Pseudomonas fluorescens and biological preparation and application in preventing and controlling sugarcane smut
A technology of Pseudomonas fluorescens and biological agents, which is applied in the direction of chemicals, biocides, and microorganism-based methods for biological control, and achieves low requirements for culture conditions, good development and application prospects, and strong inhibitory effects
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Embodiment 1
[0026] Example 1: Isolation, purification and preservation of Pseudomonas fluorescens
[0027] (1) The configuration of LB medium: The configuration of LB medium: Weigh 10 g tryptone (Tryptone, Oxoid LTD LP0042, England), 5 g yeast extract (Oxoid LTD LP0021, England), sodium chloride (NaCl) , Sinopharm Chemical Reagent Co., Ltd., 10019318) 10g, add 1000mL of water and stir evenly, then add 15g of agar, fully heat to dissolve and then distribute, sterilize at 121°C for 20min and store for later use.
[0028] (2) Isolation and purification of Pseudomonas fluorescens:
[0029] Weigh 10g of sugarcane rhizosphere soil sample from Yuejin Farm of South China Agricultural University, add it to a triangular flask containing 90mL sterile water, shake at 28℃, 200rpm for 1h, let stand for 1~2h, suck 1mL of soil for immersion The clear solution was diluted with 0.9% saline to 10 -6 Draw 100μL 10 -4 , 10 -5 , 10 -6 Wait for the solution of 3 concentrations to be evenly spread on the LB plate to i...
Embodiment 2
[0035] Example 2: Determination of the activity of Pseudomonas fluorescens by the confrontation culture method
[0036] (1) The preparation of LB medium is the same as in Example 1. The preparation of LB liquid medium is the same as that of LB solid medium except that agar is not added. The weighed reagents are fully dissolved and then dispensed into conical flasks (each Bottle of 100mL culture solution), stoppered and bandaged, sterilized at 121°C for 20min, cooled and stored for later use.
[0037] (2) Preparation of PDA culture medium: Weigh 200g potato and cut into small pieces, add water and boil for 20-30 minutes until it can be punctured by a glass rod, filter with eight layers of gauze; heat the filtrate, add 20g glucose, 20g agar powder, and stir. Evenly, after a little cooling, add water to 1000mL, dispense into Erlenmeyer flasks (100mL culture solution per bottle), stoppered and bandaged, sterilize at 121°C for 20min, and store after cooling.
[0038] (3) Preparation of a...
Embodiment 3
[0046] Example 3: Pseudomonas fluorescens to control sugarcane whip smut
[0047] (1) The preparation of LB medium is the same as in Example 1. The preparation of LB broth was the same as in Example 2.
[0048] (2) The preparation of Pseudomonas fluorescens biological preparation is the same as in Example 2.
[0049] (3) Use seed dressing method to inoculate. 1) Streak the biocontrol strain HN58 on the LB plate and activate it. After 1 to 2 days, pick a single colony and shake it to OD with LB liquid medium at 28℃, 200rpm 600 It is about 1.5; 2) Add 0.25g of pathogenic smut spores (see Example 2) per 1kg of soil, and then set three treatments of adding 5mL, 20mL, and 100mL biocontrol bacteria culture solution, each with 3 repeat. For each repetition, 30 segments of cane stem nodes are selected, so 4kg of soil containing 1 g of smut fungus and 20 mL, 80 mL, and 400 mL of biocontrol bacteria need to be prepared separately; 3) Set up a total of 2 controls with smut fungus and water. ...
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