Detection of lethal yellowing phytoplasma in embryos from coconut palms infected with Cape St Paul wilt disease in Ghana

This study investigated the potential of seed transmission of Cape St. Paul wilt disease (CSPWD) in coconuts. PCR amplification was used to assess the distribution of phytoplasmas in parts of West African Tall (WAT) palms infected with CSPWD. Employing phytoplasma universal primer pair P1/P7 in standard PCR, or followed with a nested PCR using CSPWD–specific primer pair G813f/AwkaSR, phytoplasma infection was detected in the trunks, peduncles, spikelets, male and female flowers of four infected WAT coconut palms. Through nested PCR, phytoplasma was also detected in four of 19 embryo DNA samples extracted individually from fruits harvested from three of the four infected palms and was confirmed as CSPWD by cloning and sequencing. Subsequently, CSPWD phytoplasma was again detected in five of 33 embryos from nine infected palms, and in one of eight fruits from two symptomless palms. Fruits from infected palms recorded higher percentage germinations in two field nurseries (average of 71·0%) compared to fruits from healthy palms (average of 57·6%), and matured fruits that had dropped from infected palms showed the same levels of germination as those harvested directly from the palms. This indicates that infected fruits retain the ability to germinate whether harvested or dropped. No phytoplasmas were detected in any of the resulting seedlings and plantlets obtained through embryo in-vitro culture. Therefore, although phytoplasma DNA can be detected in embryos, there is as yet no evidence that the pathogen is seed transmitted through to the seedling to cause disease in progeny palms.     

Transmission of a mycoplasma-like organism from cocos nucifera with root (wilt) disease to Catharanthus roseus by Cassytha filiformis

A mycoplasma-like organism (MLO) was successfully transmitted through dodder laurel (Cassytha filiformis) from root (wilt) diseased coconut to periwinkle (Catharanthus roseus) plants. Symptoms caused by the organism in periwinkle were chlorotic spots in the interveinal areas and at vein endings of fully opened leaves, later coalescing into yellow patches, and premature leaf abscission. Light and electron-microscopic examination revealed the presence of MLOs in diseased coconut palms, dodder laurels and periwinkles.Samenvatting Door de als warkruid groeiende lauraceeCassytha filiformis werd van kokosnootpalm (Cocos nucifera), aangetast door root(wilt), een mycoplasma-achtig organisme (MLO) overgebracht naar de toetsplantCatharanthus roseus. Hierop werden op de bladeren tussennervige, later ineenvloeiende vlekken en vroegtijdige bladval waargenomen. Uit onderzoek met de lichtmicroscoop en de elektronenmicroscoop kon de aanwezigheid van MLO's inCocos nucifera, Cassytha filiformis enCatharanthus roseus worden vastgesteld.CPCRI contribution No 622     

Comparison of Secreted in Xylem (SIX) genes in two fusarium wilt pathogens of ornamental palms

Fusarium oxysporum ff. sp. canariensis (Foc) and palmarum (Fop) cause lethal fusarium wilt on ornamental palms. Foc infects Phoenix canariensis and has a worldwide presence, whereas Fop primarily infects Washingtonia robusta and Syagrus romanzoffiana and is currently primarily restricted to Florida (USA). A Secreted in Xylem (SIX) gene profile was obtained for Fop and Foc isolates from the USA. SIX1, SIX7, SIX10 and SIX12 were detected in most Foc isolates and, with the exception of SIX12, with minimal sequence polymorphism. SIX8 and SIX9 were detected in all Fop isolates, with no sequence polymorphism. SIX10 was also present in seven of the Fop isolates. Phylogenetic trees were constructed using EF-1α-, SIX1, SIX7 and SIX10 gene sequences from this study and previously derived sequences from Foc isolates obtained in Australia, the Canary Islands and Japan. The topology of the housekeeping EF-1α gene indicated Foc is polyphyletic, with the Australian isolates separating into a distinct group from all other Foc isolates. However, the topologies of SIX1, SIX7 and SIX10 phylogenetic trees for Foc indicate monophyly. As has been proposed previously, this suggests the core genome of Foc evolved into a separate lineage after the SIX genes were conserved within the whole genome. This study is the first to generate SIX12 sequences for Foc. Interestingly, the topology of the SIX12 phylogenetic tree suggests polyphyly. For Fop, the topologies of the EF-1α, SIX8 and SIX9 phylogenetic trees support monophyly. The distinct SIX gene profile and EF-1α sequence of Fop demonstrates an independent evolutionary origin from Foc.     

万寿菊根的提取物对西瓜枯萎病反应的抗性研究

 本文研究了万寿菊根的提取物对西瓜枯萎病菌和西瓜幼苗的影响。结果表明,万寿菊根的提取物对供试的西瓜枯萎病菌菌丝生长有明显的抑制作用;同时,经对西瓜幼苗根和叶的酶活性测定和过氧化物同工酶电泳分析的结果表明,该提取物能诱导西瓜保护酶如超氧化物歧化酶(SOD)和过氧化物酶(POD)活性提高,减轻西瓜枯萎病菌粗毒素对西瓜幼苗的毒害作用。     

铵态氮和硝态氮对香蕉枯萎病发生的比较研究

 为寻找降低香蕉枯萎病发生的防治措施,通过室内盆栽接种试验,研究了铵态氮(NH₄⁺-N)和硝态氮(NO₃^--N)对香蕉枯萎病发生及其植株叶绿素含量、气体交换参数、病原菌在植物体内的数量分布和植物钙(Ca)、镁(Mg)、铁(Fe)、钼(Mo)、可溶性糖和木质素含量的效应。结果表明:不接种病原菌的条件下,不同氮素处理对香蕉幼苗生长影响无差异;接菌情况下,与NH₄⁺-N处理相比,NO₃^--N处理显著降低植株各器官的病原菌数量、发病率和发病严重程度。病原菌侵染后,不同氮素处理下植株光合作用均显著下降:NO₃^--N处理香蕉苗保持比NH₄⁺-N处理更高的光合速率;病原菌侵染后NH₄⁺-N处理的植株Ca、Mg、Fe和Mo含量相对于侵染前没有显著差异,但NO₃^--N处理下此4种元素含量均显著升高。病原菌侵染后的植株叶片可溶性糖含量在不同氮素处理中都没有显著变化,但在根系中,NO₃^--N处理的侵染植株可溶性糖含量显著降低。与此同时,病原菌侵染后,木质素含量在NH₄⁺-N处理植株中变化不显著,但其含量在NO₃^--N处理侵染后显著上升。综上所述,NO₃^--N处理可增加植株抗病相关矿质元素的吸收,诱导香蕉苗木质素形成,使其木质化程度增加,从而维持较高的光合作用,保持较高的抗病水平。     

健康与感染青枯病烟株根际土壤与茎秆真菌群落结构与多样性

为研究感染青枯病后烟株根际土壤与茎秆真菌群落结构与多样性的变化,对健康和感染青枯病烟株的根际土壤、病株茎秆发病组织和健株茎秆健康组织等样品中真菌ITS区的rDNA进行了PCR扩增、用Illumina MiSeq测序技术对扩增DNA片段进行高通量测序,并分析不同样品的真菌群落组成与多样性。结果表明,所有烟株根际土壤中优势门为子囊菌门Ascomycota和接合菌门Zygomycota;所有茎秆样品中优势门为担子菌门Basidiomycota和子囊菌门。在属水平,被孢霉属Mortierella、镰刀菌属Fusarium和隐球菌属Cryptococcus为所有土壤中的主要菌属,Boeremia主要存在于发病烟株根际土壤中,而木霉属Trichoderma主要存在于健康烟株根际土壤。发病茎秆病害组织中优势属为小画线壳属Monographella、隐球菌属、鬼伞属Coprinopsis和赤霉属Gibberella;发病茎秆病健交界处组织中优势属为隐球菌属、红酵母属Rhodotorula和小画线壳属。健康烟株茎秆组织中优势属为隐球菌属、链格孢属Alternaria和红酵母属;健康烟株中与发病茎秆病健交界处组织等高茎秆中优势属为镰刀菌属、隐球菌属、链格孢属和Gibellulopsis。青枯菌侵染烟株后根际土壤、发病茎秆病害组织和发病茎秆病健交界处组织的真菌群落中物种丰富度与多样性均显著提高,且发病茎秆病害组织与发病茎秆病健交界处组织真菌群落的变化大于根际土壤。研究结果为烟草青枯病的生物防治提供了参考。     

小麦类病变LSD基因全基因组鉴定与转录组分析

类病变(lesion simulating disease, LSD)基因编码锌指蛋白家族, 在细胞程序性死亡(PCD)和其他生物过程中发挥作用。本研究鉴定了普通小麦Triticum aestivum和其3种亚基因组供体乌拉尔图小麦T.urartu、野生二粒小麦T.dicoccoides和节节麦Aegilops tauschii中LSD基因家族。结果表明, 普通小麦、乌拉尔图小麦、野生二粒小麦和节节麦中分别含有24、4、10个和5个LSD基因; 同源性分析表明, 普通小麦相关的同源物共70对, 有38对旁系同源物(54%)。蛋白质特征显示, 13个(54%)TaLSD基因含有3个zf-LSD1结构域, 大部分TaLSD为碱性蛋白质、不稳定蛋白质和非亲水蛋白质; 转录组分析表明, 小麦LSD基因的表达量与禾谷镰刀菌、白粉菌和条锈菌的侵染紧密相连。     

茄青枯病菌引起的新病害-罗汉果青枯病

 罗汉果为葫芦科(Cucurbitaceae)罗汉果属[Siraitia grosvenorii(Swingle) C.Jeffrey]植物。罗汉果中含有0.8%~1.0%的罗汉果甜甙,其甜度为蔗糖的300倍,是肥胖症、高血压、高血脂、糖尿病等患者最理想的甜味剂与保健品。罗汉果甜贰远销美国、日本等国外市场,产品供不应求。     

Malformation and cracking of nuts in coconut palms (Cocos nucifera) due to the interaction of the eriophyid mite Aceria guerreronis and Botryodiplodia theobromae in Tamil Nadu,India

Journal of Plant Diseases and Protection - In 1998, a severe outbreak of malformation and cracking of nuts in coconut palms (Cocos nucifera) was reported in Tamil Nadu State, India. Concentrated...     

Biological and molecular characterization of tomato spotted wilt virus (TSWV) resistance-breaking isolates from Argentina

Tomato spotted wilt virus (TSWV) has been present in Argentina since 1938 and had limited sweet pepper and tomato production until the introduction of resistant cultivars bearing Tsw and Sw-5b genes. However, the wide use of TSWV-resistant pepper plants in La Plata Horticultural Belt (LPHB) triggered the emergence of resistance-breaking isolates (RB), increasing the economic impact of TSWV in pepper. This work characterized 11 natural RB pepper isolates from LPHB that have overcome the Tsw resistance gene in Capsicum sp. but are unable to break the Sw-5b-mediated resistance in tomato. Phylogenetic analysis of the N gene showed that the LPHB isolates are most closely related to isolates from Asia, indicating that Argentine TSWV isolates might have emerged from the Asian continent. The NSs sequence analysis reinforces the hypothesis that the appearance of an RB phenotype is a consequence of a number of different single amino acid substitutions spread along the NSs gene that lead to multiple independent evolutionary events. These results provide information on the current situation of the tospovirus–pepper/tomato pathosystems in LPHB, which represents a fundamental prerequisite to include these RB isolates in future screening programmes in order to select new and durable sources of resistance to TSWV in pepper.     

Diversity among agrobacteria isolated from diseased plants of blueberry (Vaccinium corymbosum) in Argentina

The aim of this study was to isolate, identify and analyze the diversity of the causative agents of crown galls and hairy roots from symptomatic plants of Vaccinium corymbossum by means of biological, biochemical and molecular tools. All the bacteria isolated from blueberries (n?=?78) were found to be Agrobacterium since they grew on three differential media, provoked cell and/or root proliferation on Kalanchoe, and contained a 730?bp partial sequence that codes for virulence genes within the virC operon found on Ti and/or Ri plasmids. Isolates were highly variable considering the ERIC-PCR patterns as well as biochemical reactions and were all represented by 7 different restriction patterns of the 16SrDNA. While most of the isolates belonged to Agrobacterium bv. 1 (n?=?33) or Agrobacterium bv. 2 (n?=?31) only fourteen were Agrobacterium rubi. A representative isolate of each of these three groups was further identified by sequencing the approximately 400?bp 16SrDNA. We concluded that Vaccinium plants are particularly susceptible to Agrobacterium bv. 1, Agrobacterium bv. 2, and also to Agrobacterium rubi. To our knowledge this is the first survey of Agrobacterium affecting blueberries in Argentina.     

从越南入境的椰子树苗中截获椰心叶甲初报

椰心叶甲 [Brontispa longissima ( Gestro) ]是我国禁止进境的二类检疫危险性害虫。我国仅 1 999年度南海口岸 6次从台湾进口的华盛顿椰子 ( Wash-ingtonia filifera )和光叶加州蒲葵 ( W.robusta )中截获此虫。我们于 2 0 0 0年 1 0月 2日在凭祥市浦寨边贸点对一批 5 0株来自越南的椰子树苗实施检疫时 ,截获此虫 ,这是我国口岸首次从越南入境的树苗中截获此虫。由于椰心叶甲对椰子等棕榈植物造成的危害十分严重 ,如果传入扩散 ,将对我国南方的椰子产业及越来越重要的观赏棕榈的种植业造成巨大的损失。现将此次截获椰心叶甲的检疫情况介…     

Occurrence,genetic diversity and pathogenicity characteristics of Pseudomonas viridiflava inducing alfalfa bacterial wilt and crown root rot disease in Iran

Alfalfa (Medicago sativa), the world’s most influential forage crop, is infected by many diseases such as alfalfa bacterial wilt disease. The causal agent of bacterial crown and root rot and wilt disease is Pseudomonas viridiflava, which is a substantial pathogen of alfalfa worldwide. This pathogen spreads through the xylem and under field conditions, plants show growth stunting, chlorosis and wilting symptoms not previously reported. In this study- the first on Pseudomonas viridiflava on alfalfa from Iran, we have investigated the pathogenicity and genetic diversity of Pseudomonas viridiflava in some parts of Iran. To survey the causal agent of the disease, symptomatic plants were collected from the main alfalfa growing area. Pathogenicity of the collected strains was confirmed on alfalfa plants under green-house conditions using a completely randomized design. Determination of bacterial strains was done based on standard bacteriological methods and PCR assay using specific primers. Effects of bacterial strains on wet weight, dry weight, stem length and root length of infected plants were measured and the data were analyzed by SAS software and Duncan’s assessment. The diversity of liquid cellular proteins of bacterial strains was examined on Polyacrylamide gel. To delineate of genetic diversity the total DNA was drawn out. Fourteen random primers were used in a RAPD test. To sketch the dendrogram, RAPD fragments were used to calculate genetic diversity with NTSYS software. This data showed pathogenicity and genetic diversity of Pseudomonas viridiflava in Iran.     

厚朴根腐病的发生特点与防治方法

厚朴(M agnolia officinalis)又称川厚、紫油厚朴,属木兰科植物。以皮、花、果实入药,具有温中、下气、燥湿、消痰功能,是一种常用的具有较高经济价值的名贵中药材。主产于四川、湖北、浙江、江西、福建、江苏等省。本县每年种植面积近70hm2。近两年来,由于厚朴苗期根腐病发生较为严重,造成大量死苗,损失达30%～50%。2004年本站对此病的症状及发生特点进行了观察,开展了药剂防治试验,提出了综合防治措施。现总结如下:1病原与症状此病系土传真菌性病害,由镰孢属菌尖孢镰孢菌(Fusarium oxysporum Schl.)侵染引起。病株的地上部生长衰弱,由下…     

Comparative host reactions and Frankliniella occidentalis transmission of different isolates of tomato spotted wilt tospovirus from Spain

The abilities of different isolates of tomato spotted wilt tospovirus (TSWV) collected from northeastern and eastern Spain to infect 10 host species, and to be acquired and transmitted by the western flower thrips (WFT), Frankliniella occidentalis were compared. Two isolates of TSWV from a single source plant could be separated according to the different type of local lesions they induced in Nicotiana glutinosa. Host ranges of the studied TSWV isolates were very similar, but differences were found in the symptoms induced and in their capacity to infect specific hosts systemically. Lycopersicon esculentumDatura stramonium were evaluated for their potential as virus acquisition host species. The proportion of transmitter adult thrips obtained from WFT larvae fed on L. esculentum was greater than from D. stramonium. No differences were detected between TSWV isolates in their ability to be acquired and transmitted by WFT. No evidence was obtained of alterations in TSWV particles which could affect WFT transmissibility due to the repeated mechanical transfers used to clone the isolates. Our findings do not support the existence of pathological effects of TSWV on WFT.     

Collar rot and wilt: a new disease of Java citronella (Cymbopogon winterianus) caused by Fusarium moniliforme Sheldon

Commercial plantations of Java citronella ( Cymbopogon winterianus ) in Lucknow, Pantnagar and their adjoining areas were affected by a collar rot and wilt disease. The causal organism was identified as Fusarium moniliforme , anamorph of Gibberella fujikuroi. Isolates of the pathogen originating from Lucknow and Pantnagar differed in their pathogenicity on the host plant under glasshouse conditions. There were also differences in growth rates, pigment production and sporulation between isolates.     

Dynamic root exudation of phenolic allelochemicals from Johnson grass (Sorghum halepense)

The phenolic allelochemicals from the root exudates of Johnson grass (Sorghum halepense) were collected at different developmental stages and analyzed using high performance liquid chromatography‐ultraviolet/photo‐diode array. The results revealed that three simple phenolic components, p‐hydroxybenzoic acid, p‐hydroxybenzaldehyde and ethyl p‐hydroxybenzoate, along with two flavonoids, diosmetin (3′,5,7‐trihydroxy‐4′‐methoxyflavone) and tricin (5,7,4′‐trihydroxy‐3′,5′‐dimethoxyflavone), were found in the root exudates of Johnson grass. However, the major phenolic compounds varied and the quality of each compound was dynamic at different developmental stages: the only phenolic compound that was detected in the seedling stage was p‐hydroxybenzoic acid; three simple phenolic allelochemicals, p‐hydroxybenzoic acid, p‐hydroxybenzaldehyde and ethyl p‐hydroxybenzoate, were found in the jointing stage; and five compounds, including the two flavonoids, tricin and diosmetin, appeared in the reproductive stage along with the other three simple phenolic allelochemicals. The highest diversity in phenolic constituents was in the reproductive stage, with five allelochemicals. Only p‐hydroxybenzoic acid was found in the whole life cycle of Johnson grass, with a range of 2.9 nmol to 6.8 nmol per plant per day. p‐Hydroxybenzaldehyde had the highest quality among all the compounds, with the highest releasing rate of 26.8 nmol per plant per day. The lowest‐content compounds were the two flavonoids, with the highest quality of 1.8 nmol per plant per day. The results suggest that the temporal dynamic of phenolic allelochemicals in the root exudates of Johnson grass corresponds with the dynamic of development.     

Allelopathic activity of root exudates from Leonurus sibiricus L. (Raktodrone)

Root exudates (RE) of Leonurus sibiricus L. show the stimulatory effect of a bioassay on rice, wheat and mustard. Root exudate shows concentration-dependent activity. A stock solution (one seedling in 200 mL of 0.1% Knop's solution for 4 days) is denoted as (1). A 10-fold higher concentration (10) was made by using a rotary vacuum evaporator; for example, 100 mL stock solution was reduced to 10 mL. For bioassay, (25), (50), (75) and (100)-fold concentrations have been used. The stimulatory effect increases with concentration (peak at 10), decreases thereafter, and at (50) (75) and (100), the effect is inhibitory (almost 100% at 75 and 100). Paper chromatography and staining suggest the presence of phenolics including caffeic acid in RE. Caffeic acid also shows the same concentration-dependent activity on rice seeds. An ether extract of acid hydrolyzed leaves shows a number of phenolics on thin layer chromatography. Among these phenolics, three are common with RE. One highly fluorescent band occurs in a much larger amount (both RE and acid hydrolyzed leaf extract), and coincides with standard caffeic acid. On ultraviolet spectrometer analysis after elution, the putative caffeic acid compound shows two characteristic peaks similar to that of caffeic acid at 277 and 231 nm, respectively. Gas liquid chromatography retention time of both standard caffeic acid and the fluorescent spot is the same.     

Identification of allele specific AFLP markers linked with bacterial wilt [Ralstonia solanacearum (Smith) Yabuuchi et al.] resistance in hot peppers (Capsicum annuum L.)

Even though the bacterial wilt is identified as the most destructive disease in hot peppers world-wide, robust molecular markers that facilitate marker assisted selection are absent till date. Kerala Agricultural University (India) has released two hot peppers named Ujwala and Anugraha which show high level field resistance to this pathogen. The variety Anugraha was developed through backcross breeding between a high yielding but highly susceptible variety Pusa Jwala with the highly resistant Ujwala, using Pusa Jwala as a recurrent parent. Thus, Pusa Jwala and Anugraha are near isogenic lines (NILs) differing for the resistance to bacterial wilt only and the resistance is governed by a homozygous recessive (rr) gene action. The F₁s of Anugraha × Pusa Jwala were selfed to generate the segregating F₂ population. The F₂ population has been field screened, 10 highly susceptible and 10 most resistant plants were identified and DNA from these plants were bulked separately. Bulked segregant analysis with AFLP primer combination EcoACT + MseCAC was done using the DNA from donor parent Ujwala, susceptible parent Pusa Jwala, resistant parent Anugraha, bulked susceptible F₂ and bulked resistant F₂ plants. On resolution using capillary electrophoresis system in genetic analyzer, the AFLP products have yielded three polymorphic bands (103, 117, and 161 bp) which were linked with the resistant recessive allele and three polymorphic bands (183, 296, 319 bp) linked with the dominant susceptible allele of the bacterial wilt resistance gene. The results were confirmed through co-segregation analysis in most resistant and susceptible plants of F₂ segregating population.     

32份木薯种质对疫霉根腐病的抗性评价和农艺性状分析

通过田间自然发病和室内离体接种方法对32份木薯种质进行了棕榈疫霉根腐病抗病性评价,结果表明,供试的32份木薯种质中,高抗种质7份,抗病种质7份,中感种质8份,感病种质7份,高感种质3份。其中高抗种质为‘H360’、‘华南11号’、‘华南8号’、‘H588’、‘桂热3号’、‘H873’和‘H971’;抗病种质为:‘F556’、‘H502’、‘GR911’、‘C-4’、‘F10’、‘南植188’和‘H47’。对其中14份抗棕榈疫霉根腐病木薯种质的农艺性状进行了鉴定和评价,结果表明‘华南8号’、‘GR911’、‘F556’、‘C-4’、‘H360’可以进一步加以利用。