An endophytic Burkholderia gladiolus strain of litchi and its application in the control of litchi anthracnose and litchi frost blight
A technology for Burkholderia spp. and litchi anthracnose, which is applied in the field of biological control of plant diseases, can solve problems such as improving the resistance of pathogenic bacteria, reducing fruit quality, and destroying the ecological environment, achieving good development and application prospects and ecological environment impact Effects of small size and low requirements for culture conditions
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Embodiment 1
[0027] Example 1: Isolation, Purification and Preservation of Endophytic Burkholderia Gladiolus in Litchi
[0028] (1) Configuration of LB medium: Weigh 10 g of tryptone (Tryptone, Oxoid LTD LP0042, England), 5 g of yeast extract (Yeast extract, Oxoid LTD LP0021, England), sodium chloride (NaCl, Sinopharm Chemical Reagent Co., Ltd. Company, 10019318) 10g, add 1000mL of water and stir evenly, then add 15g of agar, fully heat and dissolve, then subpackage, sterilize at 121°C for 20min, and store for later use. The preparation of LB liquid medium is the same as that of LB solid medium except that agar is not added. The weighed medicine is fully dissolved and then divided into Erlenmeyer flasks (100mL per bottle), stoppered, bandaged, and sterilized at 121°C for 20 minutes. , and store for later use after cooling.
[0029] (2) Isolation and purification of endophytic Burkholderia gladiolus in litchi:
[0030] In June 2019, samples were collected at the litchi base of Shenzhen Xi...
Embodiment 2
[0036] Embodiment 2: Determination of the activity of Burkholderia gladioli by confrontation culture method
[0037] (1) The preparation of LB medium and LB liquid medium is the same as in Example 1;
[0038] (2) Preparation of PDA medium: Weigh 200g of potatoes, cut into strips and boil, filter with 16 layers of gauze, make up the water to 1000mL with the filtrate, add 15g of agar, fully heat and dissolve, then pack into conical flasks (200mL per bottle) , sterilized at 121°C for 20 minutes, cooled and stored for later use.
[0039] (3) Preparation of PDA+carrot culture medium: Weigh 200g of carrots, squeeze the juice fully with a juicer and filter it with 16 layers of gauze, add water to 1000mL with the filtrate, add 15g of agar, fully heat and dissolve it, and then culture it on the PDA prepared above The bases were mixed with equal volumes, and then divided into Erlenmeyer flasks (200mL culture solution per bottle), sterilized at 121°C for 20min, cooled and stored for lat...
Embodiment 3
[0049] Embodiment 3: Litchi endophytic Burkholderia gladiolus SZPT16 controls litchi anthracnose and frost blight under indoor conditions
[0050] (1) The preparation of LB medium and LB liquid medium is the same as in Example 1. The preparation of PDA medium and PDA+carrot medium is the same as in Example 2.
[0051] (2) Cultivation of seed bacteria liquid: inoculate Burkholderia gladiolus SZPT16 into LB liquid medium, culture at 28°C with shaking at 200rpm for 24h, and after 16h, place in ultra-clean workbench every 3h Sampling was taken to measure the OD value at 600nm. During the measurement process, the uninoculated culture solution was used to adjust to zero until the OD value of the seed bacteria solution was between 0.5-0.8, and the cultivation was completed to obtain the seed bacteria solution;
[0052] (3) Preparation of Burkholderia gladiolus bacterial suspension (biological agent): ferment and culture the seed liquid prepared in step (2) in LB liquid medium at a v...
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