Comparison of flag leaf and ear photosynthesis with biomass and grain yield of durum wheat under various water conditions and genotypes

Photosynthetic activity of cereals has traditionally been studied using leaves, thus neglecting the role of other organs such as ears. Here, we studied the effects of water status and genotypes on the photosynthetic activity of the flag leaf blade and the ear of durum wheat. The various parameters related to the photosynthetic activity were analysed in relation to the total above-ground plant biomass and grain yield at maturity. Four local varieties plus two cultivars adapted to the semiarid areas of South Morocco were grown in pots in a greenhouse. Five different water treatments were maintained from the beginning of stem elongation to maturity, when shoot biomass and grain yield were recorded. The net photosynthesis (A), stomatal conductance (gs) and transpiration (T) of the ear and the flag leaf were measured at anthesis. In both organs these factors decreased significantly with water deficit, whereas the A/T and A/gs ratios increased. The genotype effect was also significant for all traits studied. Whole-organ photosynthesis was much higher in the ear than in the flag leaf in well-watered conditions. As water stress developed, photosynthesis decreased less in the ear than in the flag leaf. Whole-ear photosynthesis correlated better than flag leaf photosynthesis with biomass and yield. Nevertheless, the relationships of the whole flag leaf with biomass and yield improved as the water stress became more severe, suggesting a progressive shift of yield from sink to source limitation. For all water regimes the ratios A/gs and A/T of the ear also showed a higher (negative) correlation with both biomass and yield than those of the flag leaf. The results indicate that the ear has a greater photosynthetic role than the flag leaf in determining grain yield, not only in drought but also in the absence of stress.

Enchimento de sementes em linhas quase-isogênicas de trigo com presença e ausência do caráter "stay-green"

O caráter "stay-green" (permanência da cor verde nas plantas) tem sido utilizado para reduzir a senescência em trigo, resultando no aumento funcional da área foliar, e, conseqüentemente, na elevação da taxa e da duração do enchimento da semente. O objetivo deste trabalho foi analisar a taxa de acúmulo de matéria seca e a duração do período de enchimento da semente de linhas quase-isogênicas de trigo diferenciadas quanto à presença e ausência do caráter "stay-green". O experimento foi conduzido no Centro Experimental da Palma, Pelotas, RS. Foram utilizadas quatro linhas quase-isogênicas: F6-SG e F6-SZ, no ano de 2000 e F7-SG e F7-SZ no ano 2001, com cinco repetições. O caráter "stay-green" permitiu maior deposição de reservas na semente na fase próxima do ponto de maturidade fisiológica. A duração do período de enchimento da semente esteve associada ao acúmulo de matéria seca na semente, promovendo um maior peso médio da semente. Assim, o progresso do caráter taxa de enchimento da semente poderá ser obtido por meio de seleções de plantas com maior peso médio da semente.

Diallel analysis for frogeye leaf spot resistance in soybean

Seven soybean cultivars (Bossier, Cristalina, Davis, Kent, Lincoln, Paraná and Uberaba), with different levels of resistance to Cercospora sojina, were crossed in a diallel design to determine the general (GCA) and specific (SCA) combining abilities relative to the inheritance of the resistance. Race 04 of the fungus was inoculated in the parents and in the 21 F1 hybrids in a greenhouse in a completely randomized design, with 12 replications. The reactions to the disease were evaluated 20 days after the inoculation, always on the most infected leaflet. Both GCA and SCA were significant for all the evaluated characters, being inferred that, for the expression of the characters, the additive, dominant and, possibly, epistatic genic actions were important. The largest values of estimated SCA effect (ij) were observed in the hybrid combinations where at least one parent presented high GCA. Cristalina, Davis and Uberaba cultivars showed the largest estimates for GCA effect (i), and from the analysis of ii, the contribution of these parents to heterosis of their hybrids will be towards the reduction of the disease symptoms. Therefore, these cultivars are indicated as parents in breeding programs that seek the development of soybean cultivars with resistance to frogeye leaf spot.

Biometrical analysis and selection of tetraploid progenies of Panicum maximum using mixed model methods

The objectives of this work were to estimate the genetic and phenotypic parameters and to predict the genetic and genotypic values of the selection candidates obtained from intraspecific crosses in Panicum maximum as well as the performance of the hybrid progeny of the existing and projected crosses. Seventy-nine intraspecific hybrids obtained from artificial crosses among five apomictic and three sexual autotetraploid individuals were evaluated in a clonal test with two replications and ten plants per plot. Green matter yield, total and leaf dry matter yields and leaf percentage were evaluated in five cuts per year during three years. Genetic parameters were estimated and breeding and genotypic values were predicted using the restricted maximum likelihood/best linear unbiased prediction procedure (REML/BLUP). The dominant genetic variance was estimated by adjusting the effect of full-sib families. Low magnitude individual narrow sense heritabilities (0.02-0.05), individual broad sense heritabilities (0.14-0.20) and repeatability measured on an individual basis (0.15-0.21) were obtained. Dominance effects for all evaluated characteristics indicated that breeding strategies that explore heterosis must be adopted. Less than 5% increase in the parameter repeatability was obtained for a three-year evaluation period and may be the criterion to determine the maximum number of years of evaluation to be adopted, without compromising gain per cycle of selection. The identification of hybrid candidates for future cultivars and of those that can be incorporated into the breeding program was based on the genotypic and breeding values, respectively. The prediction of the performance of the hybrid progeny, based on the breeding values of the progenitors, permitted the identification of the best crosses and indicated the best parents to use in crosses.

A new green fluorescent protein-based bacterial biosensor for analysing phenanthrene fluxes.

The polycyclic aromatic hydrocarbon (PAH)-degrading strain Burkholderia sp. RP007 served as host strain for the design of a bacterial biosensor for the detection of phenanthrene. RP007 was transformed with a reporter plasmid containing a transcriptional fusion between the phnS putative promoter/operator region and the gene encoding the enhanced green fluorescent protein (GFP). The resulting bacterial biosensor--Burkholderia sp. strain RP037--produced significant amounts of GFP after batch incubation in the presence of phenanthrene crystals. Co-incubation with acetate did not disturb the phenanthrene-specific response but resulted in a homogenously responding population of cells. Active metabolism was required for induction with phenanthrene. The magnitude of GFP induction was influenced by physical parameters affecting the phenanthrene flux to the cells, such as the contact surface area between solid phenanthrene and the aqueous phase, addition of surfactant, and slow phenanthrene release from Model Polymer Release System beads or from a water-immiscible oil. These results strongly suggest that the bacterial biosensor can sense different phenanthrene fluxes while maintaining phenanthrene metabolism, thus acting as a genuine sensor for phenanthrene bioavailability. A relationship between GFP production and phenanthrene mass transfer is proposed.

First detection of Waddlia chondrophila in Africa using SYBR Green real-time PCR on veterinary samples.

Waddlia chondrophila is a strict intracellular microorganism belonging to the order Chlamydiales that has been isolated twice from aborted bovine fetuses, once in USA and once in Germany. This bacterium is now considered as an abortigenic agent in cattle. However, no information is available regarding the presence of this bacterium in Africa. Given the low sensitivity of cell culture to recover such an obligate intracellular bacterium, molecular-based diagnostic approaches are warranted. This report describes the development of a quantitative SYBR Green real-time PCR assay targeting the recA gene of W. chondrophila. Analytical sensitivity was 10 copies of control plasmid DNA per reaction. No cross-amplification was observed when testing pathogens that can cause abortion in cattle. The PCR exhibited a good intra-run and inter-run reproducibility. This real-time PCR was then applied to 150 vaginal swabs taken from Tunisian cows that have aborted. Twelve samples revealed to be Waddlia positive, suggesting a possible role of this bacterium in this setting. This new real-time PCR assay represents a diagnostic tool that may be used to further study the prevalence of Waddlia infection.

Ultrastructural changes of the internal limiting membrane removed during indocyanine green assisted peeling versus conventional surgery for an idiopathic epimacular membrane

Purpose:To evaluate the histological features of cellular retinal fragments on the internal limiting membrane (ILM) removed during epiretinal membrane peeling surgery with and without the aid of ICG diluted in 5% glucose Methods:ILM specimens removed from 88 eyes undergoing vitrectomy and membrane peeling surgery for idiopathic epiretinal membrane between 1995 and 2003 were reviewed retrospectively. Surgery was performed in all cases by the same surgeon using the same technique. ICG was diluted with 5% glucose. Histological analysis focused on the presence and characteristics of retinal structures on the retinal surface of the ILM. Statistical analysis compared the results between group I (conventional surgery without ICG) and group II (ICG-assisted peeling) Results:Seventy-one eyes underwent EMM surgery without the aid of ICG (group I) and seventeen underwent EMM ICG-assisted surgery assisted using ICG (group II). The amount of Muller cell debris on the retinal surface of the ILM was more significant in the group I (no ICG) than in the group II (ICG) (40.8 versus 11.8; p = 0.024). Large fragments of Muller cells were more frequently observed in the group I (no ICG) than in the group II (ICG) (63.4% versus 23.5%; p= 0.003).The presence of larger retinal elements such as neural axons and vessels were observed attached to retinal face of the ILM in 5 (7%) cases of the no-ICG group. No such retinal elements were detected in any of the histological ILM specimens of the ICG-assisted group Conclusions:The use of ICG diluted with 5% glucose in the aid of ILM removal during epiretinal membrane surgery was associated with less retinal debris attached to retinal face of the ILM compared to surgery in which ICG was not used. Our findings contradict previous reports in the literature, in which use of ICG diluted with balanced salt solution (BSS) was associated with more retinal fragments attached to the retinal face of the ILM. According to our results, we hypothesize that diluting ICG with 5% glucose may decrease the adhesion of the ILM to the underlying retinal layers such that less retinal debris is removed with peeling of the ILM.

Differentially phased leaf growth and movements in Arabidopsis depend on coordinated circadian and light regulation.

In contrast to vastly studied hypocotyl growth, little is known about diel regulation of leaf growth and its coordination with movements such as changes in leaf elevation angle (hyponasty). We developed a 3D live-leaf growth analysis system enabling simultaneous monitoring of growth and movements. Leaf growth is maximal several hours after dawn, requires light, and is regulated by daylength, suggesting coupling between growth and metabolism. We identify both blade and petiole positioning as important components of leaf movements in Arabidopsis thaliana and reveal a temporal delay between growth and movements. In hypocotyls, the combination of circadian expression of PHYTOCHROME INTERACTING FACTOR4 (PIF4) and PIF5 and their light-regulated protein stability drives rhythmic hypocotyl elongation with peak growth at dawn. We find that PIF4 and PIF5 are not essential to sustain rhythmic leaf growth but influence their amplitude. Furthermore, EARLY FLOWERING3, a member of the evening complex (EC), is required to maintain the correct phase between growth and movement. Our study shows that the mechanisms underlying rhythmic hypocotyl and leaf growth differ. Moreover, we reveal the temporal relationship between leaf elongation and movements and demonstrate the importance of the EC for the coordination of these phenotypic traits.

Somatic embryogenesis and in vitro plant regeneration from pejibaye adult plant leaf primordia

The aim of this work was to evaluate a protocol for plant regeneration by means of somatic embryos obtained from isolated adult pejibaye leaf primordia, and to describe histological origin of embryos and morphogenetic response. Explants were cultivated in modified MS medium. Mesophyll parenchymatous cells originated meristemoids (preembryonic complex formation) induced with 7.1 µM BAP in the first two subculture periods. After polarized structures with 12.9 µM NAA and 3.55 µM BAP were formed in the third subculture, somatic embryos developed and regenerated normal plants. The mesophyll parenchymatous cells display high capacity of direct response to the auxin and cytokinin.

Green manure incorporation timing for organically grown broccoli

The objective of this work was to determine the effect of incorporation timing of the velvet bean (Stizolobium cinereum) (GM) on both organic broccoli yield and N status. Mineral N content in the soil, biologically fixed N recovery by broccoli, GM biomass decomposition and N release kinetics were also determined. Plots were fertilized with 12 Mg ha-1 of organic compost and received GM either at 0, 15, 30 or 45 days after transplant. Other treatments were compost (12 or 25 Mg ha-1), GM, mineral fertilizers and control (no fertilizer). The data were collected in four completely randomized blocks. GM decomposition increased mineral N content in soil as rapidly as mineral fertilizer or the supply of 25 Mg ha-1 of compost. The N half-life in GM (24 days) is smaller than the mass half-life (35 days) and the biological fixation contributed with 23.6% of N present in the aboveground biomass of broccoli. The result suggests a higher synchrony between the crop relative growth rate and N release from the GM when incorporated at crop early growth stage. The incorporation of GM until 15 days after transplanting replaces 50% of the highest compost dose, without yield loss.

Study of simple sequence repeat markers from coffee expressed sequences associated to leaf miner resistance

The objective of this work was to identify expressed simple sequence repeats (SSR) markers associated to leaf miner resistance in coffee progenies. Identification of SSR markers was accomplished by directed searches on the Brazilian Coffee Expressed Sequence Tags (EST) database. Sequence analysis of 32 selected SSR loci showed that 65% repeats are of tetra-, 21% of tri- and 14% of dinucleotides. Also, expressed SSR are localized frequently in the 5'-UTR of gene transcript. Moreover, most of the genes containing SSR are associated with defense mechanisms. Polymorphisms were analyzed in progenies segregating for resistance to the leaf miner and corresponding to advanced generations of a Coffea arabica x Coffea racemosa hybrid. Frequency of SSR alleles was 2.1 per locus. However, no polymorphism associated with leaf miner resistance was identified. These results suggest that marker-assisted selection in coffee breeding should be performed on the initial cross, in which genetic variability is still significant.

Recovery of a persistent Canine distemper virus expressing the enhanced green fluorescent protein from cloned cDNA.

Wild-type A75/17-Canine distemper virus (CDV) is a highly virulent strain, which induces a persistent infection in the central nervous system (CNS) with demyelinating disease. Wild-type A75/17-CDV, which is unable to replicate in cell lines to detectable levels, was adapted to grow in Vero cells and was designated A75/17-V. Sequence comparison between the two genomes revealed seven nucleotide differences located in the phosphoprotein (P), the matrix (M) and the large (L) genes. The P gene is polycistronic and encodes two auxiliary proteins, V and C, besides the P protein. The mutations resulted in amino acid changes in the P and V, but not in the C protein, as well as in the M and L proteins. Here, a rescue system was developed for the A75/17-V strain, which was shown to be attenuated in vivo, but retains a persistent infection phenotype in Vero cells. In order to track the recombinant virus, an additional transcription unit coding for the enhanced green fluorescent protein (eGFP) was inserted at the 3' proximal position in the A75/17-V cDNA clone. Reverse genetics technology will allow us to characterize the genetic determinants of A75/17-V CDV persistent infection in cell culture.

Risk analysis of climate change on coffee nematodes and leaf miner in Brazil

The objective of this work was to assess the potential impact of climate change on the spatial distribution of coffee nematodes (races of Meloidogyne incognita) and leaf miner (Leucoptera coffeella), using a Geographic Information System. Assessment of the impacts of climate change on pest infestations and disease epidemics in crops is needed as a basis for revising management practices to minimize crop losses as climatic conditions shift. Future scenarios focused on the decades of the 2020's, 2050's, and 2080's (scenarios A2 and B2) were obtained from five General Circulation Models available on Data Distribution Centre from Intergovernmental Panel on Climate Change. Geographic distribution maps were prepared using models to predict the number of generations of the nematodes and leaf miner. Maps obtained in scenario A2 allowed prediction of an increased infestation of the nematode and of the pest, due to greater number of generations per month, than occurred under the climatological normal from 1961-1990. The number of generations also increased in the B2 scenario, but was lower than in the A2 scenario for both organisms.