Analysis of the gacS-gacA regulatory genes of spontaneous mutants of Pseudomonas fluorescens biocontrol strain PfG32R

Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS. The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system. The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365.     

Distribution and diversity of leaf-cutting ants in Northeastern Argentina: species most associated with forest plantations

Abstract

Leaf-cutting ants (LCA) are considered one of the main herbivores and one of the most destructive pest insects of the Neotropics. Northeastern Argentina harbors the greatest species richness of these ants and in turn comprises the highest surface with forest plantations. Our aim was to establish which species of leaf-cutting ants are most commonly associated with forest plantations by analyzing their geographic distribution using published and unpublished species occurrence data. Also, estimate their potential areas of distribution along a latitudinal gradient that entirely encompasses northeastern Argentina using Ecological Niche Modeling. Only seven of the 20 species recorded were strongly associated with productive systems along the gradient, but only 2–3 species in each region could be considered high-risk species for forest plantations. High-risk species composition shows a turnover between regions. Our models show the potential distribution areas where LCA could become more abundant and dominant, and possibly causing a detrimental effect on the forest plantations in the studied region. We find that ecological niche models are useful tools to assess the environmental suitability of important LCA.     

黑龙江省大豆疫霉根腐病调查与病原分离

１９９６年对黑龙江省东部和中部大豆产区２３个市、县的大豆苗期疫霉根腐病进行了调查、研究,应用ＰＢＮＩＣ疫霉选择性培养基分离大豆疫霉根腐病病原菌,从牡丹江、穆棱、林口和佳木斯豆田具疫霉根腐症状的大豆植株上分离到大豆疫霉根腐病菌,并从根腐病株上单独或与大豆疫霉菌同时分离到终极腐霉菌,研究进一步证实我国黑龙江省有大豆疫霉根腐病。调查发现,大豆疫霉根腐病和终极腐霉根腐病主要发生在土质粘重、土壤含水量高或易积水的田块。     

Storage performance of clementines treated with hot water,sodium carbonate and sodium bicarbonate dips

Summary

The potential of using hot water (2.5 min at 45°C), 2% sodium carbonate or 2% sodium bicarbonate solutions alone or combined with hot water, for control of Penicillium digitatum (green mould) was investigated on commecially ripe clementines. The effect of such treatments on the post-harvest storage life of clementines was determined during two months analysing weight loss, increase in rind deformation and internal maturity. Carbonate and bicarbonate solutions effectively control green mould during storage but hot water does not. Compared with the control and hot-water treated fruits, clementines treated with carbonate and, to a lesser extend with bicarbonate, showed higher weight loss and lower firmness during storage. In contrast, lower increase in ripening index was found when the fruits were treated with carbonate or bicarbonate. Fruits treated with hot water alone or combined with carbonate exhibited no change in ripening during the first month of storage. Rinsing the fruit significantly reduces weight and firmness loss in the fruits treated with carbonate and bicarbonate, but did not change the ripening index. The primary finding of this work was that carbonate and bicarbonate treatments might significantly increase the loss in weight and firmness during storage. Rinsing the fruit immediately after treatment appeared to be a simple solution to avoid these losses in quality.     

Effects of shading and NO3:NH4 ratio on the yield,quality and N metabolism in strawberry

The effect of 50% shading and NO₃:NH₄ ratio (0:100, 75:25, 50:50, and 25:75) in the nutrient solution on growth, yield, quality and N metabolism in hydroponically grown strawberry (Fragaria × ananassa var Camarosa) was evaluated. Both fresh and dry weights of leaves were significantly lower when a high concentration of either NO₃ (100%) or NH₄ (75%) was the sole N source in the nutrient solution. In unshaded plants, increasing of both NH₄ and NO₃ ratio in the nutrient solution reduced photosynthetic (Pn) rate, however in shaded plants the reduction of Pn became more pronounced at a higher ratio of NH₄ in the nutrient solution. The yield in terms of fresh and dry weight of fruit per plant was significantly increased at the 75:25 and 50:50 (NO₃:NH₄) treatments. Fruit size was significantly affected by the treatments, so that the biggest fruits in both shaded and unshaded plants were obtained under the 75:25 and 50:50 (NO₃:NH₄) treatments. Total soluble solid (TSS) in unshaded plants was increased with increasing NH₄ ratio in the nutrient solution, however in shaded plants it was reduced at high NH₄ ratio in the nutrient solution. In both shaded and unshaded plants, higher concentration of NH₄ significantly reduced the post-harvest life of the fruits. The increase of tissue N concentration was nearly proportional to the NH₄ concentration in the nutrient solution. The activity of nitrate reductase (NR) was increased by increasing NH₄ from 0 to 50% and then reduced at a higher ratio of NH₄ in the solution. Shading increased NH₄ concentration so that the shaded plant had nearly twice as high NH₄ concentration in the leaves. The increase of NH₄ concentration induced by shading could be partially the reduction of NH₄ assimilate because of the shortage of carbohydrate.     

德国核桃‘No.120'幼胚胚轴与子叶体细胞胚胎发生及其植株再生

核桃 (Juglansregia)‘No . 12 0’品系授粉 7～ 8周的幼胚胚轴与子叶在含不同植物生长调节剂组合的诱导培养基上培养 4周后转移至无植物生长调节剂的继代培养基上培养 ,不同发育时期的体细胞胚直接形成于胚轴与子叶表面。子叶与胚轴分别在KT 2mg/L BA1mg/L IBA 0 .0 1mg/L与 2 ,4 D 2mg/L KT 1mg/L的组合的诱导下体细胞胚胎发生频率最高。体细胞胚经 3～ 5℃低温处理 2个月 ,31.6%的体细胞胚能发育成正常生长的植株     

Regional scale analysis of denitrification in north temperate forest soils

Large scale analyses of biogeochemical processes are necessary for understanding anthropogenic effects on global climate and environmental quality. Regional scale estimates of denitrification from forest soils in southern lower Michigan USA were produced by stratifying the region into landscape experimental units using soil texture and natural drainage classes, and extrapolating data to larger areas using a geographic information system (GIS). Previous landscape-scale research established relationships between soil texture and drainage and denitrification and quantified annual denitrification N loss in nine soil texture/drainage groups. All forest soils within the region (64 series) were assigned to one of these nine groups based on their texture and drainage characteristics and were assigned an annual denitrification N loss value. A regional estimate of denitrification was produced by multiplying the areal extent of each of the nine soil groups by their annual denitrification N loss value. Loam-textured soils underlie 47% of the regional forest and accounted for 73% of the forest denitrification. Sandy soils were found under 44% of the regional forest but produced only 5% of the regional denitrification. Clay loam soils underlie 9% of the regional forest and produced 22% of the denitrification. Annual denitrification N loss for the region was estimated as 1.4×10⁷ kg N/yr. We used denitrification enzyme activity (DEA) as a proxy for annual denitrification N loss to determine if the relationship between denitrification and soil texture and natural drainage that we observed at the landscape scale held up at the regional scale. DEA was measured in 22 soils across the region and was strongly related to soil texture and natural drainage (r²=0.61), suggesting that extrapolation of data from the landscape to the regional scale was justified.     

Removal of flowers or inflorescences affects ‘Barnea’ olive fruitlet post-anthesis abscission

A typical olive (Olea europaea L.) inflorescence consists of about 20 flowers. However, in many cultivars, only one fruit develops. This is due to massive abscission of flowers and fruitlets, which occurs during the first month after anthesis. In this study, we used the olive cultivar 'Barnea' to characterize the abscission mechanism and to try to increase fruit set by increasing the number of developed fruit per inflorescence. Removing the lateral flowers 3 weeks before anthesis increased fruit set by more than 50%. Removing all inflorescences but one from a branch increased the number of developed fruits from 0.93 to 2.8 during 2017 and from 0.91 to 3.34 fruits per inflorescence in 2018. Sugar quantification in the pistil revealed that starch level is high on the day of anthesis and low 25 days later in abscised as well as in developed fruit. Soluble carbohydrates are low on the day of anthesis, low in abscised flowers/fruitlets 25 days after anthesis and high in developed fruit. Screening the natural variation found in the Israeli germplasm collection revealed that in most cultivars less than one fruit per inflorescence has developed. However, there are unique cultivars with a higher fruit set.     

Evaluation of non-chemical seed treatment methods for the control of Alternaria dauci and A. radicina on carrot seeds

The current study was initiated to evaluate the efficacy of physical methods (hot water, aerated steam, electron treatment) and agents of natural origin (resistance inducers, plant derived products, micro-organisms) as seed treatments of carrots for control of Alternaria dauci and A. radicina. Control of both Alternaria species by seed treatment with the resistance inducers was generally poor. Results were also not satisfactory with most of the formulated commercial micro-organism preparations. Based on the average of five field trials, one of these, BA 2552 (Pseudomonas chlororaphis), provided a low but significant increase in plant stand. Among the experimental micro-organisms, the best results were obtained with Pseudomonas sp. strain MF 416 and Clonostachys rosea strain IK726. A similar level of efficacy was provided by seed treatment with an emulsion (1%) of thyme oil in water. Good and consistent control was generally achieved with the physical methods aerated steam, hot water and electron treatment. Aerated steam treatment was, apart from the thiram-containing chemical standard, the best single treatment, and its performance may at least partially be due to extensive pre-testing, resulting in dosages optimally adapted to the respective seed lot. In some of the experiments the effect of the hot water treatment, which was tested at a fixed, not specifically adapted dosage, was significantly improved when combined with a Pseudomonas sp. MF 416 or C. rosea IK726 treatment. The results are discussed in relation to the outcome of experiments in which the same seed treatment methods and agents were tested in other seed-borne vegetable pathosystems.     

Intra-specific variation and inheritance of BYDV-PAV transmission in the aphid <Emphasis Type="Italic">Sitobion avenae</Emphasis>

Vector efficiency of 44 clonal lines (clones) of Sitobion avenae belonging to 31 different genotypes (distinct patterns for five microsatellite loci) originating from Western France was evaluated by transmitting the isolate PAV4 of BYDV-PAV to barley seedlings. Variation in transmission rates from 3.7% to 92.5% was observed, with significant effects of the aphid clone, of the plant species on which clones were collected, and of the reproductive mode of the clones. When genotypes are considered instead of clones, a continuum in transmission rates was observed. A subset of S. avenae clones was tested for transmission of one (10 clones) and 13 (4 clones) other BYDV-PAV isolates, and a clear clone effect modulated by an isolate effect was observed. Crosses were made between clones with different vectoring phenotypes and their F1 progeny were tested for PAV4 transmission. The narrow sense heritability of the PAV transmission character was rather high in the F1 families (h²=0.5) and the segregation analyses suggested an oligo/polygenic determinism of this character. The possibility of generating new transmission variants by sexual reproduction and its consequences on transmission mechanism studies and on BYD epidemics are discussed.