A convenient method for extracting oospores of Phytophthora litchii from solid medium
A technology of litchi downy mildew and solid medium, which is applied in the field of bioengineering, can solve the problems of long formation period, harsh germination conditions, and difficult medium extraction, and achieve the effect of improving the possibility
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0022] Example 1: Growth rate of Phytophthora lychee on carrot solid medium not covered with film / covered with different types of film
[0023] Preparation of carrot culture medium: Weigh 200g carrots, fully squeeze the juice with a juicer, filter it with 16 layers of gauze, make up the water to 1000mL with the filtrate, add 15g agar, fully heat and dissolve, and distribute it in conical flasks (100mL per bottle culture medium), sterilized at 121°C for 20 minutes, cooled and stored for later use.
[0024] Activated Phytophthora lychee SHS-3 (NCBI, BioProject ID: PRJNA290406) was cultured in carrot medium at 25°C for 4 days, punched holes at the edge of the colony with a puncher, and inoculated the bacteria discs on uncovered film, cellophane-covered, Cover with nitrocellulose membrane and cover with Hybond TM N + Membraned carrot solid medium was cultured at 25°C in the dark for 10 days. With no membrane covered as the control, each membrane treatment was repeated 3 times....
Embodiment 2
[0026] Example 2: Growth of oospores in different types of film-covered carrot media
[0027] The cultivation of litchi downy mildew is the same as embodiment 1. After the film was torn off, 0.5 cm × 0.5 cm bacterial blocks were cut near the inoculation point of the control group and different film treatment groups, pressed into pieces and counted the number of oospores. The experiment was repeated 3 times.
[0028] It was found that the number of oospores in the carrot medium treated with cellophane and nitrocellulose membrane was significantly reduced compared with the control without membrane, while Hybond TM N + There was no significant difference in the number of oospores in the film-treated carrot medium (see image 3 ).
Embodiment 3
[0029] Example 3: Overriding Hybond TM N + Determination of viability of oospores in film-treated carrot medium
[0030] The cultivation of litchi downy mildew is the same as embodiment 1. The viability of oospores was determined by MTT (tetrazolium bromide, thiazolium blue) staining. Prepare a concentration of 0.1% MTT with sterilized deionized water, filter and sterilize it, mix it with an equal volume of oospore suspension, incubate at 36°C for 2 days, and check the results under a microscope. Those that were stained pink or blue were viable, and those that were not stained or black were nonviable (Sutherland et al., 1983). It turns out that covering Hybond TM N + There was no significant difference in the proportion of viable oospores in the film-treated carrot medium compared with the control without film (see Figure 4 , Figure 5 ).
[0031] The above results show that the method of the present invention can effectively and quickly separate and extract oospore...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More