In June 2011, tomatoes (Solanum lycopersicum) in major growing areas of the Cameron Highlands and the Johor state in Malaysia were affected by a leaf spot disease. Disease incidence exceeded 80% in some severely infected regions. Symptoms on 50 observed plants initially appeared on leaves as small, brownish black specks, which later became grayish brown, angular lesions surrounded by a yellow border. As the lesions matured, the affected leaves dried up and became brittle and later developed cracks in the center of the lesions. A survey was performed in these growing areas and 27 isolates of the pathogen were isolated from the tomato leaves on potato carrot agar (PCA). The isolates were purified by the single spore technique and were transferred onto PCA and V8 agar media for conidiophore and conidia production under alternating light (8 hours per day) and darkness (16 hours per day) (4). Colonies on PCA and V8 agar exhibited grey mycelium and numerous conidia were formed at the terminal end of conidiophores. The conidiophores were up to 240 μm long. Conidia were oblong with 2 to 11 transverse and 1 to 6 longitudinal septa and were 24 to 69.6 μm long × 9.6 to 14.4 μm wide. The pathogen was identified as Stemphylium solani on the basis of morphological criteria (2). In addition, DNA was extracted and the internal transcribed spacer region (ITS) was amplified by universal primers ITS5 and ITS4 (1). The PCR product was purified by the commercial PCR purification kit and the purified PCR product sequenced. The resulting sequences were 100% identical to published S. solani sequences (GenBank Accestion Nos. AF203451 and HQ840713). The amplified ITS region was deposited with NCBI GenBank under Accession No. JQ657726. A representative isolate of the pathogen was inoculated on detached 45-day-old tomato leaves of Malaysian cultivar 152177-A for pathogenicity testing. One wounded and two nonwounded leaflets per leaf were used in this experiment. The leaves were wounded by applying pressure to leaf blades with the serrated edge of a forceps. A 20-μl drop of conidial suspension containing 105 conidia/ml was used to inoculate these leaves (3). The inoculated leaves were placed on moist filter paper in petri dishes and incubated for 48 h at 25°C. Control leaves were inoculated with sterilized distilled water. After 7 days, typical symptoms for S. solani similar to those observed in the farmers' fields developed on both wounded and nonwounded inoculated leaves, but not on noninoculated controls, and S. solani was consistently reisolated. To our knowledge, this is the first report of S. solani causing gray leaf spot of tomato in Malaysia. References: (1) M. P. S. Camara et al. Mycologia 94:660, 2002. (2) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiversity Series 6:775, 2007.
In August 2008, 30% of tomato (Solanum lycopersicum) plants in plots in Lubbock County, Texas showed yellowing, lateral stem dieback, upward leaf curling, enlargement of stems, adventitious roots, and swollen nodes. Yellowing in leaves was similar to that seen with zebra chip disease (ZC) of potato that was confirmed in a potato field 112 km away in July 2008 and was associated with a 'Candidatus Liberibacter' species (1), similar to findings earlier in 2008 in New Zealand and California (2,3). Tissue from four symptomatic plants of cv. Spitfire and two of cv. Celebrity were collected and DNA was extracted from midribs and petioles with a FastDNA Spin Kit (Qbiogene, Inc., Carlsbad, CA,). PCR amplification was done with 16S rRNA gene primers OA2 and OI2c, which are specific for "Ca. Liberibacter solanacearum" from potato and tomato and amplify a 1.1-kb fragment of the 16S rRNA gene of this new species (1,3). Amplicons of 1.1 kb were obtained from all samples and these were sequenced in both orientations (McLab, San Francisco, CA). Sequences of the 16S rRNA gene were identical for both Spitfire and Celebrity and were submitted to the NCBI as GenBank Accession Nos. FJ939136 and FJ939137, respectively. On the basis of a BLAST search, sequence alignments revealed 99.9% identity with a new species of 'Ca. Liberibacter' from potato (EU884128 and EU884129) in Texas (1); 99.7% identity with the new species "Ca. Liberibacter solanacearum" described from potato and tomato (3) in New Zealand (EU849020 and EU834130, respectively) and from the potato psyllid Bactericera cockerelli in California (2) (EU812559, EU812556); 97% identity with 'Ca L. asiaticus' from citrus in Malaysia (EU224393) and 94% identity with both 'Ca. L. africanus' and 'Ca. L. americanus' from citrus (EU921620 and AY742824, respectively). A neighbor-joining cladogram constructed using the 16S rRNA gene fragments delineated four clusters corresponding to each species, and these sequences clustered with "Ca. L. solanacearum". A second PCR analysis was conducted with the CL514F/CL514R primer pair, which amplifies a sequence from the rplJ and rplL ribosomal protein genes of "Ca. L. solanacearum". The resulting 669-bp products were 100% identical to a sequence reported from tomato in Mexico (FJ498807). This sequence was submitted to NCBI (GU169328). ZC, a disease causing losses to the potato industry, is associated with a 'Candidatus Liberibacter' species (1-3) and was reported in Central America and Mexico in the 1990s, in Texas in 2000, and more recently in other states in the United States (4). In 2008, a "Ca. Liberibacter solanacearum" was detected on Capsicum annuum, S. betaceum, and Physalis peruviana in New Zealand (3). Several studies have shown that the potato psyllid, B. cockerelli, is a potential vector for this pathogen (2,4). To our knowledge, this is the first report of "Ca. Liberibacter solanacearum" in field tomatoes showing ZC-like foliar disease symptoms in the United States. References: (1). J. A. Abad et al. Plant Dis. 93:108, 2009 (2) A. K. Hansen et al. Appl. Environ. Microbiol. 74:5862, 2008. (3) L. W. Liefting et al. Plant Dis. 93:208, 2009. (4) G. A. Secor et al. Plant Dis. 93:574, 2009.
Endophytic fungi are those living inside the host plant without causing any apparent negative effect on the host plant. Two
isolates endophytic fungi from leaves and two isolates from root at Universiti Teknologi MARA (UiTM) Reserve Forest,
Negeri Sembilan were successfully isolated and identified by morphology and molecular characteristic. Samples were
surface sterilized and sub-cultured to obtain a pure culture. Characteristics of the isolates such as colony appearance,
mycelial texture, conidia/spores and pigmentation were studied to explore their morphology. Isolates were also subjected to
a PCR-based genotyping test. There were noticeable differences in morphological characteristics among the four isolates.
Microscopic analysis showed four isolates consist of septa and conidia/spores. The pigmentation result showed that
colony in A1leaf samples demonstrated an orange color on potato dextrose agar (PDA) media, colony in A1root demonstrate
a black texture in PDA media while hairy colonies in the others two isolates showed a white color on PDA media. Based on
molecular analyses the fungal genera showed 99-100% similarity with the related fungi recorded in the GenBank. Both
morphology and molecular sequencing of internal transcribed spacer (ITS) regions of endophytic fungi showed that three
isolates (A1root, C2leaf, and C3root) were grouped in Basidiomycota while one isolate (A1leaf) belonged to Ascomycota. The
endophyte funguses were identified as Daldinia sp. (A1leaf), Polyporales sp. (A1root,) Lentinus sp. (C2leaf,) and Rigidoporus
sp. (C3root). Overall, the new discoveries of isolated endophyte fungal have dyeing potential of fungal pigments which
offer a viable alternative to natural vegetable and harmful synthetic dyes.
An experiment was conducted with the objective to enhance mucosal immunity against ovalbumin (OVA) by co-administration of OVA with an aqueous extract from the fruit of Solanum torvum (STE). Five groups of female ICR mice aged approximately 8 weeks at the commencement of the experiment were caged in groups of eight and received various treatments. The treatments included OVA alone, OVA with cholera toxin (CT), and OVA with various doses of STE. Mice were primed intraperitoneally with 500 microg of OVA alone or co-administered with 0.1 microg CT, or with 1 microg STE. All mice were boosted orally via gastric intubation 14 days after priming with 10 mg OVA alone, or co-administered with 10 microg CT or with 10 mg, 1 mg or 0.1 mg STE. One week later all mice were killed and organs obtained for analysis of the immune response. Intestinal, faecal and pulmonary OVA-specific sIgA concentration was significantly increased (p<0.05) in mice that received booster combinations of OVA/CT and OVA with all extract doses (p<0.05). Specific serum IgG titres did not differ significantly between groups. It is concluded that STE can significantly enhance secretory immunity in the intestine to OVA with mucosal homing to the lungs. The adjuvant effect of STE is comparable to that of CT.
A polysaccharide fraction from Solanum nigrum, SN-ppF3 was shown previously to have an immunomodulatory activity where it could possibly be used to enhance the host immune response in fighting cancer. The non-toxic SN-ppF3 was fed orally to breast tumor bearing-mice with concentrations of 250 and 500mg/kg for 10days. During the treatment period, size of the tumor and weight of the mice were monitored. At the end of the treatment, blood, tumor, spleen and thymus were harvested for physiological and immunological analyses. After the treatment, the tumor volume and tumor weight were significantly inhibited by 65% and 40%, respectively. Based on the histological observation, the treatment of SN-ppF3 resulted in the disruption of tumor cells morphology. The increase in infiltrating T cells, NK cells and macrophages were observed in tumor tissues of the treated mice, which partly explained the higher apoptosis tumor cells observed in the treated mice. Moreover, the level of TNF-α, IFN-γ and IL-4 were elevated, while the level of IL-6 was decreased significantly, in serum of the treated mice. These results suggested that tumor suppression mechanisms observed in SN-ppF3-treated mice were most probably due through enhancing the host immune response.
A preliminary study was conducted to determine the level of oxidative DNA damage, fruits and vegetables intake among 50 breast cancer patients (cases) as compared to 50 healthy women (controls) with no known medical history of breast cancer in Klang Valley. Both groups were matched for age and ethnicity. Data on socio-demographic, health status and medical history, fruits and vegetables intake, and supplements intake were obtained through an interviewbased questionnaire. Anthropometry measurements included weight, height, and waist and hip circumference were also carried out on subjects. A total of 3mL fasting venous blood was drawn to assess lymphocytes oxidative DNA damage using Alkaline Comet Assay. Results indicated that the mean intake of fruits and vegetables was lower in cases (4.09 ± 1.17 servings/d) than controls (4.77 ± 0.90 servings/d)(p < 0.05) The intake of fruits and vegetables from family groups of solanaceae, myrtaceae, caricaceae, apiaceae, brinjal, rutaceae, broccoli, orange, carrot, watermelon were 0.5 - 1 servings/week significantly higher among controls as compared to cases (p < 0.05 for all parameters). However, the intake of fruits from rosaceae family and apple was higher among controls than cases (p < 0.05). The estimated intake of ß-carotene, carotenoids, vitamin A, vitamin C (p < 0.001), a-carotene and lycopene (p < 0.05) from fruits and vegetables were higher among controls than cases. Mean DNA damage level of cases (4.55 ± 1.78 % DNA in tail, %TD; 0.35 ± 0.21 tail moment, TM) were 3.5 and 3.9 times higher than the value of controls (1.3 ± 0.70% TD; 0.09 ± 0.09 TM) (p < 0.001) and the damage increased with higher values of waist hip ratio (% TD, r = 0.396, p < 0.05; TM, r = 0.349, p < 0.05) and waist circumference (% TD, r = 0.334, p < 0.05; TM, r = 0.360, p < 0.05). There was an inverse relationship between oxidative DNA damage with intake of total fruits and vegetables, cauliflowers and water convolvulus and also consumption from rutaceae and solanaceae families. Similar trend was noted for estimated intake of vitamin A, carotenoids, vitamin C, ß-carotene and lycopene. In conclusion, the intake of fruits and vegetables of five servings/d and the consumption of specific families and types of fruits and vegetables might protect against oxidative DNA damage and further reduce breast cancer risk.