基于qPCR和LAMP技术的马铃薯晚疫病菌快速检测方法

马铃薯晚疫病菌（Phytophthora infestans）能侵染多种茄科植物,它引起的马铃薯晚疫病,是马铃薯生产中的第一大病害。为了开发能在田间快速检测马铃薯晚疫病病原的方法,利用P. infestans T30-4基因组测序数据的contig 1.18131,设计qPCR和LAMP引物,优化扩增条件后得到引物的特异性和灵敏度,最后通过检测田间收获薯块,比较形态学传统方法、qPCR及LAMP的差异。特异性检测结果发现,qPCR和LAMP仅在含有P. infestans DNA模板的体系有阳性扩增,在寄主和其他微生物DNA中均无扩增;在优化的条件下,qPCR和LAMP的检测下限可达1×10 ^-6ng/μL,在有寄主和其他微生物DNA存在的条件下,引物的灵敏度没有显著差异。利用两种快速方法对在大理、丽江及昆明3个地区田间收获薯块上检测发现,qPCR和LAMP方法得到的检出率差异极为不显著（P=0.420）,两种快速检测方法和形态学鉴定方法检出率差异极显著（P=0.009）。在大理、丽江及昆明3个地区的薯块中,两种分子检测方法检出率均比形态学方法高。其中,qPCR检测方法比形态学方法分别提高了12.00%、2.00%、8.70%;LAMP检测方法比形态学方法分别提高了11.30%、2.00%、8.70%。     

滴灌下生物质改良材料对盐渍土水盐氮运移的调控效应

为探究生物质改良材料对滴灌盐渍土水、盐、肥运移过程的调控效应，采用土箱模拟试验，研究了水肥一体化滴灌条件下，生物炭和腐殖酸两种改良材料对盐渍土水、盐、氮运移和再分布过程及其时空分布特征的影响规律。结果表明：在滴灌条件下，盐渍土壤水盐的时空动态变化表现出明显的水分入渗驱动的盐分运移过程和蒸发扩散驱动的水盐再分布过程；铵态氮含量在时间上表现出先增大、后减小的变化趋势，在空间上的运移再分布特征较弱；硝态氮含量初始时空分布表现出与水盐相似的运移特征，受铵态氮硝化作用的多重影响，后期空间分布与铵态氮空间分布相似；生物炭通过提高土壤饱和导水率，增大了入渗阶段土壤水、盐、氮的运移速率和分布范围；腐殖酸通过提高土壤田间持水率增大了再分布过程土壤水、盐、氮的分布范围和强度，同时其对尿素的水解和硝化过程表现出更强的抑制效果。应用生物质改良材料在改变土壤物理性状进而调控滴灌土壤水盐运移的同时，还影响土壤氮素转化运移过程及其分布，这为水肥一体化滴灌盐渍农田的节水、控盐、减肥治理提供了理论基础。     

基于土地利用变化的河北省坝上地区景观生态风险评价

[目的]对河北省坝上地区近40 a来的土地利用动态变化和生态风险进行分析评价并对未来趋势作出预测,为该地区生态建设和治理、可持续发展提供科学依据。[方法]基于坝上地区1980—2018年5期土地利用数据以及通过土地转移矩阵、空间相关性分析等方法揭示和预测该区1980—2026年的土地利用变化特征并评估该区生态风险水平。[结果](1)整个研究期间,坝上地区土地利用类型以耕地为主,所占比例近50%,其中,1980—2018年,耕地、林地扩张面积均超过300 km~2,草地减少近616.60 km~2,水域面积缩减36.04%,其中耕地、林地、草地之间的互相流转程度较为剧烈,空间变化上表现为各地类的重心在2000—2010年明显迁移。(2)1980—2026年,坝上地区6个时期内生态风险值全局空间自相关Moran’s I指数均在0.500左右,其空间分布表现出较高的趋同集聚性。(3)近40 a来,坝上生态风险水平升至为高风险级,其区域增加了123.22 km~2,较高风险区域分布在城镇地区,据CA-Markov模型预测,未来坝上地区中等及中等以上风险区域持续扩张,丰宁县和围场县将分别出现小规模高风险区和较高风险区。[结论](1)近40 a来坝上地区草地退化严重,水域面积显著减少,原因系安固里淖干涸所致。(2)该区生态风险水平与土地格局分布具有较强相关性,且在未来会继续升高。     

扬州市稻田高效种养模式应用现状及发展对策

近年来,扬州市根据国家农业供给侧结构性调整要求,充分利用自然资源禀赋,开展了多种形式稻田种养模式的示范与推广。本文阐述了扬州市稻鸭共作、稻虾共作以及稻田改作荷藕-虾共作等高效种养模式的应用现状,以单纯种稻为对照,比较了各种模式的生产效益;分析了稻田高效种养模式示范推广过程中存在的主要问题,并提出了促进稻田高效种养模式可持续发展的若干对策。     

油菜覆膜打孔穴播机打孔装置设计与试验

为实现油菜等小粒径作物覆膜种植中膜上均匀打孔的功能,针对传统膜上成穴装置结构庞大复杂、工作时易黏土挑种及撕挑地膜等问题,设计了一种法兰式滚轮与螺纹式圆锥型锥钉组合式结构的打孔装置,确定了其主要结构参数范围;构建了打孔装置运动学模型,分析了打孔锥钉关键点的运动轨迹,确定了膜上打孔过程,并基于轨迹方程分析了膜孔尺寸参数;运用ADAMS运动学仿真,采用四因素三水平正交试验方法,以打孔锥钉顶角、打孔锥钉直径、打孔滚轮半径、机组前进速度为试验因素,以膜孔长度、膜孔间距偏差为试验考核指标,进行了打孔装置结构和运动参数的仿真试验。仿真结果表明:影响膜孔长度的因素主次顺序为打孔滚轮半径、打孔锥钉顶角、打孔锥钉直径、机组前进速度;影响膜孔间距偏差的因素主次顺序为打孔滚轮半径、机组前进速度、打孔锥钉顶角、打孔锥钉直径;基于参数优化,获得较优参数组合为:打孔锥钉顶角53°、打孔锥钉直径16 mm、打孔滚轮半径65 mm、机组前进速度4 km/h。以打孔装置较优结构参数组合进行了田间验证试验,结果表明:打孔装置所打膜孔形状较规则,普遍呈类圆形状,膜孔长度均在18 mm以上,膜孔间距较为均匀,与仿真结果基本一致;各行膜孔长度一致性变异系数为4.98%,各行膜孔间距均匀性变异系数为3.44%。结果表明试验参数组合选取合理,打孔装置符合设计要求。     

Comparison of the microbial communities of alpacas and sheep fed diets with three different ratios of corn stalk to concentrate

The objective of this study was to investigate the characteristics of ruminal microbial communities of alpacas (Lama pacos) and sheep (Ovis aries) fed three diets with varying ratios of roughage (corn stalk) to concentrate, 3:7 (LS), 5:5 (MS) and 7:3 (HS). Six alpacas (one-year-old and weighing 29.5 ± 7.1 kg) and six sheep (one-year-old and weighing 27.9 ± 2.7 kg) were used in this study, in a replicated 3 × 3 Latin square experiment. Total protozoa concentration was determined under the microscope; total fungi and methanogens were assessed using quantitative polymerase chain reaction and expressed as a percentage of total bacterial 16S rRNA gene copies; bacterial communities were investigated by targeted 16S rRNA gene (V3–V4 region) sequencing. The percentage of fungi was significantly higher in alpacas than in sheep under the LS diet, while the concentration of protozoa was significantly lower in alpacas under HS, MS and LS diets. The alpha diversity including Shannon, Chao l and ACE indices of bacterial communities was higher in alpacas than in sheep, under the LS diet. A total of 299 genera belonging to 22 phyla were observed in the forestomach of alpaca and sheep, with Bacteroidetes and Firmicutes dominating both animal species. Phyla Armatimonadetes and Fusobacteria, as well as 64 genera, were detected only in alpacas, whereas phyla Acidobacteria and Nitrospira, as well as 44 genera, were found only in sheep. The abundance of cellulolytic bacteria, including Butyrivibrio and Pseudobutyrivibrio, was higher in alpacas than in sheep under all three diets. These differences in the forestomach microbial communities partly explained why alpacas displayed a higher poor-quality roughage digestibility, and a lower methane production. Results also revealed that the adverse effects of high-concentrate diets (70%) were lesser in alpacas than in sheep.     

水稻OsUBA基因的表达及其在促进种子萌发和开花中的功能

自噬是将功能异常或不需要的胞内组分降解的细胞学过程,广泛参与真核生物的生长发育过程、对营养缺乏的响应及生物/非生物胁迫反应。NBR1 (Next to BRCA1 gene 1, NBR1)是在植物中发现的最重要的自噬受体,但有关植物NBR1类自噬受体的研究较少,水稻中此类蛋白的研究还是空白。本文通过RT-PCR方法,从水稻日本晴幼苗的cDNA中克隆到一个含有泛素相关结构域(Ubiquitinassociated,UBA)的基因,将其命名为OsUBA。OsUBA的开放阅读框长2538 bp,编码845个氨基酸残基。OsUBA属于水稻中的NBR1类蛋白。OsUBA的启动子区有多个与光、逆境胁迫及激素反应相关的元件; OsUBA基因在水稻花药、正在萌发的种子以及根中的表达量较高,在茎和叶中也有表达; 200μmol L~(–1) ABA处理显著抑制OsUBA的表达,100μmol L~(–1) GA处理后OsUBA的表达略有升高。对OsUBA过表达水稻株系的研究表明,转基因水稻种子的萌发比野生型更快, ABA (3μmol L~(–1))处理显著抑制OsUBA过表达水稻株系种子的萌发, GA (100μmol L~(–1))处理对OsUBA过表达水稻株系种子的萌发略有促进;OsUBA过表达水稻株系的开花时间较野生型明显提前。这些结果表明,水稻NBR1蛋白基因OsUBA的表达和功能可能与对开花时间和种子萌发的调控以及生物/非生物胁迫反应有关。     

食品安全国家标准GB 2763—2016对葡萄的新要求

《食品安全国家标准 食品中农药最大残留限量》(GB 2763—2016)对食品中农药残留的种类、最大残留限量及检测方法均提出了较多的新要求。对新标准中涉及葡萄或浆果类水果规定的变化进行了归纳,并与国际上的相关标准进行对比,旨在为有关产业的发展提供参考。     

山下黑猪与鲁莱黑猪正反交F1代杂种优势的估计

旨在估计山下黑猪和鲁莱黑猪正反交F₁代杂种优势，筛选出合适的杂交方式，以提高优质肉猪的生产效率，满足人们对于优质猪肉的需求。本研究在山下黑猪（164头）和鲁莱黑猪（69头）及其正（6头山下黑猪♂×25头鲁莱黑猪♀）、反（3头鲁莱黑猪♂×35头山下黑猪♀）交4个群体中测定了生长肥育、体尺外貌（75～110 kg）、繁殖和胴体肉质（90～115 kg）4大类共43个性状，比较了这些性状在群体间的差异，并估计了正反交F₁代的杂种优势。结果表明，山下黑猪的生长肥育和体尺性状较好，鲁莱黑猪的繁殖和肉质性状较好，正反交群体介于它们之间。正交群体的繁殖性能优于反交群体，但生长肥育性能比反交差。由于这4个群体的母猪都是纯种，因此繁殖性状没有表现出明显杂种优势。除膘厚、胸椎数、45 min pH和剪切力无显著的杂种优劣势外，大部分胴体性状和肉质性状有明显的杂种劣势。生长肥育性状的杂种优势在正反交群体中的表现不一致，反交群体表现出显著杂种优势，而正交则无明显的杂种优势。本研究估计了山下黑猪和鲁莱黑猪正反交的杂种优势，为筛选山下黑猪和鲁莱黑猪的最佳杂交方式奠定基础。     

Effect of miRNA-33 on ABCA1/ABCG1 expression in RAW264.7 macrophage-derived foam cells after Hcy treatment

AIM:To study whether homocysteine (Hcy) inhibits the expression of ATP-binding cassette transporter A1 (ABCA1) and ATP-binding cassette transporter G1 (ABCG1) by microRNA-33 (miRNA-33) signaling, and reduces the efficiency of reverse cholesterol transport (RCT).METHODS:RAW264.7 macrophages were induced by oxidized low-density lipoprotein (ox-LDL) to establish foam cell model. Oil red O staining was used to determine whether the model was established successfully. miRNA-33 mimics and miRNA-33 inhibitor were transfected into the cells by Lipofectamine 2000, and the cells were exposed to Hcy at concentration of 5 mmol/L for 24 h. The intracellular lipid droplets were observed by Oil red O staining. The expression of ABCA1 and ABCG1 at mRNA and protein levels was determined by real-time PCR and Western blot. The cellular cholesterol content was analyzed by HPLC, and effluent rate of cholesterol was detected by the method of liquid scintillation counting.RESULTS:Compared with blank control group, the lipid content in miRNA-33 mimics group was increased, and the expression of ABCA1 and ABCG1 at mRNA and protein levels was decreased (P<0.05). The intracellular cholesterol content was increased gradually (P<0.05), and the cellular cholesterol efflux rate was gradually decreased (P<0.05) in miRNA-33 mimics group. Compared with blank control group, the testing results in miRNA-33 inhibitor group were the opposition of those in miRNA-33 mimics group (P<0.05). No diffe-rence of the above indexes among blank control group, miRNA-33 mimics-NC group and miRNA-33 inhibitor-NC group was observed.CONCLUSION:Hcy inhibits the mRNA and protein expression of ABCA1 and ABCG1 through miRNA-33 signaling, and reduces the efficiency of RCT in RAW264.7 macrophage-derived foam cells.