86 results about "Fungal gene" patented technology

The invention discloses a lilium regale WRKY transcription factor gene LrWRKY1. The lilium regale WRKY transcription factor gene LrWRKY1 has a nucleotide sequence shown as SEQ ID NO: 1, and codes a protein with a nucleotide sequence shown as SEQ ID NO: 2. According to the lilium regale Wilson WRKY transcription factor gene LrWRKY1, functional genomics related technical research verifies that the LrWRKY1 gene has the function of increasing the fungus resistance of plants; when the antifungal LrWRKY1 gene is constructed to a plant expression vector and is transferred to tobaccos for overexpression, transgenic tobacco plants have strong in-vitro antifungal activities, and experimental results show that the LrWRKY1 overexpressed transgenic tobaccos have obvious inhibiting effects on the growth of four fungi including botryosphaeria, sclerotinite, botrytis cinerea and fusarium oxysporum.

The invention discloses a lilium regale WRKY transcription factor gene LrWRKY4. The nucleotide sequence of the lilium regale WRKY transcription factor gene LrWRKY4 is described as in the SEQ ID NO:1,and the coded protein of the lilium regale WRKY transcription factor gene LrWRKY4 corresponds to the amino acid sequence described in the SEQ ID NO:2. According to the lilium regale WRKY transcriptionfactor gene LrWRKY4, it is proved that the LrWRKY4 gene has the function of improving the antifungal ability of plants through the technical research related to the functional genomics, the antifungal LrWRKY4 gene is constructed to a plant expression vector and transferred into tobacco for overexpression, the transgenic tobacco has very high antifungal activity, and the experimental result showsthat the tobacco with the LrWRKY4 gene overexpressed is highly resistant to infestation of nigrospora oryzae, fusarium graminearum, fusarium graminearum, botryosphaeria and fusarium solanum.

A reporter system reflecting the transport process that transports GPI-anchored proteins to the cell wall was constructed and compounds inhibiting this process were discovered. Further, fungal genes conferring resistance to the above compounds were identified and methods of screening for compounds that inhibit the activity of the proteins encoded by these genes were developed. These genes encode proteins participating in fungal cell wall synthesis. Therefore, through the novel compounds, the present invention showed that antifungal agents having a novel mechanism, i.e. inhibiting the process that transports GPI-anchored proteins to the cell wall, could be achieved.

The invention discloses a Lilium regale Wilson WRKY transcription factor gene LrWRKY11. The Lilium regale Wilson WRKY transcription factor gene LrWRKY11 has a nucleotide sequence shown as SEQ ID NO: 1, and encodes a protein having an amino acid sequences shown as SEQ ID NO: 2. According to the invention, genomic studies have confirmed that the LrWRKY11 gene can improve anti-fungal functions of plants. Being constructed onto a plant expression vector and transferred into tobacco so as to be subjected to overexpression, the anti-fungal LrWRKY11 gene disclosed by the invention is capable of endowing the transgenic tobacco with very strong resistance to fungal pathogens. Experimental results have shown that transgenic tobacco with overexpression of the LrWRKY11 gene has very high level of resistance to infection of Nigrospora oryzae, Fusarium verticillioides, Botryosphaeria dothidea, Fusarium solani and Alternaria panax.

The invention discloses a lilium regale WRKY transcription factor gene LrWRKY3 and a preparation method thereof. A nucleotide sequence of the gene is shown as SEQ ID NO: 1, and a protein with an aminoacid sequence shown as shown in SEQ ID NO: 2; according to the invention, functional genomics related technical researches prove that the LrWRKY3 gene has the function of improving the antifungal performance of plants; the antifungal LrWRKY3 gene disclosed by the invention is constructed on a plant expression vector and is transferred into tobacco for overexpression; the transgenic tobacco plantshave very strong fungus infection resistance, and experimental results show that transgenic tobacco leaves with over-expression of LrWRKY3 have very strong resistance to infection of five pathogenicfungi such as alternaria oryzae, Botryosphaeria dothidea, fusarium graminearum, Fusarium rotunda and fusarium solani.

The invention discloses a lilium regale gene Lr14-3-3 with antifungal activity. The gene Lr14-3-3 has the nucleotide sequence as shown in SEQ ID NO:1 and encodes protein 14-3-3. A relevant technology of functional genomics proves that the gene Lr14-3-3 has a function of improving the plant antifungal activity. The antifungal gene Lr14-3-3 is constructed to a plant expression vector and is transferred into tobacco to perform overexpression; the transgenic tobacco plant has strong in vitro antifungal activity; and the transgenic tobacco expressing the Lr14-3-3 has obvious inhibition effects on the growth of botryosphaeria dothidea, phomopsis fungi, fusarium oxysporum and alternaria.

The invention discloses a pseudo-ginseng WRKY transcription factor gene PnWRKY9 and the application thereof. The nucleotide sequence of the PnWRKY9 gene is as shown in SEQ ID NO: 1, and the PnWRKY9 gene encodes a WRKY transcription factor; according to the application, molecular biology and functional genomics related technical researches prove that the PnWRKY9 gene has the function of improving the fungal disease resistance of plants, the antifungal gene PnWRKY9 is constructed on a plant expression vector and transferred into tobacco for overexpression, and the transgenic tobacco plant has very strong in-vitro antifungal activity; according to the present invention, the PnWRKY9 overexpressed transgenic tobacco has the significant inhibition effect on the growth of Alternaria Compacta, Fusarium solani, and Nigrospora oryzae, and the PnWRKY9 overexpressed transgenic tobacco has the significant inhibition effect on the growth of the Alternaria Compacta, the Fusarium solani, and the Nigrospora oryzae, such that the PnWRKY9 overexpressed transgenic tobacco has the significant inhibition effect on the growth of the Nigrospora oryzae,.

The invention discloses an application of a lilium regale wilson pathogenesis-related protein 10 gene LrPR10-5. A nucleotide sequence of the LrPR10-5 is as shown in SEQIDNO:1, and the pathogenesis-related protein 10 gene is encoded. The research proves that the LrPR10-5 gene has the function of improving the plant antifungal capability by functional genomics-related technologies, the antifungal LrPR10-5 gene is built on a plant expression vector and is switched into tobacco to perform overexpression, the transgenic tobacco plant has strong in-vitro antifungal activity in the result, and the experiment result shows that the transgenic tobacco for overexpression of the LrPR10-5 has an obvious inhibiting effect on growth of a plurality of fungi, such as botrytis cinerea, rhizoctonia solani and sclerotinia sclerotiorum.

The invention discloses a radix notoginseng mitogen-activated protein kinase kinase gene PnMAPKK1.A nucleotide sequence of the gene PnMAPKK1 is as shown in SEQ ID NO:1, and the gene PnMAPKK1 codes mitogen-activated protein kinase kinase.According to functional genomics related technical researches, the gene PnMAPKK1 has a function of improving pathogenic fungus resistance of plants.After the antifungal gene PnMAPKK1 is constructed to a plant expression vector and transferred into tobaccos to realize overexpression, transgenic tobacco plants have extremely high in-vitro antifungal activity.The transgenic tobacco plants with PnMAPKK1 overexpressed have an evident inhibition effect on growth of fusarium solani, colletotrichum gloeosporioides, verticillium fusarium and botryosphaeria.

A reporter system reflecting the transport process that transports GPI-anchored proteins to the cell wall was constructed and compounds inhibiting this process were discovered. Further, fungal genes conferring resistance to the above compounds were identified and methods of screening for compounds that inhibit the activity of the proteins encoded by these genes were developed. These genes encode proteins participating in fungal cell wall synthesis. Therefore, through the novel compounds, the present invention showed that antifungal agents having a novel mechanism, i.e. inhibiting the process that transports GPI-anchored proteins to the cell wall, could be achieved.

The invention relates to regulators of fungal gene expression and their use in commercial and medical applications. More particularly, the invention relates to regulators of fungal genes involved in production of enzymes, secondary metabolites and other useful products, as well as to regulators of genes involved in fungal invasion. The invention provides novel regulators of fungal gene expression, and methods for using regulator genes in commercial and medical applications.

The inventing discloses a Lilium regle bZIP transcription factor LrbZIP1. According to the invention, the nucleotide sequence of the gene LrbZIP1 is as shown in SEQ ID No:1, and the gene LrbZIP1 encodes a protein of a nucleotide sequence as shown in SEQ ID No:2. Functional genomics related technical research proves that LrbZIP1 has a function for improving the antifungal property of plants, after the antifungal LrbZIP1 gene is constructed in a plant expression vector and transferred to tobacco for overexpression, the result shows that the transgenic tobacco plant has strong in-vitro antifungal activity, so the experiment shows that the LrbZIP1-overexpressed transgenic tobacco has an obvious inhibiting effect on growth of various fungus such as ascomycetes and fusarium oxysporum.

The invention discloses agrobacterium tumefaciens mediated RNA (Ribonucleic Acid) interfering method suitable for various filamentous fungi gene study. A recombinant vector PCB309-pfgrt suitable for transfecting agrobacterium tumefaciens is obtained through a genetic recombination and Interference system screening method, and the transformation system of agrobacterium tumefaciens is optimized. According to the method, the problem that the RNA interference vector of filamentous fungi is hard to construct and the interference efficiency is low is solved. The method has wide application prospect in gene function and genetic transformation of filamentous fungi.

The invention discloses a primer, molecular beacon and kit for fast detecting various clinically common fungi and identifying strains and belongs to the technical field of microbiological detection. The sequences of the primer is as shown in SEQ ID No. 1-4, and the sequences of the detecting molecular beacon is as shown in SEQ ID No. 5-7. The primer and the molecular beacon have the advantages that the specific primer and the molecular beacon are designed at conserved sites by comparing the gene sequences of various clinically common fungi, and the primer and the molecular beacon can fast detect and identify 8 clinically common candida, 1 cryptococcus, aspergillus, mucor and penicillium. The kit for detecting various fungi contains the primer and the molecular beacon and can fast and accurately detect various clinically common fungi.

A method for higher fungi gene disruption includes that a ganoderma lucidum strain capable of stably expressing Cas9 protein by transferring an expression vector containing codon optimized Cas9 gene into haploid ganoderma lucidum (CGMCC NO.5.26); gRNA going through in-vitro transcription is transferred into the ganoderma lucidum expressing the Cas9 protein, a targeted gene is recognized through gRNA, and disruption of specific loci of the targeted gene is realized; endogenous transcription of gRNA is mediated by U6snRNA promoter, and disruption of the specific loci of the targeted gene is realized. Ganoderma lucidum is used as a mode species, the expression vector containing the codon optimized Cas9 gene and a host cell are provided, and an editing method of constructing CRISPR / Cas9 genomethrough two modes of exogenous transcription synthesis of gRNA or endogenous U6small nuclear RNA (U6snRNA) promoter transcription synthesis of gRNA is provided.

The invention relates to a fungus colony PCR method and a pathogenic fungus identification method. The fungus colony PCR method aims to achieve the high positive rate of amplification, and the pathogenic fungus identification method aims to achieve short detection time and reliable results. The fungus colony PCR method comprises the following steps of: inoculating fungus cultures onto a culture medium for culture; taking fungus colonies which begin to appear on the culture medium as experimentally available colonies; and preparing a DNA template for colony PCR amplification. The pathogenic fungus identification method comprises the following steps of: inoculating clinical sample cultures or pathogenic fungus cultures onto the culture medium for culture; taking the colonies which begin to appear on the culture medium as the experimentally available colonies; preparing the DNA template for the PCR amplification; and detecting amplification products. The fungus colony PCR method and the pathogenic fungus identification method have the advantages of improving the positive rate of the fungus colony PCR amplification, solving the problems of complex steps, wall-breakage difficulty, longtime and the like in fungus genomic DNA extraction, performing colony identification at the very beginning of the growth of pathogenic fungi, and achieving reliable results and identification time remarkably shorter than conventional fungus phenotype identification time.

The invention discloses a susceptible fungal gene LrWRKY-S1 of lily and application thereof. A nucleotide sequence of the LrWRKY-S1 gene is shown by SEQ ID NO.1 and an amino acid sequence coded thereby is shown by SEQ ID NO.2. Biological functions of the protein are studied by a bacteriostasis experiment. Experimental results show the protein makes plants more susceptible to pathogenic fungi suchas Botrytis cinerea, fusarium oxysporum and Colletotrichum orbiculare. As found as well, the LrWRKY-S1 gene has a low expression level in lilium regale with high resistance, and a homologous gene thereof has high expression levels in lilium davidii, lilium longifolorum and lilium lancifolium with important economic values. The gene and the application thereof provide an efficient susceptible genefor genetic engineering reconstruction, lays a foundation for further cultivation of fungus-resistant / susceptible lily or new materials or new species of other plants, can provide theoretical and technical support for later application of the gene in plant genetic improvement, broadens ideas for prevention and control of plant diseases and has very extensive application values.

The invention discloses a Yangbi walnut germin-like protein gene JsGLP1 and an application thereof. The nucleotide sequence of the JsGLP1 gene is as shown in SEQ ID NO:1, and is capable of encoding germin-like protein. Through functional genomics related technological research, the gene JsGLP1 is proved to have the function of enhancing plant fungal infection resistance; the antifungal gene disclosed by the invention is constructed on a plant expression vector and is transferred into tobacco for over expression; the transgenic tobacco plant is strong in the in-vitro antifungal activity; and JsGLP1 over-expression transgenic tobacco has a significant inhibitory effect on the growth of sclerotinia sclerotiorum, beaded gibberella, colletotrichum gloeosporioides and fusarium oxysporum.

The invention discloses a pathogenesis-related protein 1 family gene PnPR1-2 of panax notoginseng. The nucleotide sequence of the pathogenesis-related protein 1 family gene PnPR1-2 is as shown in SEQ ID No. 1. The pathogenesis-related protein 1 family gene PnPR1-2 comprises protein encoded with an amino acid sequence as shown in SEQ ID No. 2. Related technological researches of functional genomics prove that the PnPR1-2 gene is capable of increasing the fungus resistance of plants; when the PnPR1-2 gene is built onto a plant expression carrier and transferred into tobacco for overexpression, the transgenic tobacco plant is high in in-vitro antifungal activity, and the overexpression PnPR1-2 transgenic tobacco has an evident inhibiting effect on the growth of various fungi such as Fusarium solani, Verticillium Fusarium, beaded gibberella and Alternaria panax.

The invention relates to a kit for extracting fungal genome DNA and belongs to the technical field of biology. The kit for extracting the fungal genome DNA comprises the specific steps of crushing fungal cells, dissociating DNA and removing impure proteins, adsorbing DNA by using a DNA adsorbing column, removing impurities and eluting DNA, finally, so as to obtain a liquid which is the fungal genome DNA which is subjected to electrophoresis detection in 1% sepharose gel. The conventional steps for extracting the fungal genome DNA are tedious, consume a long time, and can extract the fungal genome DNA in about 18 hours generally. The method provided by the invention, compared with the conventional fungal genome extraction method, only needs about 7 hours to extract the fungal genome DNA, so that a lot of extraction time is saved, and the fungal genome DNA can be extracted more effectively, quickly and economically. Meanwhile, the kit is suitable for commercial production and has a certain economic value.

The invention discloses notoginseng pathogenesis-related protein gene PnPRlike and application. The nucleotide sequence of the PnPRlike gene is shown in SEQ ID NO: 1, which is an encoding pathogenesis-related protein; the related technical research of functional genomics proves that the PnPRlike gene has the function of improving the resistance of plants to pathogenic fungus, and the antifungal gene PnPRlike is constructed onto a plant expression vector and is converted into tobacco for overexpression, transgenic tobacco plant has extremely high in vitro anti-fungal activity, and experimentalresults show that the over-expression transgenic tobacco for PnPRlike has an obvious inhibiting effect for the growth of Fusarium solani, Botryosphaeria dothidea and Nigrospora oryzae.

The nucleotide sequence of the lilium regale chitinase gene LrCHI2 is shown as SEQ ID NO: 1, and the lilium regale chitinase gene LrCHI2 encodes a protein with an amino acid sequence shown as SEQ ID NO: 2. Functional genomics related technical researches prove that the LrCHI2 gene has the function of improving the resistance of plants to pathogenic fungi. The antifungal gene LrCHI2 disclosed by the invention is constructed on a plant expression vector and transferred into tobacco for overexpression, and the transgenic tobacco for overexpression of LrCHI2 has relatively strong capability of resisting infection of alternaria oryzae, aspergillus oryzae and fusarium oxysporum.

The invention discloses a double-fluorescence screening method for fungal gene knockout, and relates to the technical field of fungal gene knockout. The method comprises the steps that a 2.9-kb URA3-2micro2_origin fragment in a pYES2 carrier DNA template is amplified, the fragment is inserted into a pCAMBIA1300 carrier, and a pUM carrier is obtained; then, a tef1 promoter, an eGFP gene, a PgpdA promoter, a ble resistance gene, an RFP gene, a TtrpC terminator, an upstream homologous arm of a target gene x and a downstream homologous arm of the target gene x are amplified respectively; finally,a knockout carrier pKO-x of the target gene x is established; the pKO-x is transformed into agrobacterium competent cells, fungi is transformed through an agrobacterium mediated method, antibiotic screening is carried out to obtain a resistance transformant, then fluorescence microscope detection is carried out on the resistance transformant, the resistance transformant is a target transformant ifonly red fluorescence is emitted, the resistance transformant is an ectopic integration transformant if both green fluorescence and red fluorescence exist, and the transformant is a wild type if no fluorescence is emitted. The method can be used for effectively distinguishing the target transformant from the ectopic integration transformant.

The invention relates to regulators of fungal gene expression and their use in commercial and medical applications. More particularly, the invention relates to regulators of fungal genes involved in production of enzymes, secondary metabolites and other useful products, as well as to regulators of genes involved in fungal invasion. The invention provides novel regulators of fungal gene expression, and methods for using regulator genes in commercial and medical applications.

The invention discloses application of a Yangbi big-bubble walnut transcription factor gene JsWRKY1. The JsWRKY1 gene is proved to have a function of enhancing the fungal disease resistance of a plant through technical research related to functional genomics. The antifungal gene JsWRKY1 is established to a plant expression vector and transferred into a tobacco for overexpression, and a transgenic tobacco plant has very high in-vitro antifungal activity, and a transgenic tobacco subjected to JsWRKY1 overexpression achieves obvious inhibiting effect on the growth of botryosphaeria dothidea, moniliform gibberella, colletotrichum gloeosporioides and fusarium oxysporum.

The invention relates to an application of a lilium regale germin protein gene LrGLP1. The nucleotide sequence of the LrGLP1gene is shown as SEQ ID NO:1, a germin-like protein is encoded, and according to the invention, functional genomics related technical researches prove that the LrGLP1gene has a function of improving the fungal infection resistance of plants. An antifungal LrGLP1 gene in the invention is built into a plant expression vector, and transferred into a tobacco to be subjected to excessive expression, so that a transgenic tobacco plant has an extremely strong in-vitro antifungal activity. LrGLP1overexpressed transgenic tobaccos have an obvious inhibitory effect on the growth of alternaria alternate, sclerotinia sclerotiorum and catenuliform gibberella.

The invention provides a general and facile method to obtain secondary metabolites from fungal sources. The invention is based on the discovery that the fungal gene veA and protein encoded thereby regulates the activity of multiple secondary metabolite gene clusters in fungi. Over expression of the gene veA provides increased production of secondary metabolites in engineered cells. In particular, such a method of increasing secondary metabolite production allows the production of improved yields of valuable secondary metabolite products.

The invention provides an antifungal gene, polypeptide and recombinant protein as well as a preparation method and an application thereof, the antifungal gene is a Ruditapes philippinarum antifungal Rpmacin gene, and a nucleotide sequence of the Rpmacin gene is shown as SEQ ID NO.1; the amino acid sequence of the polypeptide is as shown in SEQ ID NO. 2; the recombinant protein contains the antifungal gene and expresses the antifungal polypeptide. The antifungal gene, polypeptide and recombinant protein provided by the invention can be applied to production of antibacterial drugs, vaccines or feed additives for vegetables, livestock, poultry and aquatic products, and have broad application prospects. The gene, polypeptide and recombinant protein as well as the preparation method and the application thereof can provide a basis for further researching the immune defense mechanism of Ruditapes philippinarum, and lay a foundation for disease control, genetic breeding and feed additive research of Ruditapes philippinarum.

The invention relates to a traceless gene editing method for trichoderma fungi. According to the method, the traceless knockout mutant of the trichoderma guizhouense ura3 gene is obtained through twotimes of homologous recombination events and in combination with the resistance screening strategy of hygromycin B and the lethal strategy of 5-FOA; based on the mutant, the knockout fragment containing the ura3 gene expression cassette is inserted at the position of the target gene through the homologous recombination method, and the mutant having the first-time homologous recombination is screened by utilizing the nutrition defect feature of the ura3 traceless mutant; after the second-time homologous recombination, the ura3 gene and the target gene are removed through recombination, at the time, inverse screening is carried out by utilizing the lethal feature of 5-FOA, and thus the traceless mutant with the target gene completely deleted is obtained. The system is optimized sufficiently,the homologous recombination ratio is 15% or above, the single-gene traceless operating period is shortened to be within 15 days, meanwhile, the traceless overexpression of the target gene can be realized, and no exogenous fragments are introduced during the process.

The invention discloses a panax notoginseng thaumatin-like protein gene PnTLP2, which is the thaumatin-like protein encoding as an amino acid sequence shown in SEQ ID NO: 2 and having a nucleotide sequence shown in SEQ ID NO: 1. The applicaition adopts molecular biology and reverse genetics related techniques to confirm that the PnTLP2 gene has the function of improving fungal resistance of plants, the antifungal PnTLP2 gene of the invention is constructed on a plant expression vector and transferred into tobacco for overexpression, the transgenic tobacco plants have strong in-vitro antifungalactivity, the transgenic tobacco overexpressing PnTLP2 has significant inhibitory effects on five plant pathogenic fungi of fusarium oxysporum, fusarium solani, alternaria panax, sclerotinia sclerotiorum and nigrospora oryzae.