柑桔黄龙病毒研究初报

試驗用萊檬(Citrus aurantifolia)及麻疯柑(C.hystrix)作指示植物,用两广黃龙病区病株接穗嫁接接种,表現脉明、叶脉木栓化、茎部凹斑、及矮化、小叶等典型退化病毒(tristeza virus)所致症状,可知我国柑桔黄龙病与国外柑桔退化病(tristeza)系同一种病毒(或病毒群)所致。无病区的四川江津、湖南溆浦和北京的健康甜橙、桔类、及北京檸檬也带有黄龙病毒,可見黄龙病毒的分布远较病区为广。各种柑桔上所带黄龙病毒株系的強弱不同;在碰柑上的为强毒系,甜橙上的为强毒系或较弱毒系,桔类及北京檸檬上的各别为較弱毒系及弱毒系。     

毒源和寄主对褐色桔蚜传播柑桔衰退病毒效率的影响

褐色桔蚜是柑桔衰退病毒(Citrus tristeza virus,CTV)最有效的传播媒介,为了解不同柑桔寄主种类和毒源对褐色桔蚜传播CTV效率的影响,检测在以锦橙、凤凰柚和墨西哥来檬作毒源植株时,褐色桔蚜对10个CTV分离株的单蚜传毒率,以及蚜传前后CTV分离株p25/HinfⅠRFLP组群构成的变化,并对其中5个分离株与2个国外分离株进行了p20和p25基因相应氨基酸序列的比对分析。结果表明:褐色桔蚜传播CTV的能力受柑桔寄主种类的影响较大,以锦橙作毒源植株可以获得最高的传毒率;CTV毒株强弱以及褐色桔蚜虫态(有翅或无翅蚜)对单蚜传毒率影响不明显;褐色桔蚜传播具有p25/HinfⅠRFLP第3组群构成的CTV分离株的能力较强。     

文旦柚矮化病的初步鉴定

苏鸿基(1979、1981)报道了台湾葡萄柚和柚的矮化病,接种墨西哥莱檬引起轻脉明和轻茎陷点,尤力克柠檬无黄化症状,葡萄柚叶片显著黄化和皱缩,ELISA等试验证实为衰退病毒(CTV)的一个株系。Tsai等(1992)进一步指出,柚茎陷株系引起柚严重茎陷点,称为衰退病毒矮化株系(CTV—D),并指出文旦柚矮化病在国内华南发生。作者1990年注意到浙江省玉环县文旦柚发生矮化症状树,对此病进行了调查和鉴定,初步结果报道如下。 1 发生和症状特点     

枸头橙种子中柑桔衰退-茎陷点病毒的鉴定

 采用双抗体夹心-酶联免疫吸附测定试验(DAS-ELISA)对枸头橙种子中柑桔衰退-茎陷点病毒的带毒情况进行了鉴定。结果表明,从严重感染茎陷点病的枸头橙罹病树采集的种子可检测到柑桔衰退病毒(CTV)。种子带毒量以内种皮为较高,剥皮种子次之,外种皮呈阴性。种子在4℃冰箱内贮藏1~2年后仍带毒,但带毒量有所下降。对带毒种子繁殖的实生苗及嫁接在实生苗上的墨西哥来檬的检测结果,有部分呈阳性,种植在防虫隔离网室内的100余株2年生枸头橙实生苗,用无毒珠心系的墨西哥来檬芽嫁接进行生物鉴定,经2年观察未发现症状     

宫川温州蜜柑感染碎叶病观察简报

柑桔碎叶病是一种病毒病害，它主要为害以识及其杂种（如积澄）作砧木的柑桔树，其症状为砧穗接合处环缢和接口附近的接穗部肿大，叶脉黄化，类似环状剥皮引起的黄化和植株矮化，受台风等外力推动，病树砧格接合处易断裂，裂而光滑。目前围内发现有碎叶病的省（区）有浙江、四川、广西、广东、福建、湖南、湖北以及台湾省，感染的品种有本地平、少校本地早、银桔、早桔、米红、乳桔、木共柑、北京柠檬、兴津温州蜜柑、柳澄、蕉柑、交府红心袖、要柑、五布红心拍、古老钱沙田袖、祈会橙、水精橙、大红甜蹬、黄陵无核橙等。据悉，截止1993年底…     

柑桔碎叶病的分布鉴定

利用指示植物腊斯克枳橙对国内5个省(区)的29个地方品种76个标样和8个从日本引进的特早熟温州蜜柑品系进行鉴定,其中广东的蕉柑、雪柑、暗柳橙、新会橙:湖北的黄陵无核橙:四川的夔府红心柚、奎柑以及从日本引进的德森温州蜜柑都感染有碎叶病毒。在腊斯克枳橙上的症状是叶片上出现透明黄斑,圆形或不规则形,叶缘缺损,叶片畸形,茎上有微凹的褪绿茶斑,茎秆弯曲,严重时,整株矮化。     

香蕉束顶病毒株系的研究

 采自福建各地蕉区的不同香蕉束顶病毒株类型之间致病性和介体蚜虫的传病率有明显的差异,进一步依其在香蕉品种台湾蕉和Williams上的反应、香蕉交脉蚜的传病率、毒株类型之间的交互保护和血清学测定结果等,把香蕉束顶病毒区分为BBTV-S(重型)和BBTV-M(轻型)两个株系。
BBTV-M和BBTV-S均能引致香蕉叶片上的青筋症状,两者在血清学上有密切关系,但BBTV-M仅引致植株产生少量青筋,而BBTV-S除引致香蕉植株上有大量青筋之外,还引致严重矮化、束顶以及轻度黄化,BBTV-M的潜育期较BBTV-S明显长,香蕉交脉蚜对BBTV-S的传病率大大高于对BBTV-M的传病率,BBTV-M对BBTV-S有强的保护作用。     

柑桔黄龙病热治疗的研究

柑桔黄龙病村有“均匀黄化型”和“斑驳型”两类型的症状。两类型的橙类和蕉柑苗木经49℃湿热空气处理50～60分钟或用50℃处理50分钟,89株苗木全部恢复健康,经58～60个月后全未发病,而对照的病苗种后其病情日趋严重,陆续死亡。试验结果除进一步表明热处理可以消灭黄龙病病原类立克次体外,并进一步表明“黄化型”和“斑驳型”的病原有可能属于同一种病原或相近似的病原。     

一种蚜虫持久性传病的长豇豆黄化型病毒

 1983年6月,在南京郊区的长豇豆上采到1株表现植株矮缩症状的C-7病毒分离物。接种试验证明,它不能摩擦接种传病,但可以由豆蚜(Aphis craccivora)、棉蚜(A.gossypii)和桃蚜(Myzus persicae)以持久性方式传病。寄主范围测定的结果表明:分离物可以侵染长豇豆、豇豆、蚕豆、大豆、菜豆、豌豆、赤豆、利马豆、苜蓿、红三叶、地三叶、绛三叶、葫芦巴,紫云英和苕子等15种豆科植物和曼陀罗1种茄科植物。这些植物大都出现植株矮化,叶片扭曲,卷缩或僵缩,不能开花结实等症状。豆蚜的传病性状中,获毒饲育的最短传病时间为3小时,接毒饲育最短传病时间为10分钟,循回期是24小时左右。但是,传病率最高的获毒饲育时间是2~3天,接毒饲育时间在1天以上。接种1头蚜虫就具有传病能力,5头蚜虫能达到100%的传病率。蚜虫可以终身传毒,蜕皮不影响其传毒力,但传毒有间歇性。根据它的基本性状,病毒C-7分离物是一种豆科植物的黄化型病毒,可能是属于大麦黄矮病毒组(Luteovirus Group)的成员。     

柑桔黄梢(黄龙)病的进一步研究

柑桔黄梢(黄龙)病的研究几年来的进展不快,但仍取得了如下一些成果。柑桔树受侵染后,一般在夏、秋梢老熟期开始发病,但最近发现,亦可在春梢老熟后开始表现病状,其特征为新叶老熟后转为灰绿色。冬季发病的初期病状有时表现为老熟后的新叶中肋黄化。病树叶的澄清汁液与 KOH 或与 NaOH 混和,每有转变为橙缸色的反应,但不一致。曾经被认为可能是黄梢病原的真菌“Deuterophoma sp”已被证明与本病的发生无关。初步试验结果表示柑桔褐蚜可能是一种传染媒介。种子带病的可能性不能排除,有待进一步探讨。病树的根残留土中显然不起传病作用。黄梢病毒在病株体内似以相等的速度同时向上下运转。用13种柑桔类作为砧木,对蕉柑的抗病性皆不起显著的增强作用。不同树龄的植株表现显然不同的抗病性。苗木和老年树比较抗病,幼年树则容易发病。环境因素对植株的发病不起决定性的影响,因而采取栽培措施不能抑制发病,更不能收到治疗效果。挖除病株和喷药除虫则可在一定程度上起抑制蔓延的作用。初步试验表示,用热水或湿热空气处理病树芽条可以消除带病。     

Separation and interference of strains from a citrus tristeza virus isolate evidenced by biological activity and double-stranded RNA (dsRNA) analysis

Separation of strains of citrus tristeza virus (CTV), differentiated by their double-stranded RNA (dsRNA) profiles, was obtained by graft-inoculating citron plants from a Mexican lime that had been recently aphid- or graft-inoculated with a mild CTV isolate (T-385). Up to 24 sub-isolates with differing dsRNA profiles were obtained from the aphid-inoculated lime. Some of these sub-isolates induced stronger symptoms in several citrus species than the original T-385 isolate. One sub-isolate, T-385-33, was mild in Mexican lime, but induced stem pitting on sweet orange. Inoculation of this isolate on Mexican lime, sour orange and Eureka lemon induced mild or no symptoms when inoculum was taken from citron, but very severe symptoms when the inoculum was from sweet orange. Mexican lime and sweet orange plants co-inoculated with T-385-33 from sweet orange in combination with the other 23 sub-isolates showed mild symptoms. The results obtained suggest that there is natural cross-protection among sub-isolates in the original T-385 isolate.     

柑桔苗黄型衰退病毒的分布概况和六种酸橙类砧木对它的反应

 用尤力克柠檬和葡萄柚作指示植物,鉴定广西、广东、湖南、江西、浙江、四川等6个省(区)的873株柑桔中有691株受苗黄型衰退病毒感染,说明这种病毒的分布相当普遍。     

Investigation of biological properties, double-stranded RNA patterns and antigen concentration in citrus species infected with citrus tristeza virus

Biological properties and dsRNA patterns of one Cyprus and three Turkish isolates of citrus tristeza virus (CTV) were investigated. In addition, CTV antigen concentration and effect of tissue sampling time from naturally infected Shamouti sweet orange trees grown in the field of Icel Province, Turkey, were also determined. The Cyprus isolate showed vein clearing symptoms on grapefruit, ‘Madam Vinous’ and Mexican lime and stem pitting symptoms on Mexican lime. The three Turkish isolates showed only vein clearing symptoms on Mexican lime. All four isolates showed a full-length major double-stranded RNA (dsRNA) band of 13.3 × 10⁶ Da mol. wt in extracts from infected Madam Vinous sweet orange trees, and major or minor dsRNA bands with 2.0. 0.8 and 0.5 × 10⁶ mol.wt. All seven different citrus varieties inoculated with the Igdir (D) strain contained full-length dsRNA. The additional two dsRNA of 0.8 and 0.5 × 10⁶ mol.wt were also detected as clearly as full-length dsRNA in these hosts, but were weaker inCitrus exelsa and ‘Interdonat’ lemon. Madam Vinous, rough lemon and Mexican lime were the best hosts for dsRNA analysis. ELISA values were highest in April (OD_405nm =0.476), decreased steadily until August, and then increased gradually through December. ELISA values were lowest in July and August (OD_405nm =0.157 and 0.141, respectively). dsRNA recovery from a field tree infected with isolate Igdir D was good in March, April and May and poor in January and February. No dsRNA band was detected in August or September. http://www.phytoparasitica.org posting July 9, 2002.     

Characterization of Citrus tristeza virus isolates in northern Iran

The biological and molecular properties of four Citrus tristeza virus (CTV) isolates isolated from infected Satsuma trees imported from Japan, and growing in citrus groves in northern Iran (Mahdasht orchards, Mazandaran Province), were investigated. CTV-infected samples were collected from sweet orange trees and grafted onto Alemow (Citrus macrophylla Wester) seedlings. On indicator plants, these isolates produced various symptoms including vein clearing and stem pitting on Mexican lime, Alemow, and Citrus hystrix, and yellowing and stunting on sour orange and grapefruit seedlings. Citrus samples were also surveyed for CTV using serological tests. The coat protein (CP) gene of these isolates was amplified using specific primers, yielding an amplicon of 672 bp for all isolates. Sequence analysis showed 98%–99% sequence homology of Iranian isolates with the Californian CTV severe stem-pitting isolate SY568 and 97%–98% homology with the Japanese seedling yellows isolate NUagA. The Iranian isolates were compared by restriction fragment length polymorphism (RFLP) analysis of the CP amplicon for further classification.     

摩洛哥脐血橙植株上一种机械传播性病毒的鉴定

 本文采用叶片接种法和枝条接种法对25个柑桔种类体细胞杂种或品种进行了抗脚腐病鉴定,结果表明:体细胞杂种中枳+酸橙、枳+甜橙为高抗类型;酸橙+兰普来檬、枳+印度酸桔;蚝壳刺+哈姆甜橙、酸橙+澳洲指桔为抗性;酸橙+印度酸桔为中抗。建立了叶片接种法的抗病性分级标准。初步分析了柑桔体细胞杂种的抗性规律。     

Detection and characterization of <Emphasis Type="Italic">Citrus tristeza virus</Emphasis> stem pitting isolates in Jamaica

An island wide survey for Citrus tristeza virus (CTV) in citrus orchards across Jamaica (13 regions) was conducted over 2 years. Trees (1, 885) showing virus-like symptoms as well as asymptomatic trees were randomly sampled for testing by ELISA and 55 samples from the 6 major citrus growing regions were graft inoculated on indicator plants. Most samples (74%) reacted to polyclonal antibodies against CTV in ELISA, while 20% were positive in tests using monoclonal antibodies specific to severe CTV strains. Samples collected from the 6 major citrus growing regions produced vein clearing and stem pitting symptoms on Mexican lime indicator plants (87%). In addition, stem pitting symptoms were induced on Duncan grapefruit, sweet orange, sour orange or sweet orange grafted on sour orange. Nucleotide sequencing of the coat protein gene sequences isolated from these samples indicated high identities (88 to 95.5%) among the Jamaican isolates and previously reported stem pitting strains from Central and North America and Eurasia (88 to 100%). The results suggest a shared ancestry with isolates from other geographical locations, rather than geographical speciation, and presumably separate CTV introductions into Jamaica.     

应用改进的多克隆抗体Western blot技术研究柑桔速衰病毒蛋白

 本文利用多克隆抗体发展了一种无背景的Western blot技术并用于研究柑桔速衰病毒(CTV)的蛋白。结果表明,利用CTV兔多克隆抗体1212和1052发展的Western blot技术可以检测到感染CTV的墨西哥酸橙或Citrus excelsa植株内CTV的4种蛋白P1、P2、P3和P4。在健株或病株内杂交反应均无非特异性背景。从不同寄主上分离到的CTV不同分离物的蛋白条带是不同的。利用1212和1052抗体均可以检测到感染6个CTV分离物的墨西哥酸橙幼苗内的P1、P2和P3。利用1052抗体能检测到感染严重型分离物T36、T3和Mm2的墨西哥酸橙幼苗内微弱的P4,但感染轻型分离物T30、T26和T4的幼苗内则检测不到。利用1212抗体检测不到P4。在C. excelsa内,1212和1052抗体均能检测到感染所有分离物的病株内的P1。在感染T3、T26、T4或Mm2的病株内能检测到P2,但在感染T30和36分离物的病株内则检测不到。在感染T36、T3、T26、T4和Mm2的病株内可检测到P3,但在感染T30的病株内则检测不到。在大多数植物内,P1、P2、P3和P4的分子量分别约为25kDa,24kDa,21kDa和18kDa。在感染T36分离物的C. excelsa植株体内,P1和P3的分子量分别约为27kDa和22kDa,比感染其它分离物的C. excelsa和墨西哥酸橙内的P1和P3分子量略大。P1、P2和P3的分子量与利用单克隆抗体检测的CP,CP1和CP2的分子量相等。因此,P1、P2和P3可能是CTV的外壳蛋白CP,CP1和CP2。P4的特性不清楚。研究结果也表明,利用特异性的多克隆抗体进行的Western blot是研究CTV的一种有用的技术。应用该技术,病株内不同的CTV分离物或株系就可以通过特异性蛋白条带区分开来。     

Effect of vesicular arbuscular mycorrhiza on citrus growth and mineral composition

The vesicular arbuscular fungusGlomus mosseae (Nicol. & Gerd.) Gerd. & Trappe was retrieved from citrus trees growing in loess soil in the Negev region of Israel. Inoculation of citrus seedlings with the mycorrhizal fungus greatly increased the growth of plants in soil low in phosphorus. Rough lemon responded more than Sour orange. Vesicular arbuscular mycorrhiza caused higher concentrations of P and Cu and lower concentrations of N, K and Ca in leaves of inoculated plants.