数字PCR技术在动物疫病检测中的应用

为了支持动物疫病流行地区的控制及消除计划,以及在动物疫病非流行地区进行有效筛查,必须使用更加准确灵敏的诊断手段。数字PCR可实现绝对定量及痕量核酸的检测,与传统检测方法相比具有不依赖参考物或标准品、对抑制剂具有较高耐受性、灵敏度和精准度更高的优点。数字PCR作为一种定量分析的核酸检测新方法,在动物疫病检测中得到了应用,围绕数字PCR技术的原理、应用、存在的问题及发展趋势等进行综述及分析,为数字PCR在动物疫病检测中的进一步应用提供参考。     

浙江省食用菌产业发展历史与建议

总结回顾浙江省食用菌发展的历史和取得的成就,指出食用菌是浙江省农业十大主导优势产业之一,食用菌栽培种类全,分布区域集中,规模化生产水平较高;在科学技术进步、农业丰收、技术改造、品牌建设、国家地理标志等方面获得了诸多成果与荣誉,涌现出一大批优秀的行业先锋;最早提出并制定一系列食用菌标准化生产规范,并注重“菌文化”的传承。浙江食用菌面临的主要困难有:发展动力减弱,生产规模下滑;食用菌产业科技支撑不足,缺少自主核心技术;品牌以及消费宣传不够,缺乏正确的饮食引导。提出具体发展建议。     

重庆烟区硅酸盐细菌的分离鉴定及生理生化特性

采用土壤稀释平板法从重庆市彭水县的植烟土壤样品中分离出8株具有解钾能力的细菌,通过测定其对含钾矿石中有效钾的释放能力,筛选出两株具有较强的解钾能力的硅酸盐细菌,运用分子生物学方法对这两株硅酸盐细菌的16~23S rDNA序列进行扩增并鉴定。结果表明,一株细菌属于胶质类芽孢杆菌(Paenibacillus mucilaginosus),另一株细菌属于短短芽孢杆菌(Brevibacillus brevis),并且研究了这两株细菌的生理生化特性。     

Variations of SOC and MBC observed in an incubated brown loam soil managed under different tillage systems for 12 years

To assess changes in organic carbon pools, an incubation experiment was conducted under different temperatures and field moisture capacity (FMC) on a brown loam soil from three tillage practices used for 12 years: no‐till (NT), subsoiling (ST) and conventional tillage (CT). Total microbial respiration was measured for incubated soil with and without the input of straw. Results indicated that soil organic carbon (SOC) and microbial biomass carbon (MBC) under ST, NT and CT was higher in soil with straw input than that without, while the microbial quotient (MQ or MBC: SOC) and metabolic quotient (qCO₂) content under CT followed the opposite trend. Lower temperature, lower moisture and with straw input contributed to the increases in SOC concentration, especially under NT and ST systems. The SOC concentrations under ST, with temperatures of 30 and 35°C after incubation at 55% FMC, were greater than those under CT by 28.4% and 30.6%, respectively. The increase in MBC was highest at 35°C for 55%, 65% and 75% FMC; in soil under ST, MBC was greater than that under CT by 199.3%, 50.7% and 23.8%, respectively. At 30°C, the lower qCO₂ was obtained in soil incubated under NT and ST. The highest MQ among three tillage practices was measured under ST at 55% FMC, NT at 65% FMC and CT at 75% FMC with straw input. These data indicate the benefits of enhancing the MQ; the low FMC was beneficial to ST treatment. Under higher temperature and drought stress conditions, the adaptive capacity of ST and NT is better than that of CT.     

A型流感病毒分型基因芯片在流感病毒各亚型监测中的应用

为验证自行研制的A型流感病毒分型基因芯片方法在实际中的应用效果,了解A型流感病毒各亚型在我国不同地区和不同宿主中的分布情况,对我国20个地区、不同宿主源(鸡、鸭、鹌鹑、鸽子、猪、人等)拭子和组织样品共16 852份进行了检测。利用自行研制的A型流感病毒分型基因芯片方法,对样品进行RNA提取、多重不对称RT-PCR扩增、芯片杂交、洗涤和扫描分析;同时将基因芯片检测阳性的样品,用普通RT-PCR扩增后进行序列测定。结果显示:利用该方法检出阳性样品2 582份,总检出率为15.32%(2 582/16 852),共检测到H1N1、H2N9、H3N2、H4N6、H5N1、H7N1、H7N9、H9N2、H10N5、H16N3等10个亚型,且均与阳性样品测序结果一致。结果表明,该方法可准确检测不同地区、不同宿主中的A型流感病毒不同亚型。同时,该方法能在同一张芯片上准确鉴定A型流感病毒的多种亚型,解决了其他常规方法无法检测已发现和可能发生变异的所有亚型的棘手问题,从而为A型流感诊断和分子流行病学监测提供了一种新技术。     

Development and Application of Cotton InDel Markers Based on High Throughput Sequencing

[Objective] Dimorphic InDel markers can be used for cotton variety identification and purity detection, to improve the accuracy and efficiency of cotton seed testing, and to play a role in molecular breeding of cotton. [Method] Based on the whole genome sequencing of 121 cotton varieties from different sources, the InDel markers with high polymorphism were developed according to polymorphism information content(PIC) and were applied in the genetic distance analysis and cluster analysis by using 66 cotton varieties in China. [Result] Totally 10 967 InDel were identified based on the next generation sequencing data of 121 cotton varieties. Among the 85 pairs of InDel primers synthesized, 64 were selected including 35 from At group and 29 from Dt group. The minimum average allelic frequency(MAF) of At and Dt chromosomes were 0.45 and 0.32, respectively, while the PIC were 0.49 and 0.40, respectively. The genetic distances of the 66 cotton varieties ranged from 0.04 to 0.65 centimorgan (cM), with an average of 0.39 cM. The two varieties with the largest genetic distance were Simian 3 and CCRI 36, and the two varieties with the smallest genetic distance were Xumian 18 and Xuza 3. [Conclusion] The 64 cotton dimorphic InDel markers can effectively reveal the relationships among varieties based on the genetic distance, and distinguish cotton varieties from different sources, which has certain theoretical significance and application value.     

基于Kjeldahl与Dumas方法的农作物秸秆总氮含量分析

凯氏定氮法(Kjeldahl)与杜马斯燃烧法(Dumas)是测定农业生物质总氮含量的主要检测手段,但二者的测定结果数值存在差异。该研究获取农作物秸秆样本(水稻、小麦、玉米、油菜和棉花)共计1 179个,分别采用Kjeldahl和Dumas方法测定总氮(TKN和TCN,total Kjeldahl nitrogen and total combustion nitroyen)含量,通过多种统计与分析方法,系统分析比较了不同农作物秸秆总氮含量及其分布的异同和相关关系。结果表明:不同农作物秸秆氮含量分布均呈非正态分布,建议采用中位数统计;5种秸秆总体的TKN质量分数为(7.12±1.87) g/kg,TCN质量分数为(8.00±2.13) g/kg,TKN含量显著小于TCN含量;小麦和棉花秸秆的TKN含量和TCN含量与其他秸秆间均存在显著差异(P <0.05);不同生物质TKN含量与TCN含量关系不同,建议采用最小中位数二乘法进行拟合分析。研究结果可为农作物秸秆科学利用提供数据及方法互通性支撑。     

不同来源吸附载体大豆糖蜜对奶牛瘤胃内环境的影响

为考察不同来源吸附载体大豆糖蜜对奶牛瘤胃内环境的影响,采用3×3拉丁方试验设计,选用3头年龄相同(3胎次)、体重相近[(500±50)kg]且安装永久性瘤胃瘘管的中国荷斯坦奶牛为试验动物,分别饲喂基础日粮、豆皮糖蜜和麸皮糖蜜日粮。结果表明,豆皮糖蜜组和麸皮糖蜜组瘤胃pH均极显著高于对照组(P0.01),而豆皮糖蜜组和麸皮糖蜜组之间瘤胃pH没有显著差异(P0.05);麸皮糖蜜组瘤胃NH_3-N浓度显著低于对照组(P0.05),但与豆皮糖蜜组没有显著差异(P0.05);各处理间奶牛瘤胃各单个挥发性脂肪酸(volatile fatty acid,VFA,包括乙酸、丙酸、异丁酸、丁酸、异戊酸和戊酸)浓度、总VFA浓度和乙丙比均没有显著差异(P0.05);但与对照组相比,豆皮糖蜜组瘤胃总VFA浓度有降低趋势;而麸皮糖蜜组瘤胃总VFA浓度有提高趋势。结果显示,在奶牛饲料中可用豆皮糖蜜替代15%玉米以及用麸皮糖蜜替代6%玉米和10%麸皮,均具有调控瘤胃稳衡的作用,且麸皮糖蜜替代效果优于豆皮糖蜜。     

基于合成法的定位误差的分析与计算

为保证机床夹具设计的可靠性,定位误差的计算是机床夹具设计过程中必不可少的重要环节之一。为此从定位误差的产生机理出发,介绍了定位误差的概念、产生原因,并给出计算公式,有助于快速实现定位误差的求解。     

血吸虫疫苗研究现状和发展方向

血吸虫疫苗的研究工作已经进行了很长一段时间,发现了一批有价值的候选疫苗分子。同时通过改进和优化免疫途径,血吸虫疫苗的免疫保护力正在逐步提高。随着研究的日益深入,对血吸虫疫苗的免疫机制等了解更加透彻,距离研制出能诱导高的、稳定的免疫保护力的疫苗分子的目标将越来越近。