A kind of smut haploid strain uet1 and its application
A UET1, haploid technology, applied in the fields of application, fungi, plant cultivation, etc., can solve the problem of no haploid strains, etc., and achieve the effect of improving environmental adaptability and controlling mating time
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Embodiment 1
[0020] Embodiment 1: bacterial strain screening method
[0021] 1. Collect teliospores: from Longjiao No. 2 gray bamboo ( figure 1 A) Pick teliospores directly.
[0022] 2. Teliospore suspension: pile the picked teliospores into 5 mL sterile water, filter through 4 layers of sterile gauze to obtain a relatively pure teliospore suspension, and finally dilute the teliospore suspension with sterile water Dilute to a final concentration of 10 3 spores / mL. Teliospores form as figure 1 Shown in B.
[0023] 3. Teliospore culture: Take 100 µL of teliospore suspension and spread it on the basidiospore isolation medium, and culture it at 28°C for about 60 hours, finely dispersed single colonies can be seen with the naked eye ( figure 1 C) as standard. Basidiospore isolation medium (1 L) formula is: K 2 HPO 4 1g, MgSO 4 ·7H 2 O 0.5 g, FeSO 4 ·7H 2 O 0.01g, KCl 0.5 g, glucose 18 g, (NH 4 ) 2 SO 4 5.28 g, 10 g of agar.
[0024] 4. Single spore isolation and culture: Pi...
Embodiment 2
[0045] Embodiment 2: Experiment of artificial fertilized bamboo shoots
[0046] The vector of overexpressing EGFP green fluorescent protein was transformed into Ustilago sativa ( Ustilago esculenta ) haploid strain UET1 and its sex-compatible strains (select No. 2 and No. 3 in Example 1) to overexpress EGFP green fluorescent protein, and the obtained overexpression strains can be detected by fluorescence microscopy, and successfully overexpress EGFP The strains with green fluorescent protein can see the green fluorescent signal under the fluorescent microscope, such as figure 2 shown. Then carry out artificial inoculation experiment, concrete artificial inoculation process is as follows:
[0047] 1) Shake two haploid strains of smut to OD in YEPS liquid medium at 28°C 600 was 1.0, centrifuged to collect the bacteria, diluted with 0.5×YEPS liquid medium to a final concentration of OD 600 The bacterial solution is 3.0, and the two bacterial solutions are mixed for inoculati...
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