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。     

丘陵红黄壤区施用399对“一种两收”再生稻根系和产量的影响

[目的]研究399对"一种两收"再生稻的影响。[方法]以适宜丘陵红黄壤种植的丰两优香1号、准两优527、宜优673再生稻品种为材料,分别设置喷施399和不喷施399(CK)2种处理,研究各处理对再生稻根系和产量的影响。[结果]结果表明:施用399使水稻在生育期上延缓成熟,且施后再生稻的总根系数量、白根数量、根鲜重、根干重、根体积大幅度增加,同时在两季总产量上准两优527、丰两优香1号和宜优673品种分别比对照增产6.39%、4.86%、5.94%。[结论]施用399有利于准两优527、丰两优香1号和宜优673再生稻品种再生根的发生。     

重组猪抑制素蛋白质中毒素残留检测及其理化稳定性

为探索纯化的重组猪抑制素蛋白质毒素残留情况以及重组猪抑制素蛋白质稳定性,本研究利用鲎试剂、气相色谱和SDS-PAGE方法对重组猪抑制素蛋白质中的细菌内毒素、β-巯基乙醇的残留量及其蛋白质在不同温度、极端p H值和反复冻融条件下的稳定性进行检测。结果显示,经过3次等电点沉淀洗涤后,重组猪抑制素蛋白质中细菌内毒素和β-巯基乙醇的残留量大幅减少。通过在不同温度下的处理发现,重组猪抑制素蛋白质在4℃环境下较稳定,可以长期保存;但在37℃时,稳定性快速下降,随着温度上升而降解加速。通过极端p H值和反复冻融试验发现,重组猪抑制素蛋白质对极端p H值和反复冻融具有较好的耐受性。     

酸奶营养品质提升的加工工艺研究

牛奶中含有丰富的活性营养物质,包括乳铁蛋白、免疫球蛋白、乳过氧化物酶、α-乳白蛋白、β-乳球蛋白等。传统工艺生产的酸奶经过高温杀菌,活性营养物质几乎完全损失。为了避免这一情况,本文推出一种新的酸奶生产工艺,生产出的酸奶可保留大部分活性营养物质。首先将牛奶的乳脂进行分离,乳脂部分进行125 ℃、5 s杀菌,脱脂牛奶部分进行75 ℃、15 s杀菌,将两部分料液混合后加入菌种发酵至酸度75~85°T,破乳后冷却后熟8 h以上。经测定,新工艺酸奶中α-乳白蛋白、β-乳球蛋白、乳铁蛋白、免疫球蛋白、乳过氧化物酶含量分别为966 mg/L、1 963 mg/L、23.4 mg/L、102 mg/L、1 025 U/L,远高于传统工艺酸奶的68 mg/L、200 mg/L、0.7 mg/L、14 mg/L、0 U/L。通过对比新工艺与传统工艺酸奶保质期内的活菌数、黏度、酸度和口感,并无显著差异。     

三种融合标签对大熊猫轮状病毒结构蛋白VP6-VP7可溶性表达的影响

大熊猫轮状病毒(giant panda rotavirus,GPRV)是引起幼龄大熊猫腹泻的主要病原,对圈养大熊猫产生了较大的危害。轮状病毒结构蛋白VP6是一种载体蛋白,可介导黏膜免疫反应。VP7是轮状病毒结构蛋白中主要的中和抗原。因此,VP6-VP7的融合表达作为候选抗原对该病的防治具有重要的意义。传统大肠埃希菌原核表达存在表达量低、可溶性差以及纯度低等弊端。本研究使用醛缩酶(EDA)、谷胱甘肽S-转移酶(GST)、麦芽糖结合蛋白(MBP)3种融合标签,以实现获得表达量高和纯度高的GPRV-VP6-VP7重组表达蛋白。将扩增的VP6、VP7基因片段利用同源重组酶构建到含3种融合标签的表达载体pET21b上,将重组质粒转化至大肠埃希菌Rosetta(DE3)感受态细胞中进行低温诱导表达。用Ni-柱亲和层析法纯化目的蛋白,SDS-PAGE和Image J分析蛋白表达量和可溶性,Western blot分析得到表达的重组表达蛋白正确且具有蛋白活性。实验结果证明,EDA标签能显著促进VP6-VP7蛋白的原核可溶性表达,提高VP6-VP7蛋白表达量。     

Effects of baicalin on glial fibrillary acidic protein and nuclear factor-κB expression in juvenile rat hippocampus after status convulsion

AIM: To study the effects of baicalin (BC) on glial fibrillary acidic protein (GFAP) and nuclear factor-κB (NF-κB) expression and neuronal apoptosis in juvenile rat hippocampus after status convulsion (SC). METHODS: One hundred and ninety five juvenile male Sprague-Dawley rats were randomly divided into 3 groups: normal saline pretreatment group (NS group), SC group and SC with BC pretreatment group (BC group). Each of these 3 groups would be subdivided into 5 subgroups sacrificed at 4 h, 12 h, 24 h, 48 h and 72 h after SC. The rat SC model was prepared by lithium-pilocarpine chemical method. The protein expression of GFAP and NF-κB was detected by the method of immunohistochemistry. The mRNA expression of GFAP was detected by RT-PCR. The neuronal apoptosis was observed by TdT-mediated dUTP nick end labeling (TUNEL). RESULTS: Compared with NS group, the GFAP positive cells was increased in SC group (P<0.05). Compared with SC group, the expression of GFAP was significantly reduced in BC group (P<0.05). Compared with NS group, the NF-κB positive cells was increased in SC group (P<0.05). Compared with SC group, the expression of NF-κB was significantly reduced in BC group. RT-PCR showed that the expression trend of GFAP mRNA was similar to that of the protein. Compared with NS group, the TUNEL positive cells in the hippocampus CA1 area in SC group increased significantly 12 h after SC (P<0.01), and reached a peak at 48 h. After the intervention with BC, the TUNEL positive cells decreased significantly between 12~48 h after SC (P<0.05 or P<0.01), but the number of TUNEL positive cells remained significantly greater than that in NS group (P<0.05). CONCLUSION: The expression of GFAP and NF-κB in the hippocampus increased after SC in rats. Baicalin decreases the expression of GFAP and NF-κB in hippocampus of rats with pilocarpine-induced seizures, and reduces the number of neuronal apoptosis, suggesting that baicalin may protect against the brain damage caused by status convulsion.     

茶树小分子量热激蛋白基因CsHSP17.2的克隆与表达分析

[目的]进一步了解茶树小分子量热激蛋白基因CsHSP17.2在逆境胁迫条件下的分子生物学功能.[方法]利用RT-PCR技术从茶树‘迎霜’中克隆得到CsHSP1 7.2基因,运用生物信息学软件分析其核苷酸和编码蛋白,通过Real-timePCR分析其表达模式.[结果]该基因开放阅读框长度为453 bp,编码150个氨基酸,蛋白质相对分子质量为17.2×10a,理论等电点5.56;无信号肽位点,属于非分泌型蛋白;被定位于细胞质中.系统发育树分析表明,茶树CsHSP1 7.2与水稻(GenBank登录号:P27777)和花生(ABC41131)的进化关系较近,属于小分子量热激蛋白基因家族第Ⅰ亚族.qRT-PCR分析发现,茶树CsHSP17.2属于组成型基因;高温(38℃)处理1h能显著提高CsHSP17.2 mRNA的相对表达量(P＜0.05);干旱(100 g·L-1 PEG 6000)、高盐(200 mmol· L-1 NaCl)和外源脱落酸(200 mg· L-1 ABA)处理条件下,该基因的转录水平均出现不同程度的上调.[结论]克隆得到茶树‘迎霜’小分子量热激蛋白基因CsHSP17.2,其在花中表达量最高,且响应高温、干旱、高盐和外源脱落酸胁迫.     

饲料蛋白质和脂肪水平对亚东鲑亲鱼生长性能、消化酶活性和血清指标的影响

本试验旨在研究饲料蛋白质和脂肪水平对亚东鲑亲鱼生长性能、性腺指数、消化酶活性和血清指标的影响。试验采用5×2两因素随机设计,设5个蛋白质水平(36%、39%、42%、45%、48%)和2个脂肪水平(9%、18%),共配制10种实用试验饲料。每种试验饲料投喂3个重复,每个重复19尾鱼。初重为(462.53±45.40)g亚东鲑在室内水族箱中流水饲养,饲养周期为77 d。结果表明:同一脂肪水平下,随饲料蛋白质水平的升高,增重率有所升高,而饲料系数逐渐降低,至蛋白质水平为48%时又有所升高。饲料蛋白质和脂肪水平对雌性亲鱼性腺指数无显著影响(P0.05),雄性亲鱼性腺指数随蛋白质水平的升高而降低,但至蛋白质水平为48%时又有所回升。同一脂肪水平下,随着饲料蛋白质水平的升高,胃、肠道和幽门盲囊的脂肪酶活性逐渐增加,蛋白质水平为48%时显著高于蛋白质水平为36%时(P0.05);而胃、肠道和幽门盲囊的蛋白酶活性虽有所增加,但各组间无显著差异(P0.05)。同一蛋白质水平下,18%脂肪水平组胃、肠道和幽门盲囊的蛋白酶和脂肪酶活性均略高于9%脂肪水平组。同一脂肪水平下,血清氨含量随饲料蛋白质水平的升高而逐渐增加。饲料蛋白质和脂肪水平对血清超氧化物歧化酶、溶菌酶活性以及葡萄糖、甘油三酯、总蛋白、白蛋白和丙二醛含量均未产生显著影响(P0.05)。由此得出,亚东鲑亲鱼对饲料蛋白质(36%~48%)和脂肪(9%~18%)水平有一定的适应性反应;以增重率为指标进行折线模型分析,得出饲料脂肪水平为9%和18%时,亚东鲑亲鱼对蛋白质需要量分别为43.11%和45.69%。     

Reducing crude protein variability and maximizing savings when formulating corn-soybean meal-based feeds

Crude protein in corn and soybean meal have been documented to vary, and such inherent variability can result in under- or over-feeding of CP when feeds are formulated, leading to reduced bird growth, added input costs, and increased environmental pollution. The purpose of this study was to compare 2 grain-handling techniques and 2 feed formulation methods (linear vs. stochastic programming) to reduce CP variability in finished feeds and determine resulting costs or savings. The 2 grain-handling techniques were placing all the random batches of each delivered ingredient in to (1) a single bin (1-bin method) or (2) segregating above- and below-average samples into 2 bins (2-bin method). A fast way of estimating the composition of the ingredients is now available (near-infrared reflectance spectroscopy). Microsoft Excel workbooks were constructed to solve broiler starter feed formulation problems. Formulating feeds by linear and stochastic models based on the 2-bin method reduced CP variability by at least 50% compared with the 1-bin method. Formula cost was reduced by ˜20 cents per ton (averages of August 2012 United States ingredient prices) when the 2-bin method was used with the linear model. Formulating feed with a margin of safety increased formula cost by $3.40 per ton. Stochastic feed formulation increased formula cost to meet the specified CP level in feed at any probability of success, and formula cost was reduced substantially with the 2-bin method (up to $6.47 per ton). The magnitude of savings and reduced feed variability suggested that, regardless of the costs associated with building extra bins, the 2-bin method can be economically efficient in the long run. Therefore, it could be possible to split the batches of feed ingredients at a feed mill into above- or below-average bins before feed formulation to reduce CP variability and to maximize savings.     

基于S基因序列分析新型猪流行性腹泻病毒的遗传演化

最近，笔者实验室在青藏高原地区发现两种新亚型藏猪源猪流行性腹泻病毒（PEDV），为进一步调查新型PEDV是否在四川腹泻猪群中存在或流行，对实验室2018-2019年保存的116份猪腹泻粪便或肠组织样本进行PEDV的检测及其纤突蛋白基因（spike）分子特征研究。结果表明：腹泻样本的PEDV检出率为42.2%（49/116，95% CI=33.1%~51.8%），并获得了13条完整的S基因序列，全长为4 149~4 170 bp，序列相似性为94.2%~99.9%，其中SWUN-H3-CH-SCYA-2019的S基因与藏猪源新G1亚群PEDV的序列相似性高达97.0%~98.6%。遗传演化研究结果表明13株PEDV S基因划分为G1和G2大群，其中SWUN-H3-CH-SCYA-2019位于藏猪源新G1亚群；SWUN-19-CH-SCZY-2018、SWUN-4-CH-SCXC-2018、SWUN-1-CH-SCNJ-2019和SWUN-3CH-CH-SCZG-2019位于G2亚群中一个独立的分支，且与藏猪源新G2亚群毒株有着较近的亲缘关系。为了进一步研究13株PEDV的演化过程，以贝叶斯进化分析软件包（BEAST）进行分歧时间估算，结果表明SWUN-H3-CH-SCYA-2019的分歧时间约为2012.3年，早于藏猪源新G1亚群其余毒株的最早分歧时间（2015.7年）；SWUN-4-CH-SCXC-2018、SWUN-19-CH-SCZY-2018和SWUN-3CH-CH-SCZG-2019的分歧时间约为2014.2年，早于G2亚群的藏猪源毒株2014.7年，所有藏猪源PEDV的分歧时间均晚于四川毒株。本研究在四川地区首次发现了藏猪源PEDV，并且从毒株的分歧时间推断青藏高原的藏猪源PEDV来源于四川，为新型PEDV分子遗传进化的监测提供了依据。