma.The LCP and LCPG reduced(P<0.05)the abundance of Streptococcaceae,whereas the LCP reduced(P<0.05)Erysipelotrichaceae,and the alpha diversity.Conclusions When implementing low CP formulation,CP can be reduced by supplementation of Lys,Thr,Met,Trp,Val,and Ile without affecting the growth performance of growing-finishing pigs when NE is adjusted to avoid increased fat deposition.Supplementation of Trp above the requirement or supplementation of Glu in low CP formulation seems to benefit intestinal health as well as improved nitrogen utilization and glucose metabolism.

Antibacterial Activity of Bacillus subtilis and Properties of Protein Crude Extract

In order to get biological drugs with no resistance or toxic side effects and to reduce the use of antibiotics, a strain of Baci//us subtilis was isolated from animal intestine, and the isolate was identified by molecular biological method; in vitro an- tibacterial test of the isolate was performed using agar diffusion method; the optimal fermentation condition of the isoJate was screened by conventional culture method; the antibacterial crude protein of the isolate was extracted by saturated ammonium sulfate method; the physicochemical properties of antibacterial crude protein was de- tected by comparison method; The results showed that the isolate was B. subti/is, which had antibacterial effects on Staphy/ococcus aureus, streptococcus and swine erysipelas. The fermentation effect of the isolate was the best under the condition of temperature 30 ~C, pH 7, liquid volume 75 ml/250 ml, inoculation volume 20% and culture time 48 h. The antibacterial effect of the isolate was the best when extract- ed by 80% saturated ammonium sulfate. The antibacterial crude protein had strong resistance to heat and acid. Organic solvent and UV irradiation had some influences on antibacterial crude protein. Proteases had hydrolytic effects on antibacterial crude protein. The isolated B. subti/is can be used to prevent and control the diseases caused by S. aureus, streptococcus and swine erysipelas, and can regulate intesti- nal microecology by adding into expanded feeds.

Predicting Levels of Crude Protein, Digestibility, Lignin and Cellulose in Temperate Pastures Using Hyperspectral Image Data

Hyperspectral sensors provide the potential for direct estimation of pasture feed quality attributes. However, remote sensing retrieval of digestibility and fibre (lignin and cellulose) content of vegetation has proven to be challenging since tissue optical properties may not be propagated to the canopy level in mixed cover types. In this study, partial least squares regression on spectra from HyMap and Hyperion imagery were used to construct predictive models for estimation of crude protein, digestibility, lignin and cellulose concentration in temperate pastures. HyMap and Hyperion imagery and field spectra were collected over four pasture sites in southern Victoria, Australia. Co-incident field samples were analyzed with wet chemistry methods for crude protein, lignin and cellulose concentration, and digestibility was calculated from fiber determinations. Spectral data were subset based on sites and time of year of collection. Reflectance spectra were extracted from the hyperspectral imagery and collated for analysis. Six different transformations including derivatives and continuum removal were applied to the spectra to enhance absorption features sensitive to the quality attributes. The transformed reflectance spectra were then subjected to partial least squares regression, with full cross-validation “leave-one-out” technique, against the quality attributes to assess effects of the spectral transformations and post-atmospheric smoothing techniques to construct predictive models. Model performance between spectrometers, subsets and attributes were assessed using a coefficient of variation (CV), —the interquantile (IQ) range of the attribute values divided by the root mean square error of prediction (RMSEP) from the models. The predictive models with the highest CVs were obtained for digestibility for all spectra types, with HyMap the highest. However, models with slightly lower CVs were obtained for crude protein, lignin and cellulose. The spectral regions for diagnostic wavelengths fell within the chlorophyll well, red edge, and 2000-2300 nm ligno-cellulose-protein regions, with some wavelengths selected between the 1600 and 1800 nm region sensitive to nitrogen, protein, lignin and cellulose. The digestibility models with the highest CV’s had confidence intervals corresponding to ±5% digestibility, which constitutes approximately 30% of the measured range. The cellulose and lignin models with the highest CV’s also had similar confidence intervals but the slopes of the prediction lines were substantially less than 1:1 indicating reduced sensitivity. The predictive relationships established here could be applied to categorizing pasture quality into range classes and to determine whether pastures are above or below for example threshold values for livestock productivity benchmarks.

Effects of Powdery Mildew on 1 000-Kernel Weight, Crude Protein Content and Yield of Winter Wheat in Three Consecutive Growing Seasons

In order to clarify the impact posed by wheat powdery mildew （Blumeria graminis f. sp. tritici） on the yield and yield components in different epidemic seasons, field trials were conducted in three growing seasons, 2009-2010, 2010-2011 and 2011-2012, in Langfang City, Hebei Province, China. The relationships between 1000-kernel weight, crude protein content of grain and yield and disease index （DI）, as well as area under disease progress curve （AUDPC） were studied. The models of the percentage of loss of 1000-kernel weight, crude protein content and yield were constructed using DI at critical point （CP） of growth stages （GS） and AUDPC in the three growing seasons, respectively. The CPs for estimating 1 000-kernel weight, crude protein content of grain and yield of wheat caused by powdery mildew were GS 11.1, GS 10.5.3 and GS l 0.5.3, respectively. Models based on DI at CP to estimate the percentage of loss of 1000-kernel weight, crude protein content of grain and yield were better than models based on AUDPC. And models of the percentage of loss of 1000-kernel weight, crude protein content and yield for 2011-2012 season were significant different from these for 2009-2010 and 2010-2011 seasons. These results indicated that besides powdery mildew, weather conditions also had influence on 1 000-kernel weight, crude protein content of grain and yield loss of wheat when powdery mildew occurred.

The Effect of Varied Dietary Crude Protein Levels with Balanced Amino Acids on Performance and Egg Quality Characteristics of Layers at First Laying Phase

Four diets were formulated to study the influence of varying crude protein levels on the performance, egg quality, serum and haematological characteristics at the first phase of laying cycle of sixty Bovan Nera laying birds that were randomly allotted to dietary treatments. The four experimental diets had five replicates each and three birds per repli-cate. Diet 1 contained 14% crude protein (CP), while diet 2 contained 15% CP, diets 3 and 4 contained 16% and 17% CP respectively. The experimental birds were fed for 10 weeks and the data collected were statistically analysed. Apparent variations recorded for all the performance characteristics such as Egg number, Hen-day production, Feed Conversion Ratio (FCR) and weight gain were significantly different (p < 0.05). The best FCR value were recorded by birds fed 17% CP (3.45), while the highest weight gain mean value were also recorded by birds fed 17% CP value. Findings indicated a direct relationship between dietary crude protein values and performance.

The influence of phytase, pre-pellet cracked maize and dietary crude protein level on broiler performance via response surface methodology

Background:The reduction of crude protein levels in diets for broiler chickens may generate economic,environmental and flock welfare and health benefits;however,performance is usually compromised.Whole grain feeding and phytase may improve the utilization of reduced crude protein diets.Results:The effects of pre-pellet cracked maize(0,15%and 30%)and phytase(0,750 and 1500 FTU/kg)in isoenergetic maize-soy diets with three levels of crude protein(22%,19.5%and 17%)were evaluated via a BoxBehnken response surface design.Each of 13 dietary treatments were offered to 6 replicate cages(6 birds/cage)of male Ross 308 broiler chicks from 7 to 28 d post-hatch.Model prediction and response surface plots were generated from experimental data via polynomial regression in R and only significant coefficients were included and discussed in the predicted models.Weight gain,feed intake and FCR were all influenced by pre-pellet cracked maize,phytase and crude protein level,where crude protein level had the greatest influence.Consequently,the reduction from 22%to 17%dietary crude protein in non-supplemented diets reduced weight gain,feed intake,relative gizzard weight,relative gizzard content and relative pancreas weight but improved FCR.However,the inclusion of 30%cracked maize to 17%crude protein diets restored gizzard weight and 1500 FTU phytase inclusion to 17%crude protein diets increased relative gizzard contents and pancreas weights.Cracked maize and phytase inclusion in tandem to 17%crude protein diets increased weight gain,feed intake and FCR;however,this FCR was still more efficient than broilers offered the non-supplemented 22%crude protein diet.Broilers offered the prepellet cracked maize and phytase inclusions reduced AME in 22%crude protein diets but improved AME by 2.92 MJ(14.16 versus 11.24 MJ;P<0.001)in diets containing 17%crude protein.Ileal N digestibility was greater in broilers offered diets with 17%crude protein than those offered the 22%crude protein diet;irrespective of phytase and pre-pellet cracked maize.Conclusion:Pre-pellet cracked maize and phytase inclusions will improve the performance of broilers offered reduced crude protein diets.

Effects of Different Dietary Crude Protein Levels on Reproductive Performance in Breeder Roosters

[Objective] To investigate effects of different dietary crude protein levels on reproductive performance and find out the optimal additive proportion of crude protein. [Method] A total of 45 New Yangzhou breeder roosters at 21 weeks old were sampled and divided into three groups, namely, low-protein diet group, medium-protein diet group and high-protein diet group. They were fed with diets respectively at crude protein levels of 17.23%, 13.04% and 11.32%. The sperm volume, testicular traits and levels of reproductive hormones were analyzed during the whole repro- ductive period. [Result] The sperm volume, sperm motility and sperm density of the medium-protein diet group were higher than that of the other two groups, and the sperm deformity rate of the medium-protein diet group was lowest. The rooster＇s testicular traits including weight, volume and testis indexes were not significantly affected by different protein levels, but the testis shrank slowly with aging （ P 〈 0.05）. The levels of testosterone （T）, luteinizing hormone （LH） and follicule stimulating hormone （FSH） in plasma were respectively 6.34%, 19.64% and 3.88% higher in the lowprotein diet group than in the medium-protein diet group, and these indexes were respectively 7.20%, 17.54% and 9.84% higher in the low-protein diet group than in the high-protein diet group. But all these differences were not significant. [ Conclusion] The breeder roosters fed with the diet at the crude protein level of 13.04% show better reproductive performance and economic efficiency.

Rumen Degradation Regularity of Dry Matter and Crude Protein of Common Roughage in Beef Cattle

Three individuals of Horqin yellow cattle equipped with permanent fistula, weighed （548 ±21） kg, were sdected as the experimental animals. The ru- men degradation characteristics of dry matter （DM） and crude protein （CP） of roughage at 6, 12, 24, 48 and 72 h were measured by nylon bag method. The re- suits showed that the effective degradation rates of DM and CP of alfalfa hay were the highest, while higher contents of rapid degradation part and potential degrada- tion part of DM and CP also resulted in higher degradation rates of DM and CP. The effective degradation rates of CP and DM of roughage presented strong positive correlation with CP, but showed strong negative correlation with neutral detergent fiber （NDF）. The effective degradation rates of CP of five roughages successively were alfalfa hay 〉 alfalfa block 〉 ryegrass 〉 silage corn 〉 straw.

Influence of Varying Crude Protein Levels and Balanced Amino Acids on the Performance and Haematological Characteristics of Laying Hens at the Second Phase of Production

Varying levels of dietary crude proteins and balanced amino acids were fed to layers for a period of eight weeks starting from the twenty-sixth week of age of birds and six weeks into egg production. Effects on performance and haematological characteristics were investigated at this second phase of production. Sixty Black Nera hens were randomly allotted into four (4) dietary treatments, containing the following levels of crude protein 14%, 15%, 16%, 17% and the metabolizable energy was iso-caloric for each treatment. The results showed that there were no significant differences (P > 0.05) observed for lymphocyte, Haemoglobin (Hb), Packed Cell Volume (PCV), Red Blood Cell (RBC) and White Blood Cell (WBC), these haematological parameters were within the range for healthy birds. Thus crude protein level of 14% can be used in diets of layers at the second phase of production provided that adequate amino acids are given, without adverse effect on egg laying, feed intake and measured blood parameters.

Prediction of Crude Protein in Ramie by Near-Infrared Spectroscopy(NIR)

[Objective] This study was conducted to evaluate the feasibility of deter- mining crude protein in ramie using near-infrared （NIR） spectrometer. [Method] Par- tial least square regression （PLSR） was performed to establish a calibration model based on 50 samples for predicting the crude protein content in ramie, and the model was validated with data in the validation set consisting of 10 samples. [Result] The correlation coefficient of the model was 0.98. There was a good correla- tion between the predicted values by the near-infrared prediction model and the measured values by chemical analysis, and the relative error was 3.54% on aver- age between the predicted and the measured values. [Conclusion] The results showed that it is feasible to determine crude protein content in ramie using NIR spectroscopy-based prediction model.

Effects of Dietary Metabolizable Energy and Crude Protein Levels on Reproductive Performance of Magang Geese

[ Objective] To investigate the effects of different levels of metabolizable energy and crude protein on egg laying performance and hatch- ing efficiency in Magang geese. [Method] The healthy Magang geese at 2 years old were randomly assigned into four groups, group Ⅰ, group Ⅱ, group Ⅲ and control group. They were fed with diets at different levels of metabolizable energy and crude protein, and then their eggs were collect- ed and hatched. During the test, their health was observed, and the egg weight, egg yield and hatching rate were recorded. [ Result] The average egg laying rates and average egg weight were significantly higher in the group Ⅰ, group Ⅱ and group Ⅲ than in the control group. The hatchable egg rates and egg fertilization rates of the group Ⅰ, group Ⅱ and group Ⅲ were also increased, and significance was found between the group Ⅲ and the other group （P 〈 0.05）. The feed costs of the three test groups were lower than that of the control group, and the feed cost was higher in the group Ⅲ than that in the group Ⅰ and Ⅱ. [ Conclusion] The levels of metabolizable energy and crude protein in diet have significant effects on laying performance and hatching efficiency in Magang geese.

Effects of NaOH Treatment on Fiber, Crude Protein and Soluble Sugar Content of Corn Straw

[Objective] The paper was to study the effects of different levels of NaOH on fiber, crude protein and soluble sugar content of cornstraw. [Method] Four treatments were designed in the test, namely control group, group 1, group2 and group 3, which were added with 0, 2%, 3%and 4% NaOH according to the dry weight of corn straw, and the moisture of corn straw after treatment was 45%. After being treated at room tem-perature for 48 h, sensory evaluation and nutrient composition analysis were performed for each group, and the contents of neutral detergent fiber（NDF）, acid detergent fiber （ADF）, crude protein （CP） and soluble sugar （SS） were measured. [Result] With the increasing addition level of NaOH,the content of NDF and ADF decreased gradually, and significant differences were observed between group 3 （adding 4% NaOH） and control group（P〈0.05）. There was no significant difference in CP content among treatment groups（P〉0.05）; the SS content gradually increased, and there weresignificant differences between treatment groups and control group （P〈0.05）. [Conclusion] 4% NaOH reduced the fiber content and increased thesoluble sugar content of corn straw.

川西北高寒地区5种饲草产量与CNCPS组分分析

饲草种质资源是新品种创制的基础,本研究选择适宜在川西北高寒区栽培的中华羊茅(Festuca sinensis)、无芒雀麦(Bromus inermis)、垂穗披碱草(Elymus nutans)、圆柱披碱草(Elymus cylindricus)和老芒麦(Elymus sibiricus)饲草种质资源,测定了饲草产量、全株及不同部位(茎、叶、穗)的营养成分,利用康奈尔净碳水化合物-蛋白质体系(Cornell net carbohydrate and protein system, CNCPS)对其组分进行分析,为筛选高产优质饲草种质资源奠定基础。结果表明,鲜草产量随年份增加逐年降低,老芒麦连续四年的鲜草产量高于其他饲草。圆柱披碱草的干草产量最高,变异系数较小,稳产性较好。无芒雀麦茎、叶和穗的中性洗涤纤维(Neutral detergent fiber, NDF)、酸性洗涤纤维(Acid detergent fiber, ADF)含量低于其他饲草,非结构性碳水化合物(Nonstructural carbohydrate, NSC)含量显著高于其他饲草(P<0.05)。圆柱披碱草茎、叶和穗的ADF含量显著高于其他饲草(P<0.05)。老芒麦全株、茎、叶和穗的粗蛋白(Crude protein, CP)含量高于其他饲草,碳水化合物(Carbohydrate, CHO)含量低于其他饲草。综合分析得出,无芒雀麦和老芒麦的生产性能及营养品质表现较好,可作为当地种质资源研究和新品种选育的基础材料。

Effect of Dietary Crude Protein Level of Feed on Growth and Survival of Rohu Fry

Fry rearing is one of the important stage which aims at obtaining high growth and survival for production of fingerlings required for stocking into grow out ponds as well as rehabilitation in natural habitat. This experiment was conducted with the purpose to test the effect of dietary crude protein level (CP %) of feeds prepared from similar feed ingredients in different ratios on growth performance and survival rate of Rohu fry (Labeorohita). An initial density of 100 fry/m2 was maintained in hapa fixed in the cemented tank. The dietary CP% level of feed tested were 20% CP, 25% CP, 30% CP, and 35% CP fed at 5% body weight. The experiment ran for 53 nursing days. The results showed that there was no significant difference (P > 0.05) in the growth rate (g/day) of fry among treatments. Rather group fed with higher protein level grew comparatively better indicating possibility of increasing need of protein in diets. However, the survival rate (%) of rohu fry was significantly different (P < 0.05) in each tested CP% level of feed. Highest survival (82%) of fry was found in the feed of CP 35% and lowest (56%) in the feed of CP 25%. It was predicted that feed with increasing level of CP % in diet is essential for increasing survival rate.

CP、Cys-C、RBP及其联合检测在喀什地区维吾尔族慢性肾脏疾病中的诊断价值

目的分析尿外泌体铜蓝蛋白(CP)、血清胱抑素C(Cys-C)、视黄醇结合蛋白(RBP)及其联合检测对喀什地区维吾尔族早期慢性肾脏疾病(CKD)的诊断价值。方法选取2019年1月—2023年1月在新疆喀什地区第一人民医院就诊的225例维吾尔族CKD患者纳入CKD组,另选取同期在体检中心接受健康检查的186例维吾尔族体检者纳入健康对照组。收集两组的尿外泌体CP、血清Cys-C及RBP值。采用logistic回归分析影响早期CKD的因素,绘制受试者工作特征(ROC)曲线分析CP、Cys-C、RBP及其联合检测对早期CKD的临床诊断效能。结果与健康对照组比较,CKD组患者的血清白蛋白、C反应蛋白、血尿素、血沉均明显升高(P<0.05),肾小球滤过率降低(P<0.05)。与健康对照组比较,CKD组尿外泌体CP、血清Cys-C及RBP表达均明显升高(P<0.001)。Logistic回归分析显示,尿外泌体CP、血清Cys-C及RBP是影响早期CKD的独立因素(P<0.001)。ROC曲线分析显示,与尿外泌体CP、血清Cys-C、血清RBP比较,联合检测(并联)诊断早期CKD的AUC、特异度均明显升高。结论早期CKD患者的尿外泌体CP、血清Cys-C、血清RBP均明显升高,尿外泌体CP、血清Cys-C、血清RBP均可用于诊断早期CKD,但联合检测(并联)的诊断效能更高,为尽早诊断CKD提供参考。

Agronomic Trait and Protein Component of F_2 Hybrid Originated from Intergeneric Somatic Hybridization Between Triticum aestivum and Agropyron elongatum

Protoplasts derived from common wheat (Triticum aestivum L,. cv. Jinan 177) were fused with UV-treated protoplasts of Agropyron elongatum. (Host) Nevski by PEG method, and fertile asymmetric somatic hybrid plants resembling wheat morphology were obtained. The F-2 hybrid plants could be divided into 3 types according to their morphology. Type I hybrids had high and loosely standing stalks with big spikes and grains. Type ii hybrids were dwarf and compact in shape with high tillering ability and smaller spikes. Type III hybrids were similar to type I as a whole but had more compact and erect spikes. All the F-2 hybrid lines were superior to wheat in seed protein content, although some difference existed between themselves. Protein analysis of immature embryos and flag leaves from hybrids by two-dimensional electrophoresis showed that they possessed characteristic proteins of both parents and some new proteins. There existed also some different kinds of proteins in different lines.

中国马铃薯Y病毒的检测鉴定及CP基因的分子变异

【目的】查明马铃薯Y病毒(Potato virus Y,PVY)病在中国的发生情况,及时、准确地鉴定出PVY并对其分子变异进行分析。【方法】采用ELISA方法对采自中国14个省(直辖市)马铃薯种植区疑似受PVY感染的样品进行了检测,并对其中的部分材料镜检验证,然后根据CP基因序列设计1对简并引物针对随机选择感染PVY的14个省(直辖市)代表样品进行CP基因扩增克隆,将测序得到的序列进行分子变异分析,并使用贝叶斯法(Bayesian inference,BI)重建系统发育关系。【结果】ELISA检测结果表明,691份样品中220个样品与PVY抗体呈阳性反应,其余呈阴性反应;ELISA检测的阳性材料在透射电镜下均可观察到明显的风轮状内含体,14个PVY分离物均成功扩增出预期大小(约800 bp)的特异性片段,CP基因的核苷酸序列与已报道PVY不同株系的核苷酸序列一致性均在88%以上;在14个PVY分离物CP基因中共发现有29个多态性位点,其中6个简约信息位点,23个单一变异位点。系统发育分析结果显示,14个PVY分离物与PVYN:O株系相聚成簇,表明其在系统发育关系上,与PVYN:O株系的亲缘关系最近。【结论】PVY CP基因高度保守,但不同地区分离物也存在一定的分子变异,本研究可为今后了解PVY病毒病流行、变异趋势及其防治提供依据。

NSvc4和CP蛋白与水稻条纹病毒的致病相关

【目的】以水稻条纹病毒(Rice stripe virus,RSV)编码的NSvc4蛋白和CP蛋白的致病功能鉴定为切入点,研究RSV的致病机理。【方法】通过农杆菌介导在本氏烟中对RSV编码的6个蛋白进行细胞定位。采用双分子荧光互补(Bimolecular fluorescence complementation,BiFC)技术鉴定NSvc4蛋白与其余5个蛋白间的互作关系,并用酵母双杂交体系(Yeast two-hybrid,YTH)对NSvc4与CP蛋白间的互作再次验证。利用马铃薯X病毒(Potato virus X,PVX)系统在本氏烟叶片研究NSvc4蛋白和CP蛋白的致病性,采用实时荧光定量PCR(Real-time PCR)技术验证致病性鉴定结果。【结果】RSV NSvc4蛋白能与CP蛋白互作,且蛋白复合体在本氏烟细胞中能够定位到叶绿体。致病性鉴定显示,PVX-RSV-NSvc4侵染的本氏烟表现为花叶症状;PVX-RSV-CP能够在侵染叶上诱导花叶症状,但是系统叶却恢复健康;PVX-RSV-NSvc4与PVX-RSV-CP共侵染的本氏烟和PVX-RSV-NSvc4CP侵染的本氏烟的侵染叶、系统叶均表现出了严重病症。Real-time PCR结果显示,PVX载体在各侵染本氏烟中的累积量均较低,而RSV CP和NSvc4基因的表达量与本氏烟的发病症状紧密相关。【结论】NSvc4和CP蛋白均有致病性。CP蛋白的致病力不能持久,但是与NSvc4蛋白互作后具有持久致病力,表明二者协同在RSV致病过程中发挥作用。

转CP基因线辣椒对CMV和CMV-RNA的抗病性比较

本试验以转化 CMV- CP和 TMV- CP基因线辣椒纯合系作试材 ,比较了接种 CMV粒体和CMV- RNA后的发病特点和叶片中的病毒含量。结果表明 :转化线辣椒不仅能抵抗 CMV粒体的侵染 ,而且还能抵抗 CMV- RNA的侵染。不论接种 CMV粒体或 CMV- RNA,CP(+ )线辣椒的系统症状都延迟出现 ,显症株率和病害严重度级别大幅度降低 ,病毒增殖和运转受到抑制 ,接种叶片与新生叶片中的病毒含量明显减低。这一结果证实 CMV- RNA不能克服线辣椒由 CP基因介导的抗病性。

瞬时表达比较马铃薯X病毒CP基因3种结构对RNA沉默的诱导效果

RNA沉默是植物抵御病毒侵染的一种防卫机制,病原来源的抗性(pathogen-derived resistance,PDR)被认为是通过RNA沉默起作用的.转化的核酸片段在基因组中的位置、长短和不同排列结构等均影响RNA沉默的效率.用全长马铃薯X病毒(PVX)CP基因构建非翻译的正义、反义和反向重复结构转基因的植物表达载体,通过农杆菌渗入法瞬时表达测试了这些转基因对RNA介导抗性的诱导效率和有效性.结果表明,反向重复的PVXCP是更为有效的RNA沉默诱导因子,同时也证明让目的基因在受试植物中瞬时表达,在转化植物之前能比较快速地检测转基因的表达情况以及表达产物的功能,从而节约时间和资源,并减少盲目性.