海南热带雨林腐木上酵母菌物种多样性研究

用含木糖为唯一碳源(培养基X)和含葡萄糖及7.6%乙醇(培养基E)的两种富集培养基分别从采自海南热带雨林的56和57份腐木样品中分离到酵母菌67和75株.依据26S rDNA D1/D2区域序列分析并结合形态学特征对这些菌株进行了分类学研究,探讨了该地区腐木上的酵母菌物种多样性及其分布.从分离的142株酵母菌中鉴定出14个属63个种,其中疑似新种25个,占总种数的近40%,说明在热带雨林腐木中尚存在大量酵母菌新分类群有待被发现.从用培养基X和E分离的酵母菌中分别鉴定出7属37种和11属33种,优势属均为假丝酵母属Candida Berkhout和毕赤酵母属Pichia Hansen,但种类组成基本不同.用培养基x富集分离的菌株以Candida quercitrusa S.A.Meyer＆Phaff的地理分布最广,用培养基E富集分离的菌株以异常毕赤酵母Pichia anomala(Hansen)Kurtzman、酿酒酵母Saccharomyces cerevisiae Meyen ex Hansen和亚膜毕赤酵母Pichiasubpelliculosa Kurtzman分布最广泛.同一样品用两种不同富集培养基分离的菌株大多数属于不同的种,在对比的23份样品中,只从2份样品中分离到了同一个种的菌株.用培养基X和E分离的菌株分别属于可利用木糖和可耐受乙醇的酵母菌,用两种培养基同时分离到的菌株属于具备利用木糖和耐受高浓度乙醇两种能力的菌株.这些酵母菌在木质纤维素物质的生物乙醇转化技术中的应用价值值得进一步研究.     

Identification of white-rot and soft-rot fungi increasing ethanol production from spent sulfite liquor in co-culture with Saccharomyces cerevisiae

Aim:  To identify fungi that are capable of increasing ethanol production from lignocellulose in spent sulfite liquor.
Methods and Results:  In a batch fermentation study, the fungal mix could produce 24·61 g l⁻¹ ethanol using spent sulfite liquor as substrate. The fungal mix grew well on glucose, xylose, hemicellulose and cellulose. In addition, we were able to identify the fungal mix by use of PCR-amplification of DNA and sequencing, and they were identified as Chalara parvispora and Trametes hirsuta/T. versicolor. In a reconstitution study, the identified fungi were shown to produce equal amount of ethanol as the fungal mix. We were also able to show that C. parvispora could produce ethanol from xylose.
Conclusion:  The present study has shown that ethanol production from biomass can be increased by use of C. parvispora and T. versicolor when compared with fermentation using only S. cerevisiae .
Significance and Impact of the Study:  The study shows that refining biomass by ethanol production from spent sulfite liquor, a lignocellulose material, can be increased by adding C. parvispora and T. versicolor , and it is thus of great potential economical impact.     

Characterization and technological properties of Oenococcus oeni strains from wine spontaneous malolactic fermentations: a framework for selection of new starter cultures

Aims:  To characterize the genetic and phenotypic diversity of 135 lactic acid bacteria (LAB) strains isolated from Italian wines that undergone spontaneous malolactic fermentation (MLF) and propose a multiphasic selection of new Oenococcus oeni malolactic starters.
Methods and Results:  One hundred and thirty-five LAB strains were isolated from 12 different wines. On the basis of 16S amplified ribosomal DNA restriction analysis (ARDRA) with three restriction enzymes and 16S rRNA gene sequencing, 120 O. oeni strains were identified. M13-based RAPD analysis was employed to investigate the molecular diversity of O. oeni population. Technological properties of different O. oeni genotypes were evaluated in synthetic medium at increasing selective pressure, such as low pH (3·5, 3·2 and 3·0) and high ethanol values (10, 11 and 13% v/v). Finally, the malolactic activity of one selected strain was assessed in wine by malolactic trial in winery.
Conclusions:  The research explores the genomic diversity of wine bacteria in Italian wines and characterizes their malolactic metabolism, providing an efficient strategy to select O. oeni strains with desirable malolactic performances and able to survive in conditions simulating the harsh wine environment.
Significance and Impact of the Study:  This article contributes to a better understanding of microbial diversity of O. oeni population in Italian wines and reports a framework to select new potentially O. oeni starters from Italian wines during MLF.     

Effect of environmental stresses on the sensitivity of Enterobacter sakazakii in powdered infant milk formula to gamma radiation

Aim:  To evaluate the effect of starvation, heat, cold, acid, alkaline, chlorine and ethanol stresses on the resistance of Enterobacter sakazakii in powdered infant milk formula (PIMF) towards gamma radiation.
Methods and Results:  Stressed cells of E. sakazakii ATCC 51329 and four other food isolate strains were mixed individually with PIMF, kept overnight at room temperature, and then exposed to gamma radiation up to 7·5 kGy. The D ₁₀-values were determined using linear regression and for the stressed E. sakazakii strains these values ranged from 0·82 to 1·95 kGy.
Conclusions:  Environmental stresses did not significantly change the sensitivity of most E. sakazakii strains to ionizing radiation.
Significance and Impact of the Study:  Data obtained established that most forms of environmental stress are unlikely to significantly enhance the resistance of E. sakazakii strains to lethal, low dose irradiation treatment.     

Comparison of Lactococcus garvieae strains isolated in northern Italy from dairy products and fishes through molecular typing

Aims:  To investigate the genetic relatedness between Lactococcus garvieae strains isolated from fish and dairy samples collected in northern Italy, using random-amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR), Sau -PCR and amplified fragment length polymorphism (AFLP).
Methods and Results:  Eighty-one isolates from bovine and caprine dairy products ( n  = 53) and from diseased rainbow trouts and other fishes ( n  = 28) were examined. All methods showed a typeability of 100%, repeatability ranging from 84·4% to 97·5% and discriminatory powers from 0·798 to 0·986. Dairy and fish strains revealed a low genetic relatedness as they are often grouped into distinct clusters. RAPD analysis discriminated 52 genotypes when primer M13 was used, whereas with primer P5 only 27 genotypes were identified. When Sau -PCR was performed, 13 genotypes were detected while AFLP analysis allowed the differentiation of 32 genotypes.
Conclusion:  L. garvieae strains isolated from dairy samples are generally not related to those collected from fish lactococcosis outbreaks.
Significance and Impact of the Study:  L. garvieae strains exhibit a genetic diversity related to the specific animal host they colonize. RAPD M13 fingerprinting proved to be a molecular tool for comparing isolates, whereas Sau -PCR and AFLP analyses were useful techniques to investigate the distribution of L. garvieae populations in the environment.     

Clonal groups of high-level gentamicin-resistant Enterococcus faecium isolated from municipal wastewater and clinical samples in Tehran, Iran

Aims:  Clonality among high-level gentamicin-resistant Enterococcus faecium (HLGR-EF) isolates obtained from clinical and sewage treatment plants (STP) were investigated using PhePlate system (PhP), ribotyping and pulsed-field gel electrophoresis (PFGE).
Methods and Results:  During 1 year study (September 2005–2006), a total of 106 HLGR-EF isolates were collected from clinical ( n  = 48) and STP ( n  = 58) samples in Tehran, Iran. Biochemical fingerprinting of these isolates using the PhP showed the presence of 21 PhP types (diversity index, Di  = 0·97) among the clinical and 21 PhP types ( Di  = 0·91) among the STP isolates. Representative isolates of each PhP type ( n  = 42) were further characterized by the ribotyping method. Sixteen ribotypes were identified among the isolates with five types shared between the clinical and STP isolates. PFGE recognized 24 clonal types among these isolates with three pulsotypes shared between the clinical and STP isolates. Combination of the two techniques (PFGE and ribotyping) resulted in 24 ( Di  = 0·96) and 16 ( Di  = 0·93) types among the strains isolated from clinical and STP samples, respectively.
Conclusions:  We concluded that the combination of PhP typing, ribotyping and PFGE could be extremely discriminatory when examining HLGR-EF isolates.
Significance and Impact of the Study:  The emergence of highly diverse HLGR-EF population in Iran is of serious concern especially because of their multi-resistances.     

Detailed studies on Quercus infectoria Olivier (nutgalls) as an alternative treatment for methicillin-resistant Staphylococcus aureus infections

Aims:  To investigate the antimethicillin-resistant Staphylococcus aureus (MRSA) mechanism of Quercus infectoria (nutgalls) extract and its components.
Methods and Results:  Ethanol extract, an ethyl acetate fraction I, gallic acid and tannic acid could inhibit the growth of clinically isolated MRSA strains with minimum inhibitory concentration values between 63 and 250  μ g ml⁻¹. Clumps of partly divided cocci with thickened cell wall were observed by transmission electron microscopy in the cultures of MRSA incubated in the presence of the ethanol extract, the ethyl acetate fraction I and tannic acid. Because cell wall structure of the organism structures seemed to be a possible site for antibacterial mechanisms, their effect with representative β-lactam antibiotics were determined. Synergistic effects with fractional inhibitory concentration index ranged from 0·24 to 0·37 were observed with 76% and 53% of the tested strains for the combination of the ethanol extract with amoxicillin and penicillin G, respectively.
Conclusions:  The appearance of pseudomulticellular bacteria in the treated cells and the synergistic effect of the plant extract with β-lactamase-susceptible penicillins suggest that the extract may interfere with staphylococcal enzymes including autolysins and β-lactamase.
Significance and Impact of the Study:  Our results provide scientific data on the use of the nutgalls, which contain mainly tannin contents up to 70% for the treatment of staphylococcal infections.     

Viable ultramicrocells in drinking water

Aims:  To examine the diversity of cultivable 0·2 micron filtrate biofilm forming bacteria from drinking water systems.
Methods and Results:  Potable chlorinated drinking water hosts phylogenetically diverse ultramicrocells (UMC) (0·2 and 0·1  μ m filterable). UMC (starved or dwarf bacteria) were isolated by cultivation on minimal medium from a flow system wall model with polyvinyl chloride (PVC) pipes. All cultivated cells (25 different isolates) did not maintain their ultra-size after passages on rich media. Cultured UMC were identified by their 16S ribosomal DNA sequences. The results showed that they were closely related to uncultured and cultured members of the Proteobacteria, Actinobacteria and Firmicutes. The isolates of phylum Actinobacteria included representatives of a diverse set of Actinobacterial families: Micrococcaceae, Microbacteriaceae, Dermabacteraceae, Nocardiaceae and Nocardioidaceae.
Conclusions:  This study is the first to show an abundance of cultivable UMC of various phyla in drinking water system, including a high frequency of bacteria known to be involved in opportunistic infections, such as Stenotrophomonas maltophilia, Microbacterium sp., Pandoraea sp. and Afipia strains.
Significance and Impact of the Study:  Chlorinated tap water filtrate (0·2 and 0·1  μ m) still harbours opportunistic micro-organisms that can pose some health threat.     

Selective production of cis-9,trans-11 isomer of conjugated linoleic acid from trans-vaccenic acid methyl ester by Delacroixia coronata

Aims:  Bio-process development for isomer selective and efficient production of cis -9, trans -11-octadecadienoic acid (CLA) from trans -vaccenic acid ( t -VA, trans -11-octadecenoic acid) through microbial fatty acid Δ9-desaturation reaction.
Methods and Results:  A total of 550 strains of fungi and yeasts were screened for CLA production from t -VA through Δ9 desaturation. Delacroixia coronata IFO 8586 was selected as a potent producer of CLA from t -VA. Efficient CLA production was observed during cultivation in medium supplemented with the methyl ester of t -VA ( t -VAME). Under the optimal conditions with 33·3 mg ml⁻¹ of t -VAME as substrate, 10·5 mg ml⁻¹ CLA was produced by D. coronata IFO 8586 after 7 days of cultivation in the medium containing dextrin (5·0%), tryptone (2·0%) and thiourea (0·83 μmol ml⁻¹). The strain produced the cis -9, trans -11 isomer of CLA selectively (98% of total CLA), with a small amount of the trans -9, trans -11 isomer (2% of total CLA), mainly in the form of triacylglycerols (69% of total CLA).
Conclusions:  A practical bio-process for selective production of cis -9, trans -11 isomer of CLA using filamentous fungus D. coronata IFO 8586 was successfully established.
Significance and Impact of the Study:  Isomer selective bio-process for the practical production of cis -9, trans -11-CLA was first established. The process is benefitable for expanding the application of CLA for medicinal and nutraceutical purposes.     

Chimeric types of chromosome X in bottom-fermenting yeasts

Aim:  To determine the structure of the chimeric chromosome X of bottom-fermenting yeasts.
Methods and Results:  Eight cosmid clones carrying DNA from chromosome X of bottom-fermenting yeasts were selected by end-sequencing. Four of the cosmid clones had Saccharomyces cerevisiae (SC)-type and Saccharomyces bayanus (SB)-type chimeric ends, two had SC-type ends and two had SB-type ends. Sequencing revealed that the bottom-fermenting yeast strains in this study had four types of chromosome X: SC–SC, SC–SB, SB–SC and SB–SB. The translocation site in the chimeric chromosome is conserved among bottom-fermenting yeast strains, and was created by homologous recombination within a region of high sequence identity between the SC-type sequence and the SB-type sequence.
Conclusions:  Existing bottom-fermenting yeast strains share a common ancestor in which the chimeric chromosome X was generated.
Significance and Impact of the Study:  The knowledge gained in this study sheds light on the evolution of bottom-fermenting yeasts.     

Comparative analysis of different TaqMan real-time RT-PCR assays for the detection of swine Hepatitis E virus and integration of Feline calicivirus as internal control

Aims:  The aim of this study was to compare the performance of four TaqMan RT-PCR assays with a commonly used nested RT-PCR and to include the Feline calicivirus (FCV) as an internal control.
Methods and Results:  RNA extracted from 87 swine faecal samples and 103 swine blood samples was subjected to different detection systems. Faecal samples naturally contaminated with Hepatitis E virus (HEV) and negative samples were artificially inoculated with 3·2 × 10³ PFU of FCV. Detection results obtained on faecal and plasma samples were 35·6% and 4·9% with the nested RT-PCR assay, 8·0% and 0%, 0% and 0%, 13·8% and 0% and 36·8% and 3·9% with TaqMan systems A, B, C and D respectively. The Ct means obtained with the multiplex TaqMan assay were 30·11 and 30·43 for the detection of FCV with HEV contaminated samples and negative samples.
Conclusions:  The TaqMan system D was more suitable for the detection of swine HEV strains than the three others and FCV was integrated successfully as an internal control.
Significance and Impact of the Study:  FCV was demonstrated as an efficient control to monitor the RNA extraction process and HEV amplification procedure in a multiplex HEV/FCV TaqMan assay. This control would be helpful in limiting false negative results.     

Effects of the high pressure of homogenization on some spoiling micro-organisms, representative of fruit juice microflora, inoculated in saline solution

Aims:  This study was aimed to investigate the effects of a high-pressure homogenization (HPH) treatment on some micro-organisms, involved in the spoilage of fruit juices.
Methods and Results:  Lactobacillus plantarum , Lactobacillus brevis , Bacillus coagulans cells, Saccharomyces bayanus , Pichia membranaefaciens and Rhodotorula bacarum were separately inoculated in a saline solution (0·9% NaCl); the initial inoculum was ca. 5 log CFU ml⁻¹. Then, the samples were processed through a homogenizer at 10–150 MPa for 1, 2 or 3 times. Yeasts were completely inactivated at 50–110 MPa with a single pass treatment, while lactic acid bacteria counts were reduced to approximately 1 log CFU ml⁻¹ after a three-steps HPH processing.
Conclusions:  Yeasts were the most sensitive micro-organisms, followed by B. coagulans . On the other hand, lactic acid bacteria appeared resistant to HPH.
Significance and Impact of the Study:  The results of this study provided some useful information on the susceptibility of microflora of juices to homogenization; moreover, they suggested that HPH could be used successfully to inactivate yeasts.     

Corynebacterium pseudodiphtheriticum isolated from relevant clinical sites of infection: a human pathogen overlooked in emerging countries

Aims:  To examine the occurrence of and to determine the antimicrobial susceptibility of Corynebacterium pseudodiphtheriticum among patients with bacterial infections at a teaching hospital.
Methods and Results:  A total of 113 Coryne. pseudodiphtheriticum strains identified by conventional biochemical methods and API-Coryne System were recovered from patients from different age groups: 65·48% adults (18 to ≤59 years old), 9·73% aged (≥60 years old); 14·15% infants (<18 years old); 4·42% newborns (0–7 days). Micro-organisms were mostly related to infections in the urinary (29·2%) and respiratory tracts (27·45%) and intravenous sites (18·6%). Clinical samples were obtained only from 32·7% patients (26 adults, four aged, four infants and three newborns) presenting at least one of the predisposing conditions: end-stage renal disease; renal transplant; AIDS and Mycobacterium tuberculosis infection; cancer, hepatic cirrhosis; haemodialysis and catheter use. Antimicrobial susceptibility tests identified multiresistant phenotypes. Most strains (>50%) were resistant to oxacillin, erythromycin and clindamycin.
Conclusions:  Despite significant differences in age and functional status of patients Coryne. pseudodiphtheriticum may be implicated as a cause of respiratory and nonrespiratory human infections.
Significance and Impact of the Study:  Data are valuable for practitioners indicating the occurrence of multiresistant phenotypes and the possibility of severe infections due to Coryne. pseudodiphtheriticum , a pathogen usually overlooked in emerging countries.     

Biological control of grey mould in strawberry fruits by halophilic bacteria

Aims:  Grey mould caused by Botrytis cinerea is an economically important disease of strawberries in Tunisia and worldwide. The aim of this study was to select effective halophilic bacteria from hypersaline ecosystems and evaluate the abilities of antifungal bacteria to secrete extracellular hydrolytic enzymes, anti- Botrytis metabolites and volatiles.
Methods and Results:  Grey mould was reduced in strawberry fruits treated with halophilic antagonists and artificially inoculated with B. cinerea . Thirty strains (20·2%) were active against the pathogen and reduced the percentage of fruits infected after 3 days of storage at 20°C, from 50% to 91·66%. The antagonists were characterized by phenotypic tests and 16S rDNA sequencing. They were identified as belonging to one of the species: Virgibacillus marismortui , B. subtilis , B. pumilus , B. licheniformis , Terribacillus halophilus , Halomonas elongata , Planococcus rifietoensis , Staphylococcus equorum and Staphylococcus sp. The effective isolates were tested for antifungal secondary metabolites.
Conclusions:  Moderately halophilic bacteria may be useful in biological control against this pathogen during postharvest storage of strawberries.
Significance and Impact of the study:  The use of such bacteria may constitute an important alternative to synthetic fungicides. These moderate halophiles can be exploited in commercial production and application of the effective strains under storage and greenhouse conditions.     

Evidence of different fermentation behaviours of two indigenous strains of Saccharomyces cerevisiae and Saccharomyces uvarum isolated from Amarone wine

Aims:  To explain the role of Saccharomyces cerevisiae and Saccharomyces uvarum strains (formerly Saccharomyces bayanus var. uvarum ) in wine fermentation.
Methods and Results:  Indigenous Saccharomyces spp. yeasts were isolated from Amarone wine (Italy) and analysed. Genotypes were correlated to phenotypes: Melibiose⁻ and Melibiose⁺ strains displayed a karyotype characterized by three and two bands between 225 and 365 kb, respectively. Two strains were identified by karyotype analysis (one as S. cerevisiae and the other as S. uvarum ). The technological characterization of these two strains was conducted by microvinifications of Amarone wine. Wines differed by the contents of ethanol, residual sugars, acetic acid, glycerol, total polysaccharides, ethyl acetate, 2-phenylethanol and anthocyanins. Esterase and β-glucosidase activities were assayed on whole cells during fermentation at 13° and 20°C. Saccharomyces uvarum displayed higher esterase activity at 13°C, while S. cerevisiae displayed higher β-glucosidase activity at both temperatures.
Conclusions:  The strains differed by important technological and qualitative traits affecting the fermentation kinetics and important aroma components of the wine.
Significance and Impact of the Study:  The contribution of indigenous strains of S. cerevisiae and S. uvarum to wine fermentation was ascertained under specific winemaking conditions. The use of these strains as starters in a winemaking process could differently modulate the wine sensory characteristics.     

Anti-Helicobacter pylori activity of Lactobacillus delbrueckii subsp. bulgaricus strains: preliminary report

Aims:  To evaluate the activities of six Lactobacillus delbrueckii subsp. bulgaricus (LB) strains against 30 Helicobacter pylori strains by agar-well diffusion method.
Methods and Results:  LB cultures [4 × 10⁸–4 × 10⁹ CFU ml⁻¹) either were prepared in milk at their native pH, 3·8–5·0, or were adjusted to pH 6·4–7·7. At low and neutralized pH, LB strains inhibited the growth by 40–86·7% and 16·7–66·7% of H. pylori strains, respectively. LB activity was strain-dependent. At low and neutralized pH, one and five H. pylori strains, respectively, were not inhibited by any LB strain. LB2 and LB3, taken together, were active against most metronidazole and clarithromycin resistant strains.
Conclusions:  All LB strains inhibited a number of H. pylori strains, including also antibiotic resistant strains. LB activity was strain-dependent and better at low pH. At low pH values, the most active LB strains were LB1, LB2 and LB3, inhibiting 86·7% of H. pylori strains, while at neutralized pH values, the most active LB strains were LB2 and LB3, inhibiting 53·3 and 66·7% of H. pylori strains, respectively.
Significance and Impact of the Study:  LB could be utilized in the treatment or prophylaxis of H. pylori infection and warrants clinical investigations.     

Comparison of automated ribotyping and pulsed-field gel electrophoresis for subtyping of Vibrio cholerae

Aims:  To compare the discriminatory power of an automated ribotyping method for Vibrio cholerae subtyping with the pulsed-field gel electrophoresis (PFGE), to evaluate the possibility of automated ribotyping in use of outbreak investigations and surveillance of cholera.
Methods and Results:  Eight-one epidemiologically unrelated isolates of V. cholerae , and 19 isolates from seven cholera outbreaks were used as the panels. When comparing the two methods using the epidemiologically unrelated isolates, automated ribotyping using Pvu II distinguished 38 different ribotypes with a D -value of 0·8956. When combined with serotyping, the D -value is 0·9466. However, PFGE with Not I and Sfi I digestions had higher D -values of 0·9951 and 0·9948, respectively. PFGE could cluster the isolates from each outbreak into the same pattern, and distinguish different patterns from different outbreaks, whereas automated ribotyping had lower discriminatory ability.
Conclusions:  The automated ribotyping has lower discriminatory ability compared to PFGE, and is limited to application in V. cholerae subtyping and outbreak investigation.
Significance and Impact of the Study:  The study evaluated the limitation in subtyping of automated ribotyping for V. cholerae , and raise the question of improvement for the automated ribotyping in subtyping.     

Effect of γ-irradiation on membrane fatty acids and peptidoglycan's muropeptides of Pantoea agglomerans, a plant pathogen

Aims:  The aim of this study was to evaluate the effect of γ-irradiation on the fatty acids (FA) and muropeptides content of two strains of an Enterobacteriacea : Pantoea agglomerans .
Methods and Results:  Pantoea agglomerans strains ATCC 49174 and RL1 isolated from irradiated carrots were used for this study. Radiation treatments (1 and 3·5 kGy) were performed to study the radiotolerance. Total lipids were obtained by multiple extractions using methanol/chloroform (2 : 1) and were quantified by GC. Muropeptides were purified by successive enzymatic digestions and analysed using a reverse phase C₁₈ column in high performance liquid chromatography. A significant ( P  ≤ 0·05) modification of the bacterial wall was noticed for the membrane FA composition and the muropeptides.
Conclusion:  Effects of irradiation on the bacterial membrane are noticeable and could play an important role on the cellular response and ability to survive this harsh environment.
Significance and Impact of the Study:  To our knowledge, it is the first study to demonstrate the effects of ionizing irradiation on the modification of the FA and one of the few to confirm its effects on the muropeptides of the peptidoglycan.     

Auxin production by plant associated bacteria: impact on endogenous IAA content and growth of Triticum aestivum L.

Aims:  The aim of this study was to investigate the potential of bacterial strains of Bacillus, Pseudomonas, Escherichia, Micrococcus and Staphylococcus genera associated with wild herbaceous flora to enhance endogenous indole-3-acetic acid (IAA) content and growth of Triticum aestivum var. Inqalab-91.
Methods and Results:  Gas chromatography and mass spectrometric (GC–MS) analysis revealed that bacterial strains produced 0·6–8·22 μg IAA ml⁻¹ in the presence of L-tryptophan. Plant microbe experiments showed a significant positive correlation between auxin production by bacterial strains and endogenous IAA content of T. aestivum for GC–MS ( r  = 0·618; P  = 0.05) and colorimetric analysis ( r  = 0·693; P  = 0.01). Similarly, highly significant positive correlation for shoot length ( r  = 0·627; P  = 0.01) and shoot fresh weight ( r  = 0·626; P  = 0.01) was observed with auxin production under axenic conditions. Bacterial inoculations also enhanced shoot length (up to 29·16%), number of tillers (up to 97·35%), spike length (up to 25·20%) and seed weight (up to 13·70%) at final harvest.
Conclusions:  Bacterial strains have the ability to increase the endogenous IAA content and growth of T. aestivum var. Inqalab-91.
Significance and Impact of the Study:  Microbial strains of wild herbaceous flora can be effectively used to enhance the growth and yield of agronomically important crops.     

Optimization of medium constituents for improved chitinase production by Paenibacillus sp. D1 using statistical approach

Aims:  Statistical optimization of medium components for improved chitinase production by Paenibacillus sp. D1.
Methods and Results:  Urea, K₂HPO₄, chitin and yeast extract were identified as significant components influencing chitinase production by Paenibacillus sp. D1 using Plackett–Burman method. Response surface methodology (central composite design) was applied for further optimization. The concentrations of medium components for improved chitinase production were as follows (g l⁻¹): urea, 0·33; K₂HPO₄, 1·17; MgSO₄, 0·3; yeast extract, 0·65 and chitin, 3·75. This statistical optimization approach led to the production of 93·2 ± 0·58 U ml⁻¹ of chitinase.
Conclusions:  The important factors controlling the production of chitinase by Paenibacillus sp. D1 were identified as urea, K₂HPO₄, chitin and yeast extract. Statistical approach was found to be very effective in optimizing the medium components in manageable number of experimental runs with overall 2·56-fold increase in chitinase production.
Significance and Impact of the Study:  The present investigation provides a report on statistical optimization of medium components for improved chitinase production by Paenibacillus sp. D1. Paenibacillus species are gram-positive, spore-forming bacteria with several PGPR and biocontrol potentials. However, only few reports concerning mycolytic enzyme production especially chitinases are available. Chitinase produced by Paenibacillus sp. D1 represents new source for biotechnological and agricultural use.