滴灌施肥下水肥用量对温室土壤硝态氮残留的影响

【目的】通过水肥管理达到减少温室土壤硝态氮残留、维持土壤质量的目的,探求温室土壤硝态氮残留与水肥用量的关系。【方法】在滴灌施肥条件下,以灌水量和氮、磷、钾及有机肥用量为试验因素,根据当地日光温室番茄长季节栽培实际中的水肥用量,设计各试验因子的水肥水平,采用五元二次通用旋转组合设计进行试验。拉秧后测定耕层土壤硝态氮量,建立土壤硝态氮量与水肥因子间的数学模型,据此分析了各单因子效应及二因素的耦合效应。【结果】施氮量对土壤硝态氮残留量影响最大,施磷量、灌水量和施钾量次之,有机肥用量最小。当其他因子为0水平时,土壤硝态氮残留量随氮肥用量的增多而增加,随施磷量呈开口向上的抛物线变化,随灌水量、施钾量以及有机肥用量呈开口向下的抛物线变化。灌水量及氮、磷、钾和有机肥用量对土壤硝态氮残留产生的影响程度随其他因子的水平而变,存在明显交互作用。模型寻优显示:灌水量455.1～471.5 mm,施氮量532.3～586.5 kg/hm2,施磷量420.8～466.4 kg/hm2,施钾量646.1～723.5 kg/hm2,有机肥用量25.6～27.9 t/hm2,耕层土壤硝态氮量可维持在100～150 mg/kg的较低水平。【结论】温室菜地土壤硝态氮残留量相对较大,可以通过优化水肥用量来减少土壤硝态氮的残留,故在滴灌施肥条件下仍需严格控制水肥用量。     

温室整体性能模糊综合评判方法及软件实现

为解决温室整体性能评价受多因素影响以及不可能完全定量化评价等问题,分析归纳了影响温室整体性能的因素,提出了温室整体性能评价的指标体系.采用模糊数学方法,建立了温室整体性能评价的二级模糊综合评判数学模型,并开发了相应的计算机软件来实现整个评价过程,使该方法科学合理、简便实用.     

Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

Mitochondrial DNA deletion of the brain tissue of aged rats with learning and memory deficit

AIM and METHODS: The ratio of mitochondrial DNA (mtDNA) deletion was measured to find the relationship between mtDNA deletion and aged learning and memory deficit. The aged rats were divided into two groups, aged learning and memory deficit group and aged learning and memory normal group. The ratio of mtDNA deletion was measured by dilution polymerase chain reaction. RESULTS: There are deleted mtDNA (about 4834 bp) in the cerebral cortex, hippocampus and cerebellum of both young and aged rats. The ratios of deleted mtDNA were similar in the cerebral cortex,hippocampus and cerebellum of young rats (about 0.00018%). The ratio mtDNA of aged learning and memory normal rats had increased by five-fold in the cerebral cortex and hippocampus, or one-fold in the cerebellum over young rats. The ratio of aged learning and memory dificit rats had increased by one-fold in the cerebral cortex or 0.8-fold in the hippocampus or two-fold in the cerebellum over aged learning and memory normal rats.CONCLUSIONS: There was really the increase of mtDNA in aging rat brain. And this increase was double in amount in aged learning and memory deficit rats compared to the normal learning and memory aged rats. It is suggested that the mtDNA deletions in the brain regions associated with learning and memory may be contributed to the cellular and molecular mechanism of learning and memory deicit with aged rats.     

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。     

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.     

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.     

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。     

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。