Agrobacterium tumefaciens-mediated Transformation of CryⅠA(b) gene to Trichoderma harzianum

In this study, Cry ⅠA(b) gene was successfully transferred into the biocontrol fungus Trichoderma harzianum with an efficiency of 60-180 transformants per 10^6 spores by using Agrobacterium tumefaciens-mediated transformation. Putative transformants were analyzed to test the presence of Cry ⅠA(b) gene by Southern blot. Most transformants contained a single T-DNA copy. RT-PCR analysis showed that the Cry ⅠA(b) gene was transcribed. Antifungal activities and insecticidal activities of the transformants were examined. There was no obvious difference in antifungal activities between the transformants and their wild strains. The modified mortalities of the transformants T1 and T2 were 69.57% and 91.30%, respectively. The tranformation system mediated by A. tumefaciens proved to be a powerful tool for the filamentous fungi transformation and functional genomic study with its high transformation frequency, simplicity of T-DNA integration, and genetic stability of transformants.     

Mapping of genetic modifiers of <Emphasis Type="Italic">Plcd1</Emphasis> in scant hair mice (<Emphasis Type="Italic">snthr</Emphasis><Superscript><Emphasis Type="Italic">1Bao</Emphasis></Superscript>)

The scant hair mutant mouse (locus symbol: snthr ^1Bao ) is a recessive mutation that originated in an ethylnitrosourea chemical carcinogenesis study using the DBA/2J inbred strain. The gene responsible for the mutation was previously determined to be phospholipase C, delta 1 (Plcd1; mutant allele symbol Plcd1 ^snthr1Bao ). To map the modifiers of Plcd1, an intercross (DBA/2J-snthr ^1Bao /snthr ^1Bao × C57BL/6J+/+) was conducted. The F2 mutant progeny exhibited a variety of alopecia phenotypes; all F2 mutants (n=507) were classified into 3 groups (mild, moderate, and severe alopecia) and genotyped based on 96 microsatellites. A major QTL was identified on mouse chromosome (mChr) 15 at 12 cM with an LOD score greater than 7 (P < 0.0001). Three minor QTLs were detected on mChr 2, 5, and 7 at 40, 84 and 48 cM, respectively. The QTLs on mChr 7 and 15 were associated with minor alopecia while the QTLs on mChr 2 and 5 were associated with moderate to severe alopecia. No antagonistic or synergistic effects among or between the 4 QTLs were found. Integrating the functions of the 4 potential regulatory QTLs and mutant Plcd1 ^snthr1Bao , we found that these QTLs might contribute to variations of scant hair severity by altering the Ca²⁺ signal pathways in mouse skin.     

Characterizations of <Emphasis Type="Italic">L</Emphasis><Subscript><Emphasis Type="Italic">p</Emphasis></Subscript>(<Emphasis Type="Italic">R</Emphasis>) using tight wavelet frames

In this paper,a Littlewood-Paley function characterization of the spaces L p(R),1相似文献   

Fermentation of xylose to produce ethanol by recombinant <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> strain containing <Emphasis Type="Italic">XYLA</Emphasis> and <Emphasis Type="Italic">XKS1</Emphasis>

Fermentation of the pentose sugar xylose to produce ethanol using lignocellulosic biomass would make bioethanol production economically more competitive. Saccharomyce cerevisise, an efficient ethanol producer, cannot utilize xylose because it lacks the ability to convert xylose to its isomer xylulose. In this study, XYLA gene encoding xylose isomerase (XI) from Thermoanaerobacter tengcongensis MB4T and XKS1 gene encoding xylulokinase (XK) from Pichia stipitis were cloned and functionally coexpressed in Saccharomyces cerevisiae EF-326 to construct a recombinant xylose-utilizing strain. The resulting strain S. cerevisiae EF 1014 not only grew on xylose as sole carbon source, but also produced ethanol under anaerobic conditions. Fermentations performed with different xylose concentrations at different temperatures demonstrated that the highest ethanol productivity was 0.11 g/g xylose when xylose concentration was provided at 50 g/L. Under this condition, 28.4% of xylose was consumed and 1.54 g/L xylitol was formed. An increasing fermentation temperature from 30℃ to 37℃ did not improve ethanol yield.     

Ubi1 intron-mediated enhancement of the expression of Bt cry1Ah gene in transgenic maize （Zea mays L.）

The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutinl gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors （pUUOAH）. The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of CrylAh protein in the construct containing the ubil intron （pUUOAH） was 20% higher than that of the intronless construct （pUOAH）. Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubil intron was higher than that of the intronless construct. These results indicated that the maize ubil intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently     

Phytosterol of <Emphasis Type="Italic">Potamogeton maackianus</Emphasis> and its change under the stress of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis>

The phytosterol in Potamogeton maackianus was identified and quantified. From the hexane extracts analysis of unsaponifiable fraction of P. maackianus by gas chromatography-mass spectrometry(GC-MS), campesterol, stigmasterol and β-sitosterol were the main phytosterols in P. maackianus with the contents of 0.368, 1.17 and 0.824 mg·g⁻¹, respectively. After 7 days’ exposure under M. aeruginosa, the contents of campesterol, stigmasterol and β-sitosterol in P.maackianus increased by 23.1%, 40.4% and 40.8%, respectively. Both the contents of 24-ethyl sterols (sitosterol and stigmasterol) and 24-methyl sterol (campesterol) in macrophytes increased, but the percentage of total sterol composition did not change significantly. This result indicated that the content of 4-demethyl sterols and the second alkylation of the phytosterol side-chain at C-24 in P. maackianus were affected by M. aeruginosa. Biography: ZHANG Shenghua(1979–), female, Ph.D. candidate, research direction: environmental biology.     

Rate constants for the reaction of ozone with <Emphasis Type="Italic">n</Emphasis>-butyl, <Emphasis Type="Italic">s</Emphasis>-butyl and <Emphasis Type="Italic">t</Emphasis>-butyl methyl sulfides

The rate constants for the ozone reactions with n-butyl methyl sulfide （n-BMS, CHaCH2CH2CH2SCH3）, sec-butyl methyl sulfide （s-BMS, CH3CH2（CH3）CHSCHa） and tert-butyl methyl sulfide （t-BMS, （CH3）3CSCH3） were measured using our smog chamber under supposedly pseudo-first-order conditions at 30002 K and 760 Torr. The experimental determined rate constants for n-butyl, s-butyl and t-butyl methyl sulfide are （1.23 ± 0.06）×10-19, （5.08 ± 0.19）×10-20 and （2.26 ± 0.14）×10-20 cm3 molecule-1· s-1, respectively. The reactivity-structure relationship of the reactions was discussed and used to illustrate the mechanism of the ozone reaction with thioethers. The results enrich the kinetics data of atmospheric chemistry.     

Comparative proteomics analysis of <Emphasis Type="Italic">OsNAS1</Emphasis> transgenic <Emphasis Type="Italic">Brassica napus</Emphasis> under salt stress

Salt stress is one of the major abiotic stresses in agricultural plants worldwide. We used proteomics to analyze the differential expression of proteins in transgenic OsNAS1 and non-transformant Brassica napus treated with 20 mmol/L Na₂CO₃. Total protein from the leaves was extracted and separated through a high-resolution and highly repetitive two-dimensional electrophoresis (2-DE) technology system. Twelve protein spots were reproducibly observed to be upregulated by more than 2-fold between transgenic and non-transformant B. napus. These 12 spots were digested in-gel with trypsin and characterized by matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to obtain the peptide mass fingerprints. Protein database searching revealed that 5 of these proteins are involved in salt tolerance: dehydrogenase, glutathione S-transferase, peroxidase, 20S proteasome beta subunit, and ribulose-1,5-bisphosphate carboxylase/oxygenase. The potential functions of these identified proteins in substance and energy metabolism, stress tolerance, protein degradation, and cell defense are discussed. The salt tolerance of the transgenic rapeseed was significantly improved by the introduction of the OsNAS1 gene from Brazilian upland rice of Oryza sativa (cv. IAPAR 9).     

Deletion analysis of oligomycin PKS genes （olmA） in Streptomyces arermitilis

Gene deletion vector pXL05(pKC1139::△olmA1 △olmA4) was used to disrupt oligomycin PKS encoding genes (olmA ) in Streptomyces avermitilis CZ8-73, the producer of anthelmintic avermectins B and the cell growth inhibitor oligomycin, olmA gene cluster in the chromosome was displaced by deletion allele on the plasmid via double crossover. Four of disruptants were confirmed by Southern blotting. Shaking flask experiments and HPLC analyses showed that the four mutants no longer produced the toxic oligomycin, but only made four components of avermectins B, which were avermectin Bla, Blb, B2a, B2b. The yields of avermectins B in these mutants were separately equal to those in CZ8-73. This revealed that olmA genes deletion did not affect the biosynthesis of avermectins. The deletion mutants were proved to be genetically stable, and thus might be promising strains in industrial production of avermectins B.     

Asian origin for Polystichum （Dryopteridaceae） based on rbcL sequences

Chloroplast rbcL sequences of 60 species of Polystichum sensu lato (s.l.), including 23 new sequences from southwest China, were used to assess the phylogenetic relationships within the genus. On the basis of estimated evolution rate of rbcL gene and the genetic distance data that passed relative-rate tests, we further estimated the divergence times between some clades of the genus. The phylogenetic relationships were inferred using the neighbor-joining and maximum-parsimony methods, both methods producing trees with completely congruent topology. These trees reveal that all species of Polystichum s.l. in this study (including Cyrtomium and Cyrtomidictyum) form a monophyletic group. The basal split in Polystichum s.l. separates a clade with all Asian members from a clade containing other species from all over the world. The phylogenetic and divergence time estimation results lead us to suggest that Polystichum s.l. originated in Asia in the late Late Cretacous (≈76 Ma) and migrated into other places in the world in early Eocene(≈46 Ma).     

Southern rice black-streaked dwarf virus: A new proposed <Emphasis Type="Italic">Fijivirus</Emphasis> species in the family <Emphasis Type="Italic">Reoviridae</Emphasis>

For the past several years, a novel dwarf disease has been observed on rice （Oryza sativa） in some regions of Guangdong Province and Hainan Province, southern China. Infected plants showed stunting, dark leaf and small enations on stem and leaf back. Typical Fijivirus viroplasma containing crystalline arrayed spherical virons approximately 70--75 nm in diameter and tubular structures were detected in ultrathin sections by an electron microscope in parenchyma phloem cells of the infected plants. The virus was transmitted to rice seedlings by white-backed planthoppers, Sogatella furcifera （Hemiptera： Delphacidae）, collected in the diseased fields. Analysis of dsRNA extracts from infected plants revealed ten linear segments, which were similar to the electrophoretic profile of Rice black-streaked dwarf virus （RBSDV）. RT-PCR with a single primer which matched to a linker sequence ligated to both 3＇ ends of the viral genomic dsRNAs resulted in amplification of genome segments 9 （S9） and 10 （S10） cDNA products. The complete nucleotide sequences of S9 and S10 were obtained from clones of the RT-PCR amplicon exhibited characteristic properties of Fijivirus including low GC content （34.5% and 35.6%）, genus conserved 5＇ and 3＇ termini sequences and similar genome organization. Blast searches indicated that the sequences of S9 and S10 shared 68.8%--74.9% and 67.1% --77.4% nucleotide identities with those of viruses in the Fijivirus group 2, respectively. These values were similar to those among other viruses in the Fijivirus group 2 and considerably lower than those among RBSDV isolates. Phylogenetic trees based on S9 and S10 nucleotide sequences and their putative amino acid sequences showed that this virus represented a separate branch among other Fijiviruses. The virus was also detected by a nested RT-PCR assay in corn （Zea mays）, barnyard grass （Echinochloa crusgalll）, Juncellus serotinus and flaccidgrass （Pennisetum flaccidum） in and/or adjacent to the infected rice fields. I     

Effect of acetazolamide on stable carbon isotope fractionation in <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> and <Emphasis Type="Italic">Chlorella vulgaris</Emphasis>

The effect of extracellular carbonic anhydrase (CAex) on stable carbon isotope fractionation in algae is still unclear. The stable carbon isotope composition and algal growth in the presence and absence of the membrane-impermeable CA inhibitor acetazolamide were compared in Chlamydomonas reinhardtii and Chlorella vulgaris. The CAex of both algal species contributed about 9‰ of the stable carbon isotope fractionation and exhibited a dosage effect. Therefore, evidence in vivo that CAex leads to a larger carbon isotope fractionation of algae is presented.     

On reverses of the <Emphasis Type="Italic">L</Emphasis><Subscript><Emphasis Type="Italic">p</Emphasis></Subscript>-Busemann-Petty centroid inequality and its applications

In this paper,the reverse forms of the L p-Busemann-Petty centroid inequality are shown. As the applications of the reverse forms,we obtain the reverse forms of the L p-centroid-affine inequality and an upper bound of the isotropic constant for convex bodies.     

Enhanced anti-sunward flow near local noon during a period of horizontal IMF and high solar wind velocity <Emphasis Type="Italic">V</Emphasis><Subscript><Emphasis Type="Italic">Y</Emphasis></Subscript>

It is believed that a southward interplanetary magnetic field (IMF) is mainly responsible for the energy input from solar wind into the magnetosphere. This paper presents an unusual case of strong anti-sunward plasma flow (up to 2 km/s) in the polar cap ionosphere and large cross-polar cap potential (CPCP) during a period of horizontal IMF (|B _Z | < 2 nT) observed by both ACE (at the L1 point) and Geotail (on the dusk flank of the magnetosheath). The CPCP is even higher than that under preceding B _Z ≈ −23 nT. Furthermore, GOES8 observed that the magnetosheath field turns northward as the anti-sunward plasma flow and CPCP start to increase, which implies that the magnetosheath field interacting with the Earth’s magnetopause has significantly rotated and differs from the IMF observed by ACE and Geotail. In accordance with previous theoretical work, we suggest that the magnetic field line draping produces a southward magnetosheath field and enhances anti-sunward plasma flow and the CPCP.     

Properties of a class of groups on <Emphasis Type="Italic">Z</Emphasis><Subscript><Emphasis Type="Italic">m</Emphasis></Subscript>

In this paper, we obtain the factorization of direct production and order of group GL(n, Z _m ) in a simple method. Then we generalize some properties of GL(2, Z _p ) proposed by Huppert, and prove that the group \(GL\left( {2,{Z_{{z^\lambda }}}} \right)\) is solvable. We also prove that group GL(n, Z _p ) is solvable if and only if GL(n, Z _p ) is solvable, and list the generators of groups GL(n, Z _p ) and SL(n, Z _p ). At last, we prove that PSL(2, Z _p )(p > 3) and PSL(n, Z _p )(n > 3) are simple.     

Improved zero-sum distinguisher for full round <Emphasis Type="Italic">Keccak-f</Emphasis> permutation

Keccak is one of the five hash functions selected for the final round of the SHA-3 competition,and its inner primitive is a permutation called Keccak-f.In this paper,we observe that for the inverse of the only nonlinear transformation in Keccak-f,the algebraic degree of any output coordinate and the one of the product of any two output coordinates are both 3,which is 2 less than its size of 5.Combining this observation with a proposition on the upper bound of the degree of iterated permutations,we improve the zero-sum distinguisher for the Keccak-f permutation with full 24 rounds by lowering the size of the zero-sum partition from 21590 to 21575.     

Antibacterial activity and structure-activity relationships of Schiff bases on <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> by microcalorimetry

The growth of Staphylococcus aureus under the action of six kinds of Schiff bases(A,B,C,D,E,F) was studied by means of microcalorimetry. Growth constant k and inhibition ratio I were calculated. Also,the antibacterial activity,the action characteristics and the structure-activity relationships of the compounds on S. aureus were analyzed. Results indicate that B,F show good antibacterial activity(IC50:293.6 mg · L-1 and 307.8 mg · L-1) ,D,E show moderate antibacterial activity(IC50:966.3 mg · L-1 and 1 126.7...     

Engineering <Emphasis Type="Italic">Deinococcus radiodurans</Emphasis> into biosensor to monitor radioactivity and genotoxicity in environment

Based on a genetically modified radioresistant bacteria Deinococcus radiodurans, we constructed a real time whole cell biosensor to monitor radioactivity and genotoxicity in highly radioactive environment. The enhanced green fluorescence protein （eGFP） was fused to the promoter of the crucial DNA damage-inducible recA gene from D. radiodurans, and the consequent DNA fragment （PrecA-egfp） carried by plasmid was introduced into D. radiodurans R1 strain to obtain the biosensor strain DRG300. This engineered strain can express eGFP protein and generate fluorescence in induction of the recA gene promoter. Based on the correlation between fluorescence intensity and protein expression level in live D. radiodurans cells, we discovered that the fluorescence induction of strain DRG300 responds in a remarkable dose-dependent manner when treated with DNA damage sources such as gamma radiation and mitomycin C. It is encouraging to find the widely detective range and high sensitivity of this reconstructed strain comparing with other whole cell biosensors in former reports. These results suggest that the strain DRG300 is a potential whole cell biosensor to construct a detective system to monitor the biological hazards of radioactive and toxic pollutants in environment in real time.     

The composition and distribution of metal clusters in the MoFe protein from a nifZ deletion strain （DJ 194） of Azotobacter vinelandii

Through the anaerobic chromatography on the columns of DEAE 52, Q-Sepharose and Sephacryl S-200, a nitrogenase MoFe protein (△nifZ Av1) was obtained from a nifZ deleted mutant of Azotobacter vinelandii (stain DJ194).The results of Western blotting after anoxic native electrophoresis and SDS-PAGE showed that △nifZ Av1 was similar to wild type MoFe protein (OP Av1) at the electrophoretic mobility, molecular weight and subunit composition. Furthermore, △nifZ Avl was also similar to OP Av1 at the molybdenum content, EPR signal (g≈4.3, 3.65 and 2.01), and the molar extinction coefficient (△ε) of circular dichroism (CD)at 660 nm region. All of these indicated that, besides having the same α2β2 composition as OP Av1, the △nifZ Av1 also contained equal amount of reductive FeMoco in the spin state of S=3/2 to OP Av1. However, the iron content and substrate (C2H2, H^ and N2)-reduction activity of △nifZ Av1 were 74% and 46%-50% of those of OP Av1, respectively. Furthermore, the △ε at around 450 nm, which reflects P-cluster in Av1, was obviously lower than that of OP Av1. It suggested that the difference between △nifZ Avl and OP Av1 resulted from P-cluster rather than FeMoco, and from the half number of P-cluster in △nifZ Av1, but the composition or redoxstate of P-cluster in △nifZ Av1 were not changed. Thus it could propose that △nifZ Av1 is composed of two different αβsubunit pairs. One is a FeMoco-and P-cluster-containing pair, and the other is a P-cluster-deficient but FeMoco-containing pair. Since the deletion of nifZ gene leads to the deficiency of only one of two P-clusters in a α2β2 tetramer, the assembly of P-cluster may not simply depend on one gene product, and so a possible mechanism of NifZ is supposed here.