环介导等温扩增联合横向流动试纸条可视化检测海豚链球菌方法的建立

海豚链球菌(Streptococcus iniae)是一种革兰氏阳性球菌,呈β溶血,可感染多种淡水和海洋鱼类。本研究利用环介导等温扩增技术(loop-mediated isothermal amplification,LAMP)进行核酸扩增,通过横向流动试纸条方法 (lateral flow dipstick,LFD)实现检测,建立了一种可应用于海豚链球菌快速检测的LAMP-LFD技术。该技术以海豚链球菌促旋酶B亚单位(gyrase subunit B,gyr B)基因为检测靶标,设计3对引物进行由生物素标记的LAMP扩增反应,产物经异硫氰酸荧光素(fluorescein isothiocyanate,FITC)标记的探针杂交后,在LFD上完成检测。经优化的核酸扩增最适条件为65℃反应30 min,在此条件下,阳性扩增起始时间与模板浓度之间呈典型的线性相关性。从核酸扩增反应开始到LFD显色,整个检测时程只需40 min左右,比常规PCR技术缩短约2 h。LAMP-LFD能特异性地检出海豚链球菌,针对病原纯培养物的检测灵敏度为8.70×101cfu/m L,是LAMP检测的10倍、常规PCR检测的100倍。以灵敏度浓度(8.70×101cfu/m L)的海豚链球菌基因组DNA为模板进行的LAMP-LFD结果显示该方法具有良好的重复性。针对人工污染花鲈(Lateolabrax japonicus)肝组织的检测灵敏度为4.35×103cfu/m L,同样为常规PCR检测方法的100倍。利用本方法可成功从患病花鲈的组织样品中检测出海豚链球菌,检测结果与常规的细菌分离鉴定方法结果一致。因此,利用LAMP-LFD能特异、准确、高效地检测出海豚链球菌,而且操作简单、费用低、耗时短,有望成为海豚链球菌的常规检测方法。     

适宜气调包装延缓冷藏鲳鱼品质变化延长货架期(英文)

该文主要研究了不同气体比例组分的气调包装处理对冷藏鲳鱼贮藏期间品质变化及货架期的影响,从中找出能使鲳鱼冷藏货架期延长的适宜气体比例。将新鲜鲳鱼以冰藏方式快速运至实验室,将样品经去头、去尾、去内脏处理后,分别置于3种气体组分条件(MAP1:20%CO2/80%N2,MAP2:50%CO2/50%N2,MAP3:80%CO2/20%N2)的气调包装袋内冷藏(4±1)℃保鲜,以空气包装样品作为冷藏对照组(AIR),其中包装袋内的气体与样品体积比为3∶1,在12 d内隔天测定各组样品的感官指标、微生物指标(菌落总数)与化学指标(挥发性盐基氮值、K值、三甲胺值与p H值)变化。冷藏后的第4天,对照组与气调处理组的感官分值、菌落总数、挥发性盐基氮值与三甲胺值差异显著(P0.05),冷藏对照组样品的各项指标首先达到新鲜限值,其冷藏货架期仅为4-5d。经过气调处理后的鲳鱼样品,其菌落总数达到二级鲜度的速度明显缓于冷藏对照组,其他各项指标的变化也证实了气调包装的潜在优点。其中,80%CO2+20%N2处理组能明显延缓样品p H值、挥发性盐基氮值、菌落总数、三甲胺值与K值的升高(P0.05),较冷藏对照组相比,其冷藏货架期能延长6~8 d。同时,气调处理组贮藏期间的p H值未呈现先降后升的变化规律,表明p H值不适用于评价气调包装鲳鱼贮藏期间的品质变化。气调包装样品的菌落总数在贮藏前期略有下降,表明CO2对样品菌落总数的升高有较好的抑制作用。在CO2≤80%条件下,各项微生物与理化指标的测定结果显示出不同气体比例的气调包装处理对冷藏鲳鱼的保鲜效果,CO2浓度越高,其保鲜效果越显著,气调处理能使鲳鱼样品的冷藏保鲜期由4 d延长至6-12 d,其中80%CO2+20%N2的气调处理组对鲳鱼样品的冷藏货架期延长效果较佳。     

马尔尼菲青霉菌内源启动子驱动的苯菌灵抗性基因盒构建及其应用

马尔尼菲青霉菌是一种区域性流行于东南亚及中国南部地区的条件致病性温度依赖性双相型病原真菌,它能对免疫功能低下者造成致命的全身性感染,25℃时呈现菌丝状生长,而在37℃或在宿主体内为酵母相生长。为深入研究马尔尼菲青霉菌致病性和双相转化的分子机制,原有的遗传转化筛选标记基因已不能满足研究需要。为此,我们通过用马尔尼菲青霉菌微管蛋白β亚基基因启动子替换粗脉孢菌苯菌灵抗性基因的启动子,构建了一个苯菌灵抗性基因盒,并成功地将其运用于马尔尼菲青霉菌的遗传转化研究中。     

Utilization of biochar impregnated with anaerobically digested slurry as slow‐release fertilizer

We examined the possibility of an environment‐friendly slow‐release fertilizer (SRF) made of biochar impregnated by anaerobically digested slurry. The biochar materials were produced from three types of feedstocks (orange peel, residual wood, water‐treatment sludge) at different temperatures of 300°C, 500°C, and 700°C via pyrolysis. The release behaviors of the water‐soluble K⁺, Ca²⁺, and Mg²⁺ were similar for all impregnated biochars and the commercial SRF used. The water‐retention capacity was greatly improved by mixing the biochar‐SRF with the soil. The yield of lettuce was lower for the biochar‐SRF applications of 3.7 to 34.2 t ha^–1 than for the commercial SRF application of 51.4 t ha^–1. This might be due to excessive increase of soil pH for the biochar‐SRF application. Based on these results, the authors concluded that the biochar impregnated with nutrients could become an effective slow‐release K⁺ fertilizer.     

Tomato fruit quality as influenced by the interactions between agricultural techniques and harvesting period

Tomato (Solanum lycopersicum Mill) is an important crop in terms of its economic and nutritional value. Many factors, including cultivar, climate, geography, geochemistry, and agricultural practice, can affect its nutrient concentrations. An HJ‐biplot study was performed to examine the effects of cultivar (Dorothy, Boludo, Dominique, Thomas, and Dunkan), agricultural practices, climatic factors, and their interactions. Significant differences were analyzed using a one‐way ANOVA. All samples were collected and assayed at the same degree of ripeness. In the conventional and organic tomato samples, those harvested from December to April had the highest concentrations of fructose, glucose, citric acid, malic acid, ascorbic acid, protein, Na, and Mg, while those harvested in October had the highest concentrations of lycopene and hydroxycinnamic acid. There were high concentrations of Ca, P, Zn, and Cu in the no‐soil tomatoes. Conventional and organic cultivation practices showed similar results with respect to the collection period, both presenting high organic compound concentrations, while high mineral concentrations seemed to correspond to the no‐soil practice. No clear pattern was observed among the different cultivars, perhaps due to all the samples having been collected at the same degree of ripeness.     

Effect of mineral nitrogen fertilizer forms on N2O emissions from arable soils in winter wheat production

Nitrogen fertilizers are supposed to be a major source of nitrous oxide (N₂O) emissions from arable soils. The objective of this study was to compare the effect of N forms on N₂O emissions from arable fields cropped with winter wheat (Triticum aestivum L.). In three field trials in North‐West Germany (two trials in 2011/2012, one trial in 2012/2013), direct N₂O emissions during a one‐year measurement period, starting after application of either urea, ammonium sulfate (AS) or calcium ammonium nitrate (CAN), were compared at an application rate of 220 kg N ha^?1. During the growth season (March to August) of winter wheat, N₂O emission rates were significantly higher in all three field experiments and in all treatments receiving N fertilizer than from the non‐fertilized treatments (control). At two of the three sites, cumulative N₂O emissions from N fertilizer decreased in the order of urea > AS > CAN, with emissions ranging from 522–617 g N ha^?1 (0.24–0.28% of applied fertilizer) for urea, 368–554 g N ha^?1 (0.17–0.25%) for AS, and 242–264 g N ha^?1 (0.11–0.12%) for CAN during March to August. These results suggest that mineral nitrogen forms can differ in N₂O emissions during the growth period of winter wheat. Strong variations in the seasonal dynamics of N₂O emissions between sites were observed which could partly be related to weather events (e.g., precipitation). Between harvest and the following spring (post‐harvest period) no significant differences in N₂O emissions between fertilized and non‐fertilized treatments were detected on two of three fields. Only on one site post‐harvest emissions from the AS treatment were significantly higher than all other fertilizer forms as well as compared to the control treatment. The cumulative one‐year emissions varied depending on fertilizer form across the three field sites from 0.05% to 0.51% with one exception at one field site (AS: 0.94%). The calculated overall fertilizer induced emission averaged for the three fields was 0.38% which was only about 1/3 of the IPCC default value of 1.0%.     

蜡样芽孢杆菌及其生物被膜对有机酸及乙醇的抗性比较

为比较蜡样芽孢杆菌及其生物被膜对环境压力的抗性,该文主要研究了4种有机酸（乙酸、柠檬酸、乳酸和苹果酸）和乙醇对蜡样芽孢杆菌及其生物被膜存活率的影响。结果表明：生物被膜态蜡样芽孢杆菌对乙酸的抗性高于浮游态菌。扫描电镜结果显示,经乙酸处理后,浮游态蜡样芽孢杆菌的细胞表面严重受损,而生物被膜态菌的表面形态未发生明显变化。在柠檬酸、乳酸、苹果酸和乙醇存在的条件下,生物被膜态蜡样芽孢杆菌比浮游态菌表现出更强的压力抗性,特别是在高浓度（有机酸16%～20%,乙醇50%～60%）的情况下,该现象尤为显著。因此,在食品工业中控制被膜态蜡样芽孢杆菌对预防和阻止食品腐败非常重要。该研究结果为实际生产中有效控制蜡样芽孢杆菌及其生物被膜的形成提供参考。     

小麦耐热及热敏感基因型在高温胁迫下膜透性及膜脂组分的差异

高温逆境下,植物膜透性的增加导致细胞代谢紊乱并诱导热激基因表达.脂肪酸是构成生物膜的主要物质,饱和脂肪酸的含量及饱和程度高,有利于保持膜在高温时的流动性和稳定性.本研究利用小麦(Triticum aestivum)耐热基因型TAM107和热敏感基因型中国春(CS)为对象,研究了高温胁迫对膜透性、叶绿素荧光参数Fv/Fm、膜脂组分及其相关脂肪酸去饱和酶基因(FAD7)表达的影响,结果表明,高温胁迫下,两基因型膜透性均增加,膜脂中的三烯脂肪酸含量下降,但TAM107变化幅度明显高于中国春,在脂肪酸水平上说明耐热小麦品种的膜系统较热敏感品种对高温逆境有较强的耐受性;中国春比TAM107中FAD7的表达对高温更加敏感,这直接影响了热胁迫前后膜脂中三烯脂肪酸含量的变化,在转录水平上说明了耐热小麦基因型较热敏感基因型对高温逆境有较强的耐受性.三烯脂肪酸含量在热胁迫前后的变化可作为一种新的耐热鉴定指标.     

外源NO对Ca(NO3)2胁迫下黄瓜幼苗生长和抗氧化系统的影响

本研究以黄瓜为试材,采用营养液水培,研究了外源NO对Ca(NO3)2胁迫下黄瓜幼苗生长和抗氧化酶系的影响.结果显示,在70mmol·L-1的Ca(NO3)2胁迫下,植株生长受到抑制,干重和叶绿素含量显著下降,叶片和根系抗氧化酶SOD、POD、CAT和APX活性均升高,膜脂过氧化产物MDA及可溶性 蛋白含量增加.而叶面喷施NO供体SNP显著缓解了Ca(NO3)2胁迫对黄瓜幼苗生长的抑制,植株干重显著增加.同时,抗氧化酶活性、可溶性蛋白和叶绿素含量也有不同程度的上升,MDA含量显著下降.以上表明,NO能够增强黄瓜幼苗对Ca(NO3)2胁迫的耐性.喷施SNP对正常营养液培养的黄瓜幼苗无显著影响.     

中国区域畜禽粪便能源潜力及总量控制研究

为了评估中国畜禽粪便资源总量及其对环境的影响,以环保部、统计局和农业部发布的区域畜禽产排污系数为基础,利用2010年的统计数据,研究了中国及各省的畜禽粪便资源总量、能源潜力及农地的氮磷负荷,并以欧盟的农地氮磷施用标准对中国畜禽养殖的环境容量和污染风险进行了初步评估。结果表明,2010年,中国畜禽粪便总量达22.35亿t,可产沼气1072.75亿m3,山东等6省市粪便资源超过1.00亿t;全国单位面积农地氮磷平均负荷为43.73kg/hm2(TN)和9.16kg/hm2(TP),北京等6省市农地氮磷负荷超标;全国畜禽养殖环境容量为129.56亿头猪当量(以N为基准),159.74亿头猪当量(以P为基准),实际养殖总量约占环境容量的1/4,考虑化肥施用的影响,约有20个省超过本省50%环境容量。研究结果为区域畜禽养殖总量控制、合理布局和粪污的综合利用提供决策依据。