Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant

Conditions affecting Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merr.], including seed vigor of explant source, selection system, and cocultivation conditions, were investigated. A negative correlation between seed sterilization duration and seed vigor, and a positive correlation between seed vigor and regenerability of explants were observed in the study, suggesting that use of high vigor seed and minimum seed sterilization duration can further improve transformation efficiency. Selection schemes using glufosinate or bialaphos as selective agents in vitro were assessed. Glufosinate selection enhanced soybean transformation as compared to bialaphos. The use of 6 mg L^-1 glufosinate during shoot induction and shoot elongation stages yielded higher final transformation efficiency ranging from 2.0% to 6.3% while bialaphos at 4 to 6 mg L^-1 gave 0% to 2.1% efficiency. Including cysteine and DTT during cocultivation increased the transformation efficiency from 0.2–0.9% to 0.6–2.9%. This treatment also improved T-DNA transfer as indicated by enhanced transient GUS expression. Shoot regeneration and Agrobacterium infection were attained in twelve soybean cultivars belonging to maturity groups I-VI. These cultivars maybe amenable to genetic transformation and may provide a valuable tool in soybean improvement programs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of different pesticides on microbial populations,nitrogen mineralisation,and thiosulphate oxidation in the rhizosphere of jute (Corchorus capsularis L. cv.)

Summary A field experiment was carried out to investigate the effects of three pesticides (a herbicide, Basalin, and two fungicides, Dithane M-45 and Bengard), separately or in combination, on jute rhizosphere microflora (total bacteria, ammonifying bacteria, aerobic non-symbiotic N₂-fixing bacteria, thiosulphate-oxidising bacteria, actinomycetes, and fungi), N mineralisation, and thiosulphate oxidation in soil. The pesticide treatments had significantly different effects on the rhizosphere microflora and their activities, depending on the kind of pesticide, the mode of application (individually or in combination), and the stage of growth of the jute plants. Although the pesticides hindered growth in the early stages, later stages were not affected because the effects on the rhizosphere microflora subsided. In addition, the combination of the herbicide Basalin (a dinitroaniline compound) and the fungicide Bengard (a benzimidazole derivative) virtually cancelled out each other's effects.     

Sedimentary records and luminescence chronology of Late Holocene palaeofloods in the Luni River, Thar Desert, northwest India

River Luni is the only well-integrated river system in the Thar Desert of India. This river catastrophically flooded due to unusually heavy rainfall in the catchment area during July 1979. In order to establish whether floods of this magnitude have occurred in the recent geological past, sedimentary records of palaeofloods occurring in the Sindari Gorge, in the lower Luni Basin were investigated. The principal slackwater flood deposits were observed in a back-flooded tributary near Bhuka. Analysis of the slackwater flood deposits preserved in the back-flooded tributary and their luminescence dating suggests that the Luni River has experienced at least 17 extreme floods during the past millennium. Evidence from the Bhuka site also suggests that no floods comparable in magnitude to the July 1979 megaflood have occurred during this period. This observation is in conformity with the palaeoflood record of central India. Comparison of the long-term monsoon rainfall series for the Luni Basin and the Indian region reveals a clear link between the two, and indicates that the clustering of large floods in the last few decades and during the Medieval warming period is a regional phenomenon associated with wetter conditions. This correlates with a regionally extended episode of landscape stability denoted by stabilization of dunes in this region.     

Phosphate-solubilizing rhizobacteria enhance the growth and yield but not phosphorus uptake of canola (Brassica napus L.)

The ability of phosphate-solubilizing rhizobacteria to enhance the growth and phosphorus uptake of canola (Brassica napus L., cv. Legend) was studied in potted soil experiments in the growth chamber. One hundred and eleven bacteria isolated from the rhizosphere of field-grown plants, and a collection of nine bacteria known to be effective plant growth-promoting rhizobacteria (PGPR), were screened for P-solubilization in vitro. All rhizobacteria were identified using whole-cell fatty acids methyl ester (FAME) profiles. The best P-solubilizing isolates were two Bacillus brevis strains, B. megaterium, B. polymyxa, B. sphaericus, B. thuringiensis, and Xanthomonas maltophilia (PGPR strain R85). The P-solubilizers were tested for their effects on growth and P-uptake of canola plants in a P-deficient soil amended with rock phosphate. Although some of the P-solubilizing rhizobacteria significantly increased plant height or pod yield, none increased P-uptake. The most effective inoculant was a B. thuringiensis isolate which significantly increased the number and weight of pods and seed yield without rock phosphate. Xanthomonas maltophilia increased plant height, whereas the other bacilli increased the number on weight of pods. These results demonstrate the potential use of these P-solubilizing rhizobacteria as inoculants for canola, but indicate that P-solubilization was not the main mechanism responsible for positive growth response. Received: 8 February 1996     

Multiresidue pesticide analysis of agricultural commodities using acetonitrile salt-out extraction, dispersive solid-phase sample clean-up, and high-performance liquid chromatography-tandem mass spectrometry

A multiresidue method analyzing 209 pesticides in 24 agricultural commodities has been developed and validated using the original Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) procedure and high performance liquid chromatography-positive electrospray ionization-tandem mass spectrometry (LC-MS/MS) analysis. Using solvent-only calibration standards (SOCSs) and matrix-matched calibration standards (MMCSs), it was demonstrated that a minimal concentration of 5-10 μg/kg (part per billion, ppb) of analytes in matrix is required for the consistent identification of targeted pesticides with two MRM transitions. Method performance was validated by the precision and accuracy results obtained from fortification studies at 10, 25, 100, and 500 ppb and MMCSs. The method was demonstrated to achieve an average recovery of 100 ± 20% (n = 4) for >75% of evaluated pesticides at the low fortification level (10 ppb) and improved to >84% at the higher fortification concentrations in all 24 matrices. Matrix effects in LC-MS/MS analysis were studied by evaluating the slope ratios of calibration curves (1.0-100 ng/mL) obtained from the SOCSs and MMCSs. Principal component analysis (PCA) of LC-MS/MS and method validation data confirmed that each matrix exerts its specific effect during the sample preparation and LC-MS/MS analysis. The matrix effect is primarily dependent on the matrix type, pesticide type and concentration. Some caution is warranted when using matrix matched calibration curves for the quantitation of pesticides to alleviate concerns on matrix effects. The QuEChERS method with LC-MS/MS was used to identify and quantitate pesticides residues, with concentrations ranging from 2.5 to >1000 ppb in a variety of agricultural samples, demonstrating fitness for screening and surveillance applications.     

Dissipation and distribution behavior of azoxystrobin, carbendazim, and difenoconazole in pomegranate fruits

The dissipation behavior and degradation kinetics of azoxystrobin, carbendazim, and difenoconazole in pomegranate are reported. Twenty fruits/hectare (5 kg) were collected at random, ensuring sample-to-sample relative standard deviation (RSD) within 20-25%. Each fruit was cut into eight equal portions, and two diagonal pieces per fruit were drawn and combined to constitute the laboratory sample, resulting in RSDs <6% (n = 6). Crushed sample (15 g) was extracted with 10 mL of ethyl acetate (+ 10 g Na(2)SO(4)), cleaned by dispersive solid phase extraction on primary secondary amine (25 mg) and C(18) (25 mg), and measured by liquid chromatography tandem mass spectrometry. The limit of quantification was ≤0.0025 μg g(-1) for all the three fungicides, with calibration linearity in the concentration range of 0.001-0.025 μg mL(-1) (r(2) ≥ 0.999). The recoveries of each chemical were 75-110% at 0.0025, 0.005, and 0.010 μg g(-1) with intralaboratory Horwitz ratio <0.32 at 0.0025 μg g(-1). Variable matrix effects were recorded in different fruit parts viz rind, albedo, membrane, and arils, which could be correlated to their biochemical constituents as evidenced from accurate mass measurements on a Q-ToF LC-MS. The residues of carbendazim and difenoconazole were confined within the outer rind of pomegranate; however, azoxystrobin penetrated into the inner fruit parts. The dissipation of azoxystrobin, carbendazim, and difenoconazole followed first + first order kinetics at both standard and double doses, with preharvest intervals being 9, 60, and 26 days at standard dose. At double dose, the preharvest intervals extended to 20.5, 100, and 60 days, respectively.     

Speciation of Metals in Sewage Sludge and Sludge-amended Soils

This study is aimed at ascertaining the chemical partitioning ofCu, Zn, Pb, Ni, Cr and Cd in sewage sludge and agricultural soils repeatedly amended with sludge. The operationally determined speciation was investigated by using selective five steps sequential extraction method for partitioning these metals into different chemicalforms, likely to be released in solution under various environmental conditions. It provides qualitative evidence regarding the form of the association of metals and indirectly of their bioavailability. It can help to explain the process by which metals are eliminated from sewagesludge and also indicate the impact of the use of sludge on agriculturalsoils, as amendments. Data obtained by extraction procedure showed different metal distribution trend among the fractions in sewage sludgeand sludge-amended soils. Comparison of distribution pattern of metals in sludge and sludge-applied soils shows that there is possible redistribution of metals among the different phases. The sum of the metal content in the fractions compared well with the total metal content in sewage sludge, the ratios averaging 1.02, 1.04, 1.07, 0.94, 1.06, and 1.12 for Cu, Zn, Pb, Ni, Cr, and Cd, respectively. The metal recovery efficiency in sludge-amended soils was found to be: 108±6%, 102±5%, 115±8%, 111±4%, 104±7%and 124±28% for Cu, Zn, Pb, Ni, Cr and Cd, respectively.     

Efficient genomic selection using ensemble learning and ensemble feature reduction

Journal of Crop Science and Biotechnology - Genomic selection (GS) is a popular breeding method that uses genome-wide markers to predict plant phenotypes. Empirical studies and simulations have...