Bacillus thuringiensis - A New Perspective for an Induced Protection to Coffee Leaf Rust*

Susceptible coffee plants (Coffea arabica cv. ‘Mundo Novo’) treated with a suspension of Bacillus thuringiensis or with three of commercial products (Thuricide HD, Bactimos and Bactospeine PM) of B. thurngiensis were protected against a later inoculation with Hemileia vastatrix, the causal agent of coffee leaf rust. Thuricide HD was effective at concentrations from 5 to 20 mg/ml; at this concentration, protection reached 90 %, lasted for at least 5 weeks, and was systemic. Induced resistance was determined by the reduction of the average number of lesions per leaf in treated leaves when compared with non-treated controls. A decrease in lesion size as well as a delay in sporulation time were also observed in protected plants treated with Thuricide HD.     

Aggressiveness of Strains and Inoculation Methods for Resistance Assessment to Bacterial Halo Blight on Coffee Seedlings

Characterization of resistance response to pathogens is a fundamental strategy in plant breeding programmes. Thus, in this study, we developed an effective inoculation method for resistance tests of bacterial halo blight (BHB), caused by Pseudomonas syringae pv. garcae on coffee. Firstly, aggressiveness of eight selected bacterial strains as well as seven mixed strains was assessed on coffee seedlings of Mundo Novo IAC 376‐4 cultivar. Two experiments were conducted comparing three inoculation techniques: initially, we compare the methods of sprinkling and multiple needles on four cultivars of Coffea arabica; later, four cultivars and seven genotypes were compared for the multiple needles method and the abrasion technique. Severity was evaluated according a disease rating scale (DRS), considering either leaf surface area affected for sprinkling, or affected inoculated area for inoculation by multiple needles and abrasion. The area under the disease progress curve of disease (AUDPC) was calculated considering weekly evaluations of disease from seven until 42 days postinoculation (DPI). The standard deviation (SD), coefficient of variation (CV) and confidence interval (CI) variables were used to denote the effectiveness of the methods. According to the results, strain IBSBF 1197 was the most aggressive, and three other strains showed high aggressiveness. All inoculation methods were able to discriminate the resistance response to BHB, wherein the sprinkling method was less efficient than multiple needles and abrasion technique was more efficient than multiple needles. In addition, disease evaluations at 14 DPI showed a high correlation coefficient with area under AUDPC at 42 DPI, validating the early selection to BHB. Results of inoculation methods indicated the abrasion on first pair of leaves, together with evaluations on fourteenth DPI, as the most promising technique for early selection on coffee breeding to BHB.     

Effects of Pratylenchus brachyurus and P. coffeae on Seedlings of Coffea arabica

Two experiments were carried out to evaluate the effects of Pratylenchus brachyurus and P. coffeae on Coffea arabica. The first experiment was conducted in a greenhouse to determine the effects of Pratylenchus brachyurus and P. coffeae on seedlings of Coffea arabica cv. Mundo Novo. Both Pratylenchus spp. reduced the growth of coffee seedlings. Higher contents of soluble sugars were detected in the leaves of infected plants. The reproduction rate of P. brachyurus was very low on cv. Mundo Novo, indicating an intolerance to this nematode. In a second experiment, C. arabica cultivars Mundo Novo and Catuaf both were intolerant hosts of P. brachyurus.     

Susceptibility of six Arabic coffee cultivars to fruit flies (Diptera: Tephritoidea) under shaded and unshaded organic management in Valença, State of Rio de Janeiro, Brazil

The infestation indices by fruit flies were determined for six cultivars of Coffea arabica L. in shaded and unshaded systems under organic management. The experiment was set in a completely randomized design with a split-split-plot arrangement and four replicates. A 250g-sample of maturing fruits per plot was harvested in May 2005. The cultivars Icatu Amarelo and Catucaí Amarelo were the least susceptible to attack by tephritids in both systems. As for lonchaeids, Oeiras, Catucaí Amarelo and Catuaí Vermelho were the least susceptible cultivars in the shaded system, and there was no difference among the cultivars in the unshaded system. The following tephritid species were obtained: Ceratitis capitata (Wiedemann), Anastrepha fraterculus (Wiedemann) and A. sororcula Zucchi (Tephritidae). Lonchaeids were represented by Neosilba bifida Strikis & Prado, N. certa (Walker), N. glaberrima (Wiedemann), N. pendula (Bezzi), N. pseudopendula (Korytkowski and Ojeda), Dasiops rugifrons Hennig, Neosilba n.sp.10 and Neosilba n.sp.14.     

Biochemical and genomic analysis of sucrose metabolism during coffee (Coffea arabica) fruit development

Sucrose metabolism and the role of sucrose synthase were investigated in the fruit tissues (pericarp, perisperm, and endosperm) of Coffea arabica during development. Acid invertase, sucrose phosphate synthase, and sucrose synthase activities were monitored and compared with the levels of sucrose and reducing sugars. Among these enzymes, sucrose synthase showed the highest activities during the last stage of endosperm and pericarp development and this activity paralleled closely the accumulation of sucrose in these tissues at this stage. Carbon partitioning in fruits was studied by pulse-chase experiments with (14)C-sugars and revealed high rates of sucrose turnover in perisperm and endosperm tissues. Additional feeding experiments with (14)CO(2) showed that leaf photosynthesis contributed more to seed development than the pericarp in terms of photosynthate supply to the endosperm. Sugar analysis, feeding experiments, and histological studies indicated that the perisperm plays an important role in this downloading process. It was observed that the perisperm presents a transient accumulation of starch which is degraded as the seed develops. Two full-length cDNAs (CaSUS1 and CaSUS2) and the complete gene sequence of the latter were also isolated. They encode sucrose synthase isoforms that are phylogenetically distinct, indicating their involvement in different physiological functions during cherry development. Contrasting expression patterns were observed for CaSUS1 and CaSUS2 in perisperm, endosperm, and pericarp tissues: CaSUS1 mRNAs accumulated mainly during the early development of perisperm and endosperm, as well as during pericarp growing phases, whereas those of CaSUS2 paralleled sucrose synthase activity in the last weeks of pericarp and endosperm development. Taken together, these results indicate that sucrose synthase plays an important role in sugar metabolism during sucrose accumulation in the coffee fruit.     

Electrolyte leakage and lipid degradation account for cold sensitivity in leaves of Coffea sp. plants

Five Coffea genotypes differing in their sensitivity to low positive temperatures were compared with regard to the effects of chilling on membrane integrity, as well as their ability to recover from cold-induced injury upon re-warming. Membrane damage was evaluated through electrolyte leakage, changes in membrane lipid composition and malondialdehyde (MDA) production in control conditions (25/20 degrees C, day/night), after a gradual temperature decrease period to 15/10 degrees C, after chilling treatment (3 nights at 4 degrees C) and upon re-warming to 25/20 degrees C during 6 days (recovery). C. dewevrei showed the highest electrolyte leakage at 15/10 degrees C and after chilling. This was due mainly to lipid degradation observed at 15/10 degrees C, reflecting strong membrane damage. Furthermore, MDA production after chilling conditions indicated the occurrence of lipid peroxidation. A higher susceptibility of C. dewevrei to cold also was inferred from the complete absence of recovery as regards permeability, contrary to what was observed in the remaining plants. Apoat? and Piat? presented significant leakage values after chilling. However, such effects were reversible under recovery conditions. Exposure to cold (15/10 degrees C and 3 x 15/4 degrees C) did not significantly affect membrane permeability in Catuaí and Icatú. Furthermore, no significant MDA production was observed even after chilling treatments in Apoat?, Piat?, Catuaí and Icatú, suggesting that the four genotypes had the ability to maintain membrane integrity and/or repair membrane damage caused by low temperatures. Apoat?, Piat? and, to a lower extent, Catuaí, were able to cope with gradual temperature decrease through an enhanced lipid biosynthesis. After acclimation, Piat? and Catuaí showed a lowering of digalactosyldiacylglycerol to monogalactosyldiacylglycerol ratio (MGDG/DGDG) as a result of enhanced DGDG synthesis, which represents an increase in membrane stability. The same was observed in Apoat? after chilling, in spite of phospholipids decrease. The studied parameters clearly indicated that chilling induced irreversible membrane damage in C. dewevrei. We also concluded that increased lipid synthesis, lower MGDG/DGDG ratio, and changes in membrane unsaturation occurring during acclimation to low temperatures may be critical factors in maintenance of cellular integrity under chilling.     

A Comparison of Leek (Allium porrum) and Onion (Allium cepa) Seed Development

Leek and onion seed dry weight increased exponentially for thefirst 31 days after flowering (DAF) but thereafter the increasein dry weight was slower. Before maximum seed dry weight wasreached at 45 DAF in onion and 59 to 66 DAF in leek, seed moisturecontent, seed oxygen uptake and conductivity of the seed steepwater fell from initially high levels. Although some seeds germinated31 DAF in both species, full germination in both was not reacheduntil 66–80 DAF. Tolerance of the seed to artificial dryingimmediately after harvesting occurred 45 DAF in onion and 74DAF in leek. Free nuclear division continued in the endospermuntil 17–22 DAF in onion and until 31–35 DAF inleek but it was not until 45 DAF in onion and 66 DAF in leekthat the embryo and endosperm filled the cavity formed by thepericarp. After formation of cell walls in the endosperm thepattern of change in cell number in both species was similar.The shrunken appearance of the seed coat in leek, which occurredearly in seed development, was associated with the period offree nuclear division in the endosperm and, in addition, thepericarp was thinner than in onion. There was no evidence thatthe shrunken seed coat early in development was associated withself as opposed to open-pollination. Allium porrum, Allium cepa, seed development, endosperm, embryo, cell number, germination, respiration, seed leachates     

Microbiological and chemical-sensory characteristics of three coffee varieties processed by wet fermentation

This work evaluated the bacterial diversity during coffee wet fermentation of the three coffee varieties—Mundo Novo (MN), Ouro Amarelo (OA), and Catuaí Vermelho (CV). Isolates were identified by polyphasic techniques: biochemical tests, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and DNA sequencing. Chemical compositions were determined by high (HPLC) and gas chromatography-mass spectrometry (GC-MS) and the roasted beans were sensorial evaluated using the cupping test. Thirty-six mesophilic bacteria and six lactic acid bacteria were identified. Lactobacillus plantarum and Leuconostoc mesenteroides were often found in all varieties. Citric acid was the acid detected in higher concentrations. The volatile profile of the green coffee beans changed during the fermentation in the tank, but more significantly, during the roasting process. These volatiles belonged to the classes of acids, alcohols, aldehydes, and hydrocarbons. Temporal dominance of sensations analysis showed sensorial sensations of acidity (OA and CV), bitterness, chocolate, nuts (MN), and sweetness (CV). The characteristics of each coffee variety were distinct, mainly in relation to total bacteria population, volatile compounds, and sensorial profile. In conclusion polyphasic methodology was efficiently done for bacteria identification; the dominant bacteria might be used for starter cultures and the chemical and sensory analyses helped to understand the changes in coffee fermentation. Our findings are relevant to future select starter bacteria for coffee processing to improve quality and standardization of quality.     

The Potential Use of a Silicon Source as a Component of an Ecological Management of Coffee Plants

Coffee is one of the most important agricultural export commodities in the world and it represents the main export from some developing countries. Therefore, the development of new methods of coffee management that improves production without causing any damage to the environment is an attractive alternative for producers. Much effort has been invested towards understanding the mode of action of compounds that can induce resistance against several pathogens without injuring the environment. Many researches have considered silicon efficient in avoiding plant pathogen penetration and development. Our aim was to verify the effect of potassium silicate and calcium/magnesium silicate in the development of coffee seedlings ( Coffea arabica cv. Mundo Novo ) as well as to evaluate the incidence of coffee leaf rust development under greenhouse conditions. The experiment was a completely randomized design with 12 treatments with 10 plants per treatment. The treatments were 0, 0.25, 1.25, 2.5, 4 and 5  μ m of Si for each source of silicon incorporated into the soil. The seedlings were inoculated with a urediniospores suspension of Hemileia vastatrix (2 mg/ml) at the seventh month after planting (six pair of leaves). Evaluations were performed by counting the number of lesions per leaf. The statistical analysis showed that the number of lesions reduced by up to 66% at the highest silicon dose when compared to the number of lesions in control plants. Infected plants were found to have a linear decrease of lesions with the increase of silicate concentration. The lowest number of lesions per leaf area was observed in plants that received 5  μ m of Si from potassium silicate. This result indicates the use of silicon as an alternative for an ecological management system for coffee disease protection.     

基于Penman-Monteith-Leuning遥感模型的西南喀斯特区域蒸散发估算

西南喀斯特地区地形起伏大、土壤保水持水性差、生态环境脆弱.准确估算区域尺度蒸散发对研究喀斯特地区植被恢复、水资源管理等具有重要意义.本文以桂西北喀斯特植被恢复区为例,基于野外实测气象和蒸散发数据,采用最小二乘法对Penman-Monteith-Leuning(PML)模型中气孔导度土壤湿度指数进行参数优化,并结合MOD15A2叶面积指数进行空间外推,实现了区域尺度上长时序蒸散发的估算.结果表明: 研究区蒸散发模拟值与实测值有较好的一致性,模型确定系数、纳什系数和均方根误差分别为0.85、0.75和1.56 mm·d^-1.蒸散发表现出季节变化特征,并与植被生长季物候特征一致,夏季蒸散量达到峰值.研究区年蒸散量在534～1035 mm,陆面蒸散发的空间分布呈现差异性,受降水的空间分布影响较大.年蒸散发可能不仅受到降水因素的影响,还与人类活动和土地利用类型有关.     

Expression of a modified ADP-glucose pyrophosphorylase large subunit in wheat seeds stimulates photosynthesis and carbon metabolism

ADP-glucose pyrophosphorylase (AGP) is the rate-limiting step in seed starch biosynthesis. Expression of an altered maize AGP large subunit (Sh2r6hs) in wheat (Triticum aestivum L.) results in increased AGP activity in developing seed endosperm and seed yield. The yield phenotype involves increases in both seed number and total plant biomass. Here we describe stimulation of photosynthesis by the seed-specific Sh2r6hs transgene. Photosynthetic rates were increased in Sh2r6hs-expressing plants under high light but not low light growth conditions, peaking at roughly 7 days after flowering (DAF). In addition, there were significant increases in levels of fructose, glucose, and sucrose in flag leaves at both 7 and 14 DAF. In seeds, levels of carbon metabolites at 7 and 14 DAF were relatively unchanged but increases in glucose, ADP-glucose, and UDP-glucose were observed in seeds from Sh2r6hs positive plants at maturity. Increased photosynthetic rates relatively early in seed development appear to be key to the Sh2r6hs enhanced yield phenotype as no yield increase or photosynthetic rate changes were found when plants were grown in a suboptimal light environment. These findings demonstrate that stimulation of biochemical events in both source and sink tissues is associated with Sh2r6hs expression.     

Mobilization of Endosperm Reserves and Uptake of Sucrose and Valine by the Cotyledons of Euphorbia lathyris L.

Upon germination, the endosperm triacylglycerols and proteinswere converted to sucrose and amino acids. During early postgerminativegrowth, the rate of sucrose and amino acid production exceededthe rate of uptake by the cotyledons. As a result, the levelsof total amino acid and sucrose in the endosperm increased;maximum levels were reached at 7 d and 10 d after imbibition(DAI), respectively. Intact seedlings were used to measure thedevelopment of valine, arginine, glutamic acid, and sucroseuptake rate throughout the course of endosperm depletion. Maximumamino acid uptake rates were measured at around 9 DAI, the highestuptake rate for sucrose was obtained at 12 DAI (just beforedepletion of the endosperm). The daily increase of sucrose andamino acid uptake could be manipulated, by replacing the endospermwith a pre-incubation solution during 1 d. The increase in sucroseuptake in vitro was equal to that measured with intact seedlingswhen the cotyledons were pre-incubated in 10 mol m^–3 sucrose.Higher sucrose concentrations reduced the increase of sucroseuptake; at 300 mol m^–3 sucrose (corresponding to the meanendosperm sucrose concentration) sucrose uptake after pre-incubationwas even lower than before. This reduction was largely counteractedwhen the pre-incubation solution was supplemented with minerals.The development of the valine uptake was hardly affected bysucrose, but was inhibited by several amino acids. Key words: Euphorbia lathyris seedling, sucrose uptake, amino acid uptake, reserve mobilization