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.

     

Dynamics of porcine circovirus type 2 infection in a herd of pigs with postweaning multisystemic wasting syndrome

OBJECTIVE: To determine the pattern of infection for porcine circovirus type 2 (PCV2) in a herd of pigs with postweaning multisystemic wasting syndrome (PMWS). ANIMALS: 29 sows and 250 pigs. PROCEDURE: Blood samples were collected from all 3-, 7-, and 12-week old pigs and 59 pigs at 28 weeks of age. Pigs that died during the study were necropsied. Porcine parvovirus and PCV2 antibodies were assayed. A polymerase chain reaction (PCR) was used to detect PCV2 genome in serum of selected pigs. RESULTS: The PMWS started when pigs were 8 weeks old, with a prevalence of 30% in 8- to 10-week-old pigs. Eighty-three pigs died during the period between 3 and 12 weeks of age. Microscopic lesions consistent with PMWS were observed, and PCV2 nucleic acid was detected (50 of 68 pigs). Antibodies to PCV2 decreased from 3 to 7 weeks of age, increased at 12 weeks of age, and were maintained until 28 weeks of age. One sow had a positive result for PCR of serum. Nine, 37 and 8 pigs had PCV2 genome in serum obtained at 7, 12, and 28 weeks of age, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Infection with PCV2 coincided with severe clinical signs; however, infected 28-week-old pigs did not have evidence of disease. Immunity declined over time in young pigs. A long duration of PCV2 viremia was apparent in a high percentage of infected pigs, which may affect transmission and persistence of the virus in a herd.     

Histomorphological changes between short and long days in ovary guanacos (Lama guanicoe)

This is the first morpho-histological comparison of guanaco ovaries between reproductive (long-days) and non-reproductive (short-days) seasons, and oestrogen receptor-alpha (ERα) and beta (ERβ) detection. Different stages of follicle development were found in the cortical area, but no corpus luteum was detected. The size and frequency of antral follicles and large atretic follicles were higher in long-day ovaries than short-days, consistent with ovarian activity in this season. Differential expression of ERα and ERβ was observed in follicles at different stages of development between short and long days. These data reveal histological and molecular differences between reproductive and non-reproductive seasons of guanaco ovaries.     

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.     

Influence of amount and frequency of protein supplementation to ruminants consuming low-quality cool-season forages: efficiency of nitrogen utilization in lambs and performance of gestating beef cows

We evaluated the influence of amount and crude protein (CP) supplementation frequency (SF) on nitrogen (N) use by wethers and the performance of late-gestation beef cows. In exp. 1, seven Western whiteface wethers (31.8 ± 1.4 kg) were used in an incomplete 7 × 4 Latin square to evaluate intake and N use. Wethers received one of the seven treatments in a 2 × 3 factorial design containing two levels of supplemental soybean meal offered at a rate of 100% (F) or 50% (H; 50% of F) of the estimated CP requirement daily, once every 5, or once every 10 d, plus a non-supplemented control (CON). Low-quality cool-season forage (4.9 % CP; dry matter [DM] basis) was provided daily for ad libitum intake. Experimental periods lasted 30 d. In exp. 2, 84 Angus × Hereford cows (560 ± 35 kg) were stratified by age, body condition score (BCS), and expected calving date and allocated to 1 of the 21 feedlot pens (three pens per treatment). Pens were randomly assigned to receive the same treatments as in exp. 1 and cows had free access to low-quality cool-season forage (2.9% CP; DM basis). Cow body weight (BW) and BCS were measured every 14 d until calving and within 24 h after calving. In exp. 1, supplementation did not alter total DM and organic matter (OM) intake (P ≥ 0.26), but both parameters linearly decreased as SF decreased (P = 0.02). Supplementation increased DM, OM, and neutral detergent fiber (NDF) digestibility (P ≤ 0.02). Additionally, F feeding linearly increased DM, OM, and NDF digestibility as SF decreased (P ≤ 0.04). Digestibility of N, N balance, and digested N retained were greater with supplementation (P < 0.01), and N digestibility linearly increased as SF decreased (P = 0.01). Mean plasma urea-N concentration was not only greater (P < 0.01) for supplemented vs. CON wethers but also greater (P = 0.03) for F vs. H. In exp. 2, pre-calving BCS change was greater (P = 0.03) for supplemented cows. A linear effect of SF × supplementation rate for pre-calving BCS change was noted (P = 0.05), as F-supplemented cows lost more BCS compared with H as SF decreased. When considering supplementation intervals greater than 5 d, reducing the quantity of supplement provided, compared with daily supplementation, may be a feasible management strategy to maintain acceptable nutrient use and animal performance while reducing supplement and labor costs.     

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.