A kind of salt-tolerant growth-promoting bacterial strain y4 and its application
A technology that promotes growth and salt tolerance, applied in the field of microorganisms, to achieve the effects of improving salt tolerance, increasing plant type, and slowing down the phenomenon of salt damage
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Embodiment 1
[0015] Example 1 Isolation, screening and identification of salt-tolerant growth-promoting bacterial strain Y4
[0016] Weigh 2.0g of Yancheng rapeseed rhizosphere soil sample (with plant roots) and place it in a conical flask filled with 100mL of double-distilled water, seal it with a parafilm, and shake it in a shaker at 200r / min at 30°C for 24h to obtain soil suspension. Take 1mL of soil suspension and transfer it to 100mL LB liquid medium containing 2% NaCl, culture at a constant temperature of 30°C at 200r / min for 24h, then transfer again to 4% NaCl content Cultivate in LB medium for 24h, shake culture at 30°C for 24h, then transfer to LB medium with 6% NaCl content, shake culture at 30°C for 24h, and incubate the obtained bacterial culture solution for 10 6 Dilute and spread, and pick a single colony that grows well, that is, the salt-tolerant growth-promoting bacteria strain Y4 is screened.
Embodiment 2
[0017] Example 2 Identification of salt-tolerant growth-promoting bacterial strain Y4
[0018] 1. Bacterial DNA Extraction
[0019] The bacteria isolated and screened in Example 1 were cultivated to the logarithmic phase, and 1mL of bacterial liquid was drawn to a 1.5mL centrifuge tube, first inactivated in a 30min boiling water bath, and then placed in a centrifuge to centrifuge at 1000rpm for 5min, and the resulting supernatant was And for bacterial DNA extraction.
[0020] 2. PCR amplification of strain 16s rDNA
[0021] The bacterial DNA was used as a template, and the amplification primers used bacterial universal primers, namely: forward primer U8-27(F) 5'-AGAGTTTGATCCTGGCTCA-3'; reverse primer L1494-1514(R) 5'-GGTTACCTTGTTACGACTT-3 '. 50 μL PCR amplification system: 21 μL dd water, 1 μL each of forward and reverse primers, 25 μL R-Taq mix, 2 μL of the above DNA extract. Add each substance in the above system in order to a 0.5mL PCR tube, mix well, and centrifuge at ...
Embodiment 3
[0025] Example 3 Salt-tolerant growth-promoting test of tomato strain Y4
[0026] Preparation of tomato growth promoter: Y4 was inoculated in LB liquid medium, cultivated at 180rpm and 30°C until OD600 was 1 (i.e. 10 8 CFU / mL), to obtain the bacterial solution, centrifuge the bacterial solution at a speed of 6000rpm for 10min, discard the supernatant, then inject an equal volume of dd water, shake and resuspend, centrifuge at a speed of 6000rpm for 10min, discard the supernatant, repeat 2 After resuspension, the tomato growth promoter is obtained.
[0027]Experimental design: select plump tomato seeds of the same size and sterilize the surface with 5% NaClO solution for 10 minutes, rinse the seeds repeatedly with deionized water for several times, soak the seeds in deionized water for 24 hours, and then place them in a culture medium covered with moist gauze. Germinate on a dish at 27°C for 2 days. The seeds with the same bud length were selected and sown in plastic pots (po...
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