Influence of Nitrogen and Sink Competition on Shoot Growth of Poplar

Terrestrial and oceanic biomass carbon sinks help reduce anthropogenic CO2 emissions and mitigate the long-term effect of increasing atmospheric CO2. Woody plants have large carbon pools because of their long residence time, however N availability can negatively impact tree responses to elevated CO2. Seasonal cycling of internal N in trees is a component that contributes to fitness especially in N limited environments. It involves resorption from senescing leaves of deciduous trees and storage as vegetative storage proteins (VSP) in perennial organs. Populus is a model organism for tree biology that efficiently recycles N. Bark storage proteins (BSP) are the most abundant VSP that serves as seasonal N reserves. Here I show how poplar growth is influenced by N availability and how growth is influenced by shoot competition for stored N reserves. I also provide data that indicates that auxin mediates BSP catabolism during renewed shoot growth. Understanding the components of N accumulation, remobilization and utilization can provide insights leading to increasing N use efficiency (NUE) of perennial plants.

Proteomic and phosphoproteomic analysis of polyethylene glycol-induced osmotic stress in root tips of common bean (Phaseolus vulgaris L.)

Previous studies have shown that polyethylene glycol (PEG)-induced osmotic stress (OS) reduces cell-wall (CW) porosity and limits aluminium (Al) uptake by root tips of common bean (Phaseolus vulgaris L.). A subsequent transcriptomic study suggested that genes related to CW processes are involved in adjustment to OS. In this study, a proteomic and phosphoproteomic approach was applied to identify OS-induced protein regulation to further improve our understanding of how OS affects Al accumulation. Analysis of total soluble proteins in root tips indicated that, in total, 22 proteins were differentially regulated by OS; these proteins were functionally categorized. Seventy-seven per- cent of the total expressed proteins were involved in metabolic pathways, particularly of carbohydrate and amino acid metabolism. An analysis of the apoplastic proteome revealed that OS reduced the level of five proteins and increased that of seven proteins. Investigation of the total soluble phosphoproteome suggested that dehydrin responded to OS with an enhanced phosphorylation state without a change in abundance. A cellular immunolocalization analysis indicated that dehydrin was localized mainly in the CW. This suggests that dehydrin may play a major protective role in the OS-induced physical breakdown of the CW structure and thus maintenance of the reversibility of CW extensibility during recovery from OS. The proteomic and phosphoproteomic analyses provided novel insights into the complex mechanisms of OS-induced reduction of Al accumulation in the root tips of common bean and highlight a key role for modification of CW structure.

Alteration of cell-wall porosity is involved in osmotic stress-induced enhancement of aluminium resistance in common bean (Phaseolus vulgaris L.)

Aluminium (Al) toxicity and drought are the two major abiotic stress factors limiting common bean production in the tropics. Using hydroponics, the short-term effects of combined Al toxicity and drought stress on root growth and Al uptake into the root apex were investigated. In the presence of Al stress, PEG 6000 (polyethylene glycol)-induced osmotic (drought) stress led to the amelioration of Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1. PEG 6000 (>> PEG 1000) treatment greatly decreased Al accumulation in the 1 cm root apices even when the roots were physically separated from the PEG solution using dialysis membrane tubes. Upon removal of PEG from the treatment solution, the root tips recovered from osmotic stress and the Al accumulation capacity was quickly restored. The PEG-induced reduction of Al accumulation was not due to a lower phytotoxic Al concentration in the treatment solution, reduced negativity of the root apoplast, or to enhanced citrate exudation. Also cell-wall (CW) material isolated from PEG-treated roots showed a low Al-binding capacity which, however, was restored after destroying the physical structure of the CW. The comparison of the Al(3+), La(3+), Sr(2+), and Rb(+) binding capacity of the intact root tips and the isolated CW revealed the specificity of the PEG 6000 effect for Al. This could be due to the higher hydrated ionic radius of Al(3+) compared with other cations (Al(3+) >> La(3+) > Sr(2+) > Rb(+)). In conclusion, the results provide circumstantial evidence that the osmotic stress-inhibited Al accumulation in root apices and thus reduced Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1 is related to the alteration of CW porosity resulting from PEG 6000-induced dehydration of the root apoplast.

First report of Didymella americanaon baby lima bean (Phaseolus lunatus)

A new foliar disease was observed on baby lima bean (Phaseolus lunatus) in fields across western New York State, USA. The disease occurred in 10 fields with variable incidence and severity. Symptoms were initially necrotic, tan spots on leaves with red to reddish brown irregular margins that coalesced to encompass the entire leaf and cause abscission. Pycnidia were observed within the lesions. Isolations from diseased leaves yielded several pycnidial forming fungi, including a Didymella species. These isolates were characterized by morphology and sequencing of multiple reference genes (internal transcribed spacer (ITS), partial actin, β- tubulin (tub2), translation elongation factor 1-α (TEF), 28S rDNA large subunit (LSU), rpb2, and calmodulin). A four gene phylogeny (ITS, tub2, LSU, and rpb2) showed that the isolates from baby lima bean belonged to a well-supported clade that contained the type culture of Didymella americana. Pathogenicity of the isolates on three commonly grown cultivars of baby lima bean was confirmed. Symptoms that developed on inoculated plants were similar to those observed on diseased plants in the field. This is the first report of D. americana on baby lima bean.

Response of some chickpea ( Cicer arietinum L.) genotypes to Orobanche foetida Poir. parasitism

In Tunisia, broomrape ( Orobanche foetida Poir.) causes major drawbacks especially in faba bean ( Vicia faba L.) Chickpea ( Cicer arietinum L.) suffers little damage compared to faba bean, but with the winter sowing chickpea cultivars, broomrape might become a serious problem for chickpea cultivation. The development of resistant cultivars remains the most efficient way to solve this problem. The behavior of six chickpea genotypes to O. foetida was studied under field natural infestation and artificial inoculation in pots and petri dishes in greenhouse conditions. During the cropping seasons 2010-2011 and 2012-2013 the level of infection was very low. The number of emerged parasites per host plant varied from 0.18 to 0.43 and the incidence from 6.5% to 23%. Among the six tested genotypes, G1, G2, and G4 showed partial resistance to O. foetida with low number and dry weight of emerged parasite and high grain yield compared to the other genotypes, although no significant differences were recorded. In pot experiments, the number and total dry weight of broomrape per plant were lower for G1 and G2 genotypes than the other genotypes. Parasitism does not affect significantly the shoot dry weight and number of pods of these genotypes. The total chlorophyll content was significantly reduced under infestation in all genotypes. In Petri dishes experiments, results showed that percent germination of O. foetida seeds varied from 49% to 65% and does not play a role in the resistance of chickpea genotypes. In contrast, broomrape attachment was lower and slower for the genotypes G1, G2, and G4 than the other genotypes. Resistance in chickpea genotypes was characterized by few parasite attachments on roots and a limited growth of established tubercles. No necrosis of attached tubercles was observed in the different experiments.

EFFECT OF TEMPORARY DROUGHT AT DIFFERENT GROWTH STAGES ON SNAP BEAN POD QUALITY AND YIELD

High quality snap bean ( Phaseolus vulgaris L. ) can be produced under rain-fed conditions, provided that adequate moisture is available. However, drought may occur at any stage of growth of snap bean. The objective of this study was to evaluate the effect of drought stress at different growth stages on pod physical quality and nutrient concentrations. An experiment was conducted at the Horticulture Greenhouse, Hawassa University in Ethiopia. Drought stress (50% of field capacity [FC]) was applied at the unfolding of the fourth trifoliate leaf, flowering and pod formation, against a control with no drought stress. The drought stress treatments and eight cultivars were arranged as a factorial experiment in a completely randomised design, with three replications. Drought stress (50% FC) during reproductive stages significantly (P<0.05) reduced pod texture, appearance, and pod curvature. Drought stress increased protein and zinc concentrations by 41 and 15%, respectively; but reduced iron concentration by 15% in snap bean pods. All the tested cultivars had relatively similar responses to drought stress.

NUTRITIONAL AND AMINO ACID ANALYSIS OF RAW, PARTIALLY FERMENTED AND COMPLETELY FERMENTED LOCUST BEAN ( Parkia biglobosa ) SEEDS

The nutritional and amino acid analysis of raw and fermented seeds of Parkia biglobosa were carried out. Parameters investigated include moisture, crude protein, crude fat, ash, crude fibre and mineral contents; and the effect of the degree of fermentation on these parameters was also investigated. The amino acid compositions of all the samples were evaluated and amino acid quality determined by calculating amino acid scores and the predicted protein efficiency ratio (P-PER). Results showed that the proximate composition was significantly affected by fermentation, although there was little difference between the parameters for the partially fermented and completely fermented samples. Based on dry matter percentage, protein content was in the 39.77 – 43.74 % range while crude fibre ranged between 5.55 – 7.42 %. The ash content was lowest in the raw sample (2.34 %), while the fermented samples had ash contents between 4.27 and 8.33 % for the fully fermented and the partially fermented seeds, respectively. The fat content increased from 8.65 % in the raw seed to 24.4 % and 27.6 % for the partially and completely fermented samples, respectively. Results of the amino acid analysis showed that the partially fermented sample had the lowest quantities of all amino acids determined and had lysine as the limiting amino acid, whereas the raw and completely fermented samples had very similar amino acid profile with amino acid scores of 100, indicating that there are no limiting amino acids. All the samples were rich in essential amino acids. The P-PER also showed that the partially fermented sample had the lowest protein efficiency while the raw seed had the highest. Mineral contents generally increased from the raw, through the partially fermented, to the completely fermented seeds and results showed the samples to be good sources of potassium (K), calcium (Ca), manganese (Mn) and copper (Cu) in addition to being complementary sources of other metals. Locust bean seed does not accumulate lead and is, therefore, safe for consumption without the potential of food poisoning.

Development of Locust Bean Gum Nanoparticles for protein delivery

Dissertação de Mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2014

Inhalable antitubercular therapy mediated by locust bean gum microparticles

Tuberculosis remains a major global health problem and alternative therapeutic approaches are needed. Considering the high prevalence of lung tuberculosis (80% of cases), the pulmonary delivery of antitubercular drugs in a carrier system capable of reaching the alveoli, being recognised and phagocytosed by alveolar macrophages (mycobacterium hosts), would be a significant improvement to current oral drug regimens. Locust bean gum (LBG) is a polysaccharide composed of galactose and mannose residues, which may favour specific recognition by macrophages and potentiate phagocytosis. LBG microparticles produced by spray-drying are reported herein for the first time, incorporating either isoniazid or rifabutin, first-line antitubercular drugs (association efficiencies >82%). Microparticles have adequate theoretical properties for deep lung delivery (aerodynamic diameters between 1.15 and 1.67 μm). The cytotoxic evaluation in lung epithelial cells (A549 cells) and macrophages (THP-1 cells) revealed a toxic effect from rifabutin-loaded microparticles at the highest concentrations, but we may consider that these were very high comparing with in vivo conditions. LBG microparticles further evidenced strong ability to be captured by macrophages (percentage of phagocytosis >94%). Overall, the obtained data indicated the potential of the proposed system for tuberculosis therapy.

Physical and chemical properties of starch and flour from different common bean (Phaseolus vulgaris L.) cultivar.

The physical, chemical and pasting properties of the flour and isolated starches from six different bean cultivars (Phaseolus vulgaris L.) were investigated in order to obtain information for application in new products. The protein and total starch contents of the bean flours ranged from 17.72 to 20.27% and from 39.68 to 43.78%, respectively. The bean starches had low amounts of proteins, lipids and ash and showed an amylose content ranging between 45.32 and 51.11% and absolute density values between 1.55 and 1.78 g.cm?3. The bean starch granules were round to oval with a smooth surface. Results viscoamylographic profiles of the starches and flours showed the possibility of selecting cultivars for specific applications according to these characteristics.

Drought impact on rainfed common bean production areas in Brazil.

Common bean production in Goiás, Brazil is concentrated in the same geographic area, but spread acrossthree distinct growing seasons, namely, wet, dry and winter. In the wet and dry seasons, common beansare grown under rainfed conditions, whereas the winter sowing is fully irrigated. The conventional breed-ing program performs all varietal selection stages solely in the winter season, with rainfed environmentsbeing incorporated in the breeding scheme only through the multi environment trials (METs) wherebasically only yield is recorded. As yield is the result of many interacting processes, it is challengingto determine the events (abiotic or biotic) associated with yield reduction in the rainfed environments(wet and dry seasons). To improve our understanding of rainfed dry bean production so as to produceinformation that can assist breeders in their efforts to develop stress-tolerant, high-yielding germplasm,we characterized environments by integrating weather, soil, crop and management factors using cropsimulation models. Crop simulations based on two commonly grown cultivars (Pérola and BRS Radi-ante) and statistical analyses of simulated yield suggest that both rainfed seasons, wet and dry, can bedivided in two groups of environments: highly favorable environment and favorable environment. Forthe wet and dry seasons, the highly favorable environment represents 44% and 58% of production area,respectively. Across all rainfed environment groups, terminal and/or reproductive drought stress occursin roughly one fourth of the seasons (23.9% for Pérola and 24.7% for Radiante), with drought being mostlimiting in the favorable environment group in the dry TPE. Based on our results, we argue that eventhough drought-tailoring might not be warranted, the common bean breeding program should adapttheir selection practices to the range of stresses occurring in the rainfed TPEs to select genotypes moresuitable for these environments.

Major loci controlling resistance to the angular leaf spot of common bean.

Based on the evidences presented in this paper, results from classical genetic studies, fine-mapping information and physical position analysis using the reference genome sequence of P. vulgaris, the BIC Genetic Committee has formally accepted the proposed new gene symbols.

Resistance to Fusarium wilt in common bean cultivars and lines in pre-commercial stage.

The objective of this study was to identify common bean cultivars with resistance to Fusarium wilt.

Phenotypic and molecular characterization of common bean genotypes for resistance to anthracnose.

This study aimed to perform phenotypic and molecular characterization of cultivars and breeding lines of common bean for resistance to anthracnose.

Cooking time of common bean: genetic parameters and selection of elite lines.

The objectives of this study were to investigate the genetic variability and select elite lines for CT, since these lines aggregate essential agronomic traits.