大豆小肽螯合铁的制备工艺及光谱分析研究

通过铁源筛选比较得知,氯化亚铁比较适合与大豆小肽进行螯合反应制备大豆小肽螯合铁,利用响应面法优化了大豆小肽螯合铁的制备工艺,优化结果为:小肽与亚铁盐质量比4∶1,反应pH5.0,反应温度40℃,得到离子螯合率平均值为56.81%,经中试车间生产试制得到大豆小肽螯合铁的得率是78.3%,螯合率为82.39%,检测大豆小肽螯合铁的主要成分中的蛋白含量为78.94%,铁的含量为10.87%。红外和紫外光谱分析检测结果显示:大豆小肽和大豆小肽螯合铁(Fe~(2+))红外吸收峰的强度在不同波长位置上有明显的变化,大豆小肽螯合铁(Fe~(2+))在紫外波长上发生了明显的位移且宽化,表明大豆小肽螯合铁(Fe~(2+))形成了络合物。同时对大豆小肽螯合铁的结构进行了预测。     

不同行株距机插对双季稻产量的影响

引入25 cm行距的插秧机,与生产上常用的30 cm行距插秧机进行小区对比试验,以探讨该型插秧机对乐清市早、晚稻的适应性及配套高产栽培技术。试验结果表明,早稻应用25 cm行距插秧机具有比较明显的增产优势;而连晚杂交稻应用行距25 cm插秧机的增产效果不明显。生产上应根据不同类型水稻的特征特性,正确掌握插秧密度,充分发挥各类插秧机的增产优势。     

Mechanism of JAK2-STAT3 signaling pathway in ischemic postconditio-ning neuroprotection after cerebral ischemia in tree shrews

AIM: To investigate the regulatory effect of JAK2-STAT3 signaling pathway on the neuroprotection of ischemic postconditioning (IPoC) in tree shrews, and to explore the mechanisms of cerebral injury deterioration after inhibiting the JAK2-STAT3 pathway. METHODS: The model of thrombotic cerebral ischemia was induced by photochemical reaction in tree shrews and the IPoC was established at 4 h after ischemia followed by clipping ipsilateral common carotid artery on the ischemia side for 5 min (3 times). After IPoC and intracerebroventricular injection of AG490 (JAK2 inhibitor), the changes of cerebral infarction area were detected by TTC staining, and the histological and ultrastructural changes of cortical neurons were observed under light and electron microscopes, respectively. The protein levels of t-STAT3 and p-STAT3 in the cortical tissue were determined by Western blot. RESULTS: The neuronal pycnosis, mitochondrial swelling and vanish of the mitochondrial cristae were found in cortical cortex, and the infarction area was (24.78±3.30)% at 24 h after cerebral ischemia. Meanwhile, the phosphorylation level of STAT3 protein in the cortical tissue was significantly increased (P<0.01). The cortical neuronal damage and mitochondrial swelling were decreased after IPoC, the area of cerebral infarction was significantly reduced to (17.67±1.83)% (P<0.01), and the phosphorylation level of STAT3 protein was further increased (P<0.01). However, the neuronal damage was aggravated, the infarction area was expanded to (23.85±2.77)%(P<0.05) after treatment with AG490, and the phosphorylation level of STAT3 protein was also significantly reduced (P<0.05). CONCLUSION: IPoC may reduce cerebral injury by regulating the phosphorylation of STAT3 protein, and inhibition of JAK2-STAT3 signaling pathway may counteract the cerebral protective effect of IPoC and aggravate brain injury.     

Effect of GYY4137 on cytosolic lipid decomposition in mouse primary hepatocytes

AIM: To evaluate the effect of GYY4137, a novel hydrogen sulfide (H₂S) donor, on cytosolic lipid decomposition in mouse primary steatosis hepatocytes. METHODS: Oleic acid (OA) was used to induce hepatic steatosis model in vitro. The C57BL/6 mouse primary hepatocytes isolated and cultured by 2-step in situ perfusion were divided into 4 groups:the cells in control group were incubated with normal medium for 54 h; the cells in model group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 for 6 h; the cells in H₂S group or DL-propar-gylglycine (PAG; an inhibitor of cystathione γ-lysase, inhibiting H₂S synthesis) group were incubated with OA at 1.2 mmol/L for 48 h followed by serum-free phenol red-free RPMI-1640 which contained 1 mmol/L GYY4137 or 200 μmol/L PAG for 6 h. The glycerin release and the protein expression of hormone-sensitive lipase (HSL) in the cells were mea-sured. RESULTS: Compared with model group, the glycerin release and the protein expression of phosphorylated HSL (p-HSL) in H₂S group decreased significantly, while those increased significantly in PAG group. CONCLUSION: In steatosis hepatocytes, exogenous H₂S possibly decreases cytosolic lipid decomposition by decreasing the protein level of p-HSL.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

海南水稻生育期的时空变化特征及对气候变暖的响应

基于海南6个农业气象观测站点的水稻生育期资料和1961~2014年历史气象数据,分析了海南不同地区水稻生育期的时空分布特征,以及气候变暖对海南水稻生育期的影响。结果表明,近54 a海南水稻生长季内平均气温呈上升趋势,早稻气候倾向率为0.21~0.3℃/10 a,以苗期最为明显,晚稻为0.18~0.24℃/10 a,以成熟期最为明显。不同地区水稻生育期差异较大,东部地区早于西部地区,早稻播种时间最大相差55 d,早稻收获和晚稻播种最大间隔42 d,降水和高温可能是造成这一差异的主要原因。随着气候变暖,海南大部分地区早稻生育期提前,晚稻生育期延迟,苗期和成熟期持续时间缩短,与该阶段平均气温呈负相关,但全生育期持续时间变异较大,部分站点略有延长。在未来气候变暖背景下,海南早稻播期可适当提前,晚稻播期可适当延迟,并选用耐高温和抗干旱品种,以减轻高温的不利影响。     

基于改进欧拉算法的双吸离心泵泥沙磨损特性研究

固液两相流算法对双吸离心泵泥沙磨损模拟精度有直接影响。采用改进的固液两相流欧拉算法,考虑了相间阻力和泥沙扩散系数两方面因素,对典型悬移质泥沙粒径条件下的双吸离心泵流场进行了数值计算。研究发现,叶片表面湍流强度在头部和尾部较大,可达6%~10%;叶片头部和尾部的颗粒动态尺度大于中部。由湍流强度和颗粒动态尺度组成的湍动尺度效应,在叶片头部和尾部表现强烈,湍动尺度效应使固液相间阻力增大,更有利于颗粒的扩散,避免了颗粒聚集,对大颗粒的作用强于小颗粒。湍动尺度效应导致叶片表面固相体积分数分布范围减小,大颗粒的变化值大于小颗粒,叶片头部和尾部的改变值大于中部,叶片表面的严重磨损部位为叶片工作面尾部的块状磨损区,这比采用传统算法得到的带状磨损区和偏磨区计算结果,更符合离心泵实际磨损情况,考虑湍动尺度效应后得到的磨损率也有所增大。在此基础上,提出了双吸离心泵叶片水力设计和表面喷涂防护原则,为提高双吸离心泵抵抗泥沙磨损能力奠定了基础。     

马铃薯卷叶病毒PLRV RT-LAMP检测方法优化

马铃薯卷叶病毒Potato leafroll virus(PLRV)是目前严重影响马铃薯产量与品质的主要病毒之一,给马铃薯产业造成巨大损失。本研究采用环介导等温核酸扩增(loop-mediated isothermal amplification, LAMP)技术建立PLRV的RT-LAMP检测方法。采取单因素变化试验,对RT-LAMP反应体系中多个因素包括引物组合、温度条件及Mg~(2+)、betaine、Bst 3.0 DNA聚合酶、dNTPs、UNG、SYBR GreenⅠ和引物组合的浓度进行一系列试验和优化。采用RT-PCR检测方法进行平行比对试验,对优化后的RT-LAMP反应体系进行了验证。结果表明,最佳引物组合为P3,最适反应温度62℃,25μL反应体系中,Mg~(2+)、betaine、Bst 3.0 DNA聚合酶和UNG的最佳终浓度分别为4 mmol/L、0 mmol/L、0.64 U/μL和0.08 U/μL,dNTPs的最佳用量为1μL(dATP、dGTP、dCTP各0.4 mmol/L,dUTP 1.2 mmol/L),SYBR GreenⅠ(20×)的最佳用量1μL,primer mix的最佳用量2.5μL(PLRV-FIP/BIP、PLRV-F3/B3和PLRV-LF/LB的浓度分别为0.8、0.2μmol/L和0.6μmol/L),RNA模板1μL(2 ng/μL),加DEPC-H_2O至25μL,反应时间50 min。优化后的RT-LAMP检测结果与RT-PCR一致,且可视化判读结果。因此,建立的PLRV RT-LAMP检测方法为进一步开发RT-LAMP检测试剂盒及其实际应用奠定了基础。     

填闲作物腐解过程及其对后茬冬小麦产量的影响

研究了渭北旱塬地区不同填闲作物（长武怀豆（S）、黑麦草（R）及两者1∶1混合（M））翻压和氮肥水平（0、60、120 kg N·hm^-2）双因素处理下,填闲作物的腐解规律、碳氮释放动态及对后茬冬小麦产量的影响,并对腐解速率与Olsen模型进行了拟合。结果表明：各填闲作物翻压后腐解规律及碳氮释放特征均表现为“前期快-中期慢-后期加快”,填闲作物腐解规律符合Olsen模型,在第276天各处理累积腐解率均达70%以上。在第0~35天,同一施氮处理下,累积腐解率和腐解速率均表现为S>M>R（P<0.05）;第35天,S、M和R各处理干物质累积腐解率分别达到61.9%、55.5%和47.5%;在0~35 d,施氮对S、M的腐解影响不显著,对R影响显著,35 d后氮肥效应逐渐减弱;填闲作物的腐解同时伴随其碳、氮的快速释放,在第21天,S、M和R碳氮残留率分别达到40%、50%和60%左右。平均来看,S的碳氮释放速率显著高于R,与M无显著差异。与裸地对照相比,翻压填闲作物能够显著提高后茬冬小麦产量,其籽粒产量增加10%~35% （P<0.05）,其中翻压长武怀豆低氮处理和混合翻压低氮处理效果最佳。     

氯虫苯甲酰胺和氟虫腈对黏虫细胞色素P450基因表达的影响

为筛选出黏虫Mythimna separata参与杀虫剂解毒代谢的主效细胞色素P450基因,采用叶片浸渍法测定了用于处理黏虫3龄幼虫的亚致死浓度,通过构建转录组测序文库并结合数字基因表达(digital gene expression,DGE)对不同处理的黏虫进行测序,并运用实时荧光定量PCR(realtime quantitative polymerase chain reaction,RT-qPCR)技术验证12个P450基因的表达情况。结果表明,用于处理黏虫的氯虫苯甲酰胺和氟虫腈亚致死浓度LC_(10)分别为0.15、13.66 mg/L;对照样品、氟虫腈处理样品和氯虫苯甲酰胺处理样品分别获得59 521 504、64 838 148和41 722 990个原始序列数据,分别获得57 441 216、62 368 912和40 285 164个过滤后的序列数据;过滤后的序列长度分别为8.62、9.36和6.04 G;碱基错误率均为0.02%;Phred数值大于20、30的碱基占总碱基的百分比均高于90.59%;鸟嘌呤+胞嘧啶(guanine cytosine,GC)含量分别为47.16%、48.94%和47.55%,表明转录组测序质量较高;黏虫受氯虫苯甲酰胺胁迫后,29个P450基因表达量上调,27个P450基因表达量下调;黏虫受氟虫腈胁迫后,23个P450基因表达量上调,26个P450基因表达量下调;12个P450基因表达量的RT-qPCR技术检测结果与DGE测序文库显示的结果基本一致。