Zopfiella sp. and application thereof
A technology of sporosporium and stipe, which is applied in the field of endophytic podosporium in Boju, which can solve the problems of insecticide resistance, ecological balance damage, and non-target organism damage.
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Embodiment 1
[0032] Embodiment 1, the screening of endophytic Podosporum BJF10 of Boju chrysanthemum
[0033] The test pathogens used in this embodiment: Fusarium moniliforme, Fusarium oxysporum, Curvularia lunata, and Pythium. The pathogenic bacteria to be tested were respectively activated on PDA medium at 28°C for 4-5 days for later use.
[0034] The screening steps of the present embodiment mainly include:
[0035] (1) Acquisition of endophytic fungi of Boju: Rinse the healthy plants of Boju with running water and dry them, cut them into several parts and put them in the ultra-clean workbench, soak them in 75% alcohol for 3 minutes, and then wash them with sterile water Rinse 3 times, then rinse with 0.1% mercury liter for 30 sec, and then rinse with sterile water 3 to 4 times. The PDA and LB mediums were used for culture respectively, and the bacterial culture temperature was 37°C, and the fungal culture temperature was 28°C. After culturing for 3-4 days, pick the colonies accord...
Embodiment 2
[0042] Embodiment 2, Morphological and molecular identification of endophytic Podosporum BJF10 of Boju chrysanthemum
[0043] (1) Morphological identification of endophyte BJF10
[0044] Macroscopic traits such as colony size, color, decoration on the surface of the colony, and edge of the colony were observed by plate culture method. The microscopic properties such as hyphae and spore morphology of the active fungal cultured on inserts were observed under the low-power lens and 40-fold objective lens by microscopy.
[0045] After culturing and observing, the specific colony morphology of the endophytic fungus BJF10 is shown in Table 2 and figure 2 , fungal hyphae and spore morphology see image 3 .
[0046]
[0047] (2) Molecular biology identification of endophyte BJF10
[0048] Reagents involved in the embodiments of the present invention:
[0049] (1) DNA extraction reagents
[0050] TE solution: 10mmol / L Tris·HCl (pH8.0), 1mmol / L EDTA;
[0051] 3M NH 4 AC: Wei...
Embodiment 3
[0069] Example 3, the test of the endophytic Podosporum BJF10 of Boju chrysanthemum on the degradation function of cellulose
[0070] The medium involved in this example: 1) CMC-Na medium (g / L): NaCl 6.0g, MgSO 4 ·7H 2 O 0.1 g, KH 2 PO 4 0.5g, CaCl 2 0.1g, K 2 HPO 4 2.0g, CMC-Na 10.0g, (NH 4 ) 2 SO 4 2.0 g, 15.0 g agar, pH value 7.0~7.4. 2) CMC enzyme activity detection liquid fermentation medium (g / L): peptone 10.0 g, beef extract 10.0 g, CMC-Na 10.0 g, NaCl 1.5 g, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O 0.3g, distilled water, pH 7.0. 3) Filter paper enzyme activity detection liquid fermentation medium (g / L): peptone 10.0g, beef extract 10.0g, Xinhua filter paper 0.05g, NaCl 1.5g, KH 2 PO 4 1.0g, MgSO 4 ·7H 2 O 0.3g, distilled water 1000mL, pH value 7.0g. All the above mediums were sterilized at 0.1MPa, 121°C for 25 min, and cooled to 40-50°C for later use.
[0071] Solutions related to identification of cellulose degradation: 1) 1mg / mL Congo red solution: wei...
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