上海口岸从旅客携带进境的柠檬树苗中分离到1种半穿刺属线虫的2龄幼虫及雄虫,通过形态特征和分子特征分析,鉴定为柑橘半穿刺线虫(Tylenchulus semipenetrans Cobb,1913).该线虫的主要形态鉴别特征为:雄虫虫体细,头部光滑、锥圆,口针和食道退化,排泄孔位于虫体中后部,单精巢、前伸,交合刺细、弓状,无交合伞,尾长圆锥形、端圆.2龄幼虫无直肠和肛门,排泄孔位于虫体中后部,尾部渐细、钝圆.这是全国口岸系统首次截获该线虫.

剪股颖粒线虫(Anguina agrostis)是牧草和草坪上重要的有害生物,被我国列入检疫性有害生物名录.2018年,上海口岸从来自美国的细弱剪股颖种子中截获1种粒属线虫,由于该样品中仅分离得到幼虫,无法进行准确鉴定.本文采用形态学与分子生物学相结合的方法进行检测.采用28S通用引物扩增该线虫序列,测序获得741 bp的序列,与GenBank中登录的Anguina agrostis相似性达99.85％～100.00％;对ITS区扩增并测序,获得776bp的序列,与GenBank中登录的Anguina agrostis的相似性达99.60％～100.00％.基于28S及ITS区序列构建的系统进化树均显示,该线虫与Anguina agrostis位于同一进化枝上.进一步采用剪股颖粒线虫鉴定国标方法推荐的特异引物对线虫进行PCR扩增,出现目的条带.综合形态学特征及分子鉴定结果,该粒属线虫群体鉴定为剪股颖粒线虫.

Anguina wevelli is a pathogenic grass parasitic nematode, however it is difficult to identify based simply on morphology. This study developed a loop-mediated isothermal amplification (LAMP) assay to detect A. wevelli. The LAMP method developed could specifically detect A. wevelli in 45 min, and the detection limit was 1/80000 of the total DNA extracted from a single juvenile (J2), an equivalent of 2.5 pg of DNA. This is the first report of the detection of Anguina spp. by using a LAMP-based method.

The second-stage juveniles (J2s) of one Heterodera population were intercepted from seedlings carried by passenger at Shanghai port.Based on the morphological characters and molecular identification,the population was identified as Heterodera fici Kirjanova,1954.The morphological characters were mainly as follows:body length 409.9-454.4 μm;labial region with three annuli;stylet well developed,basal knob rounded,directed slightly anteriorly,22.6-24.0 μm long;lateral field with four incisures;tail long,tapering,terminus rounded;tail hyaline length 24.7-30.5 μm,about half of the tail length.The analysis of ITS region sequence showed 99.48％-99.90％ sequence identity with H.fici in GenBank.The phylogenetic tree based on ITS sequences revealed that the sample clustered with H.fici in GenBank.This is the first report of H.fici intercepted in China.

One species of Paratrichodorus nematode was isolated from rhizosphere of cherry-apple tree imported from Germany at Shanghai Port.Female nematode was didelphic,lack of spermatheca.Its sperm was dispersed throughout the uterus and pore-like vulva rounded triangular or oval,with small vaginal sclerotized pieces.Male nematode had one ventromedian cervical papillae in front of excretory pore,large sperm cells with large sausage-shaped nucleus,distal end of spicule nearly straight,spicules shaft with striation,arrow bursa and three ventromedian precloacal supplements.PCR amplification was positive with specific primers for P.pachydermus.Sequence analysis revealed that its 18S-sequence had 99.30％-100％ identity with P.pachydermus in GenBank.Phylogenetic tree of the 18S sequences showed this species clustered with P.pachydermus.Based on the morphological characteristics,PCR detection and sequence analysis,this kind of nematode was identified as P.pachydermus.This is the first report of interception of P.pachydermus in China.

To realize rapid isolation of anguinid nematode galls from grass seed,this paper used a fast separation method.The results showed that there was no significant difference in the detection rate of gall between the fast separation method and direct picking method.The working time decreased from 25 min to 5 min on average,and the working time was reduced largely.The method has the advantages of low cost and simple operation,and is suitable for the daily detection of anguinid nematode galls from grass seeds.

对2011～2014年上海口岸进境植物种苗贸易情况以及截获疫情进行了分析,并对我国进境植物种苗检疫对策进行了探讨,提出了理顺检疫审批制度、开展风险分级管理、加强部门协作以及严格对邮寄和携带植物繁殖材料的检疫等对策建议.

本文介绍了目前粮食检验检疫及监管工作所面临的问题,分析了这些问题带来的风险.同时也介绍了对于解决这些问题所做的探索和研究工作以及最终的方案,即利用计算机信息化手段,建立一个涵盖粮食检验检疫及监管全流程闭环式管理的网络系统,用于规范操作、统一标准、明确职责,确保粮食在检验检疫及调运过程的各个环节中得到有效的管理.最终为国家局“数字化动植检”战略的推进提供了参考.

本研究从欧洲进境水曲柳原木上分离获得了检疫性病原真菌苹果壳色单隔孢溃疡病菌(Botryosphaeria stevensiiShoemaker),分别测定了烘箱热处理和炭化烘房热处理对该病菌存活的影响.结果发现苹果壳色单隔孢溃疡病菌可以侵染至原木的木质部组织,原木中心处理温度达到60℃并持续6h可杀灭原木中携带的病菌.研究结果对进境原木携带苹果壳色单隔孢溃疡病菌及其它重要病菌的检疫处理有较好的参考价值.

2013年,上海口岸从来自美国的画眉草种子中检测到一种疑似粒属线虫的幼虫,并从草籽中收集大量虫瘿.序列分析显示截获的维氏粒线虫ITS序列与GenBank中维氏粒线虫(登录号为AF396317和AM888393)的序列相似性为99.85％,序列差异为1bp.系统进化树中截获线虫样品与维氏粒线虫处于一个聚类组内.经形态学特征观察,并结合ITS序列分析,将截获的虫瘿及线虫鉴定为维氏粒线虫(Anguinawevelli).这是我国口岸首次报道截获该种线虫.

上海口岸从一批进境美国针叶原木上截获活体松材线虫,为全国口岸首次从进境原木上截获活体松材线虫.开展专项松材线虫监测工作后,上海口岸又连续多次从进境美国针叶原木上截获活体松材线虫.为了对染疫原木进行除害处理,上海口岸成功研发出"锯板热处理法"和"原木直接热处理法"2种除害处理方法.本研究对进境针叶原木松材线虫的现场检疫及除害处理方法进行初步探索,为今后开展相关工作提供参考.

Lily anthracnose is one of the most common diseases infecting lily bulbs caused by Colletotrichum lilii.Specific primer pairs were designed by comparing rDNA ITS sequences of C.lilii,C.gloeosporioides and other three Colletotrichum species.By the specific assessment of PCR detection,two pairs of primer pairs,BT73/BT-R510 and BT-F98/BT-R510 were selected to amplify the target DNA sequence of C.lilii,which produced 438 bp and 413 bp specific fragments of C.lilii,respectively,and no any DNA fragments of other four Colletotrichum species and two species,Botrytis cinerea and Fusarium oxysporum f.sp.lilii from the lily bulb were amplified.PCR with the above two primer pairs could detect 50pg DNA of C.lilii at least,but the nested PCR with BT73/BT-R510 as the outer primers and BT-F98/BT-R510 as the inner primers didn′t improved the sensitivity of detection.

为了解决带松材线虫的进口原木的处理问题,利用卧式碳化炉,蒸汽烘房对进口带松材线虫的原木进行热处理试验.研究表明:高压过热蒸汽保持碳化炉内90℃下3h,可100%杀死直径25～45cm原木中松材线虫;3.8cm的板材在90℃的蒸汽烘房处理,且在此温度下保持7h,也可100%杀死3.8cm板材中的松材线虫.

本文简述了台北花博会概况,通过对上海赴台参展植物情况、台湾进境植物检疫要求等进行分析,针对性地开展检疫监管,达到便捷通关的目的,并对出境参展植物检疫监管便捷通关提出建议.

烟草霜霉病是我国禁止进境的检疫性有害生物,我国每年均从进口烟叶中截获此病菌.笔者从事该领域检疫几十年,多次截获此病菌,本文就我国对该病菌的检疫审批措施和实验室检疫方法和应注意的问题进行了介绍.

我国用于鲜切花生产的百合种球进口量逐年增加,而种球作为繁殖材料,因其携带有害生物而造成的传播风险也在不断增加.本文对国内外报道的危害百合球茎的有害生物种类,在病原菌分类地位、危害症状、传播途径等方面研究进展进行了综述.

Liquichip is a new type of biochip with 100 different kinds of fluorescent beads as carriers of different probes and the reaction of biomolecules is finished in suspenseful liquid system.Featuring many remarkable advantages such as the high-throughput,the high-sensitive,the high-accuracy,the high-precision,the great dynamic range and the easy operation.It provides an open platform for assays of protein,nucleic acid and biomolecular interaction analysis.Here we provided brief introduction of its principle and advantages,and reviewed applications of this new array technology and the prospects in plant virus quarantine.

Two isozymes from 10 populations of sweet potato stem nematode,Ditylenchus destructor,were analyzed,as well as the weight loss of sweet potato,pathogenicity,fecundity and sex ratio of these 10 populations by inoculation test in two varieties of sweet potato.The results show that among the 10 populations,there were three main bands of malate dehydrogenase(MDH),which were corresponding to two different phenotypes.Rf was about 0.21,0.26 and the other was 0.21,0.26 and 0.31,but esterase(EST) was no band. The pathogenicity,fecundity and sex ratio of the 10 populations were significant at the P=0.05 level,the highest pathogenicity population was from DeHB.There were positive relationships between fecundity and pathogenicity on the Sushu No.9 and Ningshu 9-9.Partial correlation analysis showed that there was negative relationship between fecundity and sex ratio of the 10 populations on the Sushu No.9 and significant positive relationship on the Ningshu 9-9,and their coefficients were-0.213,0.901,respectively,indicating that the sex differentiation of D.destructor was significantly affected by the different varieties of sweet potato.In addition,the Ningshu 9-9 was 33.48% higher than the Sushu No.9 in the resistant to D.destructor,and 19.86% higher in preservation.

The multiplex RT-PCR approach was developed for simultaneous detections of Arabis mosaic virus(ArMV),Strawberry latent ringspot virus(SLRSV)and Lily symptomless virus(LSV)from the imported lily bulbs.The results indicated that good specificity and sensitivity for simultaneous detection were obtained.The ultimate of RNA detection with three viruses mixture was 305 pg.The ultimate of RNA detection with ArMV,SLRSV and LSV were 156.0,13.4 pg and 1.12 ng respectively.This approach has potential to be used in quarantine of imports and exports.

ITS1 (internal transcribed spacer 1) regions were amplified from 13 Chinese populations of Ditylenchus destructor and one Dutch population of Ditylenchus dipsaci. ITS1 regions of these populations were digested by five enzymes Rsa Ⅰ, Hae Ⅲ, Msp Ⅰ, Hinf Ⅰ and Alu Ⅰ. ITS1-RFLP patterns revealed that two patterns existed in 13 Chinese populations of D. destructor with different fragments obtained by four enzymes Rsa Ⅰ, Hae Ⅲ, Hinf Ⅰ and Alu Ⅰ, and ITS1-RFLP patterns of D. dipsaci and D. destructor were different in fragments obtained by all five enzymes. 13 Chinese populations of D. destructor were divided into two genotypes, i.e. genotype A and B. Genotype A included populations of DeSD1, DeSD2, DeSD3 and DeJS1, and genotype B included populations of DeAH1, DeAH2, DeHB1, DeHB2, DeJS2, DeSX, DeSD4, DeTJ1 and DeTJ2. ITS1 regions were also sequenced and the alignments showed that the pairwise sequence divergences of four populations in genotype A were between 1% and 4%, and that of nine populations of genotype B were 0 and 1%, and the sequence divergences of D. dipsaci and D. destructor were 39% and 48%. Morphometric characters were analysed on four populations of D. destructor and one population of D. dipsai. The results demonstrated that no significant difference in most morphometric characters except c, tail, V and V’ among populations of two genotypes A and B of D. destructor. The ITS1-RFLP patterns, ITS1 sequence analysis and morphometric data supported that two genotypes of D. destructor were existed in China.