36 results about "Tilletia" patented technology

The invention relates to a preparation method of tilletia foetida protoplasts. The preparation method comprises the following steps: (1) mycelium culture: culturing teleutospores of tilletia foetida in a water agar culture medium for 6-9 days, flushing the teleutospores off from the water agar culture medium by virtue of sterile distilled water, collecting bacteria liquid, and carrying out centrifugation to remove supernatant, so as to obtain mycelia; and (2) cell wall cracking: preparing mixed enzyme liquid by taking a potassium chloride solution as an osmotic pressure stabilizer, adding the mixed enzyme liquid into the mycelia, and carrying out vibration enzymolysis at 28 DEG C for 1.5-2.5 hours, so as to obtain the tilletia foetida protoplasts, wherein the mixed enzyme liquid contains the following components in percentage by mass: 1.5% of driselase, 1.5% of lyticase and 1.5% of helicase. The method is used for preparing the tilletia foetida protoplasts, the yield of the tilletia foetida protoplasts reaches up to 17.0*10<5>unit / mL-18.0*10<5>unit / mL, and the released protoplasts are uniformly distributed in a scattered manner and are not aggregated into a pile.

The invention relates to antigens and a polyclonal antibody used for discriminating Tilletia controversa Kuehn and Tilletia foetida. A semiantigen used for preparing the polyclonal antibody for discriminating Tilletia controversa Kuehn and Tilletia foetida has an amino acid sequence as shown in SEQ ID No. 1. An immunogen used for preparing the polyclonal antibody for discriminating Tilletia controversa Kuehn is characterized in that the semiantigen mentioned above is reconstructed and coupled so as to obtain the immunogen, the immunogen is used to immunize domestic rabbits, domestic rabbits generating high-titer whole blood are screened and whole blood is acquired from a selected domestic rabbit and undergoes separating and purifying so as to obtain the polyclonal antibody. The antibody can specifically identify Tilletia controversa Kuehn without undergoing an antigen-antibody reaction with Tilletia foetida, so the polyclonal antibody can be used for discriminating Tilletia controversa Kuehn and Tilletia foetida difficult to distinguish in a field.

The invention discloses a standard molecule for detecting tilletia walkeri castebury&carris;. The standard molecule is a plasmid which can be duplicated, and the plasmid contains at least one of a sequence A (Seq ID N0.1) and a sequence B (Seq ID N0.2); the sequence A is 99.65% homologous with a 2.3kb nucleotide sequence of the mitochondrial DNA of the tilletia walkeri castebury&carris; and the sequence B is 100% homologous with the DNA sequence of a ribosomal internal transcribed spacer of the tilletia walkeri castebury&carris;. The invention also discloses a preparation method and detection method of the standard molecule. The standard molecule constructed in the invention can serve as a positive control for detecting the molecule of the tilletia walkeri castebury&carris; instead of the DNA of the tilletia walkeri castebury&carris; is suitable for qualitative and quantitative PCR (polymerase chain reaction) detection of the tilletia walkeri castebury&carris; and is suitable for the inspection and quarantine department of the port, the agricultural production department, the plant protection department and the like.

The invention relates to the field of cell biotechnologies, in particular to a method of identifying a Tilletia controversa Kuhn (TCK) teliospore and a Tilletia foetida (Walle.)Lindr. (TFL) teliospore. The method comprises the steps that a to-be-detected sample suspected to be a TCK teliospore or a TFL teliospore is placed in WGA-AF488 dyestuff for dying, washed with PBS and then placed under a confocal laser scanning microscopy (CLSM) for scanning observation; if the muri and cell wall are both dyed into green, the to-be-detected sample is the TCK teliospore; and if only the cell wall is dyedinto green, the to-be-detected sample is the TFL teliospore. Through the method, the dying result is clear, and the TCK teliospore and the TFL teliospore can be distinguished quickly and accurately.

The invention discloses a 'method for cultivating tilletia controversa kuhn disease plants in indoor environments', and belongs to the field of tilletia controversa kuhn control. The method includes steps of (1), preparing tilletia controversa kuhn germination winter spores; (2), preparing wheat seeds for tilletia controversa kuhn inoculation; (3), artificially inoculating wheat; (4), cultivating the disease plants in the indoor environments. The method for cultivating the tilletia controversa kuhn disease plants in the indoor environments has the advantages that the stable tilletia controversa kuhn morbidity can be acquired under indoor conditions, the method is beneficial to tilletia controversa kuhn biological research under the indoor isolation conditions, diffusion of germs during outdoor research can be prevented, and accordingly the method is favorable for controlling tilletia controversa kuhn diseases from various aspects.

The invention "a kit and method for detecting Tilletia dwarfis" belongs to the detection technology of plant pathogens. Including a test strip and a microporous plate, the test strip includes a bottom plate, a sample absorbent pad, a reaction film and a water absorbent pad; the sample absorbent pad, reaction film, and water absorbent pad are sequentially pasted and installed on the bottom plate; its features In that: there are freeze-dried micropore reagents in the microwells of the micropore plate, and the micropore reagents refer to gold-labeled antibodies obtained by labeling the monoclonal antibody described in claim 2 with colloidal gold; the reaction membrane is divided into A detection area and a quality control area, the detection area is coated with Tilletia tritici polyclonal antibody, and the quality control area is coated with anti-antibody. The kit of the invention has good specificity and very low detection limit, and is suitable for the prevention and control or quarantine of T. tritici.