Identification of α-amylase by random and specific mutagenesis of Texcoconibacillus texcoconensis 13CCT strain isolated from extreme alkaline–saline soil of the former Lake Texcoco (Mexico)

The alkaline α-amylase produced by Texcoconibacillus texcoconensis 13CC^T strain was identified by random mutagenesis and confirmed by directed mutagenesis. A transposon mutagenesis approach was taken to identify the gene responsible for the degradation of starch in T. texcoconensis 13CC^T strain. The deduced amino acids of the amy gene had a 99 % similarity with those of Bacillus selenitireducens MLS10 and 97 % with those of Paenibacillus curdlanolyticus YK9. The enzyme showed a maximum activity of 131.1 U/mL at 37 °C and pH 9.5 to 10.5. In situ activity of the enzyme determined by polyacrylamide gel electrophoresis showed only one band with amylolytic activity. This is the first report of a bacterium isolated from the extreme alkaline–saline soil of the former Lake Texcoco (Mexico) with amylolytic activity in alkaline conditions while its potential as a source of amylases for the industry is discussed.     

Connective Tissue Growth Factor Is Involved in Structural Retinal Vascular Changes in Long-Term Experimental Diabetes

Early retinal vascular changes in the development of diabetic retinopathy (DR) include capillary basal lamina (BL) thickening, pericyte loss and the development of acellular capillaries. Expression of the CCN (connective tissue growth factor/cysteine-rich 61/nephroblastoma overexpressed) family member CCN2 or connective tissue growth factor (CTGF), a potent inducer of the expression of BL components, is upregulated early in diabetes. Diabetic mice lacking one functional CTGF allele (CTGF^+/−) do not show this BL thickening. As early events in DR may be interrelated, we hypothesized that CTGF plays a role in the pathological changes of retinal capillaries other than BL thickening. We studied the effects of long-term (6-8 months) streptozotocin-induced diabetes on retinal capillary BL thickness, numbers of pericytes and the development of acellular capillaries in wild type and CTGF^+/− mice. Our results show that an absence of BL thickening of retinal capillaries in long-term diabetic CTGF^+/− mice is associated with reduced pericyte dropout and reduced formation of acellular capillaries. We conclude that CTGF is involved in structural retinal vascular changes in diabetic rodents. Inhibition of CTGF in the eye may therefore be protective against the development of DR.     

Enhanced dissipation of polycyclic aromatic hydrocarbons in the rhizosphere of the Athel tamarisk (Tamarix aphylla L. Karst.) grown in saline-alkaline soils of the former lake Texcoco

Remediation of polycyclic aromatic hydrocarbons (PAHs) contaminated alkaline saline soil with phreatophyte or "water loving plants" was investigated by spiking soil from the former lake Texcoco with 100 mg phenanthrene (Phen) kg(-1) soil, 120 mg anthracene (Ant)kg(-1) soil and 45 mg benzo(a)pyrene (BaP) kg(-1) soil and vegetating it with Athel tamarisk (Tamarix aphylla L Karst.). The growth of the Athel tamarisk was not affected by the PAHs. In soil cultivated with Athel tamarisk, the leaching of PAHs to the 32-34 cm layer decreased 2-fold compared to the uncultivated soil. The BaP concentration decreased to 39% of the initial concentration at a distance smaller than 3 cm from the roots and to 45% at a distance larger than 3cm, but 59% remained in unvegetated soil after 240 days. Dissipation of Ant and Phen decreased with depth, but not BaP. The biodegradation of PAHs was affected by their chemical properties and increased in the presence of T. aphylla, but decreased with depth.     

Optimization of growth regulators and silver nitrate for micropropagation of <Emphasis Type="Italic">Dianthus caryophyllus</Emphasis> L. with the aid of a response surface experimental design

This paper reports on the optimum concentrations of naphthalene acetic acid (NAA) and 6-benzyladenine (BA) to stimulate callus growth and NAA; kinetin and silver nitrate (AgNO₃) for callus redifferentiation in Dianthus caryophyllus L. Meristems were excised and placed in MS medium with 30 g l⁻¹ sucrose and 9.0 μM 2,4-d. Callus clusters were transferred to MS medium containing NAA (0, 1.7, 3.3, and 5.0 μM) and BA (0, 1.7, 3.3, and 5.0 μM) for proliferation and to MS medium with 30 g l⁻¹ sucrose, 2.5 g l⁻¹ phytagel, kinetin (0, 33, and 66 μM); NAA (0, 7.95, and 15.9 μM) and AgNO₃ (0, 23.54 and 47.08 μM) for shoot and root induction. Treatments were applied according to a Box–Behnken design. After callus growth and redifferentiation, plants were incubated in the greenhouse at 18 ± 2°C for 4 wk and at 20–26°C for 4 wk. Finally, plants were changed to near-commercial greenhouse conditions with different day (30–35°C) and night (16–24°C) temperatures. Results showed better callus growth at higher NAA concentrations. A maximum callus weight was found with 5.0 μM NAA but without BA. A maximum of 78% calluses with shoots was obtained with 15.9 μM NAA, 47.08 μM AgNO₃, and 0.74 μM kinetin and 58% with roots with 15.7 μM NAA and 47.08 μM AgNO₃, but without kinetin. The shoots obtained showed little hyperhydricity. Vigorous plants were obtained after gradual acclimatization with an 80% survival rate under nursery conditions.     

Selective production of bikaverin in a fluidized bioreactor with immobilized Gibberella fujikuroi

The best culture medium composition for the production of bikaverin by Gibberella fujikuroi in shake-flasks, i.e. 100 g glucose l^–1; 1 g NH₄Cl l^–1; 2 g rice flour l^–1; 5 g KH₂PO₄ l^–1 and 2.5 g MgSO₄ l^–1, was obtained through a fractional factorial design and then scaled-up to a fluidized bioreactor. The effects of carbon and nitrogen concentrations, inoculum size, aeration, flow rate and bead sizes on batch bikaverin production using immobilized G. fujikuroi in a fluidized bioreactor were determined by an orthogonal experimental design. Concentrations of up to 6.83 g bikaverin l^–1 were obtained when the medium contained 100 g glucose l^–1 and 1 g NH₄Cl l^–1 with an inoculum ratio of 10% v/v, an aeration rate of 3 volumes of air per volume of medium min^–1, and a bead size of 3 mm. Based on dry weight, the bikaverin production was 30–100 times larger than found in submerged culture and approximately three times larger than reported for solid substrate fermentation.     

Aggregation and C and N contents of soil organic matter fractions in a permanent raised-bed planting system in the Highlands of Central Mexico

Permanent raised bed planting with crop residue retention is a form of conservation agriculture that has been proposed as an alternative to conventional tillage for wheat production systems in the Central Highlands of Mexico. A field experiment comparing permanent and tilled raised beds with different residue management under rainfed conditions was started at El Batán (State of Mexico, Mexico) in 1999. The percentage of small and large macroaggregates and mean weight diameter (MWD) was significantly larger in permanent raised beds compared to conventionally tilled raised beds both with full crop residue retention (average for maize and wheat), while the percentages free microaggregates was lower. The percentages of small and large macroaggregates and mean weight diameter (MWD) was significantly larger in permanent raised beds with residue retention compared to permanent raised beds with removal of the residue (average for maize and wheat), while the percentages free microaggregates and silt and clay fraction was lower. Cultivation of maize significantly reduced the large macroaggregates, while wheat reduced the silt and clay fraction (average over all systems). Cultivation of maize reduced the C and N content of the free microaggregates compared to soil cultivated with wheat, while removal of plant residue reduced the C and N content of the silt and clay fraction compared to soil where residue was retained. The C and N content of the coarse particulate organic matter (cPOM) and microaggregates within the macroaggregates was significantly larger in permanent raised beds compared to conventionally tilled raised beds both with full residue retention, while C and N content of the cPOM was significantly lower when residue was removed or partially removed compared to the soil where the residue was retained. The δ ¹³C ‰ signatures of the macroaggregates, microaggregates, the silt and clay fraction, cPOM and microaggregates within the macroaggregates were not affected by tillage or residue management when wheat was the last crop, but removal of residue reduced the δ ¹³C ‰ signatures of the macro-, microaggregates and microaggregates within the macroaggregates significantly compared to soil where the residue was retained. Retaining only 30–50% of the organic residue still improved the soil structure considerably compared to plots where it was removed completely. Permanent raised beds without residue retention, however, is a practice leading to soil degradation. Kelly Lichter and Bram Govaerts contributed equally to this publication.     

A method to evaluate the isothermal effectiveness factor for dynamic oxygen into mycelial pellets in submerged cultures

Several models have been developed simulating O2 transfer in bioreactors, but three limitations are often found: (i) an inadequate kinetic representation of O2 consumption or wrong boundary conditions, (ii) unrealistic parameter values, and (iii) inadequate experimental systems. In our study we minimized those possible sources of error. Oxygen uptake rate, void fraction of the pellet, and external O2 mass transfer coefficient were experimentally obtained from bioreactor studies in which pellets of Gibberella fujikuroi were naturally formed. Michaelis-Menten kinetics and diffusion equations were used to describe the O2 consumption rate and to evaluate the effectiveness factor in dynamic mode. The nonlinear mathematical model proposed was solved by the orthogonal collocation technique. The O2 consumption rate in pellets of G. fujikuroi of 1.7-2.0 mm is only marginally inhibited by diffusion constraints under conditions tested. Simulation analysis showed that the effectiveness factor decreased as the Thiele modulus and pellet diameter increased. The proposed model was applied to experimental data reported for other fungal pellets and allowed to predict optimal conditions for O2 transfer into mycelial pellets.     

Evaluating spatial within plot crop variability for different management practices with an optical sensor?

Subsoil constraints to root growth exacerbate frequent water and nutrient limitations to crop yields in Mediterranean-type environments. Amelioration of subsoil constraints can relieve these limitations by opening root-access to subsoil water and nutrients. However, decisions in subsoil amelioration are hampered by seasonally variable yield responses in these environments. We used the APSIM model to analyse the impact of subsoil constraints on yield and yield variability. The simulated yield data were used to calculate the financial benefits of subsoil amelioration across several scenarios. There was a strong yield-dependence on accessible soil water governed by root depth. Root depth development was limited to a minimum of either the effect of subsoil constraints or the weather-dependent depth of the soil wetting front. Insufficient rainfall in dry years or in a drier region often resulted in shallow soil wetting fronts and correspondingly shallow roots even in the absence of subsoil compaction. In these situations, there is little response to subsoil amelioration. Positive yield responses and positive financial returns to subsoil amelioration are therefore greater in good rainfall years and are more likely in a wetter region. A yield response to amelioration is also greater in coarser textured sand than finer textured sandy loam in an average rainfall season because the same amount of rainfall results in a deeper wetting front in sand than in sandy loam. Hence, roots in a sand are required to grow deeper compared to a sandy loam to access the same amount of water and therefore benefited more from subsoil amelioration in an average rainfall year. In wet years, sands leach more nitrate than sandy loam, which decreases yields and the response to subsoil amelioration in sands is more than in the sandy loam. Environmental threats occur along with yield loss when roots cannot access subsoil water. These include increased nitrate leaching and deep drainage due to unused water remaining in the soil profile. By allowing roots to access deep soil water, ameliorating subsoil is expected to yield financial gains in average to good rainfall seasons and decrease the environmental risk of drainage and leaching loss. The financial gains are expected to offset potential financial losses in dry and dry finish seasons especially in coarser textured soils and wetter environment. Responsible Editor: Jan Vos.