Screening of sugar beet tissue culture clones for resistance to rhizomania disease

In this study, sugar beet tissue culture clones were used to screen rhizomania resistant genotypes. At first, explants derived from shoot tips of sugar beet seedlings were transferred to shoot tip elongation media after surface sterilization. Then, the grown shoots were transferred to media containing various hormonal combinations NAA, BA, IBA and GA3 for multiplication, growth and rooting. Later, the clones were transferred to soil-peatmoss mixture were adapted to greenhouse conditions. For screening clones against rhizomania, the genotypes of adapted clones were selected and inoculated to rhizomania-infested soil. This experiment was in a randomized complete block design with three replicates (three inoculation times) in greenhouse. Adapted plants were transferred to the soil containing rhizomania virus. All infested soils were diluted 3 to 7 with sand. After two months, infested plants were examined by DAS-ELISA test also optical densities of the samples were analyzed by SAS program. Significant differences among genotypes and blocks were observed. Genotypes were classified to few groups (ranked from completely susceptible to completely resistant). The difference between blocks was because of difference of inoculation time temperature. Use of clones of each genotype caused an increase in selection accuracy of resistant genotypes. By use of this method, chance of escaping from inoculation factor decrease and researchers can determine to be resistance of plants with high level of confidence and apply in breeding programs.     

HOTAIR Induces the Downregulation of miR-200 Family Members in Gastric Cancer Cell Lines

Background:Gastric cancer is the fourth most common human malignancy and the second reason for cancer morbidity worldwide. LncRNA HOTAIR has recently emerged as a promoter of metastasis in various cancer types, including GC, through the EMT process. However, the exact mechanism of HOTAIR in promoting EMT is unknown. Aberrant expression of the miR-200 family has been linked to the occurrence and development of various types of malignant tumors. This study investigates the correlation between the HOTAIR and miR-200 family gene expression patterns in GC cell lines. We investigated the miR-200 and HOTAIR due to their common molecular features in the EMT process. Methods:AGS and MKN45 cell lines were transfected with si-HOTAIR, along with a negative control. The effect of HOTAIR knockdown was also analyzed on cell viability and also on the expression of miR-200 family members, including miR-200a, -200b, and -200c, in cell lines using qRT-PCR. Statistical analysis was performed to find the potential correlation between the expression level of HOTAIR and miRs. Results:Our results showed significant increased miR-200 family expression level in transfected AGS and MKN45 GC cells (fold changes > 2; p < 0.001). Moreover, a negative correlation was observed between HOTAIR and miR-200 expression levels in GC cell lines (p < 0.05). Conclusion:Our findings showed a significant association between miR-200 family and HOTAIR expression levels in GC cell lines. Taken together, the HOTAIR-miR-200 axis seems to play a vital role in human GC, suggesting a potential therapeutic target in future GC treatment. Key Words: Gene expression, Long noncoding RNA, HOTAIR, MicroRNAs     

The Effect of Coenzyme Q10 and α-Tocopherol in Skim Milk–Based Extender for Preservation of Caspian Stallion Semen in Cool Condition

The purpose of this study was to determine the effects of different concentrations of coenzyme Q₁₀ (CoQ₁₀) and α-tocopherol (T) along with their interaction effects on the quality of preserved stallion semen at 5°C for a period of 48 hours. Semen was collected and diluted with skim milk–based extender that was supplemented with different antioxidants: no antioxidant (negative control [NC]), 0.9% (vol/vol) dimethyl sulfoxide (positive control [PC]), α-tocopherol (5 [T5] or 10 [T10] mM), CoQ₁₀ (1 [C1] or 2 [C2] μM), 1 μM CoQ₁₀ + 5 mM α-tocopherol (C1T5), 1 μM CoQ₁₀ + 10 mM α-tocopherol (C1T10), 2 μM CoQ₁₀ + 5 mM α-tocopherol (C2T5), and 2 μM CoQ₁₀ + 10 mM α-tocopherol (C2T10), then kept at 5°C. The results showed that C1 extender resulted in higher total motility (62.44 ± 3.82) and plasma membrane integrity (65.16 ± 3.63%) compared with NC after 48 hours of storage (P < .05). Different concentrations of α-tocopherol had no significant effects on sperm quality, with the exception of plasma membrane integrity, compared with NC and PC extenders (P > .05). Also, C1T5 extender improved total and progressive motility, plasma membrane integrity and functionality, and decreased lipid peroxidation compared with NC and C2T10 extenders over 48 hours of storage at 5°C (P < .05). The C1T5 extender was similar to C1 and T5 extenders in all semen parameters evaluated during storage time. In conclusion, between previously mentioned extenders, C1T5 could improve stallion sperm quality during 48 hours of storage. In the present study, none of extenders had effect on sperm quality until 24-hour storage.     

Prediction of macronutrients in plant leaves using chemometric analysis and wavelength selection

Purpose

Fast and real-time prediction of leaf nutrient concentrations can facilitate decision-making for fertilisation regimes on farms and address issues raised with over-fertilisation. Cacao (Theobroma cacao L.) is an important cash crop and requires nutrient supply to maintain yield. This project aimed to use chemometric analysis and wavelength selection to improve the accuracy of foliar nutrient prediction.

Materials and methods

We used a visible-near infrared (400–1000 nm) hyperspectral imaging (HSI) system to predict foliar calcium (Ca), potassium (K), phosphorus (P) and nitrogen (N) of cacao trees. Images were captured from 95 leaf samples. Partial least square regression (PLSR) models were developed to predict leaf nutrient concentrations and wavelength selection was undertaken.

Results and discussion

Using all wavelengths, Ca (R²_CV?=?0.76, RMSE_CV?=?0.28), K (R²_CV?=?0.35, RMSE_CV?=?0.46), P (R²_CV?=?0.75, RMSE_CV?=?0.019) and N (R²_CV?=?0.73, RMSE_CV?=?0.17) were predicted. Wavelength selection increased the prediction accuracy of Ca (R²_CV?=?0.79, RMSE_CV?=?0.27) and N (R²_CV?=?0.74, RMSE_CV?=?0.16), while did not affect prediction accuracy of foliar K (R²_CV?=?0.35, RMSE_CV?=?0.46) and P (R²_CV?=?0.75, RMSE_CV?=?0.019).

Conclusions

Visible-near infrared HSI has a good potential to predict Ca, P and N concentrations in cacao leaf samples, but K concentrations could not be predicted reliably. Wavelength selection increased the prediction accuracy of foliar Ca and N leading to a reduced number of wavelengths involved in developed models.

     

Effect of different safflower (Carthamus tinctorius L.)–bean (Phaseolus vulgaris L.) intercropping patterns on growth and yield under weedy and weed-free conditions

Two field experiments were carried out over two consecutive years (2010–2011) in the research field of the College of Agriculture, Shiraz University, Fars Province, southern Iran. The study was a factorial experiment based on a randomized complete block design with three replications: the first factor was the ratio of safflower (Carthamus tinctorius L. Pi cv.) to bean (Phaseolus vulgaris L. Saiad cv.) at five levels (safflower and bean sole cropping, and intercropping of safflower and bean at ratios of 1:3, 2:2 and 3:1); and the second factor was weed management at two levels: weed-free (complete weed control) and weedy (no weed control). The results showed that an intercropping system was the most appropriate method for decreasing the adverse effect of weeds on the performance of both crops. Intercropping was more suitable for weedy than weed-free conditions. According to the land equivalent ratio (LER) value, if the main crop was bean, the best intercropping treatment was one row of safflower and six rows of bean (S₁B₃) under both weedy and weed-free conditions. By contrast, if the main crop was safflower, the best treatment under weedy conditions was S₁B₃, whereas under weed-free conditions the best treatment was two rows of safflower and four rows of bean (S₂B₂). Overall, S₁B₃ can be introduced as the best intercropping method.     

Nursery performance of Pacific white shrimp (Litopenaeus vannamei Boone, 1931) cultivated in a biofloc system: the effect of adding different carbon sources

Effect of different carbon sources on nursery performance of Pacific white shrimp (Litopenaeus vannamei) cultivated in biofloc system was investigated. Shrimp postlarvae (98.47 ± 8.6 mg) were fed for 32 days in tanks with water volume of 130 L and density of 1 individual L^?1. One control treatment and four biofloc treatments (BFT1, BFT2, BFT3 and BFT4) with adding different carbon sources including molasses, starch, wheat flour and mixture of them, respectively, were considered at equal weight ratios. According to the results, salinity, dissolved oxygen and pH were not significantly different among the biofloc treatments (P > 0.05). Maximum pH (8.27) and maximum dissolved oxygen (6.35 mg L^?1) were recorded in the control. Maximum (0.43 mg L^?1) and minimum (0.09 mg L^?1) ammonia were recorded in the control and BFT2, respectively (P < 0.05). Using simple carbohydrates (molasses and starch) lowered the ammonia concentration significantly. The highest increase in body weight (1640.43 ± 231.28 mg), growth rate, specific growth rate (8.97 ± 0.42% per day) and biomass (190.29 ± 26.83 mg) were found in BFT1 and the highest survival (90 ± 0.77%) was found in BFT4. The highest feed conversion (1.52 ± 0.23) and the lowest feed efficiency (66.81 ± 7.95) were observed in the control (P < 0.05). The proximate composition analysis revealed an increase in lipid and ash in biofloc treatments. Results indicated that using biofloc technology with zero‐water exchange system and adding carbon sources could help to recycle waste and improve the water quality. Moreover, the type of carbonaceous organic matter as a substrate for heterotrophic bacteria would be effective in degradation and metabolization of ammonia and nitrite.     

Characterization of Clostridium perfringens in the feces of adult horses and foals with acute enterocolitis

Up to 60% of cases of equine colitis have no known cause. To improve understanding of the causes of acute colitis in horses, we hypothesized that Clostridium perfringens producing enterotoxin (CPE) and/or beta2 toxin (CPB2) are common and important causes of severe colitis in horses and/or that C. perfringens producing an as-yet-undescribed cytotoxin may also cause colitis in horses. Fecal samples from 55 horses (43 adults, 12 foals) with clinical evidence of colitis were evaluated by culture for the presence of Clostridium difficile, C. perfringens, and Salmonella. Feces were also examined by enzyme-linked immunosorbent assay (ELISA) for C. difficile A/B toxins and C. perfringens alpha toxin (CPA), beta2 toxin (CPB2), and enterotoxin (CPE). Five C. perfringens isolates per sample were genotyped for the following genes: cpa, cpb, cpb2 consensus, cpb2 atypical, cpe (enterotoxin), etx (epsilon toxin), itx (iota toxin), netB (necrotic enteritis toxin B), and tpeL (large C. perfringens cytotoxin). The supernatants of these isolates were also evaluated for toxicity for an equine cell line. All fecal samples were negative for Salmonella. Clostridium perfringens and C. difficile were isolated from 40% and 5.4% of samples, respectively. All fecal samples were negative for CPE. Clostridium perfringens CPA and CPB2 toxins were detected in 14.5% and 7.2% of fecal samples, respectively, all of which were culture-positive for C. perfringens. No isolates were cpe, etx, netB, or tpeL gene-positive. Atypical cpb2 and consensus cpb2 genes were identified in 15 (13.6%) and 4 (3.6%) of 110 isolates, respectively. All equine C. perfringens isolates showed far milder cytotoxicity effects than a CPB-producing positive control, although cpb2-positive isolates were slightly but significantly more cytotoxic than negative isolates. Based on this studied population, we were unable to confirm our hypothesis that CPE and CPB2-producing C. perfringens are common in horses with colitis in Ontario and we failed to identify cytotoxic activity in vitro in the type A isolates recovered.     

Molecular and histopathological characterization of Photobacterium damselae in naturally and experimentally infected Nile tilapia (Oreochromis niloticus)

Mass mortality has occurred among cultured Nile tilapia, Oreochromis niloticus, on fish farms in Manzala, Dakahlia province, Egypt, in the summer season, 2019. Moribund fish were reported with deep ulcers, septicaemic lesions and sampled for bacterial isolation. In this study, most isolates were subjected to bacteriological examination, antibiotic sensitivity test, 16S rRNA gene sequencing and histopathological examination. Following isolate identification, intraperitoneal challenge of Nile tilapia with a bacterial suspension 2 × 10⁶ CFU/ml was performed. Samples from liver, spleen and kidney were collected for histological and biochemical analysis. The results showed a high similarity (99%) to Photobacterium damselae strains using phylogenetic analysis of 16S rRNA. P. damselae exhibited resistance to amoxicillin and erythromycin, as well it was highly sensitive to chloramphenicol and doxycycline. Moreover, haemorrhage, oedema, hemosiderosis and melanomacrophage activation in the liver and head kidney of infected fish were detected by light and electron microscopy. Also, significant higher levels of CAT and SOD in the spleen and head kidney, as well as the serum levels of NO were observed in experimentally challenged O. niloticus, compared to the control fish. Our data identified P. damselae for the first time from infected Nile tilapia, describing its sensitivity to a variety of antibiotics, histopathological alterations and oxidative stress impact, and it could be useful indicators for understanding P. damselae pathogenesis, which might provide a preventive efficacy for P. damselae.     

Estimation of genetic parameters for wool fiber diameter measures.

Genetic and phenotypic correlations and heritability estimates of side, britch, and core diameters; side and britch CV; side and britch diameter difference; and clean fleece weight were investigated using 385 western white-faced ewes produced by 50 sires and maintained at two locations on a selection study. Data were analyzed using analysis of variance procedures, and effects in the final model included breed of sire-selection line combination, sire within breed-selection line, and location. Heritabilities were estimated by paternal half-sib analysis. Sires within breed-selection line represented a significant source of variation for all traits studied. Location had a significant effect on side diameter, side and britch diameter difference, and clean fleece weight. Age of ewe only affected clean fleece weight. Phenotypic and genetic correlations among side, britch, and core diameter measures were high and positive. Phenotypic correlations ranged from .68 to .75 and genetic correlations ranged from .74 to .89. The genetic correlations between side and britch diameter difference and side diameter or core diameter were small (-.16 and .28, respectively). However, there was a stronger genetic correlation between side and britch diameter difference and britch diameter (.55). Heritability of the difference between side and britch diameter was high (.46 +/- .16) and similar to heritability estimates reported for other wool traits. Results of this study indicate that relatively rapid genetic progress through selection for fiber diameter should be possible. In addition, increased uniformity in fiber diameter should be possible through selection for either side and britch diameter difference or side or britch CV.     

Leaf macro- and micro-morphological altitudinal variability of Carpinus betulus in the Hyrcanian forest (Iran)

We investigated the altitudinal variation of Carpinus betulus L. in the Hyrcanian forest using leaf macro-morphological and micro-morphological traits. We collected a total of 1600 leaves from two locations. In each location, we sampled six populations along an altitudinal gradient ranging from 100 m to 1,150 m. We found that trees in the higher elevations have smaller leaf lamina than those in the lower elevations. In contrast, leaf mass per area was high at low altitudes and increased newly at the higher ones. Stomatal dimension was negatively correlated with elevation, while stomatal density was positively correlated with elevation. We also found that two transects showed the same plasticity trend. Leaf area showed the highest plasticity, while the number of veins showed the lowest plasticity. This study shows that altitude, and related temperature and rainfall, represents an important driving force in Carpinus betulus leaf morphological variation. Moreover, our results suggest that leaf area, leaf mass per area and stomatal density could influence the species responses to different ecological conditions.