Conidia coating method for producing large number of rice blast germ sexual generations
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A technology of conidia and rice blast fungus, applied in the field of microorganisms, can solve the problems of large differences between samples, instability, and inability to support the sample size, and achieve the effect of consistent maturity and fast formation speed
Pending Publication Date: 2022-02-22
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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This method can produce mature ascothecia and ascospores, but the ascothecia bandwidth is only 2-3mm, and the total amount of ascothecia and ascospores is less, which brings inconvenience to further research, especially molecular biology research, because the useful The amount of sex spores cannot support the sample size required by molecular biology research such as nucleic acid sequencing and proteome analysis
At the same time, the above method also has disadvantages such as instability, large differences between samples, and inconsistent maturity of sexual offspring.
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Embodiment 1
[0036] Embodiment 1, 4×10 4 Individual / mL Magnaporthe grisea spore suspension to prepare sexual generation
[0037] 1. Strains and culture conditions
[0038] Pyricularia oryzae strains Guy11 (MAT1-1), TH16 (MAT1-1), 2539 (MAT1-2), TH3 (MAT1-2) were cultured on complete medium (CM). Oatmealagar medium (OMA) was used to induce sexual generations of strains with different mating types.
[0039] CM Medium Formula (1L)
[0040]
[0041] The above reagents were weighed according to the corresponding amount and then adjusted to the corresponding volume. The pH was adjusted to 6.5 with NaOH, and 15 g of agar powder was added per liter of solid medium, and sterilized by high-pressure steam at 121 ° C for 15 min.
[0042] in:
[0043] 1000×Vitamin solution (100ml):
[0044]
[0045] The above reagents were dissolved in distilled water, and the volume was adjusted to 100ml, and stored in a refrigerator at 4°C.
[0046] 1000×Trace Elements (100ml):
[0047]
[0048] The a...
Embodiment 2
[0069] Embodiment 2, 5×10 3 Individual / mL Magnaporthe grisea spore suspension to prepare sexual generation
[0070] Select Guy11+TH3 for use, the blast fungus spore suspension in embodiment one is adjusted to 5 * 10 3 individual / mL, the illumination was 5000lux, and the sexual generation ( Figure 7 ).
Embodiment 3
[0071] Embodiment three, 5×10 3 Individual / mL Magnaporthe grisea spore suspension to prepare sexual generation
[0072] Select Guy11+TH3 for use, the blast fungus spore suspension in embodiment one is adjusted to 5 * 10 3 cells / mL, the light was 10000lux, and the sexual generation was obtained.
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Abstract
The invention provides a conidia coating method for producing a large number of rice blast germ sexual generations. The conidia coating method comprises the following steps: 1) preparing conidia suspensions of different mating types of rice blast germs; 2) coating conidium suspensions of different mating types on the oat culture medium according to a ratio of 1: 1; and 3) culturing for 5-15 days under the culture conditions that the culture temperature is 12-27 DEG C and the illumination is 5000lux-15000lux, so as to obtain a large number of rice blast fungus cocysts and ascospores. According to the method, the defects that sexual generation ascocysts and ascospores generated by a confrontation culture method in the prior art are small in total amount, unstable, large in sample difference, inconsistent in sexual offspring maturity and the like are overcome, a large number of rice blast bacteria sexual generations can be obtained within a short time, the formation speed of the ascocysts and the ascospores is high, the maturity is consistent, and the obtained ascocysts and ascospores can be used for quantitative comparison of sexual reproductive capacity of strains, sexual structure observation, nucleic acid extraction and gene expression quantitative analysis.
Description
technical field [0001] The invention relates to the field of microorganisms, in particular to a new method, which relates to a method for cultivating and preparing the sexual generation of Magnaporthe grisea. The preparation method is a spore coating method (Spore coating, SC method). Background technique [0002] Rice blast is one of the most important diseases of rice, seriously affecting rice yield. Rice blast fungus, also known as rice pear spore, Latin name Pyricularia oryzae, belongs to the kingdom Fungi, Ascomycota, Sordariomycetes, Magnaporthales, Pyriculariaceae, Pyricularia. The rice blast fungus can infect the leaves, stem nodes, branches, panicle necks and grains of rice. In addition to rice, it can also infect more than 50 kinds of gramineous plants such as barley, wheat, and millet. Currently, screening and cultivating disease-resistant rice varieties is the most important measure for rice blast control. However, due to the continuous variation of blast fungu...
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