基于红外热成像的夜间农田实时语义分割

农田环境实时语义分割是构成智能农机的视觉环境感知的重要环节，夜间农田语义分割可以使智能农机在夜间通过视觉感知农田环境进行全天候作业，而夜间无光环境下，可见光摄像头成像效果较差，将造成语义分割精度的下降。为保证夜间农田环境下红外图像语义分割的精度与实时性，该研究提出了一种适用于红外图像的红外实时双边语义分割网络（Infrared Real-time Bilateral Semantic Segmentation Network，IR-BiSeNet），根据红外图像分辨率低，细节模糊的特点该网络在实时双边语义分割网络（Bilateral Semantic Segmentation Net，BiSeNet）结构基础上进行改进，在其空间路径上，进一步融合红外图像低层特征，在该网络构架中的注意力提升模块、特征融合模块上使用全局最大池化层替换全局平均池化层以保留红外图像纹理细节信息。为验证提出方法的有效性，通过在夜间使用红外热成像采集的农田数据集上进行试验，数据集分割目标包括田地、行人、植物、障碍物、背景。经试验验证，提出方法在夜间农田红外数据集上达到了85.1%的平均交并比（Mean Intersection over Union，MIoU），同时达到40帧/s的处理速度，满足对夜间农田的实时语义分割。     

Detection and quantification of Aeromonas salmonicida in fish tissue by real‐time PCR

Furunculosis, a septicaemic infection caused by the bacterium Aeromonas salmonicida subsp. salmonicida, currently causes problems in Danish seawater rainbow trout production. Detection has mainly been achieved by bacterial culture, but more rapid and sensitive methods are needed. A previously developed real‐time PCR assay targeting the plasmid encoded aopP gene of A. salmonicida was, in parallel with culturing, used for the examination of five organs of 40 fish from Danish freshwater and seawater farms. Real‐time PCR showed overall a higher frequency of positives than culturing (65% of positive fish by real‐time PCR compared to 30% by a culture approach). Also, no real‐time PCR‐negative samples were found positive by culturing. A. salmonicida was detected by real‐time PCR, though not by culturing, in freshwater fish showing no signs of furunculosis, indicating possible presence of carrier fish. In seawater fish examined after an outbreak and antibiotics treatment, real‐time PCR showed the presence of the bacterium in all examined organs (1–482 genomic units mg^?1). With a limit of detection of 40 target copies (1–2 genomic units) per reaction, a high reproducibility and an excellent efficiency, the present real‐time PCR assay provides a sensitive tool for the detection of A. salmonicida.     

蚜虫共生细菌多重PCR检测体系的建立

蚜虫中具有多种共生菌,使用常规PCR对它们进行检测,耗时耗力,而多重PCR可以更加高效地进行多种细菌的检测。沃尔巴克氏菌Wolbachia pipientis、杀雄菌属共生菌Arsenophonus和蚜虫U型共生菌Regiella insecticola是蚜虫中常见的3种共生菌。本研究针对沃尔巴克氏菌、杀雄菌属共生菌和蚜虫U型共生菌,分别选择以wsp基因、yaeT基因和gltA基因作为靶标,进行了多重PCR引物的设计和扩增体系的优化。结果显示,本研究建立的多重PCR体系在检测3种蚜虫常见共生菌时,具有较高的扩增特异性、准确性和直观性及较高的检测灵敏度,共生菌的最低检测浓度为104拷贝/μL,远低于共生菌在蚜虫1龄若虫总DNA中的浓度(108拷贝/μL),可以完全满足蚜虫共生菌检测工作的需要。     

EMS诱导水稻‘一目惚’表型变异和部分基因表达变异的比较

本研究利用甲基磺酸乙酯(Ethy methan sulfonate)试剂胁迫诱导粳稻品种‘一目惚’(Hitomebore),获得性状稳定遗传的水稻矮杆突变体(E58),对株高、分蘖、千粒重等表现与相同环境生长下的野生型‘一目惚’对照(E63)进行对比分析,结果表明,该突变体田间表现为植株矮化,分蘖增多,千粒重减少,且株高与对照相比呈极显著差异,分蘖呈显著差异,千粒重呈极显著差异。EMS诱导的序列变异可能对基因表达产生影响,进而引起水稻矮化的表型变异。根据前人试验结果,应用Real-time PCR对与胁迫及矮化相关的39对引物中筛选出24对引物进行Real-Time PCR扩增,结果表明:引物OsNR1、OsP5CS、OsGS1;1、OsGDH1、OsHKT1;5、OsNRT1;1、OsGDH3、OsAKT1、OsAMT2;3、OsGS1;2、OsSOS1、OsNHX2、OsNRT2;1、OHAK16、OsGS2、OsHAK1、OsGDH2、OsAMT3;3在E63、E58表达差异极显著;OsHAK10、OsNHX1、OsAMT1;1表达差异显著;OsNADHGOGAT、OsFd-GOGAT、OsGS1;3表达差异不显著。EMS诱导对部分水稻基因表达产生影响,从而影响水稻表现型,为进一步工作提供科学依据。     

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%。     

Intestinal Microbiota in Large Yellow Croaker,Larimichthys crocea,at Different Ages

The polymerase chain reaction–denaturing gradient gel electrophoresis (PCR‐DGGE) of 16S ribosomal RNA gene was used to investigate bacterial communities in the intestines of large yellow croaker at six different ages (12 d, 18 d, 26 d, 40 d, 3 mo, and 1 yr old) as well as within the corresponding feed and culture water. In addition, Illumina Miseq sequencing was utilized to compare intestinal microbiota between 12‐d‐old and 1‐yr‐old individuals. PCR‐DGGE results revealed that the culture water had the highest bacterial diversity, followed by the feed, while the intestines had the lowest diversity. The intestinal microbiota at six ages changed severely; however, the change did not follow any trend. The large yellow croaker intestines harbored specific bacterial communities that differed from those in both feed and water. Illumina Miseq sequencing results revealed that the diversity of intestinal bacteria in 12‐d‐old fish was higher than that in 1‐yr‐old fish, and the bacterial composition differed significantly between them. γ‐Proteobacteria and Pseudoalteromonas supplied the most abundant phylum and genus in the 12‐d‐old fish intestine. However, in the 1‐yr‐old fish intestine, Firmicutes and Clostridium were the most dominant, respectively. The study may contribute to a better understanding of gut microbiota and dynamics of the large yellow croaker and the relationship with their surrounding environment.     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

柑橘溃疡病菌LAMP实时浊度检测方法的建立及应用

以柑橘溃疡病菌特异性高的假定蛋白基因（登录号：AE008923.1）为靶标，设计并筛选4条引物来特异性识别靶标序列中的6个独立区域，建立柑橘溃疡病菌LAMP实时浊度检测方法。该方法利用实时浊度仪检测反应过程中产生的焦磷酸镁白色沉淀，实现对LAMP整个反应过程的实时监控。从镁离子浓度、甜菜碱浓度、dNTPs浓度、引物浓度和反应温度5个变量参数对 LAMP检测体系进行优化，并对方法的特异性、灵敏度、稳定性及实际样品检测效果进行评价。结果显示，该体系经优化后稳定性良好，可在66 ℃ 恒温条件下，60 min内完成检测；DNA检测下限可达0.02 ng/μL，灵敏度与荧光定量PCR方法相同，比常规PCR方法高1个数量级；能快速、准确地对田间疑似样品进行检测，与荧光定量PCR方法的检出情况一致，没有漏检。结果说明柑橘溃疡病菌实时浊度LAMP检测方法操作简便、所需时间短，特异性与灵敏度高，为柑橘溃疡病菌的快速筛查提供较好的途径。     

农用植保旋翼无人机地面监控系统的设计与实现

植保无人机凭借其低成本、高效率、精准快速作业等优点,在农业植保领域得到快速发展,成为现代农业的一种重要装备。为了能够实时远程监控农用植保旋翼无人机的飞行状态信息,提高无人机飞行作业安全和作业质量,进行更好的飞行控制管理,设计并实现了植保旋翼无人机地面监控系统,可实现与植保无人机的远距离实时通信、监测飞行姿态、显示飞行作业轨迹和飞行控制等操作。地面监控系统采用嵌入式树莓派2作为硬件平台,2.4G无线模块实现数据收发,使用跨平台C++图形用户界面应用程序框架Qt对地面监控系统软件功能和交互界面进行开发,并制定了旋翼无人机与地面监控系统之间的数据通讯协议。该系统实际测试表明:监控系统可长时间连续稳定的工作,有效实现了对农用植保旋翼无人机实时监控与操作。     

蛹虫草CmKexin基因克隆和菌丝体中表达检测

为了探讨蛹虫草类枯草杆菌蛋白酶(Subtilisin-like protease)的表达特性,以真菌蛹虫草菌丝体为研究对象,通过RT-PCR获得蛹虫草CmKexin基因的ORF序列,并进行序列分析。应用Northern杂交和Real-time PCR方法,检测蓝光照射后蛹虫草菌丝体中CmKexin基因的转录情况。结果获得蛹虫草CmKexin基因的ORF序列。对翻译蛋白质产物CmKexin进行生物信息学分析,表明该蛋白质含1个属蛋白转化酶的肽酶S8家族功能域(143~429)和1个前体蛋白转化酶P功能域(514~600),符合真菌类枯草杆菌蛋白酶的特征。蛹虫草菌丝体在黑暗预培养4 d后再蓝光照射,Northern Blotting和Real-time PCR检测均可得到相同的结果,即在持续的蓝光照射48~50 h时,出现CmKexin基因的瞬时大量转录。而其他时间内均未检测到该基因的大量转录。试验结果将为蛹虫草类枯草杆菌蛋白酶的利用提供理论依据。