灌浆温度和氮肥及其互作效应对稻米贮藏蛋白组分的影响

灌浆结实期温度与氮肥施用量是影响稻米品质的两个重要生态因子,尤其是与稻米蛋白含量及米饭食味关系密切。本文以多个水稻主栽品种为材料,通迆灌浆结实期的人工控温试验、大田长期定位点的施氮处理试验和盆栽条件下的温氮两因素复合处理试验,探讨了水稻灌浆结实期温度对稻米贮藏蛋白含量与组分影响及其有别于氮肥处理效应的差异觃律,幵分析了温度与氮肥两个因素对稻米贮藏蛋白及其组分影响的交互作用特点。结果表明,高温胁迫和增施氮肥均引起水稻籽粒总蛋白及其谷蛋白组分含量(%)的显著增加,但两者对稻米醇溶蛋白影响却存在明显差别。其中,高温处理引起醇溶蛋白含量显著下降,提高稻米谷蛋白/醇溶蛋白比值,而增施氮肥引起稻米谷蛋白和醇溶蛋白含量明显增加,但对谷蛋白/醇溶蛋白比值与贮藏蛋白各亚基的组成比例影响相对较小。在高温处理下,谷蛋白的57kD前体亚基组分含量有所提高,而37kD酸性亚基和22kD碱性亚基随温度处理的差异变化却因品种而异,且高温处理对水稻籽粒蛋白绝对含量(mg grain^–1)的影响程度也进没有其对蛋白相对含量(%)的影响明显。高氮×高温处理组合对稻米总蛋白与谷蛋白含量的影响程度显著大...     

共固定化双酶水解芝麻粕蛋白制备复合氨基酸研究

以海藻酸钠为载体、戊二醛为交联剂共固定化木瓜蛋白酶和中性蛋白酶,选择合适的固定化工艺,研究水解芝麻粕蛋白的优化条件和共固定化双酶的稳定性。结果表明：共固定化双酶的最佳条件为海藻酸钠3 g.dL^-1,戊二醛25 g.L^-1,氯化钙0.2 g.dL^-1,酶活力回收率为51.28%;水解芝麻蛋白的最佳条件为固液比1：25,pH 6.0,温度60℃,时间7 h,加酶量5 g.dL^-1,氨基氮含量最高为20.65 m g.g^-1,共固定化酶重复使用6次,酶活力仍保持50%以上。     

烟草Hsc70-2的克隆及对马铃薯Y病毒侵染烟草的促进作用

【目的】马铃薯Y病毒(Potato virus Y,PVY)是危害烟草、马铃薯、番茄、辣椒等主要农作物的重要病毒,给农业生产带来巨大损失。论文旨在克隆Hsc70-2蛋白在本氏烟(Nicotiana benthamiana)中的基因序列并分析其生物学信息,研究NbHsc70-2蛋白对PVY侵染烟草的影响,为进一步解析PVY的侵染机制提供理论依据。【方法】以本氏烟为材料,克隆NbHsc70-2蛋白的基因编码序列(coding sequence,CDS),利用MEGA 6.0进行多序列比对并构建系统进化树;利用实时荧光定量PCR(qRT-PCR)分析NbHsc70-2在本氏烟各组织中的表达水平;采用在线软件BaCeILo和SignalP 4.0对NbHsc70-2进行生物信息学分析;构建NbHsc70-2-RFP融合蛋白确定该蛋白的亚细胞定位;研究PVY处理对本氏烟叶片中NbHsc70-2的影响;利用激光共聚焦观察PVY-GFP处理对NbHsc70-2-RFP定位的影响;构建NbHsc70-2 VIGS沉默体系及瞬时表达载体,采用qRT-PCR比较NbHsc70-2在沉默/过表达后对PVY表...     

Comparison of blood electrolyte and biochemical parameters between single infections of rotavirus and Cryptosporidium parvum in diarrheic Hanwoo calves

BackgroundNeonatal calf diarrhea is a major problem in the cattle industry worldwide. Rotavirus and Cryptosporidium parvum are the primary causative agents, especially during the first three weeks of the calf’s life.ObjectivesThis study investigated the differences in acid-base, electrolytes, and biochemical parameters of diarrheic calves with infection of either rotavirus or C. parvum.MethodsA total of 61 Korean native calves (≤ 20 days old) were divided into two groups based on rotavirus or C. parvum infections: rotavirus infection (n = 44) and C. parvum infection (n = 17). The calves with at a specific blood pH range (pH 6.92–7.25) were chosen for comparison. The acid-base, electrolyte, chemistry, and serum proteins were analyzed, Further, fecal examinations were performed.ResultsCompared to C. parvum-infected calves, the rotavirus-infected calves showed lower levels of total carbon dioxide, bicarbonate (HCO₃ ⁻), anion gap, total protein, and albumin/globulin ratio, and significantly lower levels of potassium, globulin, and α2-globulin (p < 0.05). The C. parvum-infected calves (r = 0.749) had stronger correlations between pH and HCO₃ ⁻ than the rotavirus-infected calves (r = 0.598). Compared to rotavirus-infected calves, strong correlations between globulin and α2-globulin, α2-globulin and haptoglobin were identified in C. parvum-infected calves.ConclusionsThis study is the first to investigate acid-base, electrolyte, and biochemical parameters in calves in response to infections of rotavirus and C. parvum. Although rotavirus and C. parvum cause malabsorptive and secretory diarrhea in similar-aged calves, blood parameters were different. This would help establish the diagnostic and treatment strategies.     

可可毛色二孢全基因组分泌蛋白的预测及分析

【目的】可可毛色二孢(Lasiodiplodia theobromae)是一种世界性分布的重要植物病原真菌,可引起严重的葡萄溃疡病(Botryosphaeria dieback),影响果木品质并造成巨大的经济损失。本研究预测并分析可可毛色二孢基因组范围内的分泌蛋白,并明确其基本特征,为该病菌分泌蛋白致病机理的研究打下基础。【方法】依据已公布的可可毛色二孢全基因组序列,利用信号肽预测软件SignalP v5.0、跨膜结构分析软件TMHMM v2.0、细胞器定位分析软件ProtComp v9.0、GPI锚定预测软件big-PI Fungal Predictor和亚细胞器定位分析软件TargetP v2.0生物信息学软件对该菌中的典型分泌蛋白进行筛选。对分泌蛋白N端信号肽的长度、氨基酸使用频率及其切割位点进行统计分析。依据蛋白序列的同源性,应用BLASTP程序对分泌组蛋白进行功能注释分析,预测其生物学功能。采用蔗糖酶缺陷的酵母分泌系统,对所选分泌蛋白的信号肽进行活性检测。利用qRT-PCR方法检测所选分泌蛋白基因在可可毛色二孢侵染葡萄中的表达情况。【结果】在可可毛色二孢全基因组编码蛋白中共筛...     

生殖激素产品的应用和发展趋势

生殖激素是调控动物体内复杂生殖过程并影响繁殖性能的一系列的重要激素.近年来,利用生殖激素的批次化生产繁殖技术被广泛使用,提高了母猪的繁殖效率,促进了养猪业向集约化和规模化方向的发展.目前在批次化生产过程中主要采用孕马血清促性腺激素(PMSG)和人绒毛膜促性腺激素(HCG)以及化学类激素进行繁殖调控.但是这些产品的应用仍...     

砂梨扩展蛋白基因cDNA克隆及全序列分析

设计植物EXP扩展蛋白简并引物,以砂梨果实cDNA为模板,克隆得到EXP基因cDNA片段,该片段长465 bp。根据该片段序列,分别设计2条5’和3’末端扩增的特异引物,利用RACE技术,获得了该片段的5’端和3’端序列。用DNAMAN5.22软件对3个序列进行拼接和分析,获得了该基因的cDNA全长,命名为Pyp-EXP。该cDNA全长为1 395 bp,5’端起始密码子ATG始于72 bp,3’端终止子TAG止于830 bp,Ploy+(A)从1 363～1 395 bp。该基因已在GenBank基因数据库注册,注册号为EF602031。Pyp-EXP核苷酸序列有一个759 bp完整的开放阅读框,编码区与西洋梨、苹果、湖北海棠、桃的同源性分别为96%,96%,94%和86%;该cDNA推导编码252个氨基酸,含有1个组氨酸(His_Phe_Asp,HFD)功能域,与西洋梨、苹果、湖北海棠、桃中相应序列同源性分别为98%,97%,95%和93%。该基因的克隆为研究扩展蛋白的时空表达及其在果实发育和成熟过程中的作用奠定了基础。     

Expression of Cdc25a in cross-species hepatocellular carcinoma tissues and its significance

AIM：To study the expression of cell division cycle 25a protein (Cdc25a) in hepatocellular carcinoma (HCC) tissues of different species, including human, rat and tree shrew, and to verify the feasibility of the strategy of cross-species oncogenomics screening. METHODS：Real-time fluorescence quantitative PCR and Western blotting were applied to detect the expression of Cdc25a at mRNA and protein levels in the HCC tissues, corresponding HCC-adjacent liver tissues and normal liver tissues collected from humans, rats and tree shrews. RESULTS：The mRNA expression of Cdc25a in the HCC tissues of humans, rats and tree shrews was higher than that in normal liver tissues (P<0.05). The mRNA levels of Cdc25a in the HCC tissues of humans and rats were higher than those in the corresponding HCC-adjacent liver tissues (P<0.05). The mRNA expression of Cdc25a in the HCC tissues was significantly correlated with the portal vein tumor thrombus, the extrahepatic metastasis and the clinical stage, but was not correlated with the recurrence of tumor, the diameter of tumor, the number of tumor, the level of serum alpha-fetoprotein and the differentiation of tumor. The protein levels of Cdc25a in the HCC tissues of humans, rats and tree shrews were higher than those in the corresponding HCC-adjacent liver tissues and the normal liver tissues. CONCLUSION：Cdc25a may be a particularly crucial molecule for hepatocarcinogenesis. The cross-species oncogenomics screening may represent a feasible and convenient way for identifying key molecules of human HCC.     

Expression of Wnt1 and LGR5 in gastric mucosa with stress ulcer after TBI in rats

AIM To investigate the expression of Wnt1 and LGR5 in gastric mucosa with stress ulcer after traumatic brain injury （TBI） in rats, and to study the relationship between Wnt1/LGR5 expression and stress ulcer after TBI. METHODS Healthy 7-week-old male SD rats （n=30） were randomly divided into 3 groups： sham operation （sham） group, mild TBI （mTBI） group, and severe TBI （sTBI） group. The mTBI and sTBI were induced by electronic cortical contusion impactor. Neurological severity score （NSS） was calculated 24 and 48 h after modeling. Mucosal blood flow in gastric fundus, greater curvature, pylorus and cardia of anesthetized rats 48 h after injury was measured by Doppler flowmeter. All rats were sacrificed, and their gastric tissues were harvested after 48 h and then stained with hematoxylin and eosin. The protein expression of Wnt1 and LGR5 was analyzed by Western blot and immunohistochemistry. RESULTS Compared with sham group, the NSS in mTBI group and sTBI group was significantly increased （P<0.01）. Compared with sham group, the gastric mucosal blood flow of the rats after TBI was significantly decreased （P<0.01）, and the decrease in sTBI group was more significant than that in mTBI group （P<0.05）. The protein expression of Wnt1 and LGR5 in mTBI group was significantly higher than that in sham group （P<0.05）, and that in sTBI group was significantly higher than that in mTBI group （P<0.05）. Normal glandular gastric tissue was observed, and no abnormal change was found in sham group, while the infiltration of inflammatory cells in gastric lamina propria, mucosa and submucosa was obvious in mTBI group and sTBI group. CONCLUSION Traumatic brain injury activates Wnt1 and LGR5 expression to induce inflammatory changes in gastric mucosa, and then induces stress ulcer. This results provides experimental basis for the pathogenesis and treatment of stress ulcer after trauma.     

PVX病毒载体介导2种马铃薯Y病毒科病毒VPg蛋白表达诱导烟草叶片系统坏死的研究

芜菁花叶病毒（Turnip mosaic virus, TuMV）和槟榔坏死环斑病毒（Areca plam necrotic ringspot virus, ANRSV）是马铃薯Y病毒科（Potyviridae）中分别可诱导十字花科作物和槟榔产生坏死病症的2种不同属病毒。VPg（Viral protein genome linked）作为与马铃薯Y病毒科病毒基因组5′端的结合蛋白,在病毒复制、翻译和运动等侵染过程中发挥重要作用。本研究利用In-Fusion克隆策略分别构建了可表达3′端融合Strep蛋白标签序列的ANRSV和TuMV VPg全长编码基因的马铃薯X病毒（Potato virus X, PVX）载体pPVX-VPg-AR和pPVX-VPg-Tu。通过农杆菌渗透注射接种本生烟（Nicotiana benthamiana）,利用RT-PCR、Strep蛋白标签亲和层析和Western blot等方法证明了PVX病毒载体能够介导VPg-AR和VPg-Tu蛋白在本生烟中系统表达,且发现这2种VPg蛋白的过量表达诱导本生烟叶片产生了不同程度的系统性坏死病症。进一步采用DAB和NBT染色,在产生坏死症状叶片中检测到大量活性氧（reactive oxygen species, ROS）的积累。因此,推测VPg可能是TuMV和ANRSV诱发植物病症产生的一个决定因子,为后续研究这2种病毒诱导症状形成机制奠定了基础。