运用植被指数时序特征对落叶松人工林分类

为探究植被指数时序特征是否有利于落叶松人工林提取,以孟家岗林场为研究试验区域,根据落叶松人工林季相和物候特征,利用Landsat8OLI影像数据提取研究区内5种植被的归一化植被指数(I NDV)、差值植被指数(I DV)、比值植被指数(I RV)、增强型植被指数(I EV),构建相应的植被指数时序特征。采用最大似然和随机森林两种方法对单一时相影像和加入植被指数时序特征的影像进行对比试验。结果表明:影像中加入植被指数时序特征后,最大似然算法的分类总体精度为89.53%,Kappa系数为0.87,比单一时序特征的影像分类精度提高了13.35%;随机森林算法的森林类型分类总体精度为93.22%,Kappa系数为0.92,比单一时序特征的影像分类精度提高了19.8%。因此,加入植被指数时序特征后能得到更高的落叶松人工林提取精度。     

实时荧光定量PCR技术的概述及应用

实时荧光定量PCR技术是在传统PCR技术上发展而来的,不仅能判断某一基因的有无,而且还能对其进行定量分析。由于该技术与常规PCR相比,能够进行实时监测、且自动化程度高而被广大研究者青睐。本文对实时荧光定量PCR的原理、数据分析、定量方法、分类以及应用进行了系统而详细地综述。     

不同处理对柴胡种子萌发的影响

为研究不同发芽床和清水浸种时间对柴胡种子萌发的影响,以种子萌发启动日、发芽率、发芽势、发芽指标、霉烂率为测量指标,20℃恒温自然光照培养箱中开展了柴胡种子发芽试验。结果表明,不同发芽床基质处理对柴胡种子萌发的影响有显著性差异,其中以纱布处理柴胡种子的各项测量指标均显著高于其他处理;清水浸种不仅能够促使柴胡种子提早萌发,而且能够提高柴胡种子发芽率,浸种12 h柴胡种子的发芽率及发芽势最高,分别可达67.67%±3.21%和56.00%±9.54%,与未浸泡对照相比提高了约35、34个百分点。说明柴胡种子用清水浸种预处理12 h,以纱布为发芽床,在20℃恒温条件下培养,可有效提高柴胡种子萌发率,促进种子提早萌发,实际生产中可以用清水浸种后再进行播种。     

白背飞虱酚氧化酶原PPO基因特性及其免疫应答

【目的】酚氧化酶（phenoloxidase,PO）是昆虫体内的免疫蛋白,在昆虫体内起重要的调节作用,主要以无活性的酚氧化酶原（prophenoloxidase,PPO）形式存在。本研究在转录组测序获得白背飞虱（Sogatella furcifera）3条PPO序列的基础上,通过分析白背飞虱体内不同SfPPO的发育和组织表达模式,并进一步抑制SfPPO的表达以及病原菌诱导,探讨SfPPO在白背飞虱体内的生物学功能。【方法】以白背飞虱3条SfPPO序列为研究对象,利用生物信息学方法分析其蛋白结构以及与其他昆虫之间的同源性。并以注射绿色荧光蛋白（green fluorescent protein,GFP）的dsRNA作为对照组,通过注射合成的dsSfPPO后采用实时荧光定量PCR（qRT-PCR）检测基因表达抑制效果;此外,为了探究SfPPO在白背飞虱生长发育和免疫应答方面的作用,利用qRT-PCR检测SfPPO在不同发育阶段及组织中的表达情况,以及注射大肠杆菌（Escherichia coli）和枯草芽孢杆菌（Bacillus subtilis）后3个SfPPO的诱导表达情况。【结果】SfPPO1、SfPPO2-1、SfPPO2-2的开放阅读框长度分别为2 079、999、2 070 bp,分别编码692、332、689个氨基酸,预测蛋白分子量分别为79.84、37.67、79.53 kD,等电点分别为6.43、9.56、6.20。生物信息学分析表明,白背飞虱的3个PPO蛋白具有较高的同源性,且与褐飞虱（Nilaparvata lugens）的亲缘关系最近;SfPPO1、SfPPO2-1、SfPPO2-2均在成虫第3天表达量最高,其中SfPPO2-1和SfPPO2-2的发育表达模式相似;SfPPO1和SfPPO2-1在表皮和脂肪体内表达量较高,而SfPPO2-2在翅和脂肪体内表达量较高,在头、足、中肠、马氏管中表达量相对较低;与注射dsGFP组相比,注射靶标基因的dsRNA后都能够显著沉默目的基因的表达,而且当SfPPO2-1表达被沉默后,SfPPO1出现显著高表达,说明不同的PPO之间可能具有补偿功能;大肠杆菌诱导后12 h,SfPPO2-1和SfPPO2-2表达量显著升高,SfPPO1表达量则在诱导24 h后显著升高;枯草芽胞杆菌诱导24 h后,SfPPO1、SfPPO2-1和SfPPO2-2表达量均显著升高。【结论】SfPPO1、SfPPO2-1、SfPPO2-2的表达存在组织和龄期特异性,且在不同菌诱导情况下存在免疫应答差异性。研究结果有助于更加全面、深入地探索白背飞虱酚氧化酶在调控昆虫发育及免疫中的潜在分子机理。     

Evaluation of a high-throughput nucleic acid extraction method for the detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples by PCR

Johne’s disease (paratuberculosis) is an economically important disease of cattle worldwide. The disease is caused by Mycobacterium avium subsp. paratuberculosis (MAP), a fastidious gram-positive bacterium. PCR is increasingly used in diagnostic laboratories for the detection of MAP in fecal samples given the rapid test turnaround time and sensitivity and specificity comparable to fecal culture. However, efficient extraction of DNA for sensitive detection of MAP by PCR is affected by the complex lipid-rich cell wall of MAP and the presence of PCR inhibitors in feces. We evaluated a high-throughput nucleic acid extraction method (MagMAX core nucleic acid purification kit with mechanical lysis module) in conjunction with an hspX gene PCR for the detection of MAP from bovine fecal samples, which resulted in correct identification of all negative (13 of 13) and positive (35 of 35) proficiency test samples obtained from the National Veterinary Services Laboratories. In addition, all 6 negative and 50 of 51 positive diagnostic specimens tested were categorized correctly.     

咖啡湿法发酵中使用果胶酶对脱胶时间与杯品质量的影响

以云南阿拉比卡咖啡为原料,在咖啡湿法发酵环节中加入Pectinex Ultra SP-L果胶酶进行脱胶,并对加工出的咖啡豆进行理化及感官分析。结果表明：与自然发酵脱胶相比,果胶酶脱胶能缩短脱胶时间,有效快速脱除咖啡果胶。其脱胶时间与果胶酶的浓度、脱胶温度、鲜果成熟度呈正相关。与自然发酵脱胶相比,果胶酶脱胶所得咖啡豆的内含物质无显著差异,杯品质量干净稳定。就咖啡豆的杯品质量而言,0.01%果胶酶添加量脱胶处理要高于自然发酵脱胶,而添加量为1.00%、0.10%果胶酶脱胶处理略低于自然发酵脱胶。综上所述,果胶酶脱胶能保证咖啡品质的稳定性及提高咖啡加工效率,具有应用前景。     

一类具有饱和发生率的虫媒传染病模型研究

考虑虫媒传染病具有潜伏期的特征,研究了一类具有饱和发生率的时滞传染病模型的动力学行为,确定了疾病是否流行的阈值R_0.当R_01时,无病平衡点全局渐近稳定,疾病将最终灭绝;当R_01时,唯一地方病平衡点条件稳定,系统会产生Hopf分支.     

Mycobacteriosis caused by Mycobacterium marinum in reared mullets: first evidence from Sardinia (Italy)

Mycobacterium marinum is a slow‐growing non‐tuberculous mycobacterium, and it is considered the most common aetiologic agent of mycobacteriosis in wild and cultured fish. The diagnosis is principally made by histology when positive Ziehl–Neelsen stain granulomas are detected. The aim of this study was to investigate the occurrence of mycobacteriosis in extensively cultured Mugilidae of two lagoons (Cabras and San Teodoro) from Sardinia by the use of histology, microbiology, PCR and DNA sequencing. Nine of 106 mullets examined were affected by mycobacteriosis, and the spleen was the most affected organ. The histology detected higher rate (100%) of infection in spleen than the culture and PCR (75% and 62.5%, respectively). The sequencing of hsp65 gene identified M. marinum as the primary cause of mycobacteriosis in the mullets examined. Mullets affected by mycobacteriosis were mainly fished in the San Teodoro lagoon characterized by critical environmental conditions. Histology remains the most common method in detecting fish affected by mycobacteriosis, and PCR‐based methods are essential for species identification. Our finding are worthy of attention because mycobacteriosis caused by M. marinum in reared mullets was evidenced for the first time in Sardinia, suggesting that this disease may be underestimated also in other cultured fish species.     

Rapid detection of SNPs in candidate genes regulating the growth of orange‐spotted grouper,Epinephelus coioides (Hamilton, 1822), using semiconductor sequencing

Orange‐spotted grouper (Epinephelus coioides) is one of the most important marine food species in Southeast Asia and China and has been cultured for decades. In this study, we fully utilized the limited capacity of semiconductor sequencing, the high efficiency of long‐range PCR for target enrichment and a non‐indexed pooling strategy to screen single‐nucleotide polymorphisms (SNPs) in a breeding population of orange‐spotted grouper. Forty‐one genomic DNA fragments, with a total length of approximately 180 kb, including 22 candidate genes that control growth, and from a DNA pool of 20 heaviest and 22 lightest individuals of the sampled population were successfully sequenced using an Ion Torrent Personal Genome Machine. 3 503 466 clear reads were produced with a length of 192 ± 56 bp, 86.8% of which were mapped to the reference with an average coverage depth of 2567‐fold and physical coverage of 98.8%. Finally, 1623 high‐quality SNPs were adopted. Compared with Sanger sequencing of three random common regions, the sensitivity and specificity of our approach were 39.4% and 100.0% respectively. A mutation located at the third position of the previously labelled start codon of growth hormone receptor type 1 invalidated the start codon. Furthermore, comparison of the frequencies of genotypes and alleles of this site between the two extreme groups, prediction of signal peptide and identification of conservative mRNA sequences suggested that the functional start codon is likely located at the position of another downstream in‐frame ATG in the mutant. These detected SNP markers will provide important tools for the selective breeding of orange‐spotted grouper.