Influence of the stage for anther excision and heterostyly in embryogenesis induction from eggplant anther cultures

In this work we address two aspects of eggplant flower biology potentially involved on the efficiency of anther culture: the selection of the best floral stage to extract anthers for culture, and the effect of heterostyly in the identification of suitable buds and anthers. For 12 different accessions, we determined morphological criteria (length ranges) to identify buds and anthers enriched in vacuolate microspores and young bicellular pollen, the stages most responsive to embryogenesis induction. While these microspore/pollen stages were the most responsive when isolated and cultured in liquid medium, we observed that culture of anthers containing these stages is not the best choice. Instead, the highest response was found for younger anthers, containing mostly young and mid microspores. We analyzed eggplant anther walls and found that their particular thickness may be behind this apparent discrepancy, since they may delay the diffusion of inducing factors to the anther locule, reducing their effect over inducible microspores. Thus, the culture of younger anthers would allow for younger microspores to grow up to the inducible stages while factors are entering the locule. We also analyzed the embryogenic response of short and long-styled buds present in Cristal, a heterostylic cultivar. Our results demonstrated that each floral morph produced buds and anthers of different lengths, but equally useful for anther culture, since similar amounts of embryos were produced. The practical application of these results may improve the efficiency of anther culture not only in these cultivars, but in others also presenting thick anther walls and heterostyly.     

Endometrial Expression of IFNAR‐1 and Oxytocin Receptor (OTR) is not Improved by Prostaglandin Analogues when Compared to Progestagens in Ewes

The objective of this study was to investigate differences on the endometrial immunoexpression of type I IFN receptor subunit 1 (IFNAR1) and oxytocin receptor (OTR) during the time of maternal recognition of pregnancy in sheep, when oestrus is synchronized with either prostaglandin analogues (group PG) or conventional progestagens (group P). Plasma progesterone was measured from day 0 to 21 post‐coitus (pc) (day 0 = day of oestrus). Immunohistochemistry was performed in samples of uterine horns from pregnant sheep on days 9pc, 13pc, 15pc, 17pc and 21pc to locate IFNAR1 and OTR expression in different endometrial compartments. Mean levels of plasma progesterone were different between treatments, obtaining higher levels in the PG group than in the P group (p < 0.05). Comparing days of pregnancy, IFNAR1 protein expression was different in the luminal epithelium (LE) (p < 0.05), while OTR was different in the LE and in the superficial glandular epithelium (SG) (p < 0.05). Temporal variation on the expression of both proteins from day 9pc to 21pc has been evidenced. IFNAR1 and OTR expression did not show significant differences between treatments. However, the response observed in the endometrium was highly inconsistent when prostaglandin analogues were used. Therefore, the protocol based on prostaglandin analogues still needs to be optimized before being considered as a better alternative to progestagens for oestrous synchronization in sheep.     

Increase in size and nitrogen concentration enhances seedling survival in Mediterranean plantations. Insights from an ecophysiological conceptual model of plant survival

Reduction in size and tissue nutrient concentration is widely considered to increase seedling drought resistance in dry and oligotrophic plantation sites. However, much evidence indicates that increase in size and tissue nutrient concentration improves seedling survival in Mediterranean forest plantations. This suggests that the ecophysiological processes and functional attributes relevant for early seedling survival in Mediterranean climate must be reconsidered. We propose a ecophysiological conceptual model for seedling survival in Mediterranean-climate plantations to provide a physiological explanation of the frequent positive relationship between outplanting performance and seedling size and nutrient concentration. The model considers the physiological processes outlined in the plantation establishment model of Burdett (Can J For Res 20:415–427, 1990), but incorporates other physiological processes that drive seedling survival, such as N remobilization, carbohydrate storage and plant hydraulics. The model considers that seedling survival in Mediterranean climates is linked to high growth capacity during the wet season. The model is for container plants and is based on three main principles, (1) Mediterranean climates are not dry the entire year but usually have two seasons of contrasting water availability; (2) summer drought is the main cause of seedling mortality; in this context, deep and large roots is a key trait for avoiding lethal water stress; (3) attainment of large root systems in the dry season is promoted when seedlings have high growth during the wet season. High growth is achieved when seedlings can divert large amount of resources to support new root and shoot growth. Functional traits that confer high photosynthesis, nutrient remobilization capacity, and non-structural carbohydrate storage promote high growth. Increases in seedling size and nutrient concentration strongly affect these physiological processes. Traits that confer high drought resistance are of low value during the wet season because hinder growth capacity. We provide specific evidence to support the model and finally we discuss its implications and the factors that may alter the frequent increase in performance with increase in seedling size and tissue nutrient concentration.     

Biocontrol traits of plant growth suppressive arbuscular mycorrhizal fungi against root rot in tomato caused by <Emphasis Type="Italic">Pythium aphanidermatum</Emphasis>

Arbuscular mycorrhizal (AM) fungi known to cause plant growth depressions in tomato were examined for their biocontrol effects against root rot caused by Pythium aphanidermatum. The main hypothesis was that plant growth suppressive AM fungi would elicit a defence response in the host plant reducing Pythium root rot development. To test this hypothesis a fully factorial experiment was performed with AM fungi (Glomus intraradices, G. mosseae, G. claroideum or nonmycorrhizal), Pythium (± P. aphanidermatum) and harvest (7 and 14 days after pathogen inoculation (dapi)) as the main factors. Two weeks after AM fungi inoculation, roots were challenged with P. aphanidermatum. Variables evaluated at each harvest were root colonization levels of the interacting fungi, plant growth responses, and expression of a plant pathogenesis related protein gene (PR-1). All of the tested AM fungi caused marked growth suppressions, but did not affect PR-1 gene expression or the phosphorous concentration in the host plant. Plants singly inoculated with P. aphanidermatum had an increased PR-1 expression and phosphorous concentration. Among the AM fungi included in the study only G. intraradices reduced the pathogen root infection level, measured both in terms of Pythium ELISA and by recovery on selective media and only at the first harvest. Likewise, P. aphanidermatum root infection reduced colonization levels of G. intraradices, but not that of the two other AM fungi. In conclusion, plant growth suppressive AM fungi may offer plant beneficial traits in terms of biocontrol of root cortical pathogens.     

Life history parameters and scale-cover surface area of Aonidiella aurantii are altered in a mating disruption environment: implications for biological control

BACKGROUND: In recent years, environmentally safe measures to control the California red scale (CRS), Aonidiella aurantii (Maskell), have been successfully implemented. These measures include mating disruption (MD) and biological control. The goal of this study was to examine the effect of high concentrations of the CRS sex pheromone on its life history parameters and scale‐cover surface area under controlled laboratory conditions. RESULTS: The developmental time of both males and females of CRS increased with exposure to airborne pheromone. MD had an effect on both the total number of progeny and on the crawler production period for females. Accordingly, demographic parameters such as net fecundity (R₀) and intrinsic rate of increase (r_m) were significantly lower in the pheromone‐treated populations. The largest scale‐cover surface areas were observed in the CRS reared in the pheromone environment. CONCLUSION: A clear influence of airborne pheromone on the biology of CRS has been demonstrated. In addition to the classical mating disruption benefits of this technique, additional benefits, such as increase in the duration of exposure to natural enemies and increase in size, which benefits some species of parasitoids, have been confirmed. Copyright © 2012 Society of Chemical Industry     

Virulence and Molecular Diversity within Colletotrichum lindemuthianum Isolates from Andean and Mesoamerican bean Varieties and Regions

Virulence on a standard set of 12 common bean differential varieties, DNA sequence of repetitive-elements (Rep-PCR) and random amplified microsatellites (RAMS) were used to assess the genetic variability of 200 Colletotrichum lindemuthianum isolates collected from Andean and Mesoamerican bean varieties and regions. High levels of pathotypic (90 pathotypes) and genetic diversity (0.97) were identified among 200 isolates, revealing that C. lindemuthianum is a highly diverse pathogen. Although a significant number of pathotypes were common to Andean and Mesoamerican regions, many more were only found in the Mesoamerican region. Cluster analysis of virulence and molecular data did not separate isolates into groups that were structured with common bean gene pools. No genetic differentiation (G _ST=0.03) was apparent between Andean and Mesoamerican isolates of C. lindemuthianum. The diversity exhibited by C. lindemuthianum does not appear to cluster according to common bean gene pools, and the high diversity found in the Mesoamerican region seems to indicate that C. lindemuthianum originated and was disseminated from this region. Due to the high genetic variation exhibited by C. lindemuthianum, stacking major resistance genes appears to be the best option for developing cultivars with durable anthracnose resistance.     

The use of cytochemistry,immunophenotyping, flow cytometry,and in vitro differentiation to determine the ontogeny of a canine monoblastic leukemia

We evaluated the utility of cytochemistry, immunophenotyping, flow cytometry, and in vitro culture with forced differentiation of leukemic cells as diagnostic aids to identify the malignant cell ontogeny in a dog with leukemia. A tentative diagnosis of monoblastic leukemia was established by microscopic examination of Romanowsky-stained blood smears and bone marrow aspirate smears. This diagnosis also was supported by the light scatter signature that identified the blast cells as large, non-granular monocytic cells using a CellDyn 3500 automated hematology analyzer; as well as by the detection of N-butyrate esterase and the lack of choloroacetate esterase or leukocyte peroxidase by cytochemical staining. Subsequently, leukemic cells were isolated from the dog's peripheral blood and placed into tissue culture or cryopreserved. The leukemic cells grew in suspension cultures and proliferated spontaneously for up to 4 days. By day 7, proliferation was negligible. Upon culture with conditioned supernatant using mitogen-stimulated human T cells as a source of cytokines, an increased proportion of cells entered S phase by day 2 of culture; however, proliferation declined markedly by day 4, at which time the cells had apparently differentiated to adherent, vacuolated macrophages. The cytokine-stimulated leukemic cells were positive for the monocyte/macrophage specific markers alpha-1-antitrypsin, alpha-1-antichymotrypsin, lysozyme, CD14, MHC class II, and calprotectin, an antigen found in differentiated macrophages and granulocytes. Despite the strong tendency of the leukemic cells towards monocytic differentiation, our results suggested that they retained some features of a myelomonocytic precursor. These data show that cytochemistry, immunophenotyping, flow cytometry, and in vitro differentiation of canine leukemia cells are useful tools for confirming the lineage of malignant hematopoietic cells.     

Enhancement of Ovulatory Follicle Development in Maiden Sheep by Short-term Supplementation with Steam-flaked Corn

The effect of a short‐term nutritional supplementation with steam‐flaked corn on metabolism and folliculogenesis was evaluated in 14 maiden sheep. Oestrus was synchronized with two prostaglandin F_2α doses given 10 days apart. From day 11 to 15 of the oestrous cycle induced with prostaglandins, half of the ewes (group 2M) were supplemented with steam‐flaked corn, double the daily maintenance ration of the control sheep (group 1M). Body weight and condition remained unaffected, but the energetic supply increased plasma concentrations of glucose (3.6 ± 0.1 vs 4.3 ± 0.1 mmol/l, p < 0.0001) for the first 4 days and 3‐hydroxybutyrate (0.323 ± 0.58 vs 0.582 ± 0.04 mmol/l, p < 0.005) from day 2 to 4. The profile of insulin secretion was also affected by the treatment, increasing in group 2M to reach significant differences on days 13 and 14 (p < 0.05). From similar values at the start of the food supply, the treatment induced a higher follicular development in group 2M (1.1 ± 1.2 vs 7.4 ± 1.06 total follicles in day 15, p < 0.05), as evidenced by the lineal increase in the number of larger follicles (>4 mm, p < 0.005). Then, the number of follicles >4 mm in size in 2M was around 60% higher on day 16 (7.86 ± 0.45 vs 4.86 ± 0.63, p < 0.005). Thereafter, the mean number of corpora lutea per ewe was around 30% higher in group 2M (1.43 ± 0.2 vs 1.10 ± 0.1, although differences were not found to be statistically significant). These data suggest that the use of diets containing high starch sources, like the steam‐flaked corn, increases folliculogenesis and ovulation rate in sheep and can be applied in short‐term feeding practices.     

Modifications to physicochemical and nutritional properties of hard-To-cook beans (Phaseolus vulgaris L.) by extrusion cooking.

The objective of this work was to evaluate extrusion cooking as a means to improve the nutritional properties of Phaseolus vulgaris L. that had been stored either at 42 degrees C and 80% relative humidity for 6 weeks or for periods >1 year in cereal stores in tropical conditions. Storage under these conditions resulted in an increase in cooking time increased (7.7- and 12-fold, respectively) as a result of development of the hard-to-cook (HTC) defect. Single-screw extrusion of the milled beans was carried out at four barrel temperatures and two moisture contents. The extrudate bulk density and water solubility index decreased with increasing temperature, whereas the water absorption index increased due to the higher proportion of gelatinized starch in the extruded samples. Both fresh and HTC beans contained nutritionally significant amounts of lectins, trypsin, and alpha-amylase inhibitors, which were mostly inactivated by extrusion. Extrusion also caused a considerable redistribution of insoluble dietary fiber to soluble, although the total dietary fiber content was not affected. Changes in solubility involved pectic polysaccharides, arabinose and uronic acids being the main sugars involved. Stored beans subjected to extrusion cooking showed physical and chemical characteristics similar to those of extrudates from fresh beans.