A comparative study of Taxol production in liquid and solid‐state fermentation with Nigrospora sp. a fungus isolated from Taxus globosa

Aims: To determine in liquid (LF) and solid‐state fermentation (SSF) the effect of medium concentration on growth and Taxol produced by Nigrospora sp., a fungus isolated from the Mexican yew. Methods and Results: Nigrospora sp. was grown at different concentrations of the base culture medium M1D, i.e. two (2×), four (4×), six (6×) and eight times (8×) the base concentration. The titres of Taxol determined by competitive inhibition enzyme immunoassay increased with increasing medium concentration in LF and SSF but were higher in SSF in every medium concentration. The Taxol produced in SSF and LF with 8× medium was 221 and 142 ng l^?1. The SSF gave also higher biomass, growth and sugar utilization than LF in every medium. The growth and sugar consumption were modelled by the logistic and the Pirt models, respectively. However, the Luedeking–Piret model was unsuitable for Taxol. Conclusions: The SSF surpassed LF in terms of Taxol, growth and sugar utilization; thus, it has significant advantages over LF. Significance and Impact of the Study: This is the first report on Taxol production by SSF and the first contribution to evaluate the influence of the medium on Taxol production in LF and SSF.     

Influence of catclaw Mimosa monancistra on the dissipation of soil PAHs

Phytoremediation is a cost-effective biotechnology for decontamination of polycyclic aromatic hydrocarbons (PAHs)-polluted soils. A greenhouse experiment was conducted to determine the growth of Mimosa monancistra, a N2-fixing leguminous plants, and its capacity to remove phenanthrene, anthracene, and benzo(a)pyrene (BaP)from soil. The PAHs decreased shoot and root dry biomass of M. monancistra 2.7- and 3.9-fold, respectively, compared to uncontaminated soil and inhibited nodule formation. The removal of phenanthrene and anthracene was similar in vegetated and unvegetated soil, but the dissipation of BaP was significantly faster in vegetated soil as compared to unvegetated soil after 14, 56, 70, and 90 d. After 90 d, dissipation of BaP was 96% in vegetated soil and 87% in unvegetated soil. Nitrification and ammonification were not affected by the addition of PAHs as concentrations of NH4+, NO2-, and NO3- were similar in contaminated and uncontaminated vegetated soil. Growth of M. monancistra was inhibited by contamination with hydrocarbons, but removal of BaP was accelerated in the rhizosphere.     

Optimization of <Emphasis Type="Italic">Renealmia mexicana</Emphasis> (Klotzsch ex. Petersen) cultivation <Emphasis Type="Italic">in vitro</Emphasis>

Renealmia mexicana (Klotzsch ex. Petersen) is a tropical plant found in southern México with an ornamental value and a potential source of curcuminoids. Its distribution in Chiapas has decreased because of deforestation and low propagation and germination rate, so a protocol for in vitro propagation was developed. An orthogonal experimental design of L₉ (3⁴) in triplicate was used to investigate the effect of 6-benzyl adenine (BA), indole butyric acid (IBA), silver nitrate (AgNO₃), and sucrose on shoot, root, and leaf development of plantlets grown in vitro. Plantlets with well-developed shoots and roots were transferred to pots containing a mixture of peat moss and agrolite for hardening before transfer to soil. The Murashige and Skoog (Physiol. Plant. 15:473–497, 1962) mineral medium (MS) supplemented with 4.4 μM BA, 2.5 μM IBA, 11.7 μM AgNO_3y and 5.5% (w/v) sucrose gave most shoots, 8.9 μM BA, 2.5 μM IBA, 17.7 μM AgNO₃ and 5.5% (w/v) sucrose most roots, and 8.9 μM BA, 4.9 μM IBA, 11.7 μM AgNO₃ and 3.0% (w/v) sucrose most leaves, although other combinations were statistically equivalent in each case. Sucrose was the factor that most explained the variation in the promotion of shoots, roots, and leaves. The protocol developed resulted in up to 100% survival when plantlets were transferred to soil using AgNO₃, confirming that hardening of plantlets in vitro using hormonal stimulation was a suitable strategy to improve acclimatization.     

Phylogenetic analysis of the archaeal community in an alkaline-saline soil of the former lake Texcoco (Mexico)

The soil of the former lake Texcoco is an extreme environment localized in the valley of Mexico City, Mexico. It is highly saline and alkaline, where Na⁺, Cl⁻, HCO₃⁻ and CO₃²⁻ are the predominant ions, with a pH ranging from 9.8 to 11.7 and electrolytic conductivities in saturation extracts from 22 to 150 dS m⁻¹. Metagenomic DNA from the archaeal community was extracted directly from soil and used as template to amplify 16S ribosomal gene by PCR. PCR products were used to construct gene libraries. The ribosomal library showed that the archaeal diversity included Natronococcus sp., Natronolimnobius sp., Natronobacterium sp., Natrinema sp., Natronomonas sp., Halovivax sp., “Halalkalicoccus jeotgali” and novel clades within the family of Halobacteriaceae. Four clones could not be classified. It was found that the archaeal diversity in an alkaline-saline soil of the former lake Texcoco, Mexico, was low, but showed yet uncharacterized and unclassified species. César Valenzuela-Encinas and Isabel Neria-González contributed equally to this publication.     

Bacterial colonization of a fumigated alkaline saline soil

After chloroform fumigating an arable soil, the relative abundance of phylotypes belonging to only two phyla (Actinobacteria and Firmicutes) and two orders [Actinomycetales and Bacillales (mostly Bacillus)] increased in a subsequent aerobic incubation, while it decreased for a wide range of bacterial groups. It remained to be seen if similar bacterial groups were affected when an extreme alkaline saline soil was fumigated. Soil with electrolytic conductivity between 139 and 157 dS m^?1, and pH 10.0 and 10.3 was fumigated and the bacterial community structure determined after 0, 1, 5 and 10 days by analysis of the 16S rRNA gene, while an unfumigated soil served as control. The relative abundance of the Firmicutes increased in the fumigated soil (52.8 %) compared to the unfumigated soil (34.2 %), while that of the Bacteroidetes decreased from 16.2 % in the unfumigated soil to 8.8 % in the fumigated soil. Fumigation increased the relative abundance of the genus Bacillus from 14.7 % in the unfumigated soil to 25.7 %. It was found that phylotypes belonging to the Firmicutes, mostly of the genus Bacillus, were dominant in colonizing the fumigated alkaline saline as found in the arable soil, while the relative abundance of a wide range of bacterial groups decreased.     

Identification of Antibiotic Resistance Cassettes in Class 1 Integrons in Aeromonas spp. Strains Isolated From Fresh Fish (Cyprinus carpio L.)

Forty-six Aeromonas spp. strains were isolated from fresh fish and investigated for their antimicrobial susceptibility, detection of Class 1 integrons by PCR, and arrangement of gene cassettes. Selected isolates were further characterized by enterobacterial repetitive intergenic consensus-PCR. Twenty isolates were found to carry Class 1 integrons. Amplification of the variable regions of the integrons revealed diverse bands ranging in size from 150 to 1,958 pb. Sequence analysis of the variable regions revealed the presence of several gene cassettes, such as adenylyl transferases (aadA2 and aadA5), dihydrofolate reductases (dfrA17 and dfrA1), chloramphenicol acetyl transferase (catB3), β-lactamase (oxa2), lincosamide nucleotidil transferase (linF), aminoglycoside-modifying enzyme (apha15), and oxacillinase (bla _OXA-10). Two open reading frames with an unknown function were identified as orfC and orfD. The aadA2 cassette was the most common integron found in this study. Interestingly, five integrons were detected in the plasmids that might be involved in the transfer of resistance genes to other bacteria. This is a first report of cassette encoding for lincosamides (linF) resistance in Aeromonas spp. Implications on the incidence of integrons in isolates of Aeromonas spp. from fresh fish for human consumption, and its possible consequences to human health are discussed.     

Influence of permanent raised bed planting and residue management on physical and chemical soil quality in rain fed maize/wheat systems

Densely populated, intensively cropped highland areas in the tropics and subtropics are prone to erosion and declining soil fertility, making agriculture unsustainable. Conservation agriculture in its version of permanent raised bed planting with crop residue retention has been proposed as an alternative wheat production system for this agro-ecological zone. A five years field experiment comparing permanent and tilled raised beds with different residue management under rainfed conditions was started at El Batán (Mexico) (2,240 m asl; 19.31°N, 98.50°W; Cumulic Phaeozem) in 1999. The objective of this study was to determine the soil quality status after five years of different management practices. The K concentration was 1.65 times and 1.43 times larger in the 0–5 cm and 5–20 cm profiles, respectively, for permanent raised beds compared to conventionally tilled raised beds. The Na concentration showed the opposite trend. Sodicity was highest for conventionally tilled raised beds and for permanent raised beds it increased with decreasing amounts of residue retained on the surface. Permanent raised beds with full residue retention increased soil organic matter content 1.4 times in the 0–5 cm layer compared to conventionally tilled raised beds with straw incorporated and it increased significantly with increasing amounts of residue retained on the soil surface for permanent raised beds. Soil from permanent raised beds with full residue retention had significantly higher mean weight diameter for wet and dry sieving compared to conventionally tilled raised beds. Permanent raised beds with full residue retention had significantly higher aggregate stability compared to those with residue removal. A lower aggregation resulted in a reduction of infiltration. Principal component analysis (PCA) was performed using these soil physicochemical variables that were significantly influenced by tillage or residue management. The PC1 and PC2 separated the conventionally tilled raised beds from the permanent raised beds and PC3 separated permanent raised beds with at least partial residue retention from permanent raised beds with no residue retention. These clear separations suggest that tillage and residue management have an effect on soil processes. The research indicates that permanent raised bed planting increases the soil quality and can be a sustainable production alternative for the (sub)tropical highlands. Extensive tillage with its associated high costs can be reduced by the use of permanent raised beds while at least partial surface residue retention is needed to insure production sustainability.     

Natronorubrum texcoconense sp. nov., a haloalkaliphilic archaeon isolated from soil of the former lake Texcoco (Mexico)

A new haloalkaliphilic archaeon, strain B4^T, was isolated from the former lake Texcoco in Mexico. The cells were Gram-negative, pleomorphic-shaped, pink to red pigmented and aerobic. Strain B4^T required at least 2.5 M NaCl for growth, with optimum growth at 3.4 M NaCl. It was able to grow over a pH range of 7.5–10.0 and temperature of 25–50 °C, with optimal growth at pH 9 and 37 °C. Cells are lysed in hypotonic treatment with less than 1.3 M NaCl. The major polar lipids of strain B4^T were phosphatidylglycerol and methyl-phosphatidylglycerophosphate. Phospholipids were detected, but not glycolipids. The nucleotide sequence of the 16S rRNA gene revealed that the strain B4^T was phylogenetically related to members of the genus Natronorubrum. Sequence similarity with Natronorubrum tibetense was 96.28 %, with Natronorubrum sulfidifaciens 95.06 % and Natronorubrum sediminis 94.98 %. The G+C content of the DNA was 63.3 mol%. The name of Natronorubrum texcoconense sp. nov. is proposed. The type strain is B4^T (=CECT 8067^T = JCM 17497^T).     

Induction of xylanases by sugar cane bagasse at different cell densities of Cellulomonas flavigena

The effect of cell density on xylanolytic activity and productivity of Cellulomonas flavigena was evaluated under two different culturing conditions: fed-batch culture with discontinuous feed of sugar cane bagasse (SCB; condition 1) and glycerol fed-batch culture followed by addition of SBC as xylanases inducer (condition 2). The enzymatic profile of xylanases was similar in both systems, regardless of the initial cell density at time of induction. However, the xylanolytic activity changed with initial cell density at the time of induction (condition 2). The maximum volumetric xylanase activity increased with increased initial cell density from 4 to 34 g l⁻¹ but decreased above this value. The largest total volumetric xylanase productivity under condition 2 (1.3 IU ml⁻¹ h⁻¹) was significantly greater compared to condition 1 (maximum 0.6 IU ml⁻¹ h⁻¹). Consequently, induction of xylanase activity by SCB after growing of C. flavigena on glycerol at intermediate cell density can be a feasible alternative to improve activity and productivity of xylanolytic enzymes.     

Conservation agriculture, increased organic carbon in the top-soil macro-aggregates and reduced soil CO2 emissions

Background and aims

Conservation agriculture, the combination of minimal soil movement (zero or reduced tillage), crop residue retention and crop rotation, might have the potential to increase soil organic C content and reduce emissions of CO₂.

Methods

Three management factors were analyzed: (1) tillage (zero tillage (ZT) or conventional tillage (CT)), (2) crop rotation (wheat monoculture (W), maize monoculture (M) and maize-wheat rotation (R)), and (3) residue management (with (+r), or without (?r) crop residues). Samples were taken from the 0–5 and 5–10?cm soil layers and separated in micro-aggregates (< 0.25?mm), small macro-aggregates (0.25 to 1?mm) and large macro-aggregates (1 to 8?mm). The carbon content of each aggregate fraction was determined.

Results

Zero tillage combined with crop rotation and crop residues retention resulted in a higher proportion of macro-aggregates. In the 0–5?cm layer, plots with a crop rotation and monoculture of maize and wheat in ZT+r had the greatest proportion of large stable macro-aggregates (40%) and highest mean weighted diameter (MWD) (1.7?mm). The plots with CT had the largest proportion of micro-aggregates (27%). In the 5–10?cm layer, plots with residue retention in both CT and ZT (maize 1?mm and wheat 1.5?mm) or with monoculture of wheat in plots under ZT without residues (1.4?mm) had the greatest MWD. The 0–10?cm soil layer had a greater proportion of small macroaggregates compared to large macro-aggregates and micro-aggregates. In the 0–10?cm layer of soil with residues retention and maize or wheat, the greatest C content was found in the small and large macro-aggregates. The small macro-aggregates contributed most C to the organic C of the sample. For soil cultivated with maize, the CT treatments had significantly higher CO₂ emissions than the ZT treatments. For soil cultivated with wheat, CTR-r had significantly higher CO₂ emissions than all other treatments.

Conclusion

Reduction in soil disturbance combined with residue retention increased the C retained in the small and large macro-aggregates of the top soil due to greater aggregate stability and reduced the emissions of CO₂ compared with conventional tillage without residues retention and maize monoculture (a cultivation system normally used in the central highlands of Mexico).