Method for quick mycorhiza formation of azalea aseptic seedlings
A sterile seedling and mycorrhizal technology, applied in the field of botany, can solve the problem of strains growing too fast, affecting the process of seedling mycorrhiza and later growth, etc. high effect
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Embodiment 1
[0020] The isolation and identification of embodiment 1 rhododendron mycorrhizal fungus:
[0021] 1. Separation of root materials: Take the living roots of the rhododendron species ‘Purple Crane’ in Shanghai Binjiang Forest Park, put them in an ice bucket together with soil moisture, and store them in a 4°C refrigerator for later use.
[0022] 2. Separation and identification medium:
[0023] (1) Separation medium: Use Martin-Bengal red medium (referred to as MA) to adjust and improve its pH. The formula is: 10 grams of glucose, 5 grams of tryptone, 1 gram of dipotassium hydrogen phosphate, 0.5 grams of magnesium sulfate, 0.033 grams of Bengal red, 20 grams of agar, distilled water to 1000 ml, adjust the pH value to 5.0, and then 121 ℃ high pressure Sterilize for 20 minutes, add 0.03 g of streptomycin when cooled to below 60°C, mix well, and pour it into a plate for later use.
[0024] (2) Strain identification medium: use malt extract medium (MEA), the formula is: 20g of ma...
Embodiment 2
[0042] The screening of embodiment 2 suitable cultivation substrates
[0043] Choose peat, vermiculite and yellow sand three common cultivation substrates and mix them in different proportions as the ready-to-use cultivation substrate. The specific formula is peat: vermiculite 2:1, peat: vermiculite: yellow sand 2:1:1, peat: yellow sand 2:1, peat: yellow sand 3:1. After the substrate is mixed in proportion, it is sterilized at high temperature, and then poured into the sterilized MMN liquid medium, 300ml / 1000g of cultivation substrate, mixed evenly, and then packed into cultivation bottles. Transplant aseptic sowing seedlings with a height of 2-4cm, and inoculate one bacterial block (diameter 2-4mm). Any one of the fungal strains, 30 seedlings were cultivated in each substrate, and the growth of the seedlings in different substrates was counted after 12 weeks (Table 1). The growth potential of the seedlings in the cultivation substrate containing vermiculite was poor, and the...
Embodiment 4
[0046] The screening of glucose concentration in embodiment 4MMN medium
[0047] Nutrients and contents in MMN liquid medium were unchanged except for glucose.
[0048] The formula of MMN liquid medium is: calcium chloride (Cacl2 2H2O) 0.05g, magnesium sulfate (MgSO4 7H2O) 0.15g, sodium chloride (NaCl) 0.025g, ferric chloride (FeCl3) 0.012g, diphosphate Potassium hydrogen (KH2PO4) 0.5g, vitamin B10.1mg, diammonium hydrogen phosphate ((N H4)2HPO4) 0.25g, citric acid 0.2g, malt extract 2g, distilled water to 1000ml.
[0049] The amount of glucose added in 1L medium was adjusted to 0.5g (0.05%), 1g (0.1%), 3g (0.3%), 5g (0.5%), 10g (1%) and 15g (1.5%) respectively. Using peat: yellow sand 3:1 as the substrate, pour 300ml / L of MMN medium with different glucose content, mix the substrate and nutrient solution evenly, then pack into tissue culture bottles, and then sterilize at high temperature. Transplant 4 seedlings into each bottle, insert 1 bacterial block between every two se...
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