Molecular diagnostics for the sigatoka disease complex of banana

ABSTRACT The Sigatoka disease complex of banana involves three related ascomycetous fungi, Mycosphaerella fijiensis, M. musicola, and M. eumusae. The exact distribution of these three species and their disease epidemiology remain unclear, because their symptoms and life cycles are rather similar. Disease diagnosis in the Mycosphaerella complex of banana is based on the presence of host symptoms and fungal fruiting structures, which hamper preventive management strategies. In the present study, we have developed rapid and robust species-specific molecular-based diagnostic tools for detection and quantification of M. fijiensis, M. musicola, and M. eumusae. Conventional species-specific polymerase chain reaction (PCR) primers were developed based on the actin gene that detected DNA at as little as 100, 1, and 10 pg/mul from M. fijiensis, M. musicola, and M. eumusae, respectively. Furthermore, TaqMan real-time quantitative PCR assays were developed based on the beta-tubulin gene and detected quantities of DNA as low as 1 pg/mul for each Mycosphaerella sp. from pure cultures and DNA at 1.6 pg/mul per milligram of dry leaf tissue for M. fijiensis that was validated using naturally infected banana leaves.     

Dynamic Population of Gut Microbiota as an Indicator of Inflammatory Bowel Disease

Background: Inflammatory bowel disease is a chronic inflammatory disease of the gastrointestinal tract. The gut microbiota is an important factor in the pathogenesis of IBD. Due to a link between the gut microbiota and IBD, studying microbiota changes using an accurate, sensitive and rapid method for detection of the disease seems necessary. This study aimed to compare the composition of gut microbiota in three groups of people, including IBD patients, CIBD, and healthy groups. Methods:For this study, 45 stool samples (15 from each group) were collected. Using real-time PCR, the abundance of 11 bacterial 16S rRNA gene sequences was examined. Results:In the IBD group, the number of three bacterial phyla, including Firmicutes, Actinobacteria, and Bacteroidetes, decreased (p < 0.01, p < 0.01, and p < 0.001, respectively), while the population of γ-Proteobacteria increased significantly (p < 0.0001). In the CIBD group, the number of Actinobacteria enhanced (p < 0.01), but that of Bacteroidetes and Firmicutes decreased (p < 0.01, and p < 0.05, respectively). Conclusion:Findings of this study indicate that decrease in Firmicutes and increase in γ-Proteobacteria could be used as an indicator of IBD instead of employing invasive and costly detection methods such as colonoscopy and other tests. Key Words: Inflammatory bowel disease, Microbiota, Real-time polymerase chain reaction     

Seasonal,tissue and age influences on frequency and biodiversity of endophytic fungi of Citrus sinensis in Iran

A total of 4,009 endophytic fungal isolates were recovered from healthy leaves, bark and xylem of Citrus sinensis in different seasons and age‐classes. Corresponding to those factors, the majorities of fungal isolates were recovered from leaves, 2‐yr‐old trees and winter, respectively. Fungal isolates were initially categorized based on cultural and morphological characters, and representatives of each morphotype were subjected to molecular identifications based on ITS‐rDNA and β‐tubulin sequences. All isolates obtained in this study belonged to the phyla Ascomycota and Basidiomycota assigned to 30 fungal taxa. The most abundant fungal species were Alternaria spp. Species diversity indices of Margalef richness and Shannon–Wiener revealed a high diversity of fungal taxa recovered from leaf and during winter. Frequency (%) of fungal endophytes was highest in winter followed by summer, autumn, and spring significantly, whereas the corresponding sequence was winter, autumn, summer and spring for the diversity of endophytic fungi. Overall, the results of this study based on the community and diversity of endophytic fungal species in C. sinensis showed that the type of plant tissue, season of sampling and age of tree play a driving role on their abundance while their diversity was mainly dependent on the type of plant tissue, season of sampling than on the age of tree.     

Salvia officinalis hydroalcoholic extract improved reproduction capacity and behavioral activity in rats exposed to immobilization stress

This study was conducted to evaluate the Salvia officinalis hydroalcoholic extract on fertility capacity and behavioral features in rats exposed to immobilization stress. Male rats were randomly divided into five groups; Control; Stressed rats; and Stressed rats received 50, 100 and/or 200 mg/kg bw S. officinalis hydroalcoholic extract. To induce stress, rats were immobilized for 49 days and received S. officinalis extract orally. On day 56, we analyzed behavioral tests and evaluated reproduction capacity by measuring LH, FSH, and testosterone. Sperm parameters such as motility, viability, and total count were also determined. Bodyweight changes were also calculated on day 56. Male rats from different groups were mated with healthy female rats. Data showed that the use of 100 and 200 mg/kg bw S. officinalis extract in stressed rats increased bodyweight gain and improved behavioral disorders compared to control‐matched groups (p < .05). Besides, administration of 100 and 200 mg/kg bw S. officinalis extract had the potential to improve sperm parameters and fertility capacity in stressed rats (p < .05). Decreased testosterone levels were blunted in the stressed rats that received plant extract coincided with the reduction of LH and FSH compared to control‐matched stressed rats (p < .05). We found neutral effects in stressed rats that received 50 mg/kg bw plant extract. Collectively, the hydroalcoholic extract of S. officinalis could improve the fertility capacity and behavioral features under stressful conditions in a dose‐dependent manner.     

Single and competitive adsorption isotherms of some heavy metals onto a light textured calcareous soil amended with agricultural wastes-biochars

This study investigated the adsorption behavior of selected heavy metals (Pb, Cu, and Ni) under single and multi-metal conditions by a light textured calcareous soil amended with plant residue biochars (corn straw, wheat straw, rice husk and licorice root pulp each at 3% w/w). The Freundlich isotherm best described the heavy metal adsorption suggesting multilayer adsorption. For all treatments under both adsorption conditions, the heavy metal adsorption capacity followed the order of Pb > Cu > Ni, which was associated with the hydrolysis constant, ionic radius, and electronegativity of these metals. Simultaneous presence of multiple metals decreased the adsorption capacity for each metal and the sequence was in the order of Ni > Pb > Cu. The corn straw biochar (CSB) had the highest adsorption capacity (Freundlich K_f (mg g^?1) for Ni = 0.23, Cu = 1.41 and Pb = 2.73) and medium distribution coefficient (K_{d medium}(L kg^?1) for Ni = 59.30, Cu = 1961.00 and Pb = 2602.00), indicating the CSB is the best treatment for stabilization of heavy metals in the soil. This was associated with the chemical characteristics of the CSB (high amounts of CaCO₃ and P) and the greatest increase in soil pH value.     

Productivity,persistence and traits related to drought tolerance in smooth bromegrass

This study was conducted to evaluate drought tolerance and persistence in a germplasm of smooth bromegrass and association of forage productivity with different traits. Thirty‐six genotypes of smooth bromegrass were clonally propagated and evaluated under two soil moisture environments for three years (2013–2015). High genotypic variation was observed among genotypes for all the measured traits. Drought stress decreased mean values for traits related to productivity. Also, the long‐term stress for three years reduced the persistence of plants. Results indicated that indirect selection based on components of forage yield, which had high heritability and high correlation with yield, would be more effective to improve drought tolerance in this germplasm. The results of principal component analysis showed that there was a negative relationship between phenological traits with the persistence‐related traits and yield production. This suggests that selection for earliness may indirectly promote persistent genotypes. The results showed that some Hungarian genotypes are valuable gene sources for persistence. The most persistent genotypes from both groups (Iranian and foreign) were identified using the biplot method. These genotypes may be useful for the development of populations for future studies.     

Corms as a source of explants for the successful clonal propagation of <Emphasis Type="Italic">Crocus cancellatus</Emphasis>

The genus Crocus comprises plants with a potential to be developed as a new ornamental crop but to date, there are not many reports on in vitro propagation of many members of this genus. The present study involves in vitro propagation of Crocus cancellatus with ornamental and horticultural value. Two different types of corm explants (apical and basal halves of corms) were cultivated onto Murashige and Skoog’s (MS) medium supplemented with different levels of α-naphthalene acetic acid (NAA) and 6-benzylaminopurine (BAP). One to five cormlets emerged from every responding explant through direct organogenesis. Apical halves of corms were more highly responsive than basal halves and produced a maximum multiplication rate with 3.45 ± 0.06 cormlets per explant in 95.33 ± 2.33% of the explants in MS medium supplemented with 3% sucrose and 2 mg L⁻¹ NAA and 1 mg L⁻¹ BAP. The effect of cold storage temperature on in vitro cormlets sprouting was studied. Cormlets stored at 4°C for 8 weeks had more statistically significant positive effects on cormlets sprouting from the controls. In vitro rooting of cormlets was induced on MS medium without plant hormones.     

Water relations and microbial proliferation in vase solutions of Narcissus tazetta L. cv. ‘Shahla-e-Shiraz’ as affected by biocide compounds

Summary

Disruption of cut flower water relations due to the growth and proliferation of microorganisms in the vase solution is one of the most important factors causing reduced vase-life and deterioration in cut flowers. These experiments were conducted to study the effects of some biocides in controlling microbial proliferation in vase solutions and on water relations in cut Narcissus tazetta L. cv. ‘Shahla-e-Shiraz’, a native bulbous cut flower of Iran. The stems of ‘Shahla-e-Shiraz’ Narcissus cut flowers were exposed to 17 treatments: aluminium sulphate (at 100, 200 or 300 mg l^–1), citric acid (at 150, 300 or 450 mg l^–1), 8-hydroxyquinoline citrate (at 200, 300 or 400 mg l^–1), sodium hypochlorite (at 400, 600 or 800 mg l^–1), calcium hypochlorite (at 400, 600 or 800 mg l^–1), sterilised distilled water, or tap water (control). Generally, the least effective compounds in controlling microbial proliferation were low concentrations of aluminium sulphate or citric acid. 8-Hydroxyquinoline citrate treatments were most effective, and the vase solution did not contain any microbes until day-6. Vase solution contaminants were yeasts, Bacillus ssp., Staphylococcus ssp., Actinomycetes and Aspergillus ssp. Yeasts and bacilli were the most common microorganisms. Severe symptoms of phytotoxicity such as bud abortion and yellowing in aluminium sulphate-treated flowers, stem proximal-end browning and shrinking in 8-hydroxyquinoline citrate-treated flowers, and bleaching of soaked parts in sodium hypochlorite treatments, were seen. Most solution was taken up in 150 mg l^–1 citric acid, while the reduction in fresh weight was least in 200 mg l^–1 8-hydroxyquinoline citrate. The correlation between microbial counts and growth rate with solution uptake was slightly negative, but their correlation with fresh weight loss was slightly positive.