基于MAXENT的大豆南北方茎溃疡病菌在中国适生区的预测

利用MAXENT生态位模型和GIS系统,对大豆南北方茎溃疡病菌在我国的适生区进行预测.模型分析结果表明,这两种病菌在我国的潜在适生区域广泛.大豆北方茎溃疡病菌除了我国的宁夏、海南省外,其它省市均有该菌的适生分布区,其中适生等级高的地区主要在江苏、安徽、浙江、上海、江西和湖北6省.大豆南方茎溃疡病菌除了宁夏、青海、西藏外,我国的其它地区均为该菌的潜在适生区域,其中适生等级高的地区主要集中在江西、上海、江苏、安徽、浙江、河南以及陕西南部、四川东部和重庆的部分地区.     

大豆茎溃疡病菌子囊壳诱导及孢子萌发方法探索

大豆茎溃疡病是为害大豆的重要病害,世界上十大大豆生产国中已有7个国家发现大豆茎溃疡病菌的分布,对当地农业造成严重威胁[1]。该病是我国进境植物检疫性有害生物[2],目前没有发生为害的报道。大豆茎溃疡病菌以菌丝和子囊壳在大豆植株及其残体上越冬,子囊壳在干燥35 d     

大豆北方茎溃疡病菌分离鉴定技术的改进

本文对产生大豆北方茎溃疡病菌子囊壳的豆秆分别进行冷冻和水浸处理,发现2种处理均能促进成熟的子囊壳产孢,并且同一子囊壳经冷冻和水浸处理能够多次喷出子囊,释放子囊孢子,利用此特性有利于在不损失鉴定材料的前提下,多次获得分离物以完成病菌的分离鉴定,特别利于伴生其他真菌和子囊壳数量很少情况下,分离纯化大豆北方茎溃疡病菌。     

大豆茎溃疡病菌的生物学特性及其传入我国的风险

大豆南北方茎溃疡病菌是为害大豆的重要病原菌,是我国的检疫性有害生物.本文通过对这两种病菌生物学特性的了解,根据我国的作物种植特点及气候条件,对其传入我国的风险进行了评估,结果认为这两种病菌传入中国并在中国定殖、定殖后扩散等的可能性极大,风险为高.     

全国邮检口岸首次检出检疫性有害生物——大豆北方茎溃疡病菌

正2016年11月21日~2017年2月27日,浙江出入境检验检疫局杭州邮检办事处连续从英国进境邮包中截获4批次仓鼠粮,共计56.69 kg,内含大豆、小麦、玉米、葵花籽、花生、豌豆等6种种子。大豆经浙江省检验检疫科学技术研究院检测,检出检疫性大豆北方茎溃疡病菌,为全国邮检口岸首次检出,并从中检出2种抗除草剂转基因成分。     

宁波口岸连续检出大豆北方茎溃疡病菌

2005年7月,宁波检验检疫局技术中心植检实验室从北仑局送检的美国进境大豆下脚料中检测到大豆北方茎溃疡病菌（Diaporthe phaseolorum var.cauliuora）,并经浙江大学张敬泽教授复核确认.这是我国首次从进境植物中检出该病菌.不久,该实验室又从阿根廷进境的大豆中检出该病菌.这证实该病菌可以通过种子进行远距离传播.     

加拿大进境燕麦种子中大豆北方茎溃疡病病菌的检疫鉴定

从来自加拿大的燕麦种子中分离到3株可疑真菌,经形态特征鉴定、分子生物学鉴定及致病性测定,确定截获的3株真菌均为大豆北方茎溃疡病病菌(Diaporthe phaseolorum var.caulivora)。这是我国口岸首次从进境的燕麦种子中截获该病菌。     

苹果壳色单隔孢溃疡病菌的适生区预测

苹果壳色单隔孢溃疡病菌(Botryosphaeria stevensii)是一种寄主十分广泛的进境植物检疫性真菌,在我国尚无分布。近年来,我国口岸多次截获该病菌。为更好地实现对该病菌的针对性检疫防控,本研究采用DIVA-GIS软件对该病菌在我国的潜在适生区进行了预测,并对其适生区内寄主情况进行了深入分析。研究结果表明,苹果壳色单隔孢溃疡病菌在我国的适生区域较为广泛,主要分布在我国华东、华北和华中地区;该病菌在其适生区域内存在大量寄主,尤其是水果类作物,林木寄主也有较多分布。因此,该病菌一旦传入我国,极可能造成十分严重的经济损失,值得高度警惕。本文最后提出了针对苹果壳色单隔孢溃疡病菌的风险防控建议。     

气候变化情景下四川省猕猴桃溃疡病菌潜在地理分布模拟

近年来,猕猴桃溃疡病在四川各猕猴桃主产区严重发生,造成严重经济损失。本研究采用MaxEnt模型分析四川省猕猴桃溃疡病菌潜在分布,并预测2030年代、2050年代、2070年代和2080年代的RCP2.6、RCP4.5和RCP8.5等3种气候变化情景下适生区变化。预测结果运用ROC曲线评价模拟准确性。结果表明:所建立13个模型的训练数据和测试数据AUC (areas under curve)值均高于0.9,达到极高的精度。当前气候条件下,猕猴桃溃疡病菌在四川的高适生区主要位于成都市、德阳市、绵阳市、广元市、巴中市、达州市和雅安市,中适生区在四川21地市州均有分布。2030年代-2080年代,气候变化情景下,与当前情景相比,高适生区和低适生区区域均显著增加,中适生区区域先增加后减少,不同适生区几何中心位置和迁移规律均有所不同但总体上均向北移动。     

进境澳大利亚大麦夹杂油菜茎基溃疡病菌的检疫鉴定

采用常规平板分离法, 从进境澳大利亚大麦中夹杂的油菜籽上获得1株疑似油菜茎基溃疡病菌的菌株01829?通过致病性测定?形态学观察?特异性引物扩增?ITS序列比对分析, 对01829进行了种类鉴定?结果表明:菌株01829在PDA培养基上生长较慢, 菌落边缘不整齐, 产生大量分生孢子器和分生孢子; 采用特异性引物对LMR1-D和Lmb分别进行PCR检测, 结果均有预期扩增片段产生; 基于ITS序列构建的系统发育树中, 菌株01829和GenBank中其他油菜茎基溃疡病菌相关序列聚在同一分支; 菌株01829接种油菜子叶和茎基部, 在子叶和茎基部接种部位分别引起叶斑和凹陷溃疡斑?根据上述试验结果, 将菌株01829鉴定为油菜茎基溃疡病菌, 这是我国口岸首次从进境澳大利亚大麦中截获油菜茎基溃疡病菌?     

Morphologic, Molecular, and Pathogenic Characterization of Diaporthe phaseolorum Variability in the Core Soybean-Producing Area of Argentina

ABSTRACT Isolates of the Diaporthe/Phomopsis (D/P) complex were collected in the main soybean producing area of Argentina during the 1996-97, 1997-98, and 1998-99 growing seasons. Twenty-three morphologic characters related to type of colonies, stroma, pycnidia and conidia, presence of perithecia, and asci length were studied by principal component analysis (PCA). Genomic DNA were analyzed by the random amplified polymorphic DNA (RAPD) technique. From both studies, 18 isolates were identified as D/P complex and grouped in four major taxa: (i) Diaporthe phaseolorum var. meridionalis, (ii) D. phaseolorum var. caulivora, (iii) D. phaseolorum var. sojae, and (iv) Phomopsis longicolla. In addition to distinguishing interspecific and intraspecific variability, molecular markers allowed the detection of differences among isolates within the same variety. Pathogenicity was assayed in the greenhouse, by the toothpick method, inoculating the D/P isolates to soybean genotypes carrying different resistance genes (Rdc1, Rdc2, Rdc3, and Rdc4) against soybean stem canker (SSC). Pathogenic analysis distinguished two main groups: (i) the SSC-producing isolates, including D. phaseolorum var. meridionalis and D. phaseolorum var. caulivora, and (ii) the non-SSC-producing isolates, including D. phaseolorum var. sojae and P. longicolla. Cultivar RA-702 (susceptible control) was compatible with both D. phaseolorum var. meridionalis and D. phaseolorum var. caulivora isolates; meanwhile, Tracy-M (Rdc1 and Rdc 2 genes) was incompatible with D. phaseolorum var. meridionalis but compatible with D. phaseolorum var. caulivora isolates. The fact that Rdc1 and Rdc2 together (as in Tracy-M) confer an almost immune reaction to all assayed isolates of D. phaseolorum var. meridionalis but were ineffective against the D. phaseolorum var. caulivora isolates evaluated suggests that the virulence or avirulence genes in D. phaseolorum var. meridionalis and D. phaseolorum var. caulivora are different. Moreover, physiological races of D. phaseolorum var. meridionalis were detected by using differential soybean genotypes carrying distinct single Rdc genes. As far as we know, this is the first report on the existence of physiological races of D. phaseolorum var. meridionalis in South America. Selective pressure due to deployment of resistant host cultivars may have changed the frequency of the virulence or avirulence genes within the population of D. phaseolorum var. meridionalis. On the whole, our results show that pathogenic variability of D. phaseolorum in the core soybean-producing area of Argentina is higher than previously recognized.     

Molecular Identification and Phylogenetic Grouping of Diaporthe phaseolorum and Phomopsis longicolla Isolates from Soybean

ABSTRACT Diaporthe phaseolorum and Phomopsis longicolla isolates from soybean were examined using traditional mycological characteristics and molecular methods. Cultural characteristics including types of fruiting bodies and conidia were assessed for isolates collected from soybean stems and seeds. Cultures were identified as P. longicolla, D. phaseolorum var. caulivora, D. phaseolorum var. meridionalis, or D. phaseolorum var. sojae. Molecular markers for these groups were developed and analyzed using polymerase chain reaction restriction fragment length polymorphisms (PCR-RFLP) and DNA sequencing in the internal transcribed spacer (ITS) and the 5.8S ribosomal DNA. The ITS(4) and ITS(5) primers amplified PCR products for all isolates studied. Gel electrophoresis of undigested PCR products and DNA sequencing produced various fragment lengths including 604 bp for P. longicolla, 602 and 603 bp for D. phaseolorum var. caulivora, 603 bp for D. phaseolorum var. meridionalis, and from 597 to 609 bp for D. phaseolorum var. sojae. Digestion of these PCR products with enzymes AluI, HhaI, MseI, RsaI, and ScrFI resulted in distinct bands for identification of P. longicolla and the varieties of D. phaseolorum I. All P. longicolla, D. phaseolorum var. caulivora, and D. phaseolorum var. meridionalis isolates were distinguished using AluI and HhaI with RsaI or ScrFI. The banding patterns of D. phaseolorum var. sojae isolates were complex and were separated into 11 subgroups after digestion with AluI, HhaI, MseI, RsaI, and ScrFI. Phylogenetic analysis of 20 isolates of D. phaseolorum and P. longicolla based on the DNA sequence of the ITS region resolved six clades termed A, B, C, D, E, and F. Clade A included all sequenced D. phaseolorum var. caulivora isolates, two from Italy and one from the United States. Isolates in clade B were exclusively associated with D. phaseolorum var. meridionalis. Clades A and B formed a well-supported monophyletic group. Isolates in clades C, D, E, and F were morphologically defined as isolates of P. longicolla, D. phaseolorum var. sojae, and Diaporthe spp. The ITS sequences similarity of seven geographically diverse P. longi-colla isolates illustrated that P. longicolla isolates have a similar genetic background, with some affiliations to some D. phaseolorum var. sojae isolates. Morphological characteristics of the isolates along with the terminal clades of the ITS phylogeny suggest that P. longicolla is an individual species, D. phaseolorum var. caulivora and D. phaseolorum var. meridionalis are varieties of D. phaseolorum, and D. phaseolorum var. sojae is either several varieties of D. phaseolorum or possibly several distinct species.     

Molecular Detection of Diaporthe phaseolorum and Phomopsis longicolla from Soybean Seeds

ABSTRACT Species-specific detection of Diaporthe phaseolorum and Phomopsis longicolla from soybean seeds was accomplished using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and TaqMan chemistry. To use these detection systems, fungal DNA was released from soybean seed coats using an ultrasonic processor to break the cells. DNA fragment lengths ranged from 200 to 1,200 base pairs (bp), with the majority of fragments <500 bp. Based on DNA sequences of the internal transcribed spacer (ITS) regions of ribosomal DNA, three TaqMan primer/probe sets were designed. Primer/probe set PL-5 amplified a 96-bp fragment within the ITS1 region of P. longicolla, D. phaseolorum var. caulivora, D. phaseolorum var. meridionalis, and D. phaseolorum var. sojae. Set PL-3 amplified a 86-bp DNA fragment within the ITS2 region of P. longicolla. Set DPC-3 amplified a 151-bp DNA fragment within the ITS2 region of D. phaseolorum var. caulivora. TaqMan primer/probe sets were able to detect as little as 0.15 fg (four copies) of plasmid DNA. When using PCR-RFLP for Diaporthe and Phomopsis detection, the sensitivity was as low as 100 pg of pure DNA. Among 13 soybean seed lots from Italy and the United States, the total Diaporthe and Phomopsis detected using a traditional seed-plating technique ranged from 0 to 32%. P. longicolla was most prevalent, followed by D. phaseolorum var. sojae. D. phaseolorum var. caulivora, which only occurred in 0.5% of the Italian seed lots, was not detected in the U.S. seed lots. D. phaseolorum var. meridionalis was not detected in either the U.S. or Italian seed lots. Using TaqMan primer/probe set PL-3, the frequency of P. longicolla was 18% in seed lot I3, similar to the frequency obtained from PCR-RFLP and potato dextrose agar plating detection. The frequencies of D. phaseolorum and P. longicolla in each seed lot obtained by the different detection methods were comparable with respect to total infection and individual species detection. However, TaqMan detection provided the fastest results of all the methods tested.     

美国进境大豆北方茎溃疡病菌的分离与鉴定

在对美国进境大豆检疫过程中,对大豆籽粒和夹带茎秆进行病原真菌的分离培养,使用形态学和分子生物学相结合的方法对可疑菌株进行鉴定,确定该菌为Diaporthe phaseolorum(Cke.&; Ell.)Sacc.var.caulivora Athow &; Caldwell。该病原菌是进境植物检疫潜在危险性病原真菌,在国内属首次截获。     

基于高通量测序及分离培养初步研究美国大豆中真菌多样性

中国是全球最大的大豆进口国,进境大豆所携带的病原真菌传入我国的风险极高.基于高通量测序技术对6批美国大豆真菌多样性进行分析,同时采用分离培养获得单一菌株,依据菌落形态、显微结构及分子技术进行鉴定.高通量测序结果显示,大豆中所有真菌共计5门15纲35目63科112属155种,主要为链格孢属(Alternaria)、球腔菌...     

大豆南方茎溃疡病菌的分离与鉴定

在对美国进境大豆进行检疫时，从混杂的豆秆上发现并分离到了大豆南方茎溃疡病菌，通过形态学和分子生物学鉴定，确定该菌为Diaporthe phaseolorum（Cke．＆ Ell.）Sacc．var．meridionalis Morgan-jones。该病原菌是国外近年来在大豆生产中新发生的危险性病原真菌，国内未见报道。     

梨火疫病的进境风险分析

讨论了梨火疫病的世界分布、国内苹果和梨的生产状况及经济重要性、梨火疫病在国内的适用性及国内外检测技术现状；澄清了以前我国有梨火疫病分布的错误报道。梨火疫病对我国梨和苹果生产具有极大的潜在风险，同时对我国的环境美化和生物多样性亦具潜在威胁。我国大部分地区为梨火疫病的适生区，国内广布梨火疫病的寄主。针对进口苗木的风险最大及果实亦可传病，提出了相应的风险管理措施。     

噻唑膦在不同介质不同pH条件下热贮稳定性

噻唑膦加工后的稳定性是剂型选择的关键,为了提高其稳定性,延长其持效期,本研究通过噻唑膦在几种介质中的热贮稳定性试验,研究了酸碱度及介质对噻唑膦化学稳定性及稳定剂环氧大豆油对噻唑膦水乳剂热贮稳定性的影响,从而探索出噻唑膦在不同介质中稳定的最佳pH范围。研究结果发现,噻唑膦在同一pH下不同介质中的稳定性表现为硅藻土膨润土水乳剂,当pH为4.5时噻唑膦的稳定性最佳,而且加入0.2%的环氧大豆油做稳定剂可使噻唑膦在水乳剂中的分解率控制在10%以下。总之噻唑膦在酸性介质中较稳定,在硅藻土和膨润土中的稳定性要高于水乳剂中的稳定性。