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.     

Landscape genetics of the foundational salt marsh plant species black needlerush (<Emphasis Type="Italic">Juncus roemerianus</Emphasis> Scheele) across the northeastern Gulf of Mexico

Context

Common species important for ecosystem restoration stand to lose as much genetic diversity from anthropogenic habitat fragmentation and climate change as rare species, but are rarely studied. Salt marshes, valuable ecosystems in widespread decline due to human development, are dominated by the foundational plant species black needlerush (Juncus roemerianus Scheele) in the northeastern Gulf of Mexico.

Objectives

We assessed genetic patterns in J. roemerianus by measuring genetic and genotypic diversity, and characterizing population structure. We examined population connectivity by delineating possible dispersal corridors, and identified landscape factors influencing population connectivity.

Methods

A panel of 19 microsatellite markers was used to genotype 576 samples from ten sites across the northeastern Gulf of Mexico from the Grand Bay National Estuarine Research Reserve (NERR) to the Apalachicola NERR. Genetic distances (F_ST and D_ch) were used in a least cost transect analysis (LCTA) within a hierarchical model selection framework.

Results

Genetic and genotypic diversity results were higher than expected based on life history literature, and samples structured into two large, admixed genetic clusters across the study area, indicating sexual reproduction may not be as rare as predicted in this clonal macrophyte. Digitized coastal transects buffered by 500 m may represent possible dispersal corridors, and developed land may significantly impede population connectivity in J. roemerianus.

Conclusions

Results have important implications for coastal restoration and management that seek to preserve adaptive potential by sustaining natural levels of genetic diversity and conserving population connectivity. Our methodology could be applied to other common, widespread and understudied species.

     

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.     

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.

     

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.     

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.     

Seasonal Changes of Shoot Carbohydrates and Growth Characteristics of ‘Trakya İlkeren’ Grape Variety (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.)

This research was conducted to determine the changes in the seasonal carbohydrates of annual shoots and growth characteristics of 10 years old ‘Trakya ?lkeren’ grape variety. Grapevines are grafted on 5BB and 5C rootstocks and they were grown in heavy clay soil conditions. In the study, changes of sugar, starch and total carbohydrates were determined from bud burst to dormancy. While sugar, starch and total carbohydrates showed significant differences (P < 0.01) there were no differences between rootstocks. Although sugar, starch and total carbohydrates of shoots decreased from bud burst to blooming, they increased thereafter until vegetative growth stopped. Total carbohydrates of annual shoots have been found to accumulate in period from blooming to harvest. Total and mean leaf area, shoot length and diameter, total chlorophyll and chlorophyll a/b ratio showed significantly variation among rootstocks and phenological stages (P < 0.01). Whereas the highest chlorophyll contents were found in the blooming, it was found at the lowest through to harvest. Overall mean and total leaf area, shoot length and shoot diameter, total chlorophyll content and chlorophyll a/b ratio were significantly higher on 5BB grafted vines. In the study, 5BB rootstocks are found suitable in terms of shoot carbohydrate accumulation and growth characteristics for ‘Trakya ?lkeren’ grape variety.