东非(进口)植物检疫法规的明细表(续)

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(按农药登记证号)LS 200019金都尔原药批准正式登记的国外农药产品 (包括变更登记)续展临时登记的国外农药产品PD 327一2000LS 93002PD 328一2000高特克5。%悬浮剂(重量/容量)农地乐52.25%乳油(重量/容量)高特克原药精禾草克原药LS 95001PD 329一2000PD 330一2000LS 95016LS 96     

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(按农药登记证号)批准正式登记的国外农药产品 (包括变更登记)PD 320一2000伴地农原药续展正式登记的国外农药产品 (包括变更登记)WL 2000109矶2000110WL 2000111WL 2000112WL 2000113WL 2000114乳2000115WLWL讹乳WL WLWL跳WLWLPD 1 19一90倍乐霸25%可湿性粉剂续展临时登记的国     

番茄褪绿病毒(tomato chlorosis virus)的新寄主:水茄(Solanum torvum Swartz)

Tomato chlorosis virus (ToCV) is a plant virus that is mainly propagated by Bemisia tabaci in a semi-persistent and non-circulative manner.It has a wide range of host plants,and has been reported in many countries,causing serious economic losses in vegetable production.In 2019,we investigated about 10 fields,one ha each in Shouguang,Shandong province (China),and in each field we observed symptoms of interveinal chlorosis on lower leaves of the Solanum torvum Swartz,and a large number of B.tabaci gathered on the back of its leaves.To determine the presence of ToCV,total RNA of S.torvum was extracted followed by RT-PCR.The 1 074 (GenBank accessions number MN545620) and 466 bp (GenBank accessions number MN545621) fragments were gel purified and sequenced.The sequences shared 99.44% and 99.57% similarity with ToCV reference sequence tomato chlorosis virus segment RNA1 (AY903447) and RNA2 (AY903448).The results of insect transmission test confirmed that ToCV can spread from S. torvum to tomato.This study confirms S.torvum as a newly reported host of ToCV.     

番茄褪绿病毒侵染黄瓜的首次报道

番茄褪绿病毒Tomato chlorosis virus(ToCV)是严重危害世界经济作物的一种病毒,寄主范围广泛。田间调查发现黄瓜Cucumis sativus表现出叶片黄化、脉间褪绿的疑似番茄褪绿病毒感病症状,同时叶片背面聚集了大量烟粉虱。采用RT-PCR方法对样品叶片和烟粉虱进行检测,ToCV感染率为65%,且发病叶片上烟粉虱携带ToCV。为进一步确定黄瓜是否为番茄褪绿病毒的新寄主,室内利用农杆菌侵染性克隆接种健康黄瓜,结果显示:接种30 d的黄瓜新生叶片出现褪绿症状。采用ToCV HSP70基因的引物对田间黄瓜叶片、烟粉虱和室内黄瓜新生叶片进行RT-PCR,扩增出约450 bp的条带,在NCBI上BLAST显示与KC887999.1的同源性最高,为99%。这些数据表明黄瓜是番茄褪绿病毒的寄主。这是ToCV感染黄瓜的首次报道。     

番茄褪绿病毒病在山东设施西葫芦上的发生及潜在危害

番茄褪绿病毒tomato chlorosis virus (ToCV)是我国蔬菜生产的重要新发病毒,其寄主范围逐年扩大。2019年10月项目组在山东寿光西葫芦温室调查时发现部分叶片呈现黄化、脉间褪绿,类似于ToCV侵染症状,并伴有烟粉虱发生。利用特异性引物检测发现,扩增的目的片段与GenBank中登录的侵染番茄的ToCV基因序列(登录号：KC887998.1)同源性高达99.58%,充分说明西葫芦植株已被ToCV侵染。通过对病毒病发生规律和烟粉虱虫口数量调查发现,西葫芦定植后1个月表现侵染症状的植株为2.0%,定植后2个月达到4.2%,定植后4个月后高达68.2%,病毒发生呈指数增长,而烟粉虱虫口数量却维持较低密度。从济南和德州采集的西葫芦疑似病叶和烟粉虱中也检出ToCV病毒,说明该病毒可能已经在山东省设施西葫芦主要种植区普遍发生,并经烟粉虱广泛传播,需引起高度重视。     

Tomato chlorosis virus: a new whitefly-transmitted, Phloem-limited, bipartite closterovirus of tomato

ABSTRACT Tomato chlorosis virus (ToCV) is the second whitefly-transmitted, phloem-limited, bipartite closterovirus described infecting tomato. ToCV is distinct from tomato infectious chlorosis virus (TICV), based on lack of serological and nucleic acid cross-reactions and differences in vector specificity. TICV is transmitted only by the greenhouse whitefly (Trialeurodes vaporariorum), whereas ToCV is transmitted by the greenhouse whitefly, the banded-wing whitefly (T. abutilonea), and Bemisia tabaci biotypes A and B (B. argentifolii). Double-stranded (ds) RNA analyses of ToCV show two prominent dsRNAs of approximately 7,800 and 8,200 bp, with several small dsRNAs. Digoxigenin-11-UTP-labeled riboprobes derived from cDNA clones representing portions of RNAs 1 and 2 were used in Northern blot hybridizations to detect two large nonhomologous dsRNAs and a subset of smaller dsRNAs. These probes were used in dot blot hybridizations to detect ToCV in infected tomato. Inclusion bodies and cytoplasmic vesicles were consistently observed in phloem tissues of ToCV-infected Nicotiana clevelandii. Computer-assisted sequence analysis showed significant homology between ToCV clones that hybridize specifically with RNAs 1 and 2 and the lettuce infectious yellows virus methyltransferase of RNA 1 and the HSP70 heat shock protein homolog of RNA 2, respectively. Thus, ToCV is another member of the growing subgroup of bipartite closteroviruses transmitted by whiteflies.     

番茄褪绿病毒与番茄黄化曲叶病毒复合侵染对番茄褪绿病毒传播的影响

番茄褪绿病毒Tomato chlorosis virus(ToCV)是一种由烟粉虱Bemisia tabaci传播的正义单链RNA病毒,在田间常与番茄黄化曲叶病毒Tomato yellow leaf curl virus(TYLCV)复合侵染而造成番茄生产上重大的经济损失。为了明确ToCV与TYLCV的复合侵染对烟粉虱传播ToCV所造成的影响,本文采用RT-PCR以及qRT-PCR检测了复合侵染的番茄对烟粉虱获取和传播ToCV的影响。研究表明,烟粉虱取食复合侵染的番茄后对ToCV的传播效率显著提高,仅25头烟粉虱的传毒率即可达到100%,ToCV在烟粉虱以及番茄体内的累积量均显著提高。说明这种复合侵染促进了烟粉虱对ToCV的传播,在田间应当及时防控烟粉虱,警惕病毒与烟粉虱的蔓延。     

番茄褪绿病毒病预测预报模型的建立

研究烟粉虱传播番茄褪绿病毒Tomato chlorosis virus (ToCV)的发生规律,建立其预测预报模型,能够指导田间早期有效防治。本研究于2014年－2018年每年采集山东寿光蔬菜基地大棚番茄和杂草的植株叶片,并收集植株上携带的所有烟粉虱,以健康番茄、杂草叶片和室内饲养的健康烟粉虱为阴性对照;实验室ToCV侵染性克隆接种的感病叶片以及从感病叶片上获毒的烟粉虱为阳性对照,将采集的样品带回实验室进行检测和鉴定等试验。根据病情发生规律,建立了番茄褪绿病毒病的预测预报模型,模拟所得方程为:Y=2.570+0.089X4-7.548X7,其中Y为11月份番茄褪绿病毒病发生率,X4为采集的样品上平均每株所携带的烟粉虱数量,X7为采集的田间杂草的带毒率。预测模拟结果显示,烟粉虱数量以及杂草的带毒率与番茄褪绿病毒病的发生率极显著正相关,回归检测结果历史符合率为96.8%以上。明确了影响番茄褪绿病毒病发生的影响因子,基于烟粉虱数量以及杂草的带毒率,构建了病害预测预报模型。研究结果有助于及时发现番茄褪绿病毒病并采取相应预防措施。     

烟草花叶病毒丁香分离物的分离与鉴定

 从表现花叶症状的丁香病株上获得一病毒分离物,其在电镜下为约300 nm×18nm的杆状粒子;电泳分析表明感病组织中ds RNA大约为6.4kbp,而其外壳蛋白分子量约为17.6k Da。以上实验结果初步将该病毒分离物鉴定为烟草花叶病毒属(Tobamovirus)。根据该属病毒复制酶基因序列设计通用引物,进行RT-PCR检测,扩增出约1000 bp的预期特异片段(Gen Bank AY566703)。将PCR产物克隆后测序,序列分析表明,与从蚕豆中分离的TMV-B株系序列(Gen Bank AJ011933.1)同源性为99.90%。根据烟草花叶病毒(Tobacco mosaic virus,TMV)的RNA CP基因序列设计引物,进行RT-PCR,扩增出约800 bp的预期特异片段(Gen Bank AY56672),序列分析表明,与TMV-B株系序列(Gen Bank AJ011933.1)同源性达99%,上述实验结果表明,该病毒分离物为TMV。由于该分离物与TMV-B在指示植物上的症状存在明显差异,所以,作者把该分离物暂命名为TMV-S。     

Synergistic Interaction Between Tomato chlorosis virus and Tomato spotted wilt virus Results in Breakdown of Resistance in Tomato

ABSTRACT Multiple viral infections frequently are found in single plants of cultivated and wild hosts in nature, with unpredictable pathological consequences. Synergistic reactions were observed in mixed infections in tomato plants doubly infected with the positive-sense and phloem-limited single-stranded RNA (ssRNA) crinivirus Tomato chlorosis virus (ToCV) and the negative-sense ssRNA tospovirus Tomato spotted wilt virus (TSWV). Synergism in a tomato cultivar susceptible to both viruses resulted in a rapid death of plants. A pronounced enhancement of ToCV accumulation mediated by TSWV co-infection was observed with no evident egress of ToCV from phloem tissues. No consistent alteration of TSWV accumulation was detected. More remarkable was the synergism observed in tomato cultivars which carry the Sw-5 resistance gene, which are resistant to TSWV. Pre-infection with ToCV resulted in susceptibility to TSWV, whereas co-inoculations did not. This suggested that a threshold level or a time lapse is needed for ToCV to interfere or downregulate the defense response in the TSWV-resistant plants.     

陕西杨凌番茄褪绿病毒的分子检测

近年来在陕西番茄生产区出现了一种新病害,病株表现为叶片褪绿,叶脉颜色变深及叶片增厚,果实变小发白,严重影响番茄产量及经济效益,2016年发病极为严重,部分产区甚至绝收。该病疑似由番茄褪绿病毒Tomato chlorosis virus(ToCV)引起。利用ToCV特异引物对感病番茄样品进行RT-PCR检测,结果从所采集的4份病样中均检测到ToCV预期大小的特异片段。对ToCV外壳蛋白CP基因和类热激蛋白HSP70h基因进行克隆与序列分析,ToCV-YL1 CP基因774个核苷酸序列与已报道的ToCVCP基因有较高的一致性,与山东青岛和山西晋中分离物同源性为100%。ToCV-YL1 HSP70h基因1 665个核苷酸序列与已报道的ToCV HSP70h基因有较高的同源性,与中国、韩国、日本、美国、希腊分离物同源性达到99%以上。研究表明ToCV已传播至陕西地区。     

河北省番茄黄化曲叶病毒和番茄褪绿病毒复合侵染及分布

 番茄黄化曲叶病毒(Tomato yellow leaf curl virus, TYLCV) 及番茄褪绿病毒(Tomato chlorosis virus, ToCV)是危害番茄的主要病毒。2016~2017年,在河北省18个番茄主产区调查番茄病毒病危害情况,采集262份植株矮化、叶片黄化、褪绿、卷曲的番茄样品,利用分子生物学技术对病原进行鉴定。结果表明:2016~2017年河北省番茄病毒病发生普遍,所检样品中TYLCV的侵染率为68.66%,其中与ToCV复合侵染率为19.5%;除栾城、滦南、乐亭、永年4地样品暂未检测到ToCV外,其他14个采样点的样品均检测到ToCV,侵染率为19.5%,且全部表现为与TYLCV复合侵染,河北省番茄主产区暂未发现ToCV单独侵染的样品。