13results about "Bryophytes" patented technology

The invention discloses a screening method and application of a floating moss endogenous constitutive promoter, and belongs to the technical field of gene engineering. A floating moss endogenous constitutive promoter is screened out through experiments such as screening verification of candidate constitutive promoters, a plant expression vector of which a reporter gene started by the promoter is positioned in a cell nucleus is constructed, and floating moss is transformed to obtain a transgenic plant. The fluorescence intensity of the reporter gene driven by the promoter is higher than that of other common constitutive promoters, such as CaMV35S and the like, and meanwhile, when fluorescence expression of transgenic plants is observed, it is found that the reporter gene can be stably expressed in thalli, scales, various different types of cells of the thalli and the scales and the like. The invention provides an efficient and universal core tool for floating moss gene function research, genetic manipulation, biotechnology application and the like, and has a certain application prospect.

The invention discloses a stable genetic transformation method of floating moss. The genetic transformation method comprises the following steps: (1) crushing thallus tissues of the floating moss, (2) pre-culturing explants, (3) infecting and co-culturing, (4) screening and culturing for the first time, (5) screening and culturing for the second time, (6) enlarging and culturing, and (7) verifying transformation. According to the genetic transformation method, a thallus of floating moss is taken as a starting explant, an agrobacterium-mediated hygromycin and kanamycin screening transformation system of the floating moss is established for the first time, the blank of a floating moss genetic transformation system is filled, and experimental results show that after multi-generation culture, a transformed plant can still stably express a target gene. The method has the advantages of high transformation efficiency and simplicity in operation, and can be used for subsequent genetic transformation research of unknown floating moss genes. The method has a wide application prospect.

The invention discloses a construction and optimization method of a CRISPR / Cas9 system of floating moss, and belongs to the technical field of gene editing. According to the constructed and optimized system, firstly, a positive plant which is successfully knocked out in floating moss is successfully obtained on the basis of a CRISPR / Cas9 system of an existing moss plant liverwort, but the transformation efficiency of the positive plant is not high; meanwhile, based on the fact that a constitutive promoter of a floating moss endogenesis is screened out in a laboratory, in order to ensure the stability and efficiency of the system, a self RnU6 promoter RnU68 of the floating moss is screened out, and the floating moss CRISPR / Cas9 system which is high in conversion efficiency and stable is successfully constructed.

The present invention relates to a recombinant construct comprising a first expression cassette comprising: a first nucleic acid molecule functional in a plant; the first nucleic acid molecule is selected from ICAs family genes; preferably, the first nucleic acid molecule codes an amino acid sequence as shown in any one of SEQ ID NO.1-9, or an amino acid sequence having at least 60% homology with the sequence as shown in any one of SEQ ID NO.1-9; and a first promoter heterologous operably linked to the first nucleic acid molecule. The application shows that by performing genetic manipulation on the ICAs gene of the plant and the promoter thereof, the Ca < 2 + > nutrient absorption of the plant can be promoted, so that the Ca < 2 + > content of the plant is increased, the tolerance of the plant to environmental stress is enhanced, and the method has important significance for cultivating new varieties of crops with high composite stress tolerance and new varieties of crops with high calcium nutrition.

The present invention provides a high throughput method capable of screening compounds for herbicidal or plant growth regulating activity, and further allows the prediction of the mode of action of herbicidal compounds or plant growth regulators.SOLUTION: The methods provided herein also facilitate identification of mutations involved in tolerance to herbicides in plants and identification of herbicide targets. The methods further provide for the identification of mutations involved in plant growth regulation and the identification of plant growth regulator targets.SELECTED DRAWING: Figure 1

The invention discloses a subcellular localization method suitable for floating moss, and belongs to the technical field of mutation or genetic engineering and DNA or RNA and vectors related to the genetic engineering. According to the method disclosed by the invention, the tender thallus of the floating moss is used as a material, and a related protein positioning gene is transferred into a floating moss species through a floating moss stable genetic transformation system. A series of subcellular localization genes are obtained through cloning in a floating moss genome for the first time, the genes can assist GFP fusion protein to enter different subcellular organelles, and a marker line (localization strain) is obtained through stable transformation of a floating moss material. The method can be used for verifying a subcellular localization result of a candidate floating moss target gene, and is a simple, convenient and stable floating moss subcellular localization method. The method has a wide application prospect.

The invention relates to the identification and characterization of herbicides and plant growth regulators. The present application provides high throughput methods that are capable of screening for compounds with respect to herbicidal or plant growth regulating activity and further allow for prediction of the pattern of action of herbicide compounds or plant growth regulators. The methods provided herein also facilitate identification of mutations responsible for herbicide resistance in plants and identification of herbicide targets.

This invention discloses the application of the *Rhizopus dentata* ScPRT1 gene in regulating plant responses to abscisic acid (ABA), the nucleotide sequence of which is shown in SEQ ID NO:1. This invention found that *Arabidopsis thaliana* lines overexpressing the *Rhizopus dentata* ScPRT1 gene, or *Rhizopus dentata* lines transiently expressing the *Rhizopus dentata* ScPRT1 gene, exhibited significantly reduced ABA sensitivity and significantly enhanced ABA response. Therefore, the *Rhizopus dentata* ScPRT1 gene negatively regulates plant responses to ABA signals. This invention provides resources for the discovery of stress-resistance genes, improves the regulatory network of *Rhizopus dentata* in response to ABA signals, and offers a new perspective for understanding the molecular mechanisms of stress adaptation in desert bryophytes.

The invention discloses application of syntrichia caninervis ScPRT1 gene in regulation and control of plant response to abiotic stress, the nucleotide sequence of the syntrichia caninervis ScPRT1 gene is as shown in SEQ ID NO: 1, and the amino acid sequence of protein coded by the syntrichia caninervis ScPRT1 gene is as shown in SEQ ID NO: 2. It is found that the syntrichia caninervis ScPRT1 gene negatively regulates the resistance of plants to salt and drought, and as a negative regulation factor of an ABA signal channel, the syntrichia caninervis ScPRT1 gene reduces the sensitivity to ABA. According to the invention, resources are provided for excavation of the stress resistance regulation gene, and an important scientific basis is provided for comprehensive analysis of a stress resistance regulation mechanism of the ScPRT1 gene and cultivation of stable-yield and high-quality crop varieties.

The present disclosure relates to a bryophyte cell that is transformed or genetically modified to express aureusidin synthase and chalcone-4'-glycosyltransferase. The aureusidin synthase and chalcone-4'-glycosyltransferase may be expressed as a single biscistronic cassette separated by a P2A peptide or separately through insertion of two unique gene cassettes into said bryophyte cell. The expression may be driven by an exogenous promoter. The transformed bryophyte cell may be exposed to UV radiation to enhance production of exogenous flavonoids.

The invention discloses a nucleotide fragment combination for constructing a paclitaxel biosynthesis engineering strain and an engineering strain construction method, and relates to the technical field of bioengineering, the nucleotide fragment combination comprises a nucleotide fragment I, a nucleotide fragment II and a nucleotide fragment III; each fragment respectively comprises genes required by the biological synthesis pathway of 5a-hydroxylated taxadiene, baccatin III and paclitaxel; in each nucleotide fragment, a single promoter is at most used for starting four genes, and the genes started by the single promoter are connected through 2A peptides. The invention also discloses a method for constructing the paclitaxel biosynthesis engineering strain and the paclitaxel biosynthesis engineering strain. According to the invention, the nucleotide fragment combination is utilized to successfully construct the taxol biosynthesis engineering strain of the taxiphocephalum leptosphaeroides through iterative transformation, so that stable biosynthesis of the taxol in a heterologous chassis is realized, and the method has important practical application value.

A method for in vitro culturing Sphagnum is provided. The method comprises providing a vegetative fragment of Sphagnum and surface cleaning the fragment with alcohol. The method further comprises dissecting the fragment to provide a piece (106) of Sphagnum. The method further comprises initiating the piece (106) of Sphagnum in a culture medium (104).

The invention discloses a method for realizing large-fragment DNA (Deoxyribose Nucleic Acid) insertion by utilizing an IVC (Intravariant Vitamin C) donor system with a shortened homologous arm in Amplistrum canescens. Aiming at the problem of high cloning complexity caused by dependence of an existing IVC donor system on a long homologous arm (about 1kb), the invention initiatively proposes that the homologous arm is shortened to 250-510bp, and the insertion efficiency of an IVC ring donor and a linearized donor after the homologous arm is shortened is compared and evaluated for the first time, and the result shows that the insertion efficiency of the IVC ring donor is obviously higher than that of the linearized donor, and the insertion efficiency of the linearized donor is obviously higher than that of the linearized donor. And it is proved that the IVC ring donor with the shortened homologous arm can still keep about 50% of insertion efficiency. According to the method, the cloning difficulty is remarkably reduced while the relatively high insertion efficiency is maintained, the technical bottleneck that long homologous arms are needed for large-fragment DNA insertion is successfully broken through, and an efficient tool is provided for plant synthetic biology and crop improvement.