H+-ATPase as a biochemical marker for early detection of root (wilt) disease in coconut palms (Cocos nucifera L).

H+-ATPase activity in leaves and roots of coconut palms growing in 'root wilt disease-prevalent areas' was compared with that of coconut palms growing in 'disease-free areas'. The activity was found to be significantly less in the leaves and roots of palms in the disease-prevalent zone as compared to that in disease-free zone. Histochemical examination of the leaves showed results that corroborated the biochemical findings. The possible application of H+-ATPase activity as a marker for the early detection of wilt disease in coconut palms is suggested.     

Electron microprobe X-ray microanalysis of diseased coconut (Cocos nucifera) roots

Summary Electron microprobe X-ray analysis of root (wilt) diseased and healthy coconut roots were scanned for the deposition of metal ions to implicate the involvement of these metal toxicity in the root (wilt) disease of coconut. The results indicated that a high concentration of Al, Mn, Cu and Co ions localised in the disease roots compared to healthy palms. The chemical analysis of tissue samples and soils also confirmed the results of XMA.     

First record of Fusarium vascular wilt on grapevine in Egypt

During the summer season of 2003 and 2004, wilt syndromes of grapevine leaves (Cv. crimson) and vascular discolouration of roots have been observed in 2-year-old grapevine plants in the field at two sides in Gharbeia Governorate, Egypt. First, symptoms of wilt began on bottom leaves borderline as chlorosis and then these turned to necrotic spots and the leaves died. Wilt symptoms were spread to apical associated with vascular discolouration of roots and stem basal. Routine isolations of discoloured root tissue from diseased plant yielded eight isolates of Fusarium oxysporum Schlechtend only where no other fungi were developed. Microscopic examination revealed the presence of three shapes of microconidia, first is avoid shape non-septate measuring 2.5–3.0 μm × 6–10 μm, second is cylindrical with one septa measuring 2.6 μm × 17.0 μm and third shape also cylindrical with two septate measuring 3.0 μm × 20.0 μm. Macroconidia was rarely with three septate measuring 3.5– 4.0 μm × 35.0–38.0 μm, and chlamydospores were found singly or in pairs or chains. F. oxysporum isolates attacked grapevine plants (Cv. crimson) causing vascular wilt (66.7%) and root-rot syndrome (33.3%). In vitro isolates of F. oxysporum causing wilt of grapevine (Cv. crimson) varied for producing lytic enzymes, i.e. polygalacturonase (PG) and cellulase. The reactions of several grapevines (Cvs.) with a virulent isolate of F. oxysporum indicated the presence of two different symptoms, i.e. vascular wilt only on grapevine plants (Cv. crimson) and root-rot on the other grapevine (Cvs.), i.e. superior, Thompson, King robi and flame seedless. All F. oxysporum isolates caused vascular wilt of grapevine Cv. crimson, successfully reisolated from symptomatic vascular infected tissue and complete identification on the basis of colony, conidia morphology and host range at formae speciales level as F. oxysporum f. sp. herbemontis (Tochetto) Gordan. This is the first report of Fusarium wilt on grapevine in Egypt.     

PCR amplification of the spliced leader gene for the diagnosis of trypanosomatid parasites of plants and insects in methanol-fixed smears

A PCR-based method was adapted for the amplification of DNA from methanol-fixed smears of insects and plants parasitized by trypanosomatids. The PCR target was the multicopy spliced leader (SL) gene. Amplicons were hybridized with an oligonucleotide probe (SL3') specific for Phytomonas. The method has the advantage of dispensing with the cultivation of parasites, many of which are very fastidious or non-cultivable. The technique was applied to archival glass slides and to newly collected material. It proved to specific for Phytomonas spp., enabling their detection in plants and insects. Sequence comparison of the amplicons obtained revealed the existence of different strains/species of Phytomonas circulating among diseased palsms and fruit.     

Lethal bole rot disease of coconut in East Africa

A lethal bole, rot disease of coconut, caused by Marasmiellus cocophilus Pegler, sp. nov., reaches epidemic proportions in several areas along the coasts of Kenya and Tanzania. The first symptoms noticed on palms 8 or more years old are a frond wilt and a crown rot, but these follow a primary bole rot. Highest mortality is among seedlings and young palms up to 8 years old. Where the disease occurs sporophores are sometimes common on exposed roots, dead seedlings and the soil surface where diseased palms have been dug out. The fungus appears to be a persistent colonizer of coconut debris in the soil. Mycelial cultures from infected bole tissues and from sporophores were highly pathogenic to seedlings, and slowly invaded older tissue. M. cocophilus reaches the inner bole tissues only through the roots, and wounding of roots of their aerenchymatous protuberances is important in infection. Seedlings may become infected through roots damaged during transplanting from nurseries to the field; movement of seedlings from affected areas is one certain method of dispersal. Suggested control measures are: (1) selection of seedlings in nurseries and subsequent transplanting should be as early as possible; (2) seedlings should be transplanted very carefully, and damaged roots pruned and disinfected; (3) seedlings should never be taken from affected areas; (4) periodic soil sterilization of nurseries; and (5) no cultivation should be done between palms, especially where disease is present.     

我国南方花生发生一种由蕃茄斑萎病毒引起的新病害

我们于1984和1985年6月上、中旬,在广州市郊、县,湛江市郊以及广西南宁市郊、县,北海市郊和合蒲县等花生产区,调查花生病毒病时,除了发现花生轻斑驳病毒病外,还发现一种新的病毒病害。其症状特征是:病株顶端叶片上出现很多褪绿黄斑或环斑,有的环斑变     

Johnsongrass Chlorotic Stripe Mosaic and its Associated Viruslike Particles in Iran1

A mosaic disease of Johnsongrass characterized by the presence of short and long chlorotic stripes in leaves and stunting of plants was found in two locations about 300 km apart in the Fars Province of Iran. The disease occurred in patches or along the irrigation ditches. The disease agent could not be transmitted mechanically or by certain insects and mites. About 0.5% of the seedlings, grown in soil from the root zone of diseased plants, developed the disease after 1–8 months. Electron microscopy, of leaf-dip preparations showed presence of isometric viruslike particles (VLPs) of about 35 nm diameter in unusually high concentrations in diseased plants but not in healthy plants. VLPs from diseased leaves were purified by low-pH clarification followed by differential and densitygradient centrifugation. Purified VLP preparations showed UV absorption spectrum typical of nucleoproteins. A260/A280 was 1.48. High titered antisera obtained by injecting rabbits with purified VLPs reacted with sap from diseased plants but not healthy plants. Purified VLP preparations and sap from diseased plants did not react with antisera to Bermuda grass etched line, Brachypodium sylvaticum, brome mosaic, maize chlorotic dwarf, maize rayado fino, maize white line mosaic and tobacco necrosis viruses.     

Biology and Structure of Phytomonas staheli sp. n., a Trypanosomatid Located in Sieve Tubes of Coconut and Oil Palms

SYNOPSIS. Phytomonas staheli sp. n. is described from the oil palm ( Elaeis guineensis ) and the coconut palm ( Cocos nucifera ) of Surinam, South America. The phytomonad is the probable cause of "Hartrot" in the coconut palm and "Marchitez sopresiva" in oil palms. Parasites are confined to the sieve tubes in palms. Some success was obtained at cultivation of the organism from Elaeis.     

Root (wilt) disease of coconut palms in South Asia – an overview

Root (wilt) disease (RWD) caused by phytoplasma is one of the most devasting diseases of coconut palms. The major symptoms of the disease in leaves are wilting and drooping and flaccidity; ribbing, paling/yellowing and necrosis of leaflets are typical symptoms of foliar diseases. Unopened pale yellow leaflets of spindle leaves are more susceptible to leaf rot disease, which is caused by Exerohilum rostratum and Colletotrichum gloeosporioides. RWD is caused by phytoplasmas, the cell wall-less prokaryotes that are bounded by a “unit” membrane. In ultrathin sections, they appear as a complex multi-branched, beaded, filamentous or spheroidal pleomorphic bodies. The disease was transmitted by plant hoppers (Proutista moesta) and lace wing bug (Stephanitis typica). Phytoplasmas are generally present in the phloem sieve tubes and in the salivary glands of these insect vectors. Phytoplasmas cannot be cultured in vitro, and hence it is very difficult to identify them. Using polymerase chain reaction technique, group-specific primers have been applied to detect mixed-phytoplasma infections in a single host. RWD, is a non-lethal, debilitating disease, and hence an integrated approach for the management of this disease in coconut palms has been discussed in this study.     

Further studies on a spiroplasma isolated from coconut palms in Jamaica

The pathogenicity of a spiroplasma isolated from coconut palms was tested by (1) transmission experiments to palms and other plants susceptible to infection by mycoplasmas, using the suspected vector of lethal yellowing, Myndus crudus, and vectors of the agents of other yellows diseases and (2) enzyme-linked immunosorbent assay (ELISA) to detect spiroplasma antigens in diseased palm tissues. Results of both these tests were negative and, as earlier attempts to repeat the isolations from lethal yellowing diseased palms had also been unsucessful, it was concluded that this organism was not the causal agent of lethal yellowing disease. Further analysis by serological tests and by polyacrylamide gel electrophoresis (PAGE) of spiroplasma proteins confirmed that the coconut isolates were related to members of the Spiroplasma citri serogroup but were distinct from other strains tested.     

Ecology of baculovirus-infected and healthy adults of Oryctes rhinoceros (Coleoptera: Scarabaeidae) on coconut palms in the Philippines

Abstract.

• 1 Males and females of Oryctes rhinoceros (L.), a serious pest of coconut palms, congregate in decaying coconut trunks to mate, oviposit and prepare the wood for the young larvae. A baculovirus disease is often transmitted from beetle to beetle in this environment and transmission to and from larvae probably also occurs.
• 2 Young beetles are mainly found feeding on palms, where they also frequently contract the virus disease, presumably through mating with diseased partners.
• 3 Monitoring beetle populations in the Philippines with attractant traps and by collections from palms showed seasonal fluctuations which were often inversely correlated with the incidence of the virus disease. Disease prevalence matched closely the proportion of females in the population. Possible explanations for this correlation and for the seasonality of the disease prevalence are outlined.

     

Phytoplasma distribution in coconut palms affected by lethal yellowing disease

Lethal yellowing (LY), the most devastating disease affecting the coconut palm in America, is caused by phytoplasmas known to be distributed in different parts of infected plants. However, no comprehensive reports exist on the phytoplasma distribution. This study refers to the detection of LY phytoplasma DNA using PCR in different coconut plant parts, throughout the development of the disease. Sample analysis of positive palms taken at different stages of disease development (either symptomatic or symptomless) showed differences in the percentage of LY detection between plant parts. Some parts showed a very high level of LY DNA (stem, young leaves, inflorescences, stem apex and root apex), low levels were found in the intermediate leaves and roots without apex, whereas no LY phytoplasma DNA was detected in mature leaves. The detection percentage of LY phytoplasma DNA was lowest in symptomless‐infected palms for all parts, except the stem, where phytoplasma accumulations were consistently detected. This pattern of detection among parts is consistent with the hypothesis that phytoplasmas move from photosynthate source tissues to sink tissues via the phloem mass flow process. The accumulations in the (lower) stem, prior to the appearance of symptoms, suggest that this part of the palm is where phytoplasmas first move from leaves after foliar feeding by vectors and in which they probably multiply and distribute to other palm parts, including roots. Embryos from infected palms were analysed by nested‐PCR and 28% of 394 embryos were positive. Phytoplasma DNA was detected in embryos from fruit on any of the fruiting bunches regardless the age, but no pattern of quantitative distribution throughout the bunch developmental stages was observed. Germination of seeds from LY‐positive symptomatic palms was 58% and from LY‐negative symptomless palms were 71%. No phytoplasma was detected in seedlings tested from both symptomatic and non‐symptomatic palms. Seedlings tested after 2 years did not develop LY symptoms or eventually died.     

ROOT INJURY AS A FACTOR IN THE ASSESSMENT OF CHEMOTHERA-PEUTANTS BY THE TOMATO FUSARIUM WILT TEST

Studies of the tomato Fusarium wilt test for screening chemotherapeutants were made by the writer at the Connecticut Agricultural Experiment Station, U.S.A., during 1951. The test compounds were applied on ten successive days to the roots of tomato plants growing in sand. The plants were then uprooted, their roots washed and dipped in a bud-cell suspension of F. oxysporum lycopersici and the plants re-potted. A standardized assessment of wilt and vascular discoloration was made some 21 days later, by which time the control plants were usually severely diseased. Several compounds greatly reduced disease severity.     

Bacterial antagonists of Sunflower (Helianthus annuus L.) fungal pathogens

Bacteria isolated on nutrient agar and King's medium B from sunflower leaves, crown and roots inhibited in vitro growth of the leaf spot and wilt pathogens Alternaria helianthi, and Sclerotium rolfsii, respectively, and also the root rot pathogensRhizoctonia solani and Macrophomina phaseolina. Antagonistic bacteria from leaves were mainly actinomycetes and pigmented Gram-positive bacteria, while those from roots and crowns were identified asPseudomonas fluorescens-putida, P. maltophilia, P. cepacia, Flavobacterium odoratum andBacillus sp. In soil bioassays, when used as seed inoculum in the presence ofS. rolfsii, P. cepacia strain N24 increased significantly the percentage of seedling emergence. Bacterial strains which exhibited broad spectrum in vitro antagonistic activity were tested for colonisation of sunflower roots, when used as a seed inoculum. Good colonisers (10⁴ to 10⁶ bacteria/g root) were consistent in their ability to reduce disease and fungal wilt. A seedling having a primary root length < 5 cm with fewer lateral roots, necrosed cotyledons or crown and a wilted shoot indicated its diseased status. On an average, only 30% of seedlings were diseased when treated with the antagonistic strains, in the presence of the pathogen, while 60% of the seedlings were diseased in the presence of the pathogen alone. In microplots treated with strain N24, only 1 to 3% of the seedlings were wilted, while 14% of the seedlings were wilted in the presence of the pathogen alone. The results obtained show that bacterial antagonists of sclerotial fungi can be used as seed inocula to improve plant growth through disease suppression     

Preliminary Investigations about the Mode of Transmission and Spread of Xanthomonas campestris pv. graminis on Forage Grasses

Extensive studies showed that no disease was caused when seeds of different forage grasses were inoculated with Xanthomonas campestris pv. graminis. The disease could easily be induced by infecting the plants in the root system, leaves or flower. The inoculation site in the leaf proved to be of vital importance for the development of the disease. Wilting symptoms were quickly induced when the pathogen was inoculated near the leaf base. Plants in root-contact with diseased plants showed disease symptoms. It is not known whether these symptoms were caused by the bacteria or by toxins released by nearby diseased plants. Cross inoculation trials on different grass varieties revealed that different pathovars exist in the group of xanthomonads, pathogenic to forage grasses. Some have a broad host range whereas others are more limited to a single plant genus. Field trials suggest that in Belgian climatic conditions, the losses caused by bacterial wilt are rather limited.     

Rearing and transmission techniques for Haplaxius sp. (Horn: Cixiidae), a suspected vector of lethal yellowing disease of coconuts

Adult Haplaxius sp., identified from Jamaica as H. crudus or H. cocois, are common on coconut foliage in both Jamaica and Florida and are suspected vectors of lethal yellowing disease in both regions. Nymphs, which are subterranean, were mass-reared on roots of the grasses Stenotaphrum secundatum, Axenopus compressus and Cynodon spp. Transmission of the disease was tested by feeding the nymphs on roots of diseased palms, rearing them on grasses and transferring the emergent adults to foliage of test palms for infection feeds. No transmission was proven.     

Development of Diagnostic Methods for Detecting Ganoderma-infected Oil Palms

Two diagnostic methods have been applied for detecting the plant pathogenic fungus Ganoderma , a basidiomycete, causing the basal stem rot (BSR) diseaseof oil palms. One approach was the use of polyclonal antibodies (PAbs) raised against mycelial proteins of a single Ganoderma isolate and a mixture of nine different Ganoderma isolates. Both PAbs could detect Ganoderma in diseased oil palm root tissue by applying an indirect enzyme-linked immunosorbent assay. Low cross-reactions were observed with the five mainly occurring saprophytic fungi which could be isolated from diseased oil palms and were used as negative controls. The other approach was based on the polymerase chain reaction (PCR) in order to increase the sensitivity of the Ganoderma detection. The primers were generated from the internal transcribed spacer region 1 of rDNA of Ganoderma boninense and produced an PCR product of 167 bp in size. Fungal isolates and oil palm root samples were processed for PCR by three different DNA extraction methods. The most suitable extraction procedure was a simple alkaline extraction method. For a practical approach, a semiquantification was performed by assessing the PCR sensitivity limits of pure culture Ganoderma and naturally infected root samples.     

Microscopic and polyclonal antibody-based detection of yellow leaf disease of arecanut (Areca catechu L.)

Phytoplasma, the pathogen of yellow leaf disease (YLD) of arecanut (Areca catechu L.) was detected by transmission and scanning electron microscopy. Tissues of YLD affected palms contained phytoplasmas in the phloem sieve elements, but not in symptomless healthy palm tissues. Phytoplasma was purified from tissues of diseased palms employing percoll density gradient centrifugation and confirmed by transmission electron microscopy. Using the purified phytoplasma preparation, a polyclonal antiserum was raised in rabbits and used for standardisation of agar gel double diffusion (Ouchterlony) test and DAC-ELISA. Clear precipitin line was observed in Ouchterlony test between the antigen from diseased palms and the pathogen-specific antibodies after 48 h incubation and only undiluted antiserum showed best result in the test. However, in ELISA, 1:10 antigen dilution and 1:400 pathogen-specific antibodies dilution produced sensitive detection of the pathogen with a difference of >3.5 times absorption values between healthy and diseased samples. The results thus confirmed the ability of antiserum to distinguish healthy and infected plants and utility of ELISA for effective diagnosis of YLD.     

PHYSIOLOGICAL STUDIES ON THE VERTICILLIUM WILT DISEASE OF TOMATO

The water loss per unit leaf area of tomato plants was decreased after inoculation with Verticillium albo-atrum. When diseased plants began to wilt water loss temporarily increased, but then rapidly decreased to become less than that of healthy plants grown under conditions of adequate or restricted water supply.
The transpiration of excised leaves from plants grown with a restricted water supply was reduced, but not so severely as that of comparable leaves from infected plants. Water loss from leaves on infected plants was reduced irrespective of any blocking of the petiolar xylem.
The rate of water loss from turgid leaf disks on mannitol solutions, and the rate of water uptake of leaf disks on water was similar for disks cut from wilting or turgid leaves of diseased plants or healthy plants grown with an adequate or restricted water supply.
Disease or poor water supply reduced leaf growth but had no effect on the rate of leaf initiation. Although the density of stomata was higher on leaves of diseased plants the stomatal area was less than on healthy plants.
The resistance to water flow in diseased stems was high and was correlated with vessel blockage. About half the blocked vessels contained hyphae. The severity and localization of symptoms in inoculated plants growing on susceptible or resistant rootstocks was directly related to the extent of invasion by the pathogen and to vessel blockage.
Experiments on the wilting activity of cell-free filtrates from cultures of the pathogen in vitro indicated that it produced a stable substance, not an enzyme, that caused wilting in cut shoots by blocking the end of the stem. It is suggested that an increasing internal water shortage causes major symptoms of disease.