A high-yielding strain hmgim-t43 of A. oosporum and its breeding method
A technology for high-yielding strains and microbial strains, applied in microorganism-based methods, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of unstable quality, high artificial cultivation cost, low transformation efficiency, etc. Improve the effect of long cultivation cycle
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Embodiment 1
[0043] This example is the preparation of protoplast bacterial suspension and ARTP mutagenesis.
[0044] (1) Activating the starting strain HMGIM-W160136 (HMGIM-W160136 was collected from Meihua Mountain, Fujian, and was preserved in the Guangdong Provincial Microorganism Culture Collection Center, and the deposit number is GDMCC NO: 61004, which has been disclosed in the patent 202010554107.6), and then the pure The culture was inoculated on a 90mm PDA medium plate, and placed in an incubator at 25±1°C for 7 days in the dark. Use a hole punch with a diameter of 5mm at the edge of the colony to make holes, pick out the bacterial block of the developed strain, inoculate it into a 250mL conical flask with 100mL PD liquid medium, and after standing for 1 day, shake at 25±1℃. Culture in the bed at a speed of 140 r / min in the dark for 7 d to obtain mycelium prepared from the bacterial suspension.
[0045] (2) Preparation of protoplast bacterial suspension, put the cultured myceliu...
Embodiment 2
[0050] This example is the primary screening of the mycelial growth rate of the mutant strains.
[0051] (1) Same as (1) in the above-mentioned embodiment 1, take the starting strain as the control group, screen out the mutant strain with better growth, and then subculture the mutant strain for 5 generations; Stable strains were tested for cultivation.
[0052] (2) Taking the mycelial growth rate of CK (0.81cm / d) as a reference, 958 mutant strains were obtained through 4 rounds of ARTP mutagenesis, and the strains whose mycelial growth rate was more than 5% higher than the starting strain were screened, and were screened for 5 generations. , 10 dominant strains with stable growth rate were obtained (see Table 1 below).
[0053] Table 1. Determination of genetic stability of mutant strains
[0054]
[0055] Note: CK has been domesticated and cultivated, and the growth rate of mycelium has been improved after isolation and purification; data are expressed as mean ± standard...
Embodiment 3
[0057] This example is the re-screening of domestication and cultivation of mutant strains.
[0058] (1) Original seed preparation, fully mix sorghum and calcium carbonate, adjust the water content to 65%, and put it into a high temperature resistant polypropylene bag (17 cm × 35 cm), each bag is equivalent to 80-100g dry material, at 0.147 MPa atmospheric pressure, sterilized under high temperature and high pressure at 126°C for 90min. Inoculate 10 strains of A. oosporum into the sterile original culture material, and cultivate in the dark incubator at 25±1°C for 7-14 days, and the mycelium is covered with the fungus bag.
[0059] (2) For the preparation of cultivation fungus bags, carbon source raw materials such as sawdust and cottonseed husks are pre-wetted one day in advance; nitrogen sources such as bran and corn flour are added to the ingredients on the same day, and the water content is adjusted to 65%, and placed in a high temperature resistant polypropylene bag (17c...
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