Greenhouse-grown Plants Research Articles

Down-regulation of flavonoid 3′-hydroxylase gene expression by virus-induced gene silencing in soybean reveals the presence of a threshold mRNA level associated with pigmentation in pubescence

Changes in flavonoid content are often manifested as altered pigmentation in plant tissues. Two loci have been identified as controlling pigmentation in soybean pubescence. Of these, the T locus appears to encode flavonoid 3'-hydroxylase (F3'H) protein: the T and t alleles are associated with tawny and gray colors, respectively, in pubescence. We previously down-regulated F3'H gene expression by virus-induced gene silencing (VIGS) in soybean. Despite this successful VIGS, the tawny pubescence pigmentation proved to be unchanged in greenhouse-grown plants. We hypothesized that the reduced mRNA level of the F3'H gene resulting from VIGS remained high enough to induce pigmentation. To verify this hypothesis, in the present study, we performed F3'H VIGS on plants grown under controlled conditions, in which the steady-state mRNA level of the F3'H gene was reduced to approximately 5% of that of greenhouse-grown plants. This VIGS treatment resulted in the loss of tawny pigmentation in pubescence, suggesting that the sf3'h1 gene is involved in the control of pigmentation in pubescence. We detected a marked decrease in target mRNA, an accumulation of short interfering RNAs (siRNAs), and a decrease in quercetin content relative to kaempferol in leaf tissues, indicating that sequence-specific mRNA degradation of the F3'H gene was induced. These results suggest that leaf tissues have a threshold mRNA level of the F3'H gene, which is associated with the occurrence of tawny pigmentation in pubescence. The estimated threshold mRNA level for pigmentation in pubescence was approximately 3% of the steady-state mRNA level of the F3'H gene in greenhouse-grown plants.

Occurrence of Late Leaf Rust Caused by Pucciniastrum americanum in Red Raspberry (Rubus idaeus) in Buenos Aires, Córdoba, and Entre Ríos, Argentina.

In the fall of 2003, severity of late leaf rust in leaves and fruits of red raspberry (Rubus idaeus L.) reached 50% in Buenos Aires (Azul, Baradero, Capilla del Señor, Gral. Rodríguez, Mar del Plata, and Tandil), Córdoba (Villa de Las Rosas), and Entre Ríos (Concordia). The south of Argentina, Río Negro (El Bolsón), and Chubut (El Hoyo, Lago Puelo) remained rust free. The abaxial side of the field infected leaves had pustules filled with masses of yellow spores. Chlorotic areas corresponded in the adaxial side. Urediospores were vacuum harvested from field infected leaves collected in the Tandil area and placed onto a healthy 1-year-old greenhouse-grown plant (cv. Heritage). Spores from a single pustule were increased on plants of the same cultivar. Spores were studied with optic and electronic microscopy. Uredial ostiolar cells were warted, laterally free, and constricted in the middle. The obovoid, echinulate urediospores, from infected leaves averaged 24 × 16 μm (16 to 30 × 11 to 21 μm). Morphological characteristics and spore measurements agreed with those reported for Pucciniastrum americanum (1). Urediniospores were suspended in mineral oil and sprayed onto three raspberry cultivars that were maintained in a darkened mist chamber at 20°C for 48 h and the transferred to a 20°C and 12-h light cycle chamber. Control plants were inoculated with sterile water. There were three replicate plants of each treatment. After 11 days, large sporulating uredia (0.5 mm) were produced on inoculated leaves of cv. Autumn Bliss and smaller uredia (0.1 to 0.3 mm) were produced on cv. Heritage. There were necrotic flecks and the least and smallest uredia were produced on cv. Himbo Queen. No symptoms were present in control plants. To our knowledge, this is the first report of P. americanum causing disease on raspberry in Argentina. Reference: (1) G. F. Laundon and A. F. Rainbow. Pucciniastrum americanum. No. 210 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, England, 1969.

SodERF3, a Novel Sugarcane Ethylene Responsive Factor (ERF), Enhances Salt and Drought Tolerance when Overexpressed in Tobacco Plants

The molecular signals and pathways that govern biotic and abiotic stress responses in sugarcane are poorly understood. Here we describe SodERF3, a sugarcane (Saccharum officinarum L. cv Ja60-5) cDNA that encodes a 201-amino acid DNA-binding protein that acts as a transcriptional regulator of the ethylene responsive factor (ERF) superfamily. Like other ERF transcription factors, the SodERF3 protein binds to the GCC box, and its deduced amino acid sequence contains an N-terminal putative nuclear localization signal (NLS). In addition, a C-terminal short hydrophobic region that is highly homologous to an ERF-associated amphiphilic repression-like motif, typical for class II ERFs, was found. Northern and Western blot analysis showed that SodERF3 is induced by ethylene. In addition, SodERF3 is induced by ABA, salt stress and wounding. Greenhouse-grown transgenic tobacco plants (Nicotiana tabacum L. cv. SR1) expressing SodERF3 were found to display increased tolerance to drought and osmotic stress.

Production of transgenic potato exhibiting enhanced resistance to fungal infections and herbicide applications

Potato (Solanum tuberosum L.), one of the most important food crops, is susceptible to a number of devastating fungal pathogens in addition to bacterial and other pathogens. Producing disease-resistant cultivars has been an effective and useful strategy to combat the attack of pathogens. Potato was transformed with Agrobacterium tumefaciens strain EHA101 harboring chitinase, (ChiC) isolated from Streptomyces griseus strain HUT 6037 and bialaphos resistance (bar) genes in a binary plasmid vector, pEKH1. Polymerase chain reaction (PCR) analysis revealed that the ChiC and bar genes are integrated into the genome of transgenic plants. Different insertion sites of the transgenes (one to six sites for ChiC and three to seven for bar) were indicated by Southern blot analysis of genomic DNA from the transgenic plants. Expression of the ChiC gene at the messenger RNA (mRNA) level was confirmed by Northern blot analysis and that of the bar gene by herbicide resistance assay. The results obviously confirmed that the ChiC and bar genes are successfully integrated and expressed into the genome, resulting in the production of bialaphos-resistant transgenic plants. Disease-resistance assay of the in vitro and greenhouse-grown transgenic plants demonstrated enhanced resistance against the fungal pathogen Alternaria solani (causal agent of early blight).

Mass propagation and essential oil analysis of Artemisia vulgaris

Artemisia vulgaris L. (Mugwort) is a threatened and valuable medicinal plant. Attempts have been made in this research to mass propagate its plantlets through in vitro liquid culture technology using Murashige and Skoog (MS) basal medium supplemented with 6-benzyl adenine (BA) (0.44-8.88 microM). Initially, 22.6 shoots (99.9% shooting frequency) developed from shoot tip explants cultured in MS with 4.44 microM BA at 100 ml flask capacity. This was further subcultured at increasing flask capacity (150, 250, and 500 ml) for shoot proliferation. Of the different concentrations of BA and flask capacities tested, 4.44 microM BA and 500 ml flask capacity were found to produce a maximum of 85.5 shoots after 30 d of culture. Shoot proliferation was found to increase with increasing flask capacity whereas shoot number decreased with increasing BA concentration (>4.44 microM). Individual shoots were isolated and rooted on MS medium containing 8.56 microM indole-3-acetic acid (IAA). Then the plantlets were acclimatized under standard laboratory conditions and later under greenhouse conditions. Fresh leaves were collected from greenhouse-grown plants and subjected to essential oil analysis by the simultaneous distillation and extraction method. GC-MS results revealed the presence of 88 components and the extracted oil was rich in camphor (16.8%), alpha-thujone (11.3%), germacrene D (7.2%), camphene (6.5%), 1,8-cineole (5.8%) and beta-caryophyllene (5.4%). This in vitro strategy can be a reliable method for the steady production of a large number of plants for essential oil production, which is reported for the first time for A. vulgaris.

Phytophthora bisheria sp. nov., a new species identified in isolates from the Rosaceous raspberry, rose and strawberry in three continents

A homothallic semipapillate slow growing Phytophthora species associated with root rot of strawberries from greenhouse-grown plants in North Carolina, USA, root rot of roses in the Netherlands, and root rot of raspberry in Knoxfield, Australia, was identified. The main character of this organism is the production of paragynous antheridia with broad attachment to the oogonial wall. The morphology of the pathogen does not match that of any of the more than 85 described Phytophthora species. Phylogenetic analysis based on sequences of the internal transcribed spacer rDNA region (ITS1-5.8S-ITS2) of this taxon and those from other Phytophthora species from GenBank supports the conclusion that this organism is an unreported new species. In the phylogenetic tree with other reported Phytophthora species at the GenBank, the new species is more closely related to others in ITS clade 2 comprising semipapillate taxa including P. botryosa, P. citrophthora, P. colocasiae, P. meadii, P. citricola, P. inflata, P.tropicalis, P. capsici, Phytophthora sp. ‘glovera’ and P. multivesiculata. The most closely related species is P. multivesiculata isolated from Cymbidium orchid in the Netherlands. In this paper we describe the morphological characteristics and the phylogenetic relationships that support the description of this taxon as a new species Phytophthora bisheria sp. nov.

Phytophthora bisheria sp. nov., a new species identified in isolates from the Rosaceous raspberry, rose and strawberry in three continents

A homothallic semipapillate slow growing Phytophthora species associated with root rot of strawberries from greenhouse-grown plants in North Carolina, USA, root rot of roses in the Netherlands, and root rot of raspberry in Knoxfield, Australia, was identified. The main character of this organism is the production of paragynous antheridia with broad attachment to the oogonial wall. The morphology of the pathogen does not match that of any of the more than 85 described Phytophthora species. Phylogenetic analysis based on sequences of the internal transcribed spacer rDNA region (ITS1-5.8S-ITS2) of this taxon and those from other Phytophthora species from GenBank supports the conclusion that this organism is an unreported new species. In the phylogenetic tree with other reported Phytophthora species at the GenBank, the new species is more closely related to others in ITS clade 2 comprising semipapillate taxa including P. botryosa, P. citrophthora, P. colocasiae, P. meadii, P. citricola, P. inflata, P.tropicalis, P. capsici, Phytophthora sp. 'glovera' and P. multivesiculata. The most closely related species is P. multivesiculata isolated from Cymbidium orchid in the Netherlands. In this paper we describe the morphological characteristics and the phylogenetic relationships that support the description of this taxon as a new species Phytophthora bisheria sp. nov.

Predicting sublethal effects of herbicides on terrestrial non-crop plant species in the field from greenhouse data

Guidelines provided by OECD and EPPO allow the use of data obtained in greenhouse experiments in the risk assessment for pesticides to non-target terrestrial plants in the field. The present study was undertaken to investigate the predictability of effects on field-grown plants using greenhouse data. In addition, the influence of plant development stage on plant sensitivity and herbicide efficacy, the influence of the surrounding vegetation on individual plant sensitivity and of sublethal herbicide doses on the biomass, recovery and reproduction of non-crop plants was studied. Results show that in the future, it might well be possible to translate results from greenhouse experiments to field situations, given sufficient experimental data. The results also suggest consequences at the population level. Even when only marginal effects on the biomass of non-target plants are expected, their seed production and thereby survival at the population level may be negatively affected.

Spectral Response of Cotton Aphid– (Homoptera: Aphididae) and Spider Mite– (Acari: Tetranychidae) Infested Cotton: Controlled Studies

Remote sensing is a precision tool that can detect plant health. Ground-based methods in small-scale experiments were used to explore the applicability of this technology for detection of arthropod-damaged cotton and to find useful indices or wavelengths for detecting arthropod-damaged cotton. Individual leaves of greenhouse-grown cotton plants and cotton plants in the field were infested with populations of cotton aphids, spider mites, and aphids + mites. Several sets of reflectance measurements were collected from the adaxial surface of the leaves at various intervals after infestation using a portable hyperspectral spectrometer with an integrating sphere or a contact probe. Vegetation indices were calculated from the reflectance values; these indices and the raw reflectance values, represented by narrow wavelength bands, were tested to see if arthropod damaged cotton could be distinguished from healthy cotton. Results indicated that it was possible to detect cotton aphid- and spider mite-damaged leaves by tracking the spectral changes in the leaf, although the damage type of each arthropod could not be distinguished spectrally. In addition, spider mite- and aphid-infested cotton leaves increased reflectance in the near infrared wavelength at approximately 850 nm in comparison to uninfested leaves.

Redirection of the phenylpropanoid pathway to feruloyl malate in Arabidopsis mutants deficient for cinnamoyl-CoA reductase 1

Cinnamoyl-CoA reductase 1 (CCR1, gene At1g15950) is the main CCR isoform implied in the constitutive lignification of Arabidopsis thaliana. In this work, we have identified and characterized two new knockout mutants for CCR1. Both have a dwarf phenotype and a delayed senescence. At complete maturity, their inflorescence stems display a 25-35% decreased lignin level, some alterations in lignin structure with a higher frequency of resistant interunit bonds and a higher content in cell wall-bound ferulic esters. Ferulic acid-coniferyl alcohol ether dimers were found for the first time in dicot cell walls and in similar levels in wild-type and mutant plants. The expression of CCR2, a CCR gene usually involved in plant defense, was increased in the mutants and could account for the biosynthesis of lignins in the CCR1-knockout plants. Mutant plantlets have three to four-times less sinapoyl malate (SM) than controls and accumulate some feruloyl malate. The same compositional changes occurred in the rosette leaves of greenhouse-grown plants. By contrast and relative to the control, their stems accumulated unusually high levels of both SM and feruloyl malate as well as more kaempferol glycosides. These findings suggest that, in their hypolignified stems, the mutant plants would avoid the feruloyl-CoA accumulation by its redirection to cell wall-bound ferulate esters, to feruloyl malate and to SM. The formation of feruloyl malate to an extent far exceeding the levels reported so far indicates that ferulic acid is a potential substrate for the enzymes involved in SM biosynthesis and emphasizes the remarkable plasticity of Arabidopsis phenylpropanoid metabolism.

Abscisic acid relations of plants grown on tungsten enriched substrates

The effect of tungsten as a potential inhibitor of ABA biosynthesis in plants was tested both under controlled conditions in the greenhouse or laboratory and in the field at a site naturally enriched with tungsten. Although as predicted in all cases the activity of aldehyde oxidase was strongly inhibited by tungsten, in well watered, greenhouse-grown maize plants increased concentrations of tungsten did not reduce ABA concentration in tissues. Depending on the time of incubation, mostly an accumulation due to an increased net ABA synthesis could be observed. Under drought stress, however, tungsten may have an inhibitory effect on ABA accumulation. Field experiments with drought-stressed clover and dandelion plants from a habitat close to a former tungsten mine confirm those of the greenhouse and laboratory. The ineffectiveness of tungsten in decreasing ABA levels in well-watered plants and its minimal effects in inhibiting ABA accumulation of droughted plants indicate that it is unsuitable as an inhibitor of ABA biosynthesis.

Identification and Host Relations of Turnip ringspot virus, A Novel Comovirus from Ohio.

Viruslike chlorotic ring spot symptoms and line patterns of unknown origin were observed on a greenhouse-grown turnip plant. The suspected virus was mechanically transmissible to plants in the Brassicaceae. Electron microscopic analysis revealed icosahedral particles approximately 28 nm in diameter. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses suggested that the pathogen is a comovirus, an observation that was confirmed by analysis of portions of the genomic sequence. This virus was provisionally named Turnip ringspot virus (TuRSV). Based on the RNA 1 sequence, TuRSV is most similar to Radish mosaic virus, another pathogen that infects members of the Brassicaceae. Arabidopsis thaliana is susceptible to TuRSV, and 12 out of the 23 ecotypes studied showed symptoms when inoculated with the virus. TuRSV induced a variety of responses on ecotypes from death to no infection. Some ecotypes showed one or two rounds of symptom display followed by recovery when inoculated with TuRSV. About half of the ecotypes (11/23) analyzed showed no symptoms when inoculated with TuRSV. Col-0 plants showed no symptoms, and infectious virus was not recovered from systemic leaves, although it could be detected by RT-PCR. Col-0 plants harboring mutations impairing the ethylene, jasmonic acid, or salicylic acid signaling pathways did not show symptoms when inoculated with TuRSV.

Host–pathogen interaction between Phytophthora infestans and Solanum nigrum, S. villosum, and S. scabrum

Potato and tomato are the two major hosts for Phytophthora infestans causing late blight. The susceptibility of leaves and whole plants of Solanum nigrum, S. villosum, and S. scabrum to infection by P. infestans was tested under laboratory conditions. Out of 39 plants representing 38 different S. nigrum accessions, 16 were highly resistant (seven accessions did not show any symptoms of infection, nine were highly resistant showing necrotic lesions in the place of infection), and 23 plants of S. nigrum were colonized by, at least, 1 of the 2 isolates of P. infestans (17 accessions were infected with two P. infestans isolates, and 6 accessions showed different reactions depending on the isolate used for inoculation). Three accessions of S. villosum, and one accession of S. scabrum were tested and did not show any symptoms of infection. The majority of S. nigrum accessions infected by P. infestans in a detached leaf assay were also infected in the whole plant assay. The reaction of field- and greenhouse-grown plants to inoculation with P. infestans in detached leaf assays was similar, but in some cases leaves from field-grown plants reacted as resistant in comparison with the leaves from greenhouse-grown plants, which were susceptible.

Floral Biology of Physaria ludoviciana (Brassicaceae), a Plant Rare to the Midwest

ABSTRACT Physaria ludoviciana (Brassicaceae) is rare in Illinois, Minnesota, and Wisconsin. Environmental effects on floral development are unclear. Both self-compatibility and self-incompatibility occur within Physaria species. Objectives were to describe flowers, to determine how photoperiod affects flower development, and to predict whether flowers are self-compatible or self-incompatible. For photoperiods, greenhouse-grown plants were placed in either 16 or 8 hr photoperiods. Inflorescences and open flowers were counted weekly. For pollination, flowers were self-pollinated or cross-pollinated. Plants developed inflorescences after 20 and 28 d in long and short days, respectively. Inflorescences/plant increased for both photoperiods throughout the study. In short days, plants produced more inflorescences (10.8/plant) than in long days (7.1/plant). Anthesis started at 48 and 56 d for long and short days, respectively. Blooming peaked at 83 d (4.9 flowers/plant/day) for long days, and at 98 d (3.5 flower...

Rhizosphere persistence of three Pythium oligandrum strains in tomato soilless culture assessed by DNA macroarray and real-time PCR

In tomato soilless culture, plant-disease optimal control and growth promotion are achieved when the rhizosphere is heavily colonized by the biocontrol agent Pythium oligandrum. Discrepancies in performance are generally attributed to the poor persistence of P. oligandrum on roots. In this study, three selected strains of P. oligandrum were introduced into the rhizosphere of greenhouse-grown tomato plants, and their persistence was assessed by DNA macroarray hybridization and real-time PCR. The experimental data from DNA detection and plate counting were compared. PCR-based methods detected P. oligandrum throughout the 6-month growing season, whereas plate counting indicated its presence only over the first 3 months. Moreover, the DNA array method provided information about the various Pythium species present in the rhizosphere: P. dissotocum was frequently detected on roots of plants, without distinction between plants inoculated or not inoculated with the antagonist. The detection of other Pythium species was noticed sporadically (P. ultimum, P. sylvaticum and P. intermedium), independent of the treatment. Even though the yield enhancement is not significant throughout the entire growing season, data obtained from epidemiological studies demonstrate an enhancement of P. oligandrum persistence on the rhizosphere of plants and less use of mycoparasitism.

Antisense-mediated reduction in ADC activity causes minor alterations in the alkaloid profile of cultured hairy roots and regenerated transgenic plants of Nicotiana tabacum

In species of the genus Nicotiana, as in most plants, the important polyamine precursor putrescine can be derived from the amino acids ornithine and/or arginine via the activity of ornithine decarboxylase (ODC) and/or arginine decarboxylase (ADC), respectively. Nicotiana species also utilize putrescine to provide the pyrollidine ring of the defensive alkaloid nicotine and its derivatives. Previous biochemical studies, involving callus tissues cultured in vitro, suggested that the ADC-mediated route to putrescine is used preferentially to provide the putrescine that is utilized for nicotine synthesis in N. tabacum. To ascertain if this is the case in N. tabacum plants, where nicotine synthesis takes place exclusively in roots, we used an antisense approach to diminish ADC activity in transformed roots which were cultured in vitro. Several independent lines were recovered possessing markedly reduced levels of ADC transcript and ADC activity compared to controls. Transcript levels of other genes in this general area of metabolism, including ODC, were not altered as a result of the antisense-mediated downregulation of ADC. Concentrations of nicotine were comparable in antisense-ADC and control hairy root lines throughout most of their respective culture cycles, except at the latter stages of growth when the nicotine content of antisense-ADC hairy root lines was observed to be ∼20% lower than in controls. Levels of anatabine, the second most abundant alkaloid typically found in N. tabacum, which is not derived from putrescine, were slightly elevated in two antisense-ADC hairy root lines at the latter stages of their culture cycles compared to controls. Comparison of alkaloid levels in leaves of transgenic plants that were regenerated from separate antisense-ADC and control transformed root lines indicated that the former possessed slightly elevated levels of anatabine but did not contain average levels of leaf nicotine that were different from that of controls. Our overall conclusion is that the ADC mediated route to putrescine plays a role, but is not of prime importance, in providing the pyrollidine ring which is used for nicotine synthesis in cultured hairy roots of N. tabacum and in roots of healthy greenhouse-grown plants.

Estimation of the impact of leaf scorch on photosynthesis and "physiological-lesion" size in strawberry

The impact of strawberry leaf scorch, caused by Diplocarpon earlianum, on net CO2 assimilation (A) and transpiration (E) rates was determined for both greenhouse- and field-grown plants. In the greenhouse, strawberry (Fragaria ×ananassa) plants were inoculated with an aqueous suspension of D. earlianum (1 × 105 conidia/mL) and placed in environmental conditions to generate a range of disease severities. Photosynthesis measurements on individual leaflets of greenhouse-grown plants ('Kent') were taken in a biotron under saturating light conditions. Photosynthesis was also measured on leaflets in a naturally infected field planting of 'Jewel' in 2003. Leaf-scorch severity ranged from 0% to nearly 100% on individual leaflets. Strong negative linear correlations were found between the proportion of leaflet area with visual symptoms and A, E, and water-use efficiency (WUE = A/E). "Physiological-lesion" size was estimated to be 1.6 and 2.1 for greenhouse-grown and field-grown plants, respectively, meaning that the total area of leaflet tissue affected by leaf scorch was 1.6-2.1 times larger than the leaflet area showing visual symptoms of the disease. The larger physiological lesion found in field-grown plants may be due to field-associated stress factors or cultivar variability. The results have application for establishing guidelines for the management of foliar diseases of strawberry.

Enhancement of Cardenolide and Phytosterol Levels by Expression of an N-Terminally Truncated 3-Hydroxy-3-methylglutaryl CoA Reductase in Transgenic Digitalis minor

Pathway engineering in medicinal plants attains a special significance in Digitalis species, the main industrial source of cardiac glycosides, steroidal metabolites derived from mevalonic acid via the triterpenoid pathway. In this work, the Arabidopsis thaliana HMG1 cDNA, coding the catalytic domain of 3-hydroxy-3-methylglutaryl CoA reductase (HMGR1S), a key enzyme of the MVA pathway, was expressed in the cardenolide-producing plant Digitalis minor. Transgenic plants were morphologically indistinguishable from control wild plants and displayed the same developmental pattern. Constitutive expression of HMG1 resulted in an increased sterol and cardenolide production in both in vitro- and greenhouse-grown plants. This work demonstrates that transgenic D. minor plants are a valuable system to study and achieve metabolic engineering of the cardenolide pathway and in consequence for the genetic improvement of Digitalis species.

Xanthomonas campestris pv. vitians에 의한 상추 세균성점무늬병

1997년과 1998년 광주와 광명의 상추 재배 온실에서 새로운 병해가 발견되었다. 처음에는 잎에 병반이 불규칙적이고 작은 연녹색이었다가 점점 커져 지름이 2<TEX>$\sim$</TEX>5 mm 정도인 수침상의 검은색 병반이 형성되었다. 잎의 아래쪽에서 엽맥을 따라 검은색의 줄무늬 병반이 형성되기도 하고 심해지면 잎이 시들어 말라 죽었다. YDC 배지에서 병원세균을 순수 분리하였을 때 Xanthomonas속 세균의 전형적인 특징인 노란색 색소를 띤 세균이 형성되었다. 분리된 세균을 <TEX>$10^8$</TEX> cfu/ml로 현탁한 후 상추의 엽맥에 주사 접종하여 병원성을 확인하였다. 지방산 조성 및 함량과 다양한 탄소원 이용정도를 이용하여 분리세균 SL0246과 SL1352를 X. campestris pv. vitians로 동정하였으며 이 병을 상추의 세균성점무의병으로 명명하였다. During 1997 and 1998, a new disease of lettuce (Lactuca sativa) was observed on greenhouse-grown plants in Kwangju and Kwangmyung. Lesions on leaves were irregular, small, pale green to black, water-soaked, and 2 to 5 mm in diameter, Coalescing lesions sometimes caused defoliation of older leaves. Isolations made from diseased leaves on yeast extract dextrose calcium carbonate agar yielded nearly pure cultures of a yellow pigmented bacterium typical of a xanthomonad. Two bacterial strains (SL0246 and SL1352) were purified and used for further tests. Pathogenicity of strains was confirmed on 5-week-old lettuce plants injected with bacterial suspensions containing <TEX>$10^8$</TEX> cfu/ml of phosphate buffered saline. The representative Xanthomonas strains isolated from lettuce were compared with a reference strain X. campestris pv, vitians for fatty acid profiles and metabolic fingerprints using GN2 microplate, showing that all outcomes were indistinguishable between the representative and reference strains. This is the first report of bacterial leaf spot and dry rot of lettuce in Korea.

Early Detection of Leifsonia xyli subsp. xyli in Sugarcane Leaves by Real-Time Polymerase Chain Reaction.

A real-time, polymerase chain reaction (PCR) assay was developed for detecting Leifsonia xyli subsp. xyli in sugarcane leaf tissue. Real-time PCR assays were conducted on the youngest, fully expanded leaf of three cultivars collected bi-weekly from field nurseries between 11 April and 19 July 2005. L. xyli subsp. xyli infection was detected in leaves collected at all sampling dates, including those from 1-month-old plants on 11 April. Assays conducted on older, more rapidly growing plants (28 July and 21 October 2005) indicated that leaf position affects assay efficiency. Conventional PCR was less efficient than real-time PCR for detecting L. xyli subsp. xyli in leaf tissue. Real-time PCR was used to rank cultivars for susceptibility to L. xyli subsp. xyli infection based on the relative titer of L. xyli subsp. xyli in leaves of inoculated, 3- and 4-month-old greenhouse-grown plants. The ranking of cultivars by real-time PCR was in close agreement with the ranking determined by tissue-blot enzyme immunoassay performed on tissue from 7- to 9-month-old stalks.