Tomato mosaic virus (ToMV), a member of the genus Tobamovirus, infects several ornamental and horticultural crops worldwide. In this study, the nucleotide sequences of the coat protein gene of worldwide ToMV isolates were analyzed to estimate the genetic structure and diversity of this virus and the involved evolutionary forces. The phylogenetic analysis showed three clades with high bootstrap support: Clade I contained three ToMV isolates from Brazil collected from pepper, Clade II comprised one Brazilian ToMV isolate from pepper, and Clade III was composed of ToMV isolates collected from different plant hosts (pepper, tomato, eggplant, lilac, camellia, dogwood, red spruce, etc.) and water (from melting ice, lakes and streams) from different countries: USA, Brazil, Korea, Germany, Spain, Denmark (Greenland), China, Taiwan, Malaysia, Iran, and Kazakhstan. With the exception of Brazil, nucleotide diversity within and between different geographic regions was very low, although statistical analyses suggested some gene flow between most of these regions. Our analyses also suggested a strong negative selection which could have contributed to the genetic stability of ToMV.
Lycopene extraction was carried out via the ultrasonic assisted extraction (UAE) with response surface methodology (RSM). Sonication enhanced the efficiency of relative lycopene yield (enhancement of 26% extraction yield of lycopene in 6 replications at 40.0 min, 40.0 °C and 70.0% v/w in the presence of ultrasound), lowered the extraction temperature and shortened the total extraction time. The extraction was applied with the addition of oxygen-free nitrogen flow and change of water route during water bath sonication. The highest relative yield of lycopene obtained was 100% at 45.0 °C with total extraction time of 50.0 min (30:10:10) and ratio of solvent to freeze-dried tomato sample (v/w) of 80.0:1. Optimisation of the lycopene extraction had been performed, giving the average relative lycopene yield of 99% at 45.6 min, 47.6 °C and ratio of solvent to freeze-dried tomato sample (v/w) of 74.4:1. From the optimised model, the average yield of all-trans lycopene obtained was 5.11±0.27 mg/g dry weight. The all-trans lycopene obtained from the high-performance liquid chromatography (HPLC) chromatograms was 96.81±0.81% with 3.19±0.81% of cis-lycopenes. The purity of total-lycopene obtained was 98.27±0.52% with β-carotene constituted 1.73±0.52% of the extract. The current improved, UAE of lycopene from tomatoes with the aid of RSM also enhanced the extraction yield of trans-lycopene by 75.93% compared to optimised conventional method of extraction. Hence, the current, improved UAE of lycopene promotes the extraction yield of lycopene and at the same time, minimises the degradation and isomerisation of lycopene.
Tomatoes have a short shelf life thus they pose a big challenge for growers to maintain the quality of tomatoes to increase customer acceptance. In this study, fungi associated with tomato disease symptoms were isolated and the potential of kaffir lime aqueous extract was evaluated in maintaining post-harvest quality of tomatoes. For this purpose, healthy tomatoes were dipped in 10% aqueous kaffir lime extract before evaluating the post-harvest parameters namely weight loss and firmness. A fungus namely Rhizophus stolonifer was isolated from the symptomatic tomatoes. Subsequently, it was confirmed to be pathogenic on healthy tomato fruits with 100% disease severity. Application of aqueous kaffir lime extract showed that tomato fruits dipped in 10% aqueous kaffir lime extract recorded higher weight loss and higher firmness as compared to untreated tomato fruits. The results showed that treatment with this concentration of plant extract did not help to reduce the weight loss, but it retained the firmness of the tomato fruits stored at room temperature at 27+2oC. Higher transpiration process would lead to shrinkage, weight loss, changes in texture and appearance of the fruits. Therefore, this study suggested an increased concentration of aqueous kaffir lime extract as a treatment agent in order to have a better effect in maintaining the quality of tomato fruits.
Soft fleshed tomatoes are easily damaged due to mechanical injuries. Later, the wounded tissue will be exposed to fungal infection thus fasten the deterioration rate and reduce the quality of tomato. The aim of this study was to evaluate the potential aqueous ginger extract to inhibit fungal pathogen that causes tomato wilt and its potential in delaying the weight loss of tomato fruits. For this purpose, in vitro antifungal assay using poison plate technique was used to observe the inhibition of fungal pathogen. Then, healthy tomato fruits were dipped in aqueous ginger extract before evaluated for the post-harvest quality such as weight loss and firmness. The results of this study show that 10% aqueous ginger extract can inhibit the fungal pathogen (Fusarium oxysporum) that causes tomato wilt with 13.57% inhibition. Through in vivo antifungal assay, tomato fruits dipped in this plant extract showed lower weight loss (14.44%) and higher firmness (1.7 N) as compared to untreated fruit, but the data were not significantly different. Therefore, manipulation of this extract was suggested to increase its antifungal properties or as eco-friendly coating to lengthen the shelf life of agricultural produces.
The objective of this study was to investigate the influence of spent mushroom compost of Pleurotus eous strain P-31 on the growth and yield performance of pepper and tomato seedlings under greenhouse conditions. Sandy loam soil was combined with different percentages of SMC to obtain the following combinations (0, 5, 10, 15, 20, 25 and 30) %. Lower concentrations SMC5, SMC10 and SMC15 promoted vegetative growth (plant height, leaf area, chlorophyll content, number of leaves and axillary branches) of the two test plants. Tomato seedlings grown in SMC10 recorded the highest plant height (50.3 ± 7.2cm); leaf area (378.8 ± 1.2cm2); number of floral buds (51) and flowers (28) whereas SMC5 recorded the highest chlorophyll content 34.1 ± 0.9CCI though SMC15 recorded the highest number of leaves (8). Tomato seedlings grown in SMC30 produced both the maximum number of fruits (8) with corresponding high weight (34.2 ± 7.7g). Pepper seedlings grown in lower concentrations (SMC5-15) recorded the highest plant heights (29.8-30.8cm), chlorophyll content (20.3CCI) and leaf area (53.5-66.2 cm2). Although the different combinations of sandy loam soil and SMC did not significantly (p ≥ 0.05) affect the number of axillary branches developed; different combinations significantly (p ≤ 0.05) affected the number of floral bud, flower and fruit, weight of fruits formed and value of each of these increased with increasing percentage of SMC. Pepper seedlings grown on SMC30 recorded the maximum number of floral buds (32.0 ± 3.6), number of flowers (19.4 ± 1.3), number of fruits (10.8 ± 1.2) and weight of fruits (31.9 ± 3.4g). Tomato seedlings raised on SMC100 (spent mushroom compost only) and soil only did not significantly (p ≥ 0.05) differ from each other however, was statistically significant (p ≤ 0.05) from amended sandy loam soil by all criteria investigated. The study shows that SMC provide favourable soil conditioners for the cultivation of fruits, vegetables and foliage crops as it improved growth and yield of tomato and pepper seedlings.
beta-D-N-Acetylhexosaminidase, a family 20 glycosyl hydrolase, catalyzes the removal of β-1,4-linked N-acetylhexosamine residues from oligosaccharides and their conjugates. We constructed phylogenetic tree of β-hexosaminidases to analyze the evolutionary history and predicted functions of plant hexosaminidases. Phylogenetic analysis reveals the complex history of evolution of plant β-hexosaminidase that can be described by gene duplication events. The 3D structure of tomato β-hexosaminidase (β-Hex-Sl) was predicted by homology modeling using 1now as a template. Structural conformity studies of the best fit model showed that more than 98% of the residues lie inside the favoured and allowed regions where only 0.9% lie in the unfavourable region. Predicted 3D structure contains 531 amino acids residues with glycosyl hydrolase20b domain-I and glycosyl hydrolase20 superfamily domain-II including the (β/α)8 barrel in the central part. The α and β contents of the modeled structure were found to be 33.3% and 12.2%, respectively. Eleven amino acids were found to be involved in ligand-binding site; Asp(330) and Glu(331) could play important roles in enzyme-catalyzed reactions. The predicted model provides a structural framework that can act as a guide to develop a hypothesis for β-Hex-Sl mutagenesis experiments for exploring the functions of this class of enzymes in plant kingdom.
The plant hormone, ethylene, is an important regulator which involved in regulating fruit ripening and flower senescence. In this study, RNA interference (RNAi) technology was employed to silence the genes involved in ethylene biosynthetic pathway. This was achieved by blocking the expression of specific gene encoding the ACC oxidase. Initially, cDNA corresponding to ACO1 of lowland tomato cultivar (MT1), which has high identity with ACO1 of Solanum lycopersicum in GenBank, was cloned through RT-PCR. Using a partial coding region of ACO1, one hpRNAi transformation vector was constructed and expressed ectopically under the 35S promoter. Results showed that transgenic lines harboring the hpRNA-ACO1 construct had lower ethylene production and a longer shelf life of 32 days as compared to 10 days for wild-type fruits. Changes in cell wall degrading enzyme activities were also investigated in cases where the transgenic fruits exhibited reduced rates of firmness loss, which can be associated with a decrease in pectin methylesterase (PME) and polygalacturonase (PG) activities. However, no significant change was detected in both transgenic and wild-type fruits in terms of β-galactosidase (β-Gal) activity and levels of total soluble solid, titratable acid and ascorbic acid.
Background: The low consumption of fruits and vegetables among children is a global challenge. Foods recognition, nutrition knowledge and attitude are factors that influence children's dietary practices. This study aims to assess the preference, attitude, recognition and knowledge of fruits and vegetables intake among Malay children.
Methods: A cross-sectional study was conducted among Malay children from five primary schools in Kuala Lumpur using self-administered questionnaires.
Results: A total of 134 Malay children (70 males and 64 females) with a mean (SD) age of 10.3 (1.0) years were recruited. Majority of the children had a father (61.9%) and a mother (56.0%) with secondary school education and earned below RM3,900 (70.9%) per month. The most preferred fruits and vegetable were bananas (91.9%) and carrots (71.4%), while the most recognised was oranges (100.0%) and tomatoes (96.3%). The children demonstrated an overall moderate level of attitude, recognition and knowledge with mean (SD) scores of 70.3 (19.9), 76.8 (18.1) and 73.6 (17.5), respectively, towards fruits and vegetables intake. Majority of the children (53.0%) were not aware of the daily recommended servings of fruits and vegetables, while 40.0% of children expressed a low attitude towards eating a variety of fruits and vegetables. The willingness to try a new type of vegetables and consume more vegetables was lower (68.7%) compared to fruits (75.4%).
Conclusion: The preferences and recognition of fruits were higher compared to vegetables among the children. The children demonstrated a moderate level of attitude, recognition and knowledge towards fruits and vegetables consumption. Efforts to educate children on the recommended number of servings per day and improve their acceptability of vegetables should be implemented to promote the increase in fruits and vegetables consumption among children.
Optimum microclimate parameters, including air temperature (T), relative humidity (RH) and vapor pressure deficit (VPD) that are uniformly distributed inside greenhouse crop production systems are essential to prevent yield loss and fruit quality. The objective of this research was to determine the spatial and temporal variations in the microclimate data of a commercial greenhouse with tomato plants located in the mid-west of Iran. For this purpose, wireless sensor data fusion was incorporated with a membership function model called Optimality Degree (OptDeg) for real-time monitoring and dynamic assessment of T, RH and VPD in different light conditions and growth stages of tomato. This approach allows growers to have a simultaneous projection of raw data into a normalized index between 0 and 1. Custom-built hardware and software based on the concept of the Internet-of-Things, including Low-Power Wide-Area Network (LoRaWAN) transmitter nodes, a multi-channel LoRaWAN gateway and a web-based data monitoring dashboard were used for data collection, data processing and monitoring. The experimental approach consisted of the collection of meteorological data from the external environment by means of a weather station and via a grid of 20 wireless sensor nodes distributed in two horizontal planes at two different heights inside the greenhouse. Offline data processing for sensors calibration and model validation was carried in multiple MATLAB Simulink blocks. Preliminary results revealed a significant deviation of the microclimate parameters from optimal growth conditions for tomato cultivation due to the inaccurate timer-based heating and cooling control systems used in the greenhouse. The mean OptDeg of T, RH and VPD were 0.67, 0.94, 0.94 in January, 0.45, 0.36, 0.42 in June and 0.44, 0.0, 0.12 in July, respectively. An in-depth analysis of data revealed that averaged OptDeg values, as well as their spatial variations in the horizontal profile were closer to the plants' comfort zone in the cold season as compared with those in the warm season. This was attributed to the use of heating systems in the cold season and the lack of automated cooling devices in the warm season. This study confirmed the applicability of using IoT sensors for real-time model-based assessment of greenhouse microclimate on a commercial scale. The presented IoT sensor node and the Simulink model provide growers with a better insight into interpreting crop growth environment. The outcome of this research contributes to the improvement of closed-field cultivation of tomato by providing an integrated decision-making framework that explores microclimate variation at different growth stages in the production season.
Fusarium species is one of the common pathogens of post-harvest disease to cause rot on tomato and other perishable vegetable fruits. The objectives of this study were to determine the diversity of Fusarium isolated species from post-harvest diseases of tomato fruit, to identify the causal organisms by using phenotype characteristics and to verify the pathogens of Fusarium fruit of tomato based on pathogenicity test. Carnation leaf-piece agar (CLA) and potato dextrose agar (PDA) media were used for phenotype-based identification of the Fusarium isolates with emphasis for characterizations of the shapes and sizes of the macroconidia and microconidia, colony features, growth rates, conidiogenous cells and chlamydospores. A total of 180 Fusarium isolates were obtained from 13 locations throughout Selangor. Fusarium solani was most abundantly isolated (34%) followed by F. semitectum (31%) and F. oxysporum (31%), F. subglutinans (3%) while the least was F. equiseti (1%). Twenty seven isolates were tested for pathogenicity test by injecting 1 mL of the conidial suspension onto healthy tomatoes. All the tested Fusarium isolates were pathogenic on tomato with different severity levels. The non-inoculated controls showed no symptoms of fruit rot. The most virulent was F. oxysporum isolate B711T with DSI 93.75%, while the least were isolates of F. solani (B647T) and F. oxysporum (B727T) with DSI 37.5%. Majority of the isolated Fusarium species can potentially produce mycotoxins as their secondary metabolites. The potential production of mycotoxins by pathogenic isolates of Fusarium species in contaminated tomato fruits could pose health hazards when consumed.
Fusarium species are known to cause various diseases on plantations including fruits and vegetables. The most common Fusarium that can cause plant diseases are Fusarium proliferatum and Fusarium verticillioides. Ear rot disease on maize, wilt disease on cucurbits and fruit rot disease on tomato as well as banana are example of diseases caused by these two species. The objectives of this study were to identify F. proliferatum and F. verticillioides based on species-specific primers and polymerase chain reaction (PCR) amplification and to evaluate the genetic diversity of both species based on microsatellite markers. Fifty isolates of Fusarium species that were previously collected throughout Malaysia from different hosts were identified by using species-specific PCR amplification. Twenty-nine isolates were identified as F. proliferatum and 21 isolates were identified as F. verticillioides based on species-specific primer. The genetic diversity of all the fungal isolates was evaluated by using microsatellite analysis with six established primers. Five out of six primers amplified polymorphic bands with the most effective primer showing high polymorphism were (AG)7C and (TCC)5 meanwhile one primer (TTTC)4 gave negative result with no band amplified. The phylogenetic tree that was constructed showing two different clades distinguished between F. proliferatum and F. verticillioides.
Gen Proteolisis 6 (PRT6) merupakan gen yang memainkan peranan penting dalam tapak jalan N-end rule dan berfungsi
sebagai enzim E3 ligase. PRT6 berperanan dalam pengenalan protein sasaran bagi proses degradasi. Objektif utama kajian
ini adalah untuk mentransformasi konstruk RNAi PRT6 ke dalam tomato berperantarakan Agrobacterium tumefaciens.
Ini bertujuan untuk memahami peranan tapak jalan N-end rule semasa proses pemasakan buah. Beberapa faktor yang
memberi kesan kepada transformasi seperti masa ko-penanaman dan juga kepekatan antibiotik yang digunakan telah
dioptimumkan. Keputusan kajian menunjukkan pengeraman kotiledon selama 48 jam pada medium ko-penanaman dapat
meningkatkan penghasilan kalus sebanyak 61% manakala penggunaan 500 mg/L antibiotik karbenisilin dalam medium
regenerasi pucuk dapat mengurangkan kontaminasi A. tumefaciens sehingga 5.2%. Selain itu, strain A. tumefaciens
C58 merupakan strain A. tumefaciens yang paling sesuai digunakan sebagai perantara dalam kajian ini. Tindak balas
berantai polimerase (PCR) telah dijalankan pada pucuk yang terhasil untuk mengesahkan integrasi fragmen PRT6 ke dalam
genom tomato. Berdasarkan analisis PCR, kesemua tujuh pucuk putatif transgenik adalah merupakan transforman positif.
This work aims to develop corn starch/chitosan nanoparticles/thymol (CS/CNP/Thy) bio-nanocomposite films as potential food packaging materials that can enhance the shelf life of food. CS/CNP/Thy bio-nanocomposite films were prepared by the addition of different concentrations of thymol (0, 1.5, 3.0, 4.5 w/w%) using a solvent casting method. The resulting films were characterized in terms of optical, mechanical, and water vapor permeability (WVP) properties. The addition of thymol was found to reduce the tensile strength (TS), elongation at break (EAB), and Young's modulus (YM) of the films. Generally, the increment in the concentration of thymol did not significantly affect the TS, EAB, and YM values. The addition of 1.5 w/w% thymol increased the WVP of the films but the WVP reduced with the increase in thymol concentrations. CS/CNP/Thy-3% bio-nanocomposite films demonstrated the potential to lengthen the shelf life of cherry tomatoes packed with the films, whereby the cherry tomatoes exhibited no significant changes in firmness and the lowest weight loss. In addition, no mold growth was observed on the sliced cherry tomatoes that were in direct contact with the films during 7 days of storage, proving the promising application of the films as active food packaging materials.
Helicoverpa armigera (Hub.) is a destructive pest of the tomato (Lycopersicon esculentum Mill) crop in Pakistan. Although insecticides are the primary management strategy used to control H. armigera, most of them are not effective due to considerable toxic residual effects on the fruits. Nonetheless, H. armigera is rapidly evolving resistance against the available pesticides for its management. This situation calls upon the need of alternative management options against the pest. Different plant extracts have been suggested as a viable, environment-friendly option for plant protection with minimal side effects. Furthermore, the plant extracts could also manage the insect species evolving resistance against pesticides. This study evaluated the efficacy of different plant extracts (i.e., Neem seed, turmeric, garlic and marsh pepper) against H. armigera. Furthermore, the impact of the plant extracts on growth and yield of tomato crop was also tested under field conditions. The results revealed that all plant extracts resulted in higher mortality of H. armigera compared to control. Similarly, the highest plant height was observed for the plants treated with the plant extracts compared to untreated plants. Moreover, the highest tomato yield was observed in plants treated with plant extracts, especially with neem seed (21.013 kg/plot) followed by pepper extract (19.25 kg/plot), and garlic extract 18.4 kg/plot) compared to the untreated plants (8.9 kg/plot). It is concluded that plant extracts can be used as eco-friendly approaches for improving tomato yield and resistance management of H. armigera.
Entomopathogenic fungi (EPF) are natural enemies which affect insect population and have long been recognized as biological control agents against many insect pests. Some isolates have also been established as endophytes, benefiting their host plants without causing any symptoms or negative effects. Here we demonstrated two entomopathogenic fungal species, Isariajavanica (Frieder. & Bally) Samson & Hywel-jone 2005 and Purpureocillium lilacinum (Thom) Luangsa-ard, Hou-braken, Hywel-Jones & Samson (2011) as endophytes in tomato plants by using the seed inoculation method and examined their effect on plant growth, B. tabaci mortality, and adult emergence. Our study indicated that tomato seeds treated with a fungal suspension of I. javanica and P. lilacinum enabled their recovery from plant tissues (root, stem and leaf) up to 60 days after inoculation (DAI). Both endophytic isolates also caused significant mortality of adult B. tabaci on seedlings inoculated with, I. javanica (51.92±4.78%), and P. lilacinum (45.32±0.20%) compared to the control treatment (19.29±2.35). Adult emergence rates were significantly high in the control treatments (57.50±2.66%) compared to I. javanica (15.00±1.47%) and P. lilacinum (28.75±4.78%) treatments. This study provides evidence that endophytic isolates of I. javanica and P. lilacinum have a biocontrol potentials for used against whiteflies and could also explored as plant growth promoters.
Dehydration-responsive element binding (DREB) transcription factor plays an important role in controlling the expression of abiotic stress responsive genes. An intronless oil palm EgDREB1 was isolated and confirmed to be a nuclear localized protein. Electrophoretic mobility shift and yeast one-hybrid assays validated its ability to interact with DRE/CRT motif. Its close evolutionary relation to the dicot NtDREB2 suggests a universal regulatory role. In order to determine its involvement in abiotic stress response, functional characterization was performed in oil palm seedlings subjected to different levels of drought severity and in EgDREB1 transgenic tomato seedlings treated by abiotic stresses. Its expression in roots and leaves was compared with several antioxidant genes using quantitative real-time PCR. Early accumulation of EgDREB1 in oil palm roots under mild drought suggests possible involvement in the initiation of signaling communication from root to shoot. Ectopic expression of EgDREB1 in T1 transgenic tomato seedlings enhanced expression of DRE/CRT and non-DRE/CRT containing genes, including tomato peroxidase (LePOD), ascorbate peroxidase (LeAPX), catalase (LeCAT), superoxide dismutase (LeSOD), glutathione reductase (LeGR), glutathione peroxidase (LeGP), heat shock protein 70 (LeHSP70), late embryogenesis abundant (LeLEA), metallothionine type 2 (LeMET2), delta 1-pyrroline-5- carboxylate synthetase (LePCS), ABA-aldehyde oxidase (LeAAO) and 9-cis- Epoxycarotenoid dioxygenase (LeECD) under PEG treatment and cold stress (4 °C). Altogether, these findings suggest that EgDREB1 is a functional regulator in enhancing tolerance to drought and cold stress.
Tomato (Solanum lycopersicum) is a globally important crop with an economic value in the tens of billions of dollars, and a significant supplier of essential vitamins, minerals, and phytochemicals in the human diet. Shelf life is a key quality trait related to alterations in cuticle properties and remodeling of the fruit cell walls. Studies with transgenic tomato plants undertaken over the last 20 years have indicated that a range of pectin-degrading enzymes are involved in cell wall remodeling. These studies usually involved silencing of only a single gene and it has proved difficult to compare the effects of silencing these genes across the different experimental systems. Here we report the generation of CRISPR-based mutants in the ripening-related genes encoding the pectin-degrading enzymes pectate lyase (PL), polygalacturonase 2a (PG2a), and β-galactanase (TBG4). Comparison of the physiochemical properties of the fruits from a range of PL, PG2a, and TBG4 CRISPR lines demonstrated that only mutations in PL resulted in firmer fruits, although mutations in PG2a and TBG4 influenced fruit color and weight. Pectin localization, distribution, and solubility in the pericarp cells of the CRISPR mutant fruits were investigated using the monoclonal antibody probes LM19 to deesterified homogalacturonan, INRA-RU1 to rhamnogalacturonan I, LM5 to β-1,4-galactan, and LM6 to arabinan epitopes, respectively. The data indicate that PL, PG2a, and TBG4 act on separate cell wall domains and the importance of cellulose microfibril-associated pectin is reflected in its increased occurrence in the different mutant lines.
In August 2011, sweet potato (Ipomoea batatas), tomato (Solanum lycopersicum), and eggplant (S. melongena) crops from major growing areas of the Cameron highlands and Johor state in Malaysia were affected by a soft rot disease. Disease incidence exceeded 80, 75, and 65% in severely infected fields and greenhouses of sweet potato, tomato, and eggplant, respectively. The disease was characterized by dark and small water-soaked lesions or soft rot symptoms on sweet potato tubers, tomato stems, and eggplant fruits. In addition, extensive discoloration of vascular tissues, stem hollowness, and water-soaked, soft, dark green lesions that turned brown with age were observed on the stem of tomato and eggplant. A survey was performed in these growing areas and 22 isolates of the pathogen were obtained from sweet potato (12 isolates), tomato (6 isolates), and eggplant (4 isolates) on nutrient agar (NA) and eosin methylene blue (EMB) (4). The cultures were incubated at 27°C for 2 days and colonies that were emerald green on EMB or white to gray on NA were selected for further studies. All bacterial cultures isolated from the survey exhibited pectolytic ability on potato slices. These bacterial isolates were gram negative; rod shaped; N-acetylglucosaminyl transferase, gelatin liquefaction, and OPNG positive; and were also positive for acid production from D-galactose, lactosemelibiose, raffinose, citrate, and trehalose. They were negative for indol production, phosphatase activity, reducing substances from sucrose, and negative for acid production from maltose, sorbitol, inositol, inolin, melezitose, α-mathyl-D-glocoside, and D-arabitol. The bacteria did not grow on NA at 37°C. Based on these biochemical and morphological assays, the pathogen was identified as Pectobacterium wasabiae (2). In addition, DNA was extracted and PCR assay with two primers (16SF1 and 16SR1) was performed (4). Partial sequences of 16S rRNA (GenBank Accession Nos. JQ665714, JX494234, and JX513960) of sweet potato, tomato, and eggplant, respectively, exhibited a 99% identity with P. wasabiae strain SR91 (NR_026047 and NR_026047.1). A pathogenicity assay was carried out on sweet potato tubers (cv. Oren), tomato stems (cv. 152177-A), and eggplant fruits (cv. 125066x) with 4 randomly representative isolates obtained from each crop. Sweet potato tubers, tomato stems, and eggplant fruits (4 replications) were sanitized in 70% ethyl alcohol for 30 s, washed and rinsed in sterile distilled water, and needle punctured with a bacterial suspension at a concentration of 108 CFU/ml. Inoculated tubers, stems, and fruits were incubated in a moist chamber at 90 to 100% RH for 72 h at 25°C when lesions were measured. All inoculated tubers, stems, and fruits exhibited soft rot symptoms after 72 h similar to those observed in the fields and greenhouses and the same bacteria were consistently reisolated. Symptoms were not observed on controls. The pathogenicty test was repeated with similar results. P. wasabiae have been previously reported to cause soft rot on Japanese horseradish (3), and aerial stem rot on potato in New Zealand (4), the U.S. (2), and Iran (1). To our knowledge, this is the first report of sweet potato, tomato, and eggplant soft rot caused by P. wasabiae in Malaysia. References: (1) S. Baghaee-Ravari et al. Eur. J. Plant Pathol. 129:413, 2011. (2) S. De Boer and A. Kelman. Page 56 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. N. Schaad et al., eds. APS Press, St. Paul, 2001. (3) M. Goto et al. Int. J. Syst. Bacteriol. 37:130, 1987. (4) A. R. Pitman et al. Eur. J. Plant Pathol. 126:423, 2010.
Phytoplasmas (mycoplasmalike organisms, MLOs) associated with mitsuba (Japanese hone-wort) witches'-broom (JHW), garland chrysanthemum witches'-broom (GCW), eggplant dwarf (ED), tomato yellows (TY), marguerite yellows (MY), gentian witches'-broom (GW), and tsu-wabuki witches'-broom (TW) in Japan were investigated by polymerase chain reaction (PCR) amplification of DNA and restriction enzyme analysis of PCR products. The phytoplasmas could be separated into two groups, one containing strains JHW, GCW, ED, TY, and MY, and the other containing strains GW and TW, corresponding to two groups previously recognized on the basis of transmission by Macrosteles striifrons and Scleroracus flavopictus, respectively. The strains transmitted by M. striifrons were classified in 16S rRNA gene group 16SrI, which contains aster yellows and related phytoplasma strains. Strains GW and TW were classified in group 16SrIII, which contains phytoplasmas associated with peach X-disease, clover yellow edge, and related phytoplasmas. Digestion of amplified 16S rDNA with HpaII indicated that strains GW and TW were affiliated with subgroup 16SrIII-B, which contains clover yellow edge phytoplasma. All seven strains were distinguished from other phytoplasmas, including those associated with clover proliferation, ash yellows, elm yellows, and beet leafhopper-transmitted virescence in North America, and Malaysian periwinkle yellows and sweet potato witches'-broom in Asia.
Production of tomato (Lycopersicon esculentum) in Bangladesh, Malaysia, Myanmar, Vietnam, and Laos has been severely affected by yellow leaf curl disease. Tomato leaf samples were collected from symptomatic tomato plants from farmers' fields in the five countries from 1997 to 1999. DNA was extracted from all samples, four from Vietnam, two each from Malaysia, Laos, and Myanmar, and seven from Bangladesh. Virus DNA was amplified by polymerase chain reaction (PCR) using the begomovirus-specific degenerate primer pair PAL1v 1978/PAR1c 715(1), which amplifies the top part of DNA A. All samples gave the expected 1.4-kb PCR product. The PCR product of one sample per country was cloned and sequenced. Based on the sequences of the 1.4-kb DNA products amplified by the first primer pair, specific primers were designed to complete each of the DNA A sequences. Computer-assisted sequence comparisons were performed with begomovirus sequences available in the laboratory at the Asian Vegetable Research and Development Center, Shanhua, Tainan, and in the GenBank sequence database. The five DNA species resembled DNA A of begomoviruses. For the detection of DNA B two degenerate primer pairs were used, DNABLC1/DNABLV2 and DNABLC2/DNABLV2 (DNABLC1: 5'-GTVAATGGRGTDCACTTCTG-3', DNABLC2: 5'-RGTDCACTT CTGYARGATGC-3', DNABLV2: 5'-GAGTAGTAGTGBAKGTTGCA-3'), which were specifically designed to amplify DNA B of Asian tomato geminiviruses. Only the virus associated with yellow leaf curl of tomato in Bangladesh was found to contain a DNA B component, which was detected with the DNABLC1/DNABLV2 primer pair. The DNA A sequence derived from the virus associated with tomato yellow leaf curl from Myanmar (GenBank Accession No. AF206674) showed highest sequence identity (94%) with tomato yellow leaf curl virus from Thailand (GenBank Accession No. X63015), suggesting that it is a closely related strain of this virus. The other four viruses were distinct begomoviruses, because their sequences shared less than 90% identity with known begomoviruses of tomato or other crops. The sequence derived from the virus associated with tomato yellow leaf curl from Vietnam (GenBank Accession No. AF264063) showed highest sequence identity (82%) with the virus associated with chili leaf curl from Malaysia (GenBank Accession No. AF414287), whereas the virus associated with yellow leaf curl symptoms in tomato in Bangladesh (GenBank Accession No. AF188481) had the highest sequence identity (88%) with a tobacco geminivirus from Yunnan, China (GenBank Accession No. AF240675). The sequence derived from the virus associated with tomato yellow leaf curl from Laos (GenBank Accession No. AF195782) had the highest sequence identity (88%) with the tomato begomovirus from Malaysia (GenBank Accession No. AF327436). This report provides further evidence of the great genetic diversity of tomato-infecting begomoviruses in Asia. Reference: M. R. Rojas et al. Plant Dis. 77:340, 1993.