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Method for infecting wheat by inoculating tilletia foetida to roots

A technology of Tilletia clematis and wheat, which is applied in the field of root inoculation of Tilletia clematis to infect wheat, and achieves the effect of simple method, easy implementation and operation, and is conducive to popularization and application

Active Publication Date: 2016-02-17
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the research on the physiological growth and infection mechanism of Tilletia tritici, usually it needs to be inoculated to wheat plants by artificial method so that the latter infects the infected wheat plants for observation and analysis, and, only for In the research on the roots of wheat plants, it may only be necessary to obtain wheat materials that are infected at the roots and appear normal in other parts, but so far, there is no effective method for artificially inoculating T. Obtain wheat plants with high root infection rate and normal appearance in other parts for research

Method used

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  • Method for infecting wheat by inoculating tilletia foetida to roots
  • Method for infecting wheat by inoculating tilletia foetida to roots
  • Method for infecting wheat by inoculating tilletia foetida to roots

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Embodiment 1

[0042] Embodiment 1, adopt method for the present invention to artificially inoculate wheat Tilletia glabra

[0043] Specific steps:

[0044] (1) prepare Tilletia tritici teliospore suspension:

[0045] Put the galls into a centrifuge tube, add distilled water to soak for half an hour, then clip the seed coat, filter through a 150-mesh gauze, and centrifugally shake, pour out the water and then add water to cultivate at room temperature for 24 hours; pour out the water the next day, After adding 0.25% NaCLO for 1 min, pour it out, add sterile water and centrifuge and wash it repeatedly 2 to 3 times to obtain a teliospore suspension; figure 1 shown.

[0046] (2) Cultivate the mycelia of Tilletia tritici: the concentration is 1×10 5 Spray the teliospore suspension of Tilletia tritici per ml on 2% water agar medium, add 0.1-0.5ml of the suspension to each 9cm-diameter petri dish, culture at 16°C, and confirm by observation until the growth of the first bacteria Silk, when the...

Embodiment 2

[0054] The infection situation verification and infection rate statistics of the wheat seeds that embodiment 2, the inventive method obtains

[0055] (1) Verification of infestation:

[0056] The root tissues of the vernalized seedlings obtained in Example 1 were taken to detect whether the wheat was infected. Using the CTAB method to extract the DNA of wheat roots, the DNA after extraction was subjected to PCR reaction with the specific primers SC1:ACAAGTTGGAGTCAGGAG; SC2:ATCGTATGGTTTCGTCTT of Tilletiafoetida after the extraction, and the reaction product was detected by agarose gel electrophoresis, and it was found that Specific bands, such as Figure 5 shown.

[0057] At the same time, the wheat root tissue was cut into small pieces and fixed in absolute ethanol. The sample was stained with 10 μg / mL Propidiumidodide (0.5 mL), 0.02% Tween20 (0.5 mL) for 10 min, and then 20 μg / mL WGA-AF488 (0.5 mL) After staining for 10 min, wash with 10×PBS (pH7.4, 1 mL) 3 to 5 times. WG...

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Abstract

The invention relates to a method for infecting wheat by inoculating tilletia foetida to roots, which is characterized by comprising the following steps: (1) culturing hypha of tilletia foetida: coating a 2% water agar culture medium with tilletia foetida teleutospore suspension with concentration of 1*10<5> / ml, culturing at 16 DEG C, and observing and confirming until promycelium grows, wherein 0.1-0.5ml of suspension is added to the culture dish of every 9cm diameter, and the promycelium is used for inoculation in the next step when the size of the promycelium is 1 / 2 the diameter of the teleutospore; (2) inoculating: placing sprouted wheat seeds to the culture medium in which the hypha grows, placing the culture dish to a 5 DEG C incubator, and simultaneously carrying out vernalization and infection for 30-45d; and (3) after vernalization is finished, planting wheat seedlings on sterile soil, and culturing in an artificial climate incubator with day temperature 23 DEG C and night temperature 21 DEG C, wherein the sprouted wheat seeds are sprouted wheat with coleoptile not more than 1mm. By adopting the method, wheat plants with roots infected by tilletia foetida and other parts presenting normal are obtained.

Description

technical field [0001] The invention belongs to the field of agricultural microorganisms, and in particular relates to a method for inoculating roots with Tilletia glabrata and infecting wheat. Background technique [0002] Tilletia tritici, also known as Tilletia tritici (Latin name: Tilletiafoetida), belongs to the order Tilletiaceae, Tilletiaceae, and the genus Tilletia tritici. It can cause tickle smut in wheat. When the teliospores of T. tritici germinated, only one original hyphae was produced, but the inner wall was disintegrated in many places. The original mycelium contains multiple bacterial nuclei, and there are fat particles, ribosomes, glycogen and mitochondria. Eight basidiospores can be formed on the top of the original hyphae, and bridged to form "H" type hyphae. Its spores are born in the ovary, surrounded by pericarp, and are the same size as the seeds. Its spores are spherical to oval, sometimes oblong to polygonal, light grayish brown to olive brown, ...

Claims

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Application Information

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IPC IPC(8): A01G7/06C12Q1/04C12R1/645
CPCA01G7/06C12Q1/04
Inventor 高利沈慧敏蔚慧欣李超陈万权刘太国刘博
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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