青花菜锌指蛋白基因BoCCCH1的克隆和表达分析

CCCH锌指蛋白广泛存在于真核生物中,在植物的生长、发育和逆境胁迫响应中起着重要作用。为获知青花菜CCCH转录因子基因的序列特征和表达特性,从青花菜叶片中克隆到1个CCCH基因,命名为Bo CCCH1,同时,利用RT-PCR方法研究了其在不同器官及霜霉菌侵染叶片中的表达模式。序列分析结果表明,Bo CCCH1的基因组全长为1 568 bp,包含1个长度为599 bp的内含子,2个长度分别为627 bp、342 bp的外显子,编码322个氨基酸,蛋白质的分子量与等电点分别为35 296.42 Da和8.47;Bo CCCH1有2个锌指结构,类型均为C-X8-C-X5-C-X3-H。系统发育分析结果表明,Bo CCCH1与其它植物的21条同源序列在进化树上分为6组,来自十字花科的CCCH与Bo CCCH1处于同一分支。基因表达结果表明,Bo CCCH1在叶、花茎、嫩角果、花蕾和花中表达,其中花茎和嫩角果的表达量最高,但在根中未检测到表达;在霜霉菌侵染下,叶片Bo CCCH1的表达量升高,到24 h和36 h时达到最大,推测Bo CCCH1与霜霉病抗性相关。本研究为进一步探明Bo CCCH1在霜霉病抗性反应中的功能奠定了基础。     

Evaluation of two commercial PRRSV antibody ELISA kits with samples of known status and singleton reactors

Two commercial PRRSV ELISA kits (IDEXX and Bionote) were evaluated for their sensitivity and specificity using 476 PRRS-positive serum samples collected from 7 animal challenge experiments and 1,000 PRRS-negative sera. Both ELISA kits exhibited 100% sensitivity with sera collected 14 to 42 days post-infection, and the results from the kits were highly correlated (R²=0.9207). The specificity of IDEXX or Bionote kit was 99.9% or 99.7%, respectively. In addition, the Bionote ELISA kit was used to examine 100 sera that were determined to be falsely positive either by IDEXX 2XR or 3XR ELISA, and only 7 of these samples were found to be positive. These results indicate that both ELISA kits exhibited similar levels of sensitivity and specificity and would complement one another for the verification of false-positive samples.     

Influence of paricalcitol on renal tubulointerstitial fibrosis in diabetic nephropathy

AIM: To investigate the effect of paricalcitol (P) on renal tubulointerstitial fibrosis and the underlying mechanisms in diabetic nephropathy (DN).METHODS: DN rat model was induced by a single intraperitoneal injection of streptozotocin after fasting. The animals were randomly divided into 2 groups:the DN rats in paricalcitol-intervened group (group P) were injected intraperitoneally with paricalcitol dissolved in propylene glycol after the day when the model was induced successfully at a dose of 0.4 μg/kg (3 times a week); the DN rats in DN group (group D) were given isopyknic propylene glycol. Normal control group (group C) was also set up. The samples of blood, urine and renal tissue were collected after intervention of paricalcitol for 12 weeks. The biochemical indexes were measured. The renal tissues were used for pathologic observation and determining the expression of transforming growth factor-β1 (TGF-β1), Wnt-4, β-catenin and Klotho by immunohistochemistry and Western blotting. In addition, the correlation among the above indexes was analyzed.RESULTS: (1) Scr, BUN and 24 h urine protein increased significantly in group D compared with group C, while decreased in group P compared with group D (P<0.05). (2) The area of renal tubulointerstitial fibrosis increased in group D compared with group C, while decreased in group P compared with group D (P<0.05). (3) The expression of Klotho decreased, while the expression of TGF-β1, Wnt-4 and β-catenin increased in group D compared with group C (P<0.05). Compared with group D, the expression of Klotho increased, while the expression of TGF-β1, Wnt-4 and β-catenin decreased in group P (P<0.05). (4) The expression of Klotho was negatively correlated with the fibrosis area, TGF-β1, Wnt-4 and β-catenin (P<0.05).CONCLUSION: Paricalcitol inhibits renal tubulointerstitial fibrosis in DN by promoting the expression of renal Klotho, and inhibiting Wnt/β-catenin signaling pathway activation and TGF-β1 synthesis.     

红花粕和水飞蓟粕对生长獭兔的营养价值评定

本试验旨在通过饲养试验和消化试验来评定红花粕和水飞蓟粕在生长獭兔上的营养价值。选取18只60日龄左右、平均体重(1.73±0.21)kg、健康状况良好的白色獭兔,随机分为3个组,每组6个重复,每个重复1只兔。各组分别饲喂基础饲粮、红花粕饲粮(85%基础饲粮+15%红花粕)和水飞蓟粕饲粮(85%基础饲粮+15%水飞蓟粕)。预试期和正试期各5 d。采用化学分析法测定红花粕和水飞蓟粕的总能(GE)及各营养物质含量,采用全收粪法测定生长獭兔对各营养物质的表观消化率。结果表明:红花粕和水飞蓟粕中的GE、干物质(DM)、粗蛋白质(CP)、粗纤维(CF)、中性洗涤纤维(NDF)、酸性洗涤纤维(ADF)、酸性洗涤木质素(ADL)、粗脂肪(EE)、粗灰分(Ash)、钙(Ca)、磷(P)以及无氮浸出物(NFE)的含量分别为:18.81 MJ/kg、93.76%、23.94%、14.95%、19.92%、11.99%、2.19%、1.64%、4.93%、0.37%、0.57%、49.32%与17.12 MJ/kg、91.94%、23.62%、16.21%、38.57%、22.73%、4.04%、2.07%、5.31%、0.29%、0.68%、42.09%。生长獭兔对红花粕和水飞蓟粕中GE、DM、CP、CF、NDF、ADF、EE、Ash、Ca、P和NFE的表观消化率分别为62.60%、61.72%、62.39%、15.68%、26.30%、14.75%、80.69%、38.35%、59.35%、31.98%、79.61%与63.13%、61.94%、68.01%、15.74%、27.64%、14.98%、79.90%、38.20%、60.44%、32.99%、79.81%。由此可见,生长獭兔对红花粕和水飞蓟粕中的不同营养成分的表观消化率存在一定差异,结合2种原料各营养物质含量总体分析,2种原料均可作为獭兔的蛋白质饲料资源应用,并且对生长獭兔的营养价值相近。     

Bioinformatics Analysis of Bone Morphogenetic Protein 4 (BMP4) of Sheep

The assay was aimed to explore the biological characteristics of bone morphogenetic protein 4 (BMP4) of sheep,NCBI,DNAMAN DNAStar,TMHMM Server v.2.0,PsortⅡ,SignalP various bioinformatical softwares were used to speculate the physical and chemical properties,hydrophobic property,phosphorylation site,conservative structure domain,protein secondary structure of BMP4 protein.Also,the three-dimensional structure was forecasted with the SWISS-MODEL Workspace software.The results indicated that the BMP4 of sheep had high homologies with the BMP4 of various species.The encoded protein was a hydrophilic protein which was unstable.There was no transmembrane regions and it was likely to be located in the nucleus.What was more,there was signal peptide and eighteen phosphorylation sites.Through the forecast of functional domains,the protein had two functional domains,including the transforming growth factor-beta (TGF-beta) superfamily and TGF-beta propeptide superfamily.The result was consistent with the function of BMP4 gene family,it also demonstrated that BMP4 was a growth factor and it had the function of signal transduction.The amino acid homology between the predicted 3D structure of protein and template 3bmp.1.A was 88.29%.The bioinformatics analysis of BMP4 gene could provide reference for the further study in practice.     

Can additional N fertiliser ameliorate the elevated CO2‐induced depression in grain and tissue N concentrations of wheat on a high soil N background?

Elevated CO₂ stimulates crop yields but leads to lower tissue and grain nitrogen concentrations [N], raising concerns about grain quality in cereals. To test whether N fertiliser application above optimum growth requirements can alleviate the decline in tissue [N], wheat was grown in a Free Air CO₂ Enrichment facility in a low‐rainfall cropping system on high soil N. Crops were grown with and without addition of 50–60 kg N/ha in 12 growing environments created by supplemental irrigation and two sowing dates over 3 years. Elevated CO₂ increased yield and biomass (on average by 25%) and decreased biomass [N] (3%–9%) and grain [N] (5%). Nitrogen uptake was greater (20%) in crops grown under elevated CO₂. Additional N supply had no effect on yield and biomass, confirming high soil N. Small increases in [N] with N addition were insufficient to offset declines in grain [N] under elevated CO₂. Instead, N application increased the [N] in straw and decreased N harvest index. The results suggest that conventional addition of N does not mitigate grain [N] depression under elevated CO₂, and lend support to hypotheses that link decreases in crop [N] with biochemical limitations rather than N supply.     

苦瓜对白粉病的抗性与相关生理生化指标的关系

为探讨苦瓜抗白粉病的生理生化机制,以对白粉病不同抗性的4个苦瓜品系为材料,研究苗期感染白粉病菌后,苦瓜叶片生理生化指标的变化。结果表明:接种后,叶绿素和可溶性蛋白质量分数均呈先上升后降低的趋势;可溶性糖质量分数呈下降-上升-下降-上升的趋势;抗病品系的可溶性糖、叶绿素质量分数和过氧化物酶(POD)、多酚氧化酶(PPO)活性均高于或显著高于感病品系和高感品系;抗病品系的抗坏血酸(AsA)质量分数的上升和下降的幅度均小于感病品系和高感品系;接种后10~20d,抗坏血酸过氧化物酶(APX)活性表现为抗病品系感病品系高感品系。叶绿素、可溶性糖质量分数和POD、PPO活性与病情指数呈显著或极显著负相关。综上说明,白粉病菌侵染苦瓜后,抗病品系可通过保持较高的叶绿素质量分数,增加可溶性糖、可溶性蛋白、AsA质量分数及增强POD、PPO、APX活性来提高抗病性。叶绿素、可溶性糖质量分数和POD、PPO活性均可作为苦瓜对白粉病抗性早期鉴定的指标。