Endogenetic streptomycete for preventing kiwi berry bacterial canker and preparation thereof
A kiwifruit and bacterial technology, applied to the field of endophytic Streptomyces and its preparation, can solve problems such as unsatisfactory field disease prevention effect, and achieve the effects of good antibacterial activity, simple separation and screening method, and good water solubility
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Embodiment 1
[0029] Example 1: Isolation and morphological identification of the TIASA5 strain
[0030] 1) Wash the healthy wild Artemisia annua stem tissue collected from Yangling, Shaanxi with water, blot the water dry with absorbent paper, soak in 99% ethanol for 1 min, then soak in 3.125% NaClO for 6 min, and put it in again Soak in 99% ethanol for 30 sec, and finally rinse with sterile saline for 5 times;
[0031]2) Cut the sterilized Artemisia annua stem tissue to be separated into 0.2mm long pieces with a sterilized blade, place them on the Gaoshi No. 1 plate, culture them in an incubator at 28°C for 21-28 days, and count the number of actinomycete colonies And purify into a single colony by streaking method;
[0032] 3) The purified single colony is fermented as follows: use a puncher to punch the bacterial lawn plate that has been activated and cultured on the soybean flour plate for 5 to 7 days into a 6mm diameter bacterial cake, insert it into the soybean flour medium, and heat...
Embodiment 2
[0033] Embodiment two: the preparation method of the antibacterial active substance of TIASA5 bacterial strain
[0034] The TIASA5 strain that was activated and cultivated on the soybean flour plate for 5 days was punched into a bacterium cake with a diameter of 6 mm, inserted into the seed medium, and cultivated for 36 hours at 28 ° C and 150 r / min. The solution was introduced into the fermentation medium, and fermented at 30°C and 180r / min for 72h. Centrifuge the cultured fermentation broth at 10,000 r / min for 30 minutes, and take the supernatant, which is the crude fermentation broth. The crude fermentation broth was treated with 1M H 2 SO4 adjusted to pH2, 80°C water bath for 1h, then adjusted to pH7 with 1M NaOH, then centrifuged (10000r / min, 30min), collected the supernatant, and preliminarily removed the protein and CaCO in the fermentation broth 3 . Put the pretreated fermentation broth on 732 # Ion exchange resin, dynamically adsorbed, and then eluted with 1N ammo...
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