A strain of Streptomyces burdock endophyte, microbial agent containing the endophyte and its application
A technology of endophytic streptomyces and microbial agents, applied in the field of endophytic bacteria, achieving good application prospects, simple use, and easy promotion
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Embodiment 1
[0030] Isolation and identification of embodiment 1 bacterial strain WY228
[0031] 1.1 Isolation of strain WY228
[0032] Select healthy and well-growing burdock plants in the field of Hekou Town, Peixian County, Xuzhou City, Jiangsu Province, dig out the fleshy roots from the soil, shake off the floating soil on the surface, put the roots in a ziplock bag and seal them, and bring them back to the laboratory for strain isolation . Rinse the surface of the plant root sample with running water, cut the plant sample into a suitable size and soak it with sterile 0.01% Tween20 for 1 min, then soak the sample root with NaClO with 5% available chlorine content for 3 min, and then soak it with sterile 2.5% NaClO 2 S 2 o 3 Treat for 10 minutes, wash 5 times with sterile water, soak the sample with 75% ethanol for 2 minutes, and finally wash 3-5 times with sterile water. Take the sterile water cleaned in the last step of burdock root surface disinfection, spread it evenly on the me...
Embodiment 2
[0043] Example 2 Detection of growth-promoting potential characteristics of bacterial strain WY228
[0044] Detection of siderophore activity: Inoculate the strain WY228 on the CAS detection medium, and after culturing at 28°C for 5-7 days, observe whether there is a yellow transparent circle around the colony. If it appears, it proves that the strain has siderophore activity. That is positive. Qualitative detection found that strain WY228 could produce siderophore.
[0045] Detection of plant growth hormone indole acetic acid (IAA): Prepare a 2.5 mg / mL tryptophan solution, filter and sterilize with a 0.22 μm filter membrane in an ultra-clean workbench. Prepare liquid nitrogen-containing medium and divide it into test tubes, and sterilize under high pressure at 115°C for 30 minutes. Add 1 mL of tryptophan solution to the sterilized nitrogenous medium to make the concentration of tryptophan 0.5 mg / mL, insert the strain to be tested and culture it in a shaker at 28°C for 7 day...
Embodiment 3
[0052] Embodiment 3 contains the preparation method of the burdock special-purpose microbial bacterial agent of bacterial strain WY228
[0053] (1) Strain activation: Inoculate the slant of the isolated Streptomyces sp.WY228 strain or the preservation of the cryopreservation tube into solid medium containing ISP 2 (4g yeast extract, 10g malt extract, 4g glucose, 20g agar , 1000mL water, pH 7.2) on the slant, 28 ° C for 5-7 days, thereby activating the strain.
[0054] (2) Seed liquid culture: Pick a little bacteria from the activated ISP 2 solid medium slant with a bamboo stick and insert it into the ISP 2 liquid medium, and culture it on a shaker at 28°C with a speed of 150-180rpm for 2- After 3 days (logarithmic phase), the seed solution was obtained.
[0055] (3) Bacterial agent preparation: the seed culture solution cultivated to the logarithmic phase is inserted into the ISP 2 liquid fermentation medium (4g yeast extract, 10g malt extract, 4g glucose, pH 7.2, in 1000ml ...
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