Suppression subtractive hybridization and microarray analysis reveal differentially expressed genes in the <Emphasis Type="Italic">Lr39/41</Emphasis>-mediated wheat resistance to <Emphasis Type="Italic">Puccinia triticina</Emphasis>

Wheat leaf rust caused by Puccinia triticina (Pt) is one of the most severe fungal diseases threatening the global wheat production. The use of leaf rust resistance (Lr) genes in wheat breeding programs is the major solution to solve this issue. Wheat isogenic line carrying the Lr39/41 gene has shown a moderate to high resistance to most of the Pt pathotypes detected in China. In the present study, a typical hypersensitive response (HR) was observed using microscopy in leaves of the Lr39/41 isogenic line inoculated with the avirulent Pt pathotype THTT from 48 h-post inoculation. Two Lr39/41 resistance-associated suppression subtractive hybridization (SSH) libraries with a total of 6000 clones were established. Microarray hybridizations were performed on all obtained SSH clones using RNAs extracted from leaves of the Pt-inoculated and non-inoculated Lr39/41 isogenic lines, and leaves of the Pt-inoculated and non-inoculated Thatcher susceptible lines. Differentially expressed clones were analyzed by significance analysis of microarrays (SAM), followed by further sequencing. A total of 36 Lr39/41-resistance-related differentially expressed genes (DEGs) were identified, many of which had been previously reported to be involved in the plant defense response. The expression levels of eight selected DEGs during different stages of the Lr39/41-mediated resistance were further quantified by a qRT-PCR assay. Several pathogenesis-related (PR) and HR-related genes seem to be crucial for the Lr39/41-mediated resistance. In general, a brief profile of DEGs associated with the Lr39/41-mediated wheat resistance to Pt was drafted.     

Alien species of aphids (Hemiptera: Aphididae) found in Serbia,new to the Balkan Peninsula

New alien species of aphids (Illinoia liriodendri (Monell), Wahlgreniella nervata (Gillete), Takecallis arundicolens (Clarke) and Tinocallis takachihoensis Higuchi) have been found for the first time in Serbia. These records are new for the Balkan Peninsula. Data of Ericaphis scammelli (Mason), species recently mentioned in Serbia, are also presented in the text. The species have been found during the last five years at four sites (Belgrade, Zemun, Mladenovac and Bojnik) on five cultivated plants. Ericaphis scammelli is a pest of blueberry and a vector of Blueberry scorch virus. The other four develop colonies on decorative plants: I. liriodendri has been found on Liriodendron tulipifera, W. nervata on Rosa sp., T. arundicolens on Pseudosasa japonica and T. takachihoensis on Ulmus x hollandica. The origin of three of the species (E. scammelli, I. liriodendri and W. nervata) is North America and two (T. arundicolens and T. takachihoensis) have come from East Asia. Their morphology, biology and the damage they inflict are discussed here, as well as their potential economic and environmental threat. Only parthenogenetic viviparous females of all 5 species have been collected. Their biology has not been studied enough in Europe and needs further examination. Original drawings of all species are presented.     

Using operating area size and adjacency constraints to mitigate the effects of harvesting activities on boreal caribou habitat

Context

Sustained timber harvesting conflicts with the long-term viability of boreal caribou (Rangifer tarandus caribou) populations. The spatial arrangement of harvest blocks in the landscape could mitigate the impact of logging on caribou populations. For the forest industry, however, these measures represent constraints that reduce the annual allowable cut (AAC).

Objective

Estimate the long-term impacts of spatial constraints to harvesting, applied alone or in combination, on AAC and boreal caribou populations.

Methods

We divided a 30,000 km² region into 20 harvest block sizes varying from 50 to 1000 km², and modeled the implementation of spatially explicit harvest schedule plans in combination with wildfire and caribou population dynamics. We then evaluated the probability of persistence of boreal caribou populations.

Results

The probability of maintaining an AAC target declined with increasing target AAC, increasing size of operating area, and increasing adjacency constraints. In contrast, the probability of maintaining caribou populations declined with increasing AAC, decreasing size of operating areas, and decreasing adjacency constraints. An increase in operating area size from 50 to 300 km² produced a considerable gain in AAC for all adjacency constraints.

Conclusions

Because adjacency constraints led only to a small increase in the probability of maintaining caribou populations, we recommend adopting less constraining landscape management actions, such as a 70-year period between two consecutive harvests in the same ~300-km² operating area.

     

Host status and reaction of <Emphasis Type="Italic">Medicago truncatula</Emphasis> accessions to infection by three major pathogens of pea (<Emphasis Type="Italic">Pisum sativum</Emphasis>) and alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis>)

Ditylenchus dipsaci, the stem nematode of alfalfa (Medicago sativa), Mycosphaerella pinodes, cause of Ascochyta blight in pea (Pisum sativum) and Aphanomyces euteiches, cause of pea root rot, result in major yield losses in French alfalfa and pea crops. These diseases are difficult to control and the partial resistances currently available are not effective enough. Medicago truncatula, the barrel medic, is the legume model for genetic studies, which should lead to the identification and characterization of new resistance genes for pathogens. We evaluated a collection of 34 accessions of M. truncatula and nine accessions from three other species (two from M. italica, six from M. littoralis and one from M. polymorpha) for resistance to these three major diseases. We developed screening tests, including standard host references, for each pathogen. Most of the accessions tested were resistant to D. dipsaci, with only three accessions classified as susceptible. A very high level of resistance to M. pinodes was observed among the accessions, none of which was susceptible to this pathogen. Conversely, a high level of variation, from resistant to susceptible accessions, was identified in response to infection by A. euteiches.     

Influence of <Emphasis Type="Italic">Nigella sativa</Emphasis> seeds, <Emphasis Type="Italic">Rosmarinus officinalis</Emphasis> leaves and their combination on growth performance,immune response and rumen metabolism in Dorper lambs

The objective of this study was to determine the effects of dietary supplementation of Nigella sativa L. seeds, Rosmarinus officinalis L. leaves and their combination on rumen metabolism, nutrient intake and digestibility, growth performance, immune response and blood metabolites in Dorper lambs. Twenty-four entire male Dorper lambs (18.68?±?0.6 kg, 4–5 months old) were randomly assigned to a concentrate mixture containing on a dry matter basis either, no supplement (control, T1), 1% R. officinalis leaves (T2), 1% N. sativa seeds (T3) or 1% R. officinalis leaves +1% N. sativa seeds (T4). The lambs had ad libitum access to urea-treated rice straw (UTRS) and were raised for 90 days. Supplemented lambs had greater (P?< 0.05) intake of DM and UTRS than the control lambs. The T4 lambs had lower (P?< 0.05) nutrient digestibility than those fed other treatments. Total and daily weight gain was greater (P?< 0.05) in T2 lambs than those fed other diets. The T3 and T4 lambs had greater (P?< 0.05) ruminal pH than the T1 and T2 lambs. Supplemented lambs had lower (P?<?0.05) ruminal total volatile fatty acids, acetate, propionate, NH₃-N and C18:0 than the control lambs. The T4 lambs had lower (P?< 0.05) population of Fibrobacter succinogenes, Ruminococcus albus, methanogens and total protozoa compared with those fed other diets. Supplemented lambs had lower (P?< 0.05) neutrophils, basophils and serum urea and greater (P?<?0.05) serum IgA and IgG compared with the control lambs. The current results emphasised the variation in the efficacy of medicinal plants in ruminant nutrition.     

Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Studies on production performance and toxin residues in tissues and eggs of layer chickens fed on diets with various concentrations of aflatoxin AFB1

1. An experiment on 1-week-old, White Leghorn female chicks was conducted to study the effect of aflatoxin AFB1 on weight gain, feed intake, feed gain ratio, age at sexual maturity, production and quality of eggs, retention of nutrients, pathoanatomical and histopathological parameters, and also on AFB1 residues in eggs and muscles of hens. The chicks were assigned to 4 dietary treatments: D1 (without AFB1), D2 (2.50 mg/kg AFB1), D3 (3.13 mg/kg AFB1), D4 (3.91 mg/kg AFB1) up to the age of 40 weeks. 2. At the end of the experiment, the mean body weight gain and feed intake were significantly lower in all aflatoxin-fed groups compared to control. The feed gain ratios were noted as 13.41, 13.59, 13.82 and 14.71, with the group fed the highest concentration of AFB1 showing a significantly poorer ratio than other groups. 3. Age at sexual maturity was also affected adversely by dietary AFB1: 193 d for D4 as compared to as early as 148 d for D1. Hen-d egg production was recorded as 96.92, 74.67, 65.98 and 50.75 in D1, D2, D3 and D4, respectively. 4. Average egg weights at the end of the experiment were 57.77, 57.49, 57.54 and 54.66 for D1, D2, D3 and D4, respectively. Shape index was significantly lower in D4 as compared to control. Contrary to this, albumen index was significantly higher in D4 as compared to D1. The values of yolk indices and eggshell thickness did not differ significantly among treatment groups. However, colour of yolk was reduced in all aflatoxin-fed groups compared to control. 5. Retentions of dry matter, crude protein, ether extract, calcium and metabolisable energy were adversely affected at various levels of AFB1 compared to control. 6. Pathoanatomical and histopathological studies showed various adverse changes in liver, kidney, heart, ovaries and bursa of Fabricius in AFB1-fed groups. 7. Different amounts of aflatoxin residues were detected in eggs and breast muscles of hen in all AFB1-fed groups.     

Comprehensive transcriptional profiling and functional study of lymphangiogenic growth factor in placentome of early-stage pregnant water buffalo

The aim of the present study was to evaluate the expression and localization of lymphangiogenic factors (VEGF-C and VEGF-D), their receptor (VEGFR3) and lymphatic endothelial marker (LYVE1) in buffalo placenta during early pregnancy [EP], and to investigate the functional role of lymphangiogenic growth factors in placental lymphangiogenesis. The mRNA and protein expression of VEGF-C, VEGF-D, their receptor VEGFR3 and LYVE1 showed significant expression in EP1 (29–42 days) and EP2 stages (51–82 days) both in caruncle (maternal part) and cotyledon (foetal part) of the buffalo placenta. Immunoreactivity of VEGF-C, VEGF-D and LYVE1 was observed around the endometrial gland, in lymphatics and trophoblast cells, whereas VEGFR3 mainly localized in lymphatics of the caruncle and cotyledons. Cultured trophoblast cells were treated with VEGF-C/VEGF-D (50, 100 and 150 ng/ml) and combined doses of VEGF-C and VEGF-D (150 ng/ml) each for different time durations (24, 48 and 72 h). The mRNA expression of LYVE1 and PCNA was significantly (p < .001) upregulated with VEGF-C and VEGF-D and combined treatment (@150 ng/ml), as well as significantly downregulating Caspase-3 at 48 and 72 h. Thus, the present study provides evidence that lymphangiogenic factors are expressed in buffalo placental compartments and they may play a significant role in the regulation of placental function in water buffaloes.     

Zirconium-Modified Activated Sludge as a Low-Cost Adsorbent for Phosphate Removal in Aqueous Solution

A highly effective zirconium-modified activated sludge (Zr(IV)-AS) adsorbent was prepared from activated sludge and applied to remove phosphate from aqueous solutions by batch and column experiments. Characterized results revealed that zirconium was successfully loaded onto the activated sludge (AS), and the specific surface area and pore volume were substantially improved after zirconium loading on the AS. Zr(IV)-AS exhibited a high adsorption affinity for phosphate and the maximum adsorption amount was 27.55 mg P·g^?1 at 25 °C. Adsorption isotherms of phosphate could be described by the Langmuir model, and the adsorption kinetics were well described by the pseudo-second-order model. Phosphate adsorption on Zr(IV)-AS increased monotonically with decreasing solution pH. The presence of SO₄^2? in water resulted in slightly decreased phosphate adsorption on the adsorbent even at a high concentration (25 mmol/L), and a greater influence of HCO₃^? on adsorption could be ascribed to the increased solution pH with the addition of the HCO₃^?. Column adsorption experimental results showed that the adsorbent has excellent phosphate adsorption properties and that the effluent can meet the requirement of phosphorus in the national wastewater discharge standard of China. Phosphate-saturated Zr(IV)-AS can be effectively desorbed in 0.1 mol L^?1 NaOH solution, and the regenerated adsorbent still possessed the high capacity. The adsorption between the adsorbent and the phosphate is due to the electrostatic interaction and anionic exchange at the surface of the Zr(IV)-AS. Furthermore, this approach provides a possibility of treating wastewater with waste and has the potential for industrial applications for the removal of phosphate from wastewater.