Ripening-related defense proteins in Annona fruit

In order to obtain a better understanding of the active defense strategy of cherimoya (Annona cherimola Mill.) fruit, hydrolytic and antifungal activity, as well as expression of proteins functionally and immunogenically related to the pathogenesis-related proteins chitinase (PR-Q) and 1,3-β-glucanase (PR-2), were estimated in fruit at different ripening stages. Increase in expression of the 27 kDa constitutive chitinase and the induction of two new proteins, a 26 kDa chitinase and a 51 kDa 1,3-β-glucanase were associated with enhanced in vitro hydrolytic and antifungal activity of the acidic protein extract in ripe fruit. Ripening modified the expression of constitutive basic isoenzymes, with a sharp decrease in both relative accumulation and hydrolytic activity. Likewise, a new basic 33 kDa chitinase was induced in the over-ripe fruit, concomitant with accumulation of a basic constitutive 76 kDa 1,3-β-glucanase. At this stage, the basic protein extract modified in vitro growth inhibition of Botrytis cinerea. Short-term high CO₂ treatment delayed fruit ripening and maintained a similar distribution of activity and isoenzymatic pattern in both protein fractions to that in unripe fruit. These results indicate that the changes in the pattern of defense proteins and hydrolytic activity in cherimoyas appear to be associated with ripening. Moreover, unlike the constitutively expressed isoenzymes, only the transitorily induced chitinases and 1,3-β-glucanases were associated with an active defense-related response.     

Bioprospection of Microalgae and Cyanobacteria as Biocontrol Agents Against Vibrio campbellii and Their Use in White Shrimp Litopenaeus vannamei Culture

Vibrio spp. are the most common and harmful shrimp pathogenic bacteria; however, microalgae and cyanobacteria have the ability to produce antimicrobial substances against these species. In this study, the organic and aqueous extracts of 28 species of marine microalgae and cyanobacteria were screened against Vibrio campbellii M1. Two of these phytoplankton species with antibacterial activity in aqueous extracts (Dunaliella tertiolecta and Skeletonema costatum) and nontoxic to brine shrimp Artemia franciscana nauplii were used to evaluate their anti‐Vibrio effect when used as green‐water cultures in Vibrio‐challenged white shrimp Litopenaeus vannamei cultures. No differences in mortality of juvenile L. vannamei were observed between treatments tested, suggesting that the pathogenicity of V. campbellii could be related to the growth stage of shrimp. The proximal composition of D. tertiolecta and S. costatum was in the recommended range for penaeid shrimp nutrition, allowing shrimp supplemented with these microalgae to have significantly greater total length and weight than control shrimp. Shrimp supplemented with S. costatum presented the highest values of organic mass (11.48 mg/organism) and growth rate (0.31 mg/d) in comparison to D. tertiolecta. These results indicate that microalgae are not only capable of producing antibacterial compounds against Vibrio but can also help shrimp nutrition.     

Insulin binding to bovine mammary membranes: comparison of microsomes versus smooth membranes

The study was undertaken to determine if membrane preparations of bovine mammary tissue bound insulin. If binding occurred, it was also the intent to compare binding kinetics between microsomes and smooth membranes. Insulin binding to bovine mammary membranes attained equilibrium, was saturable and was specific for insulin. Additional studies showed binding to be pH sensitive and maximal at 10 mM calcium. Binding affinity of insulin to microsomes and smooth membranes was similar, with the exception that smooth membranes bound 1.8 times more insulin per unit of membrane protein than microsomes. Two different methods were used to generate data for kinetic analysis of the insulin-receptor interaction in microsomes. Competitive binding assays (.6 ng [125I]insulin plus 0 to 100 ng insulin) indicated the presence of two binding sites with dissociation constants (Kd) of .32 and 15.8 nM. Direct titration of microsomes with [125I]insulin (.02 to 10 ng/ml) revealed two binding sites with Kd of .017 and .31 nM. The affinity of the second binding site measured by the competitive binding assay method (Kd of 15.8 nM) is low and therefore may not be of physiological importance for insulin action. Insulin appears to bind to two high-affinity receptor sites in bovine mammary microsomes with Kds of .017 nM and .32 nM. These findings show that bovine mammary tissue contains receptors for insulin. In addition, isolation of smooth membranes from microsomes enriches the number of insulin receptors per unit of membrane protein without altering their binding characteristics.     

Relation between Soil Health, Wave-like Fluctuations in Microbial Populations, and Soil-borne Plant Disease Management

A healthy soil is often defined as a stable soil system with high levels of biological diversity and activity, internal nutrient cycling, and resilience to disturbance. This implies that microbial fluctuations after a disturbance would dampen more quickly in a healthy than in a chronically damaged and biologically impoverished soil. Soil could be disturbed by various processes, for example addition of a nutrient source, tillage, or drying-rewetting. As a result of any disturbance, the numbers of heterotrophic bacteria and of individual species start to oscillate, both in time and space. The oscillations appear as moving waves along the path of a moving nutrient source such as a root tip. The phase and period for different trophic groups and species of bacteria may be shifted indicating that succession occurs. DGGE, Biolog and FAME analysis of subsequent populations in oscillation have confirmed that there is a cyclic succession in microbial communities. Microbial diversity oscillates in opposite direction from oscillations in microbial populations. In a healthy soil, the amplitudes of these oscillations will be small, but the background levels of microbial diversity and activity are high, so that soil-borne diseases will face more competitors and antagonists. However, soil-borne pathogens and antagonists alike will fluctuate in time and space as a result of growing plant roots and other disturbances, and the periods and phases of the oscillations may vary. As a consequence, biological control by members of a single trophic group or species may never be complete, as pathogens will encounter varying populations of the biocontrol agent on the root surface. A mixture of different trophic groups may provide more complete biological control because peaks of different trophic groups occur at subsequent locations along a root. Alternatively, regular addition of soil organic matter may increase background levels of microbial activity, increase nutrient cycling, lower the concentrations of easily available nutrient sources, increase microbial diversity, and enhance natural disease suppression.     

Assessment of real-time PCR as a method for determining the presence of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> in different Solanaceae cultivars

Real-time PCR was used to detect and quantify Verticillium dahliae and to assess the susceptibility of four Capsicum annuum cultivars (Luesia, Padrón, SCM331 and PI201234) and the Capsicum chinense cv. C118 to this pathogen. The symptoms which developed after infection included stunting and yellowing, and were more acute in the cv. SCM331, which also suffered defoliation in later stages of the disease and in C118, which suffered severe stunting. Quantification of the pathogen DNA in roots 23 and 34 days post-inoculation (dpi) revealed that there were significantly higher amounts of Verticillium dahliae DNA in C118 than in the other cultivars, followed by SCM331, Padrón and PI201234. The lowest amounts of fungal DNA in roots were found in Luesia. In hypocotyls, the highest amounts of fungal DNA were found in SCM331, while Luesia, Padrón and PI201234 had much lower amounts, and C118 had intermediate levels. When a compatible versus an incompatible system was studied, using the near-isogenic tomato lines LA3030 (susceptible) and LA3038 (resistant to V. dahliae), we were able to detect fungal DNA in both lines. As expected, the fungus/plant DNA ratio was lower in LA3038 than in LA3030 and it decreased with time in LA3038. The amount of Verticillium dahliae DNA in the roots of LA3030 remained constant between days 23 and 34 post-inoculation, but increased 10-fold in collars. Finally, when real-time PCR was applied as a diagnostic method to samples from pepper plants, soil and water collected from farms in northwest Spain, we were able to detect V. dahliae DNA in these samples even when symptoms of the disease were not evident.     

Potato brown rot incidence and severity under different management and amendment regimes in different soil types

Ralstonia solanacearum race 3 biovar 2, the causative agent of potato brown rot (bacterial wilt), is an economically important disease in tropical, subtropical and temperate regions of the world. In view of previous reports on suppression of the disease by organic amendments, and the expansion of organic agriculture, it was timely to compare the effects of organic and conventional management and various amendments on brown rot development in different soils (type: sand or clay; origin: Egypt or the Netherlands). Brown rot infection was only slightly reduced in organically compared to conventionally managed sandy soils from Egypt, but organic management significantly increased disease incidence and pathogen survival in Dutch sandy and clay soils, which correlated with high DOC contents in the organic Dutch soils. There was no correlation between disease incidence or severity and bacterial diversity in the potato rhizosphere in differently managed soils (as determined by 16S DGGE). NPK fertilization reduced bacterial wilt in conventional Egyptian soils but not in Dutch soils. Cow manure amendment significantly reduced disease incidence in organic Dutch sandy soils, but did not affect the bacterial population. However, cow manure did reduce densities of R. solanacearum in Egyptian sandy soils, most probably by microbial competition as a clear shift in populations was detected with DGGE in these and Dutch sandy soils after manure amendment. Amendment with compost did not have a suppressive effect in any soil type. The absence of a disease suppressive effect of mineral and organic fertilization in Dutch clay soils may be related to the already high availability of inorganic and organic nutrients in these soils. This study shows that the mechanism of disease suppression of soil-borne plant pathogens may vary strongly according to the soil type, especially if quite different types of soil are used.     

Developing a dynamic growth model for teak plantations in India

Background： Tectono grandis （teak） is one of the most important tropical timber species occurring naturally in India. Appropriate growth models, based on advanced modeling techniques, are not available but are necessary for the successful management of teak stands in the country. Long-term forest planning requires mathematical models and the principles of Dynamical System Theory provide a solid foundation for these.
Methods： The state-space approach makes it possible to accommodate disturbances and avarying environment. In this paper, an attempt has been made to develop a dynamic growth model based on the limited data, consisting of three annual measurements, collected from 22 teak sample plots in Karnataka, Southern India.
Results： A biologically consistent whole-stand growth model has been presented which uses the state-space approach for modelling rates of change of three state-variables viz., dominant height, stems per hectare and stand basal area. Moreover, the model includes a stand volume equation as an output function to estimate this variable at any point in time. Transition functions were fitted separately and simultaneously. Moreover, a continuous autoregressive error structure is also included in the modelling process. For fitting volume equation, generalized method of moments was used to get efficient parameter estimates under heteroscedastic conditions.
Conclusions： A simple model containing few free parameters performed well and is particularly well suited to situations where available data is scarce.     

Optimization of experimental infection protocols for the estimation of genetic parameters of resistance to White Spot Syndrome Virus (WSSV) in Penaeus (Litopenaeus) vannamei

Two infection protocols, individual oral (IO) and waterborne infection (WB), were evaluated to challenge Penaeus (Litopenaeus) vannamei with White Spot Syndrome Virus (WSSV). Five different batches of full and half-sib families were infected experimentally and tested for growth performance under commercial growth conditions. The genetic variance for WSSV resistance was estimated using a linear sire-dam repeatability model that considers test-day survival as the dependent variable. The heritability estimates using the IO protocol ranged from 0.01 ± 0.00 to 0.02 ± 0.01, whereas the heritability estimates using the WB infection protocol were not significantly different from zero. The genetic correlations measured as the product moment correlation between full-sib family breeding values for resistance to WSSV and harvest body weight in ponds were unfavourable in three of the five batches and favourable in one of them. The dosage of WSSV was better controlled with IO oral infection than with other methods, with all animals being exposed to approximately the same risk of infection at the same time. This should improve the accuracy of the genetic parameters and hence improve the accuracy of the breeding values. It should, however, be noted that once the outbreak was established and the mortalities began, the shape and the magnitude of the slope of the mortality curves showed little difference in the infection pattern between batches irrespective of the dosage and infection protocol, and in most of the cases the cumulative mortality was greater than 80%. The main reason for this is probably the high densities of animals in the tanks needed for the genetic evaluations.     

A biologically-consistent stand growth model for loblolly pine in the Piedmont physiographic region, USA

A theory-influenced dynamical stand growth model formulation described well the behavior of thinned and unthinned loblolly pine plantations. A simplification containing few free parameters performed as well as a fully parametrized version. It seems particularly well suited to situations where available data is scarce. The stand model can be interfaced to additional climate, nutrition and carbon cycling modules for studying the effects of a changing environment.