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.     

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     

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.     

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).     

Comparative study of Cambrian lobopods <Emphasis Type="Italic">Miraluolishania</Emphasis> and <Emphasis Type="Italic">Luolishania</Emphasis>

The rare fossil Miraluolishania described by Liu et ah from the Lower Cambrian Chengjiang Lagerstatte in 2004 is regarded as an arthropod sphinx because it bears mosaic features of both Iobopods and arthropods. The discovery of this rare transitional form offers direct fossil evidence for exploring the relationship between Iobopods and arthropods. However, some scientists consider Miraluolishania to be a junior synonym of Luolishania because the former superficially resembles the latter in general appearance. Considering the significant differences between the two taxa, a thorough comparative study of Miraluolishania and Luolishania leads to the conclusion that there are definitely two different genera. Nevertheless, the ＂Luolishania＂ of the Haikou area is indeed ＂Miraluolishania＂, whereas Luolishania is most likely the typical genus of the Maotianshan area of Chengjiang County.     

Attractiveness of tobacco volatiles induced by <Emphasis Type="Italic">Helicoverpa armigera</Emphasis> and <Emphasis Type="Italic">Helicoverpa assulta</Emphasis> to <Emphasis Type="Italic">Campoletis chlorideae</Emphasis>

The attraction of Helicoverpa armigera-and Helicoverpa assulta-induced and mechanical damage-induced tobacco volatiles to Campoletis chlorideae was investigated, and the induced volatiles were analyzed. In windtunnel, C. chlorideae was strongly attracted by herbivoreinduced tobacco volatiles. Mechanically damaged tobacco leaves, whether treated with caterpillar regurgitant or water,were more attractive to the parasitoid than undamaged tobacco leaves. GC-MS analysis revealed that only 4 compounds were released from undamaged tobacco leaves,whereas 13 compounds were commonly emitted from herbivore-infested and mechanically damaged tobacco leaves.Compound β-pinene was specifically induced by the infestation of H. armigera, and (Z)-3-hexenal was only induced by the infestation of H. armigera and H. assulta, whereas hexyl acetate was only induced by mechanical damage. Tobacco leaves infested by H. armigera and H. assulta released larger amounts of volatiles than undamaged tobacco leaves did.Tobacco leaves treated with artificial damage plus caterpillars regurgitant or water emitted the same levels of volatiles,which were higher than that emitted by undamaged tobacco leaves. The emission amounts of single compounds were also different between differently treated plants. The differences were large between herbivore-induced and mechanical damage-induced compounds, and small between H. armigeraand H.assulta-induced compounds, and among compounds emitted from mechanically damaged plants treated with water or caterpillar regurgitant.     

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).     

Genetic diversity in the male-specific <Emphasis Type="Italic">SRY</Emphasis> gene of <Emphasis Type="Italic">Lepus yarkandensis</Emphasis>

Lepus yarkandensis, an endemic hare species in the Tarim Basin of China, has been suffering from habitat fragmentation due to desert expansion. To evaluate the effect of habitat fragmentation on its genetic diversity, the genetic diversity based on male-specific SRY gene marker is examined. A relatively low level of SRY genetic diversity is found compared to previous studies with mtDNA data, possibly due to the low SRY mutation rate and positive selection. Furthermore, one haplotype exists in eight populations along the Tarim River but not in many other relatively isolated populations, suggesting that habitat fragmentation may affect population divergence. Despite this, our pairwise Fst analysis shows no significant differentiation among populations, and this may be mainly caused by positive selection on the SRY gene in that 88 percent of individuals share the same haplotype. Finally, the phylogenetic analysis shows deep differentiation between L. yarkandensis and other two hare species (L. capensis and L. europaeus).     

Analysis of type II toxin-antitoxin genes <Emphasis Type="Italic">all</Emphasis> 3211-<Emphasis Type="Italic">asl</Emphasis> 3212 in <Emphasis Type="Italic">Anabaena</Emphasis> PCC 7120

The type II toxin-antitoxin genes are responsible for the phenotypic switch to a quasi-dormant state that enables cell survival under stresses, a similar function to heterocyst of cyanobacteria. In this paper, we particularly study the role of gene pair all3211-asl3212 under Spectinomycin stress to reveal how the type II toxin-antitoxin involved in environmental stress responses. Bioinformatics prediction shows that toxin protein gene All3211 is homologous to MazF, a member of mazEF family that encoding nucleases. We clone gene all3211-asl3212 into expression vectors to identify its molecular characteristics. Deletion mutant strains of all3211-asl3212 are selected in a tri-parental mating screen. Phenotype comparisons of mutant and wild type reveals no difference of single-deletion-mutants in pigment integrity, the sensitivity to antibiotics, and heterocyst formation. The results show that deletion mutation of single TAS gene pair all3211-asl3212 results in limited effects on the cellular growth of PCC 7120. Thus, we suggest that dosage compensating might be provided from redundant genes or bypass pathways to offset obvious phenotypic differences.     

Biologically active <Emphasis Type="Italic">cis</Emphasis>-cinnamic acid occurs naturally in <Emphasis Type="Italic">Brassica parachinensis</Emphasis>

The biologically active cis-cinnamic acid (cis-CA) has been perceived as a synthetic plant growth regulator for decades,However,in the present study,we found that cis-CA actually exists as a naturally occurring compound in a Brassica plant,This natural growth-regulating substance presents in both the sunlight-irradiated leaf tissue and the non-irradiated root tissue ,The concentrations of cis-CA in both tissues are comparable to the bilogi-cally effective lvels of those major plant hormones,the presence of cis-CA in root tissue suggests that it may be produced through both light-dependent and -independent path-ways or it can be transproted from a plant organ to another.     

Genetic analysis and gene fine mapping of a rolling leaf mutant (<Emphasis Type="Italic">rl</Emphasis><Subscript><Emphasis Type="Italic">11(t)</Emphasis></Subscript>) in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The exploration of new genes controlling rice leaf shape is an important foundation for rice functional genomics and plant archi-tecture improvement. In the present study, we identified a rolling leaf mutant from indica variety Yuefeng B, named rl_11(t), which exhibited reduced plant height, rolling and narrow leaves. Leaves in rl_11(t) mutant showed abnormal number and morphology of veins compared with those in wild type plants. In addition, rl_11(t) mutant was less sensitive to the inhibitory effect of auxin than the wild type. Genetic analysis suggested that the mutant was controlled by a single recessive gene. Gene Rl_11(t) was initially mapped between SSR markers RM6089 and RM124 on chromosome 4. Thirty-two new STS markers around the Rl_11(t) region were developed for fine mapping. A physical map encompassing the Rl_11(t) locus was constructed and the target gene was finally delimited to a 31.6 kb window between STS4-25 and STS4-26 on BAC AL606645. This provides useful information for cloning of Rl_11(t) gene.     

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.     

Development of <Emphasis Type="Italic">Gossypium barbadense</Emphasis> chromosome segment substitution lines in the genetic standard line TM-1 of <Emphasis Type="Italic">Gossypium hirsutum</Emphasis>

Chromosome segment substitution lines （CSSL） consist of a battery of near-isogenic lines that have been developed and cover the entire genome of some crops. With the exception of one homozygous chromosome segment transferred from a donor parent, the remaining genome of each CSSL line is the same as the recipient parent. It is an ideal material for genome research and particularly QTL mapping. In the present study, we first developed one set of CSSL lines using G hirsutum acc. TM-1 （the genetic standard）, as the recipient parent and G barbadense cv. Hai7124 as the donor parent using molecular assistedlselection in BC5S1-3 generations. The CSSL consisted of 330 different lines, in which 1-4 different lines had the same or overlapping substituted segments. The genetic length of the substituted segments covered 5271.9 cM with an average segment distance of 10.9 cM, 1.5 times the total genetic length of Upland cotton （3514.6 cM）. The substituted segments of each line varied in length, ranging from 3.5 cM for the shortest segment to 23.2 cM in the longest segment. Our CSSL have not yet to cover the entire tetraploid cotton genome, due to the absence of some donor parent interval segments.     

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.     

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.     

Effect of GbKTN1 from Gossypium barbadense on cell elongation of fission yeast（Schizosaccharomyces pombe）

The GbKTN1 gene was isolated from 10 DPA fiber cells of Gossypium barbadense using 5′RACE/3′RACE.Full-length cDNA of this gene is 2006 bp, including a 113 bp of 5′untranslated region, a 1563 bp of an open reading frame(ORF), and a 327 bp of 3′untranslated region (excluding the stop codon TAA). The ORF of GbKTN1 encodes a 521-amino acid protein with a predicted size of 55 kD. Near C-terminal of the deduced protein there is a putative ATP binding site between amino acid residues from 233 to 414. Southern blot analysis indicated that the GbKTN1 was a single copy gene in G barbadense. Combining semi-quantitative RT-PCR with Southern blot hybridization revealed that GbKTN1 expressed in all the organs detected such as roots, stems, leaves and fibers. However, the mRNA of GbKTN1 was the most abundant in fiber cells, while it was the lowest in leaves. The GbKTN1 cDNA was transformed into S. pombe to verify its function on cell elongation. Results showed that most yeast cells over expressing GbKTN1 gene were elongated dramatically with an average length increase of 2.18 times than that of the non-induced cells. Even the morphology of some yeast cells appeared irregularly. To the best of our knowledge this is the first evidence that KTN1 is correlated with cell elongation in vivo.     

Genetic and physical finemapping of the Sc locus conferring indica-japonica hybrid sterility in rice （Oryza sativa L.）

Hybrid sterility is a major hindrance to utilizing the heterosis in indica-japonica hybrids. To isolate a gene Sc conferring the hybrid sterility, the locus was mapped using molecular markers and an F2 population derived from a cross between near isogenic lines. A primary linkage analysis showed that Sc was linked closely with 4 markers on chromosome 3, on which the genetic distance between a marker RG227 and Sc was 0.07 cM. Chromosome walking with a rice TAC genomic library was carried out using RG227 as a starting probe, and a contig of ca. 320 kb covering the Sc locus was constructed. Two TAC clones, M45EI4 and M90J01 that might cover the Sc locus, were partially sequenced. By searching the rice sequence databases with sequences of the TACs and RG227 a japonica rice BAC sequence, OSJNBb0078P24 was identified. By comparing the TAC and BAC sequences, six new PCR-based markers were developed. With these markers the Sc locus was further mapped to a region of 46 kb. The results suggest that the BAC OSJNBb0078P24 and TAC M45EI4 contain the Sc gene. Six ORFs were predicted in the focused 46-kb region.