农杆菌介导的朝鲜碱茅PuP5CS基因转化紫花苜蓿的研究

为提高紫花苜蓿(Medicago sativa)的抗逆性,利用在朝鲜碱茅(Puccinellia chinampoensis)已克隆的Pu P5CS基因成功构建了p CAMBIA3300-Pu P5CS表达载体,以子叶为外植体,通过农杆菌介导共培养法转化紫花苜蓿"公农5号"(M.sativa cv.Gongnong No.5),并以2.0 mg·L-1的草铵膦进行筛选,抗性愈伤组织诱导率为22.4%,经草铵膦筛选的植株进行PCR检测和RT-PCR检测。结果显示,共获得11株转化植株,表明Pu P5CS基因已转入T0紫花苜蓿植株,且能够在RNA水平上正常表达。     

Pathogenicity of a Strain of H5N6 Subtype Avian Influenza Virus in Ducks and Mice

To investigate the pathogenicity of A/Hero/Guangdong/C1/2013(H5N6), an AIV strain isolated from Heron on ducks and mice, pathogenicity of this new virus strain and changing of histopathology as well as a preliminary study on its biological characteristics were studied by virus challenges test via intranasal and eye-drop and in chicken via jugular vein injection.Results showed that the EID₅₀ of this strain of virus was 10^-8.16/0.1 mL in embryonated chicken eggs and the intravenous inoculation of pathogenic index (IVPI) was 2.76.In ducks and mice, the 50% lethal doses (LD₅₀) of it were determined to be 10^-4.0/0.2 mL and 10^-4.67/0.05 mL, respectively.Symptoms of infection included loss of appetite, depression, swollen head and tears after being infected with 10⁶ EID₅₀ per duck by intranasal and eye-drop administration.Most of ducks died 4 to 7 days post-infection, liver, lung and kidney still eliminated toxicants at day 7 post-infection.Anatomy showed symptoms of pericardial effusion, pulmonary congestion and kidney enlargement, while pathological section showed pathological change like karyopycnosis and inflammatory cell infiltration in heart, liver, kidney and spleen.Mice developed symptoms of infection like loss of appetite, depression, shaggy coat and ruffled coat after being infected with 5×10⁵ EID₅₀ per mouse by intranasal and eye-drop administration.Most of the infected mice died 5 to 7 days post-infection and only liver still eliminated toxicants at day 7 post-infection.Although organ anatomy showed no obvious pathological changes, pathological section showed pathological changes like karyopycnosis and inflammatory cell infiltration in heart, kidney and lung.Our research demonstrated that this H5N6 subtype AIV had a strong pathogenicity and could be defined as a highly pathogenic AIV strain as its IVPI was greater than 1.2.Our work laid a theory foundation for study, prevention and control of H5N6 subtype AIV.     

Construction and Identification of PRRSV ORF5 Gene Prokaryotic Expression Vector

The present study was designed to construct recombinant plasmids,which could express porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 gene.RNA was extracted from spleen and lung samples of the suspected pigs which were infected with PRRSV.According to PRRSV ORF5 gene,a pair of primers was designed for RT-PCR amplification.The ORF5 target gene was cloned into pMD19-T vector and then the recombinant pMD19-ORF5 was achieved.According to the sequencing results and the characteristics of expression vectors,a pair of primers with NcoⅠand XbaⅠenzyme cleavage sites was designed.Target fragment dORF5 was amplified and then connected to pProEXHTb and pNZ8149 vectors,respectively.And recombinant HTb-dORF5/DE3 and pNZ8149-dORF5/NZ3900 was induced by IPTG and Nisin,respectively,and analyzed by SDS-PAGE and Western blotting.Recombinant HTb-dORF5/DE3 induced by 1.5 mmol/L IPTG was expressed in the highest quantity.There were specific band at about 22 ku with reactionogenicity when it was tested by SDS-PAGE and Western blotting.Recombinant pNZ8149-dORF5/NZ3900 induced by 20 ng/mL Nisin was expressed in the highest quantity.There were specific band at about 19 ku with reactionogenicity when it was tested by SDS-PAGE and Western blotting.The IFA result showed specific green fluorescence.This study successfully constructed recombinant plasmids HTb-dORF5 and pNZ8149-dORF5 and expressed,the result laid a solid foundation for further development of PRRS vaccines.     

Avian Influenza A(H5N1) Virus Outbreak Investigation: Application of the FAO‐OIE‐WHO Four‐way Linking Framework in Indonesia

WHO, FAO and OIE developed a ‘four‐way linking’ framework to enhance the cross‐sectoral sharing of epidemiological and virological information in responding to zoonotic disease outbreaks. In Indonesia, outbreak response challenges include completeness of data shared between human and animal health authorities. The four‐way linking framework (human health laboratory/epidemiology and animal health laboratory/epidemiology) was applied in the investigation of the 193rd human case of avian influenza A(H5N1) virus infection. As recommended by the framework, outbreak investigation and risk assessment findings were shared. On 18 June 2013, a hospital in West Java Province reported a suspect H5N1 case in a 2‐year‐old male. The case was laboratory‐confirmed that evening, and the information was immediately shared with the Ministry of Agriculture. The human health epidemiology/laboratory team investigated the outbreak and conducted an initial risk assessment on 19 June. The likelihood of secondary cases was deemed low as none of the case contacts were sick. By 3 July, no secondary cases associated with the outbreak were identified. The animal health epidemiology/laboratory investigation was conducted on 19–25 June and found that a live bird market visited by the case was positive for H5N1 virus. Once both human and market virus isolates were sequenced, a second risk assessment was conducted jointly by the human health and animal health epidemiology/laboratory teams. This assessment concluded that the likelihood of additional human cases associated with this outbreak was low but that future sporadic human infections could not be ruled out because of challenges in controlling H5N1 virus contamination in markets. Findings from the outbreak investigation and risk assessments were shared with stakeholders at both Ministries. The four‐way linking framework clarified the type of data to be shared. Both human health and animal health teams made ample data available, and there was cooperation to achieve risk assessment objectives.     

基于Faster-NAM-YOLO的黄瓜霜霉病菌孢子检测

黄瓜霜霉病由古巴假霜霉病菌孢子通过侵染引起,严重影响了黄瓜的品质和产量;病菌孢子数量与病情严重度相关,因此建立快速、简便和高效的病菌孢子定量检测方法,实现黄瓜霜霉病防治关口前移。基于YOLO v5模型提出了一种基于Faster-NAM-YOLO的黄瓜霜霉病菌孢子定量检测模型,该模型首先提出了一种特征提取模块C3＿Faster,使用C3＿Faster替换YOLO v5中的C3模块,有效降低了模型参数计算量和模型深度,提升了对黄瓜霜霉病菌孢子检测速度和精度;其次在主干网络中加入了NAM注意力模块,通过应用权重稀疏性惩罚抑制不显著权重,进而提高模型的特征提取能力和计算效率;最后实现了对黄瓜霜霉病菌孢子的定量检测。实验结果表明,Faster-NAM-YOLO模型在测试集上mAP@0.5和mAP@0.5:0.95分别达到95.80%和60.90%,对比原始YOLO v5模型分别提升1.80、1.20个百分点,较原始YOLO v5模型内存占用量和每秒浮点运算次数分别减少5.27 MB和1.49×10¹⁰;通过与YOLO v3、THP-YOLO v5、YOLO v7、YOLO ...     

低数据集下基于ASPP-YOLO v5的苋菜识别方法研究

针对田间苋菜识别存在准确率低、样本数量少等问题,通过引入扩展感受野和提取上下文信息的ASPP注意力机制改进YOLO v5苋菜识别模型,在低数据集下改进后的模型能够显著提高F1值和mAP。实验结果表明,在低数据集下引入ASPP注意力机制后苋菜识别模型F1值提高13个百分点、mAP提高18.6个百分点。采用横向录制的方式苋菜被检测到的概率提高15.4个百分点。因此,本研究为苋菜或其他杂草在低数据集下的识别提供了有效的方法,为农业领域的杂草识别和管理研究提供了参考。     

SPOT5在小流域水土保持生态建设遥感监测中的应用

在伊洛河赵沟小流域应用SPOT5卫星影像开展水土保持生态建设遥感监测,以制作流域"4D"产品为基础,基于虚拟地理信息系统构建赵沟小流域虚拟现实环境进行定义分类模板技术研究,整体分类模板定义精度可达95.48%;进而对SPOT5卫星影像实施水土保持措施遥感监测分类处理,运用ERDAS IMAG INE精度评价统计功能对虚拟现实环境参照下的水土保持措施分类结果进行评价,总体分类精度达到80.49%;对各地物类型和总体Kappa系数进行分析,总体Kappa系数达到0.769 4。研究表明,SPOT5在大规模水土保持生态建设遥感监测中具有很好的应用前景。     

Frozen bread dough: Effects of freezing storage and dough improvers

This review focuses on the effects of freezing storage on the microstructure and baking performance of frozen doughs, and provides an overview of the activities of dough improvers, including emulsifiers, hydrocolloids and other improvers used in frozen dough applications. The overall quality of bread baked from frozen dough deteriorates as the storage of the dough at sub-zero temperatures increases due to several factors which are discussed. Lipid-related emulsifiers such as diacetyl tartaric acid esters of mono and diglycerides and sucrose esters employed as anti-staling agents, dough modifiers, shortening sparing agents, and as improvers for the production of high-protein bread have also been employed in frozen doughs. Hydrocolloids are gaining importance in the baking industry as dough improvers due to their ability to induce structural changes in the main components of wheat flour systems during breadmaking steps and bread storage Their effects in frozen doughs is discussed. Other dough improvers, such as ascorbic acid, honey and green tea extract, are also reviewed in the context of frozen doughs.     

飞蝗表皮蛋白LmKnk3-5′的抗体制备及组织定位

【目的】LmKnickkopf3-5′(LmKnk3-5′)是飞蝗（Locusta migratoria）蜕皮发育中重要的表皮蛋白。本文旨在制备LmKnk3-5′特异抗体并对其进行组织定位,为LmKnk3-5′生物学功能研究提供蛋白水平的证据,同时为进一步深入探究LmKnk3-5′与其他表皮蛋白在飞蝗表皮形成过程中的协同作用打下基础。【方法】首先比对飞蝗Knickkopf(Knk)家族4个基因LmKnk、LmKnk2、LmKnk3-FL和LmKnk3-5′的氨基酸序列,选取LmKnk3-5′的3段特异抗原序列R1、R2和R3,设计包含BamH I、Hind III酶切位点的引物,以LmKnk3-5′全长cDNA为模板,PCR获得LmKnk3-5′的特异抗原片段;然后将特异抗原片段与pET-32a经双酶切、T4连接酶连接、测序验证、成功构建重组质粒后,转入BL21（DE3）表达菌株中,加入IPTG（终浓度0.5 mmol·L^-1）低温（16℃）诱导重组蛋白表达,SDS-PAGE检测重组蛋白表达情况;接着扩大培养可在菌体裂解液的上清中表达重组融合蛋白的大肠杆菌,提取蛋白并用Ni-NTA对其进行纯化,之后再用纯化后的重组特异抗原免疫小鼠4次,获得anti-LmKnk3-5′多克隆抗体,ELISA法对其进行效价检测,并用Western blot法检测其特异性。取飞蝗5龄若虫第8天表皮,制备石蜡切片,通过免疫组化法对LmKnk3-5′蛋白进行组织定位。【结果】通过氨基酸序列比对,选取LmKnk3-5′的特异抗原区域R1、R2和R3,分别包含208、147和131个氨基酸残基,蛋白理论分子量分别为24.0、17.0和14.8 kD。酶切连接后,成功获得 pET-32a-R1、pET-32a-R2 和pET-32a-R3重组质粒。诱导表达检测,发现只有含pET-32a-R2的表达菌在菌体裂解液的上清液中大量表达重组蛋白。将其扩大培养纯化后获得 R2重组融合蛋白,免疫小鼠取抗血清进行效价检测,抗体效价高达1﹕512 000,可满足抗体使用需求。Western blot结果表明,注射dsLmKnk3-5′后飞蝗体壁LmKnk3-5′的蛋白表达显著减少,表明anti-LmKnk3-5′抗体特异性良好。免疫组化试验结果表明,表皮蛋白LmKnk3-5′在飞蝗腹部体壁皮细胞及新表皮中均有分布,特别是新合成的外表皮顶端。【结论】成功获得飞蝗LmKnk3-5′多克隆抗体,证明其特异性良好。LmKnk3-5′在飞蝗腹部体壁新合成的外表皮顶端分布较多。研究结果为飞蝗表皮蛋白LmKnk3-5′功能研究提供了蛋白水平的证据。     

湿地生态系统实际蒸散发数据驱动估算模型研究

利用Fluxnet2015全球通量塔观测数据集,研究了随机森林（RF）、梯度增强回归分析（GBR）、支持向量回归（SVR）和深度学习神经网络（DNN）预测湿地生态系统的实际蒸散发（Evaporation,ET）。通过对比研究,确定了预测实际蒸散发的最佳特征变量组合,包括短波辐射、净辐射、初级生产总值、气温、土壤温度、风速、降水、经度、纬度和时间。以此为模型输入,利用Fluxnet2015站点测试数据集和ERA5-Land再分析资料提供的输入特征,对比分析了不同模型的实际蒸散发估计精度,结果表明：以站点数据为输入,SVR算法精度相对较高,其R²可达0.896,RPE最小为31.5%;以ERA5-Land再分析资料为输入,除了GBR算法以外,其余3种方法R²高于0.820,RPE小于57%。另外,模型算法估计的ET精度要明显高于ERA5-Land再分析资料提供的ET产品。