烟草品种对烟草花叶病毒病和黄瓜花叶病毒病的抗性鉴定

由烟草普通花叶病毒(TMV)和烟草黄瓜花叶病毒(CMV)引致的烟草病毒病是世界烟草主产区普遍发生且危害严重的侵染性病害,每年给烟叶生产造成了重大的经济损失。本文采用温室苗期接种鉴定的方法,对16份烟草种质进行了TMV和CMV的抗病性鉴定。结果表明:不同的烟草品种对TMV和CMV的抗病性存在较大差异。在供试种质中,对TMV表现免疫的有‘牛耳烟’、‘8301’、‘台烟7号’、‘三生-NN’共4份材料;表现抗病的有‘吉烟5号’、‘双抗70’、‘大护脖香’、‘秦烟95’共4份材料;表现中抗的有‘铁把子’、‘中烟15’、‘秦烟98’、‘中烟98’共4份材料;表现中感的有‘NC89’、‘翠碧1号’、‘云烟97’共3份材料;表现感病的只有‘秦烟97’。对CMV表现中抗的材料有1份,是‘铁把子’;表现中感的有‘秦烟95’、‘三生-NN’、‘8301’、‘牛耳烟’、‘翠碧1号’共5份材料;表现感病的有‘秦烟98’、‘云烟97’、‘中烟98’、‘NC89’、‘大护脖香’、‘双抗70’、‘秦烟97’、‘中烟15’、‘台烟7号’、‘吉烟5号’共10份材料。研究发现,‘铁把子’是兼抗这两种病毒病的材料。本研究明确了我国16个烟草品种资源的抗病性水平,为抗耐病品种的利用与品种合理布局提供科学依据,同时为烟草抗病毒病育种的亲本选择提供抗源信息。     

辽中地区西瓜花叶病病原的分子鉴定

 利用ELISA和RT-PCR方法对采自我国辽中地区的西瓜花叶病样品进行检测,表明其病原为黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)。将此分离物(CGMMV-Wcn)的外壳蛋白基因克隆后进行序列分析,结果表明其cp基因由486个碱基组成,编码161个氨基酸,与已报道的其它分离物一致;CGMMV-Wcn所致症状与西瓜株系(CGMMV-W)相同,且二者cp基因的氨基酸序列完全一致,因此该分离物应为CGMMV-W。     

云南、福建、湖南烟区烟草花叶病主要病毒种类检测及黄瓜花叶病毒亚组鉴定

 采用抗原直接包被和双抗体夹心酶联免疫吸附测定法(ELISA)对采自云南、福建、湖南烟区烟草花叶病样品进行了病毒种类检测,利用三抗体夹心ELISA对黄瓜花叶病毒(Cucumber mosaic virus,CMV)的亚组类型进行了鉴定。在云南采集的520个花叶病样品中,烟草花叶病毒(Tobacco mosaic virus,TMV)、CMV和马铃薯Y病毒(Potato virus Y,PVY)总检出率分别为71.74%、55.01%和6.35%;在福建采集的150个花叶病样品中,TMV、CMV和PVY的总检出率分别为94%、24.66%和8.00%;在湖南采集的74个花叶病样品中,TMV、CMV和PVY的总检出率分别为58.11%、51.35%和2.70%。部分样品为2种以上病毒复合侵染。云南、福建和湖南采集的64个CMV阳性样品中,属亚组Ⅰ的样品为57个,占89.1%;属亚组Ⅱ的样品为10个,占15.6%;其中3个样品为亚组Ⅰ和亚组Ⅱ的复合侵染。     

烟草品种对烟草花叶病毒和黄瓜花叶病毒的抗性鉴定

 2010-2011年采用大田人工接种鉴定的方法,对生产中大面积推广使用的24份烟草品种进行了烟草花叶病毒（TMV）和黄瓜花叶病毒（CMV）的抗性鉴定。结果表明,供试品种对TMV和CMV的抗病性存在较大差异,对TMV表现抗病的有Coker86、吉烟5号、Coker176、CV87、辽烟8号、CV91、中烟90等7份材料;表现中抗的有秦烟98、双抗70、C151等3份材料;表现中感的有秦烟96、G80、金星6007、龙江981、K326、秦烟201、NC89等7份材料;表现感病的有G28、云烟97、净叶黄、红花大金元、RG11、云烟85、云烟87等7份材料。对CMV表现抗病的有Coker86、龙江981、C151、秦烟201、云烟87等5份材料;表现中抗的材料是金星6007;表现中感的有CV91、RG11、Coker176、中烟90、K326、红花大金元、净叶黄、G80、G28等9份材料;表现感病的有秦烟98、云烟85、秦烟96、NC89、双抗70、云烟97、CV87、辽烟8号、吉烟5号等9份材料。其中兼抗TMV和CMV两种病毒病的材料有2份,分别是Coker86和C151。同时研究还发现,抗病性不同的烟草品种在受到病毒危害以后,对烟叶的产量和品质的影响也不同。明确了中国24个烟草品种的抗病性水平,为抗病品种的利用与品种合理布局提供科学依据,同时为烟草抗病毒病育种的亲本选择提供抗源信息。     

Identification of blackeye cowpea mosaic virus from germplasm of yard-long bean and from soybean,and the relationships between blackeye cowpea mosaic virus and cowpea aphid-borne mosaic virus

Two potyvirus isolates, one from germplasm of yard-long bean (Vigna unguiculata ssp.sesquipedalis) introduced into the Netherlands, and another one from soybean plants (Glycine max) in Indonesia, were compared with two virus isolates of blackeye cowpea mosaic virus (BICMV) from the USA and a Moroccan isolate of cowpea aphid-borne mosaic virus (CAMV). It is proposed that all five isolates be now considered BICMV on the basis of host ranges, symptoms and serology. From our results, and a reassessment of the literature it is suggested to drop the name CAMV in favour of BICMV.Samenvatting Twee potyvirussen, de een in Nederland ingevoerd met genenmateriaal vanVigna unguiculata ssp.sesquipedalis en de ander uit planten van sojaboon (Glycine max) in Indonesië, werden vergeleken met twee isolaten van blackeye cowpea mosiac virus (BICMV) en een Marokkaans isolaat van cowpea aphid-borne mosaic virus (CAMV). Op grond van waardplantenreeksen, symptomen en serologie stellen de auteurs voor om alle vijf isolaten te beschouwen als BICMV. Gebaseerd op de verkregen resultaten en een kritische beschouwing van de literatuur wordt de aanbeveling gedaan om de naam CAMV te laten vallen ten gunste van BICMV.     

合肥地区发生的西瓜花叶病的病原鉴定

 从合肥市郊区的西瓜病叶上分离出一病毒分离物,该分离物的热钝化温度为60~65℃,稀释终点为10^-4~10^-5,寄主体外存活期为20~23 d,电镜观察病毒粒子约750 nm×13 nm。参考已发表的小西葫芦黄化花叶病毒(Zucchini yellow mosaic virus,ZYMV) CP基因序列设计引物,RT-PCR扩增得到CP基因片段。将该CP片段克隆到pGEM-3Zf (+)中,测序结果表明,该CP基因全长840 nt,共编码279个氨基酸,与国内报道的ZYMV CP基因的核苷酸序列同源性为83.0%~97.3%,氨基酸序列的同源性为91.8%~99.8%,证明该分离物为ZYMV。     

两个马铃薯Y病毒黑龙江马铃薯分离物株系鉴定

为明确分离自黑龙江省克山县马铃薯上的2个病毒分离物KS4和KS7的分类地位,通过RTPCR扩增、克隆获得其基因组序列,利用重组分析程序包和最大似然法分别进行重组分析和系统发育分析。结果显示,分离物KS4和KS7的开放阅读框均有9 186个核苷酸,编码3 061个氨基酸,分离物KS4的核苷酸和氨基酸序列均与马铃薯Y病毒(potato virus Y,PVY)分离物Mb112一致率最高,分别为96.9%和98.4%;分离物KS7的核苷酸序列与PVY分离物12-94一致率最高,为97.4%,其氨基酸序列与PVY分离物SYR-Ⅱ-Be1一致率最高,为97.8%。重组分析表明,分离物KS4和KS7均为分离物N-605和Oz的重组体,其中KS4基因组5′-端的2 392个核苷酸来自分离物N-605,其余核苷酸来自分离物Oz;KS7基因组的第800～2 227个核苷酸和第5 637～8 950个核苷酸来自分离物N-605,其余核苷酸来自分离物Oz。系统发育分析发现,分离物KS4被聚类到N:O株系(PVY~(N:O)),分离物KS7被聚类到NTN株系(PVY~(NTN))b型。     

植物诱抗剂AHO联合茎尖培养脱除马铃薯S病毒和马铃薯Y病毒

研究了植物诱抗剂3-丙酮基-3-羟基羟吲哚(3-acetonyl-3-hydroxyoxindole, AHO)联合茎尖培养从试管苗中脱除马铃薯S病毒(PVS)?马铃薯Y病毒(PVY)的方法和效率?取PVS侵染的‘定薯3号’和‘定薯4号’以及PVY侵染的‘靖薯3号’和‘靖薯4号’的壮芽, 茎尖剥离后培养至4~5个叶片, 用100 mg/L植物诱抗剂 AHO水剂喷施试管苗, 每隔2 d喷施一次, 共3次, 末次喷施2 d后取茎尖剥离培养, 获得再生试管苗?用电子显微镜负染色?ELISA?荧光定量RT-PCR检测再生试管苗的带病毒情况?结果显示, AHO对4个品种的马铃薯试管苗生长无影响, 用AHO处理后再茎尖剥离培养, 脱毒率均高于未处理的对照; 检测结果还显示AHO处理的马铃薯再生苗的带毒量也低于未处理的对照, 且随处理次数增加带毒量下降?研究结果表明, 利用植物诱抗剂AHO联合茎尖剥离培养方法可以提高脱除PVS?PVY的效率, 获得无病毒核心苗?     

插接与劈接嫁接西瓜对黄瓜绿斑驳花叶病毒病传播的比较

为了明确西瓜嫁接对黄瓜绿斑驳花叶病毒病传播的影响,应用插接法与劈接法中刀片、竹签2种工具分别粘毒后再嫁接西瓜进行传毒能力比较,结果为：插接法嫁接传毒能力强于劈接法嫁接,粘毒竹签的传毒能力强于粘毒刀片,最初嫁接的1、2株瓜苗最容易染毒发病。     

西瓜感染黄瓜绿斑驳花叶病毒后糖的变化与倒瓤关系的研究

 By high performance liquid chromatography (HPLC), the contents of glucose, fructose and sucrose, and the sweetness were analyzed in watermelon inoculated with Cucumber green mottle mosaic virus (CGMMV).The relationship between sugar change and blood-flesh of watermelon was determined. The results indicated that the content of glucose was apparently increased compared to that of the control before maturity (within 28 days after pollination), but reduced to 24.8% of the control after maturity (at 35 days after pollination). The content of fructose was higher than that of the control within 14 days after pollination, and then decreased with a significant difference. In all cases, the content of sucrose increased with the growth of watermelon. However, compared with the control, the sucrose content of watermelon inoculated with CGMMV was lower. The ratios of glucose, fructose and sucrose in the total sugar were abnormal. Coincidence with the changes of the total sugar, the fruit sweetness before maturity was higher than that of the control, whereas decreased sharply after maturity (lower than that of the control). The inner pulp of the mature fruit appeared to be water-soaked and dirty red with no edibility. In a word, after inoculation with CGMMV, the changes of sugars and sweetness affected the watermelon quality.     

西瓜花叶病毒2号新疆昌吉分离物外壳蛋白基因核苷酸序列分析

 利用RT-PCR从新疆昌吉地区表现花叶、疱斑、扭曲等症状的南瓜病株上检测到西瓜花叶病毒2号新疆昌吉分离物(简称WMV-2-XJ-CJ),并测定了该分离物外壳蛋白(CP)基因序列。序列分析表明,新疆昌吉分离物CP基因全长850个核苷酸,编码197个氨基酸。与国内外报道的12个WMV-2CP基因相比,其核苷酸序列同源性为92.6%~98.3%,由此推导的氨基酸序列同源性为94.7%~99.3%。新疆昌吉分离物在CP N'端可变区明显不同于国内外报道的核苷酸序列。WMV-2新疆昌吉分离物与日本和郑州分离物较其它国家和地区的分离物多出6个核苷酸,但其核苷酸及其推导的氨基酸序列差异较大。新疆昌吉分离物外壳蛋白有2个氨基酸残基明显不同于其它分离物,其中蚜传株系的特征结构域DAG突变为DAE。     

Inhibition of different cowpea viruses by non-virulent cowpea mosaic virus is dependent on the type of immunity of the plant to the inhibitory virus

Samenvatting In cowpea-planten die onvatbaar zijn voor het cowpea-mozaïekvirus (CPMV) kan dit virus, mits infectieus, infectie door vier andere, onderling geheel verschillende virussen remmen. Dit vermogen tot aspecifieke remming komt tot uiting in cowpea-planten met onvatbaarheid die bepaald wordt door één dominant gen, maar niet in planten met onvatbaarheid op basis van één recessief gen. De beide typen van onvatbaarheid laten dus in verschillende mate expressie van de genetische informatie van CPMV toe.     

人工接种CGMMV后与西瓜倒瓤相关部分生理指标的研究

 本研究以辽宁省田间主栽西瓜品种“京欣”为试材,通过测定多聚半乳糖醛酸酶(PG)的活性,以及乙烯、丙酮酸及丙酮酸代谢产物—乳酸、乙醛、乙醇含量的变化,明确了西瓜感染黄瓜绿斑驳花叶病毒后上述生理指标的改变与西瓜倒瓤的关系。研究结果表明:接种处理的西瓜果实授粉后7 d和28 d PG出现2次活性高峰,PG活性过高将导致果肉软化;乙烯含量亦在授粉后21~28 d明显高于健康对照,这一变化将导致果肉水渍化败坏;无论接种早晚,西瓜果实内丙酮酸及丙酮酸代谢产物均有不同程度的积累。定植期接种处理的西瓜,授粉后21 d时西瓜果实内丙酮酸含量为健康对照的2.51倍,35 d时达健康对照4.09倍;授粉后7 d乳酸含量已显著高于对照,35 d时接种处理的西瓜果实内的乳酸含量迅速增加至43.5 μmol·g^-1FW,达健康对照的4.7倍;乙醛含量随着西瓜成熟逐渐增加,尤其是授粉后21~35 d乙醛含量增加速度极为迅速,35 d时达健康对照的26.4倍;乙醇含量授粉后7 d最高,达健康对照的10.1倍,随后有所下降。于压蔓期接种处理的西瓜果实,授粉后21 d,丙酮酸、乳酸、乙醛含量明显高于健康对照,但均低于定植期接种处理的,乙醇含量与健康对照无明显差异,也就是说,感染该病毒越早,对西瓜的生长影响越明显。     

Symptom severity of cassava mosaic disease in relation to concentration of African cassava mosaic virus in different cassava genotypes

The concentration of African cassava mosaic virus (ACMV) was assessed by enzyme-linked immunosorbent assay in relation to symptom severity among resistant, moderately resistant and susceptible cassava genotypes. Resistant genotype NR 8083 had significantly lower symptom severity scores ( P  < 0·05) than the susceptible genotype TMS 91934, but the two genotypes contained similar levels of virus concentration. The moderately resistant genotypes TMS 30572 and NR 8082 expressed significantly lower symptom severities ( P  < 0·05) than the susceptible genotypes TMS 91934 and TME 117, but they contained significantly higher virus concentrations ( P  < 0·05) than TMS 91934 and similar virus concentration as in TME 117. However, two other resistant genotypes, TME 1 and TME 8, had low symptom severity scores and virus concentrations. There was significant interaction ( P  ≤ 0·05) between cropping season and virus concentration in all the genotypes except TMS 30572. The resistant and moderately resistant genotypes that had high virus concentrations sustained storage root yield losses. The severity of symptoms expressed was not necessarily a reflection of the virus concentration in some of the genotypes. In addition to the use of symptom severity scores to group genotypes into resistant classes, it is recommended that virus concentration should also be considered. Genotypes displaying mild symptoms, but with high levels of virus accumulation, could be an important source of inoculum in the spread of ACMV by the whitefly vectors. This suggests that each genotype should be tested for virus accumulation prior to its release to the farmers.     

人工接种CGMMV的增殖及其对西瓜 光合作用的影响

为明确西瓜感染黄瓜绿斑驳花叶病毒CGMMV后病毒的增殖过程及其对光合作用的影响,本研究以辽宁省田间主栽西瓜品种‘京欣’为试材,测定了人工接种CGMMV后,西瓜植株体内病毒含量、光合色素、光合作用相关指标以及参与光合作用的关键酶活性的变化,结果显示:接种3d后,即可检测到病毒,24d植株内病毒含量达最高;叶绿素a、叶绿素b及类胡萝卜素的含量均有所降低,叶绿素a在9d后均低于健康对照,而叶绿素b和类胡萝卜素在15d后明显低于健康对照,第18天时,健康对照的叶绿素b含量达接种处理的1.33倍;ALA脱水酶活性除第15天与对照无显著差异外,整个测定期间均低于健康对照,21d时健康对照的ALA脱水酶的活性达29.9U/h,为接种处理的2.18倍;PEP羧化酶的活性显著低于健康对照,测定期间接种处理的PEP羧化酶活性变化不大;乙醇酸氧化酶活性明显升高,测定期间其活性均显著高于健康对照;西瓜叶片的光合速率、气孔导度和蒸腾速率均远小于健康对照,而细胞间隙CO_2浓度却比对照高,本研究为揭示西瓜感染CGMMV后的光合作用变化规律奠定了基础。     

马铃薯Y病毒在种薯中检出率及其株系鉴定

 马铃薯是我国重要粮食和经济作物。马铃薯Y病毒(potato virus Y,PVY)是危害马铃薯生产的重要病害。种植脱毒种薯是防治PVY最有效的途径。马铃薯种薯携带PVY问题严重,但种薯中PVY株系还不清楚。本研究利用PVY特异性抗体检测了7个马铃薯品种362个种薯,发现不同品种种薯带毒率差异较大,最高达12%。通过RT-PCR方法扩增获得了7个PVY分离物编码区全序列。重组分析发现7个分离物基因组均为重组型,根据重组位点的差异可以分为PVY^NTN-NW(SYR-II型)、Rec-1、Rec-2和Rec-3等4种重组类型,后3种为新重组类型。系统进化分析发现,分离物HQH18G3-10与PVY^NTN-NW(SYR-II型)处于同一个大的分支,但与中国PVY大田分离物聚集在一起形成一个相对独立的组,命名为PVY^NTN-NW(CN型);其余6个分离物与数据库中的中国分离物聚集在PVY^N-Wi组。这暗示PVY中国分离物具有相对独立的进化过程,PVY马铃薯大田分离物和种薯分离物进化上相近。所有分离物均能在珊西烟上引起典型叶脉坏死症状,HQH18G3-10引起的坏死症状最为严重。本研究首次报道了我国种薯内PVY发生情况,对分析病毒发生发展规律和防控具有借鉴作用。     

马铃薯Y病毒在种薯中检出率及其株系鉴定

马铃薯是我国重要粮食和经济作物。马铃薯Y病毒(potato virus Y,PVY)是危害马铃薯生产的重要病害。种植脱毒种薯是防治PVY最有效的途径。马铃薯种薯携带PVY问题严重,但种薯中PVY株系还不清楚。本研究利用PVY特异性抗体检测了7个马铃薯品种362个种薯,发现不同品种种薯带毒率差异较大,最高达12%。通过RT-PCR方法扩增获得了7个PVY分离物编码区全序列。重组分析发现7个分离物基因组均为重组型,根据重组位点的差异可以分为PVY^NTN-NW(SYR-II型)、Rec-1、Rec-2和Rec-3等4种重组类型,后3种为新重组类型。系统进化分析发现,分离物HQH18G3-10与PVY^NTN-NW(SYR-II型)处于同一个大的分支,但与中国PVY大田分离物聚集在一起形成一个相对独立的组,命名为PVY^NTN-NW(CN型);其余6个分离物与数据库中的中国分离物聚集在PVY^N-Wi组。这暗示PVY中国分离物具有相对独立的进化过程,PVY马铃薯大田分离物和种薯分离物进化上相近。所有分离物均能在珊...     

Characteristics of a resistance-breaking isolate of potato virus Y causing potato tuber necrotic ringspot disease

An Austrian isolate of potato virus Y^NTN, the causal agent of potato tuber necrotic ringspot disease (PTNRD), was serologically compared with seven Dutch PVY^N isolates. Using polyclonal and monoclonal antibodies, it was found indistinguishable from PVY^N. Determination of the nucleotide sequence of the coat protein cistron and comparison of the deduced amino acid sequence with coat protein sequences of other potyviruses revealed a high level of homology with PVY^N coat protein sequences. This confirmed the close taxonomic relationship of PVY^NTN with the PVY^N subgroup of potato virus Y. PVY^NTN is able to overcome all resistance genes known so far in commercial potato cultivars. Remarkably, transgenic PVY-protected tobacco plants are also resistant to PVY^NTN infection upon mechanical and aphid-mediated inoculation. These experiments indicate that genetically engineered resistance offers great potential in protection of potato to new aggressive strains of PVY^N.