Pichia manshurica for controlling postharvest diseases of oranges and grapes
A technology of Pichia pastoris and Pichia pastoris, applied in the direction of microorganism-based methods, biochemical equipment and methods, food preservation, etc., can solve the problems of lack of bio-control strains, fruit rot, large losses, etc., and achieve significant economic benefits and social benefits, wide antibacterial spectrum, and the effect of avoiding harm to people
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Embodiment 1
[0026] Embodiment 1: the microbiological characteristics of Pichia manshurica (Pichia manshurica) Y4 strain are as follows:
[0027] 1. Morphological characteristics
[0028] (1) NYDA culture medium (0.8% beef extract, 0.5% yeast extract, 1% glucose, 2% agar, 121°C damp heat sterilization for 20min) cultured at 28°C for 48h, the colonies were round, smooth and moist, Easy to provoke. The cells are oblong in shape and scattered individually.
[0029] (2) After culturing in NYDB liquid medium for 24 hours, no mold was formed, and the bacterial solution was turbid with precipitation. Microscopically, the yeast cells were oval and budded.
[0030] 2. Molecular biological identification
[0031] Sequence analysis of the 5.8S rDNA-ITS region of the screened strain Pichia manshurica was performed by searching on GenBank and identified as Pichia manshurica. According to the retrieved homologous strains, the Mege5.1 program of DNAStar software was used to construct the biological e...
Embodiment 2
[0033] Embodiment 2: the control effect of Pichia chastori to citrus green mold
[0034] 1) Test plan
[0035] The tested fruits have been carefully selected, and the fruit should be selected as uniform as possible, without scratches or mechanical damage on the surface and damage by diseases and insect pests. Use a sterile puncher to punch 3 holes at the equator of the fruit, and the surface wound of the citrus is 5mm (diameter) × 3mm (depth). Add 30 μL of the following treatment solution equally to each wound: (1) 1×10 8 cells / mL of Pichia chaltiformis suspension; (2) Sterile distilled water. After 2 hours, 30 μL of 1×10 5 spores / mL of Penicillium digitatum (P.digitatum) spore suspension. After drying naturally, put the fruit into a plastic basket and seal it with a plastic wrap, cultivate it in a constant temperature incubator (humidity 95%), and place it at room temperature (20°C). Each treatment was replicated 3 times, and 12 fruits were replicated each time. The who...
Embodiment 3
[0042] Embodiment 3: the control effect of Pichia crestella to Botrytis cinerea and Aspergillus niger
[0043] 1) Test plan
[0044] The activated yeast was inoculated in a conical flask filled with 50 mL of NYDB (5 g of yeast extract, 8 g of beef extract, 10 g of glucose, and 1000 mL of distilled water), and cultured on a shaking table at 28°C and 180 r / min for 20 h. Take the bacterial solution and centrifuge it at 8000r / min for 10min, discard the supernatant, wash it repeatedly with sterile water for 3 times, and count it on a hemocytometer to obtain a concentration of 1×10 8 cells / mL of bacterial suspension, and the control group was sterile water. A hole with a diameter of 3m (width)×3mm (depth) was punched at the waist of the fruit, and then 15 μL of the yeast suspension of the above concentration was injected into each hole, and 15 μL of sterile water was injected into the control fruit. After 2 hours, 10 μL of 1×10 5 spores / mL of Botrytis cinerea (B.cinerea) spore su...
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