2. Inducing plant disease resistance means that Bacillus subtilis can not only inhibit plant pathogenic bacteria, but also induce plant disease resistance mechanism, thus enhancing plant disease resistance.
3. Bacillus subtilis can synthesize various vitamins and improve the activity of interferon and macrophages in animals.
4. Bacillus subtilis can stimulate the growth and development of animal immune organs, activate lymphocytes, improve immunoglobulin and antibody levels, enhance cellular immunity and humoral immunity, and improve group immunity.
5. Bacillus subtilis can improve the utilization rate of chemical fertilizer, inhibit the absorption of nitrate nitrogen, heavy metals and pesticides by crops, purify and repair soil, reduce the occurrence of crop diseases, promote the decomposition and utilization of crop straw and municipal waste, and protect the environment. It has played an irreplaceable role in improving the quality of crop products and food safety.
/iknow-pic . cdn . BCE Bos . com/0df 43 1 adcbef 7609646 1 94 ff 23 DDA 3c cc 7 CD 99 e 63 " target = " _ blank " title = " " class = " ikqb _ img _ alink " >/iknow-pic . cdn . BCE Bos . com/0df 43 1 adcbef 7609646 194 ff 23 DDA 3c cc 7 CD 99 e 63？ x-BCE-process = image % 2f resize % 2Cm _ lfit % 2Cw _ 600% 2Ch _ 800% 2c limit _ 1% 2f quality % 2Cq _ 85% 2f format % 2Cf _ auto " esrc = "/ 0 df 43 1 adcbef 7609646 194 ff 23 DD a3 cc 7 CD 99 e 63 "/>
Extended data
(1) Formula of common culture medium for Bacillus subtilis:
1L distilled water +20g glucose+15g peptone +5g sodium chloride +0.5g beef paste +20g agar?
(2) Preparation process of Bacillus subtilis protoplast:
1. culture Bacillus subtilis:
The fresh slopes of parent strains T44 12 and TT2 were respectively connected to test tubes filled with liquid complete medium (CM), and cultured at 36℃ for 14h. The 1mL bacterial liquid was transferred to a 250mL conical flask filled with 20mL liquid complete medium, and cultured at 36℃ for 3 hours, so that the cells grew into the pre-logarithmic phase, respectively.
2. Collect cells:
Take 65438±00ml bacterial liquid, centrifuge at 4000r/min for 65438±00min, discard the supernatant, suspend the bacteria in phosphate buffer and centrifuge. After washing twice, the bacteria were suspended in 10mLSMM, and each mL contained about 108 ~ 109 viable bacteria. ?
3. Determination of the total number of bacteria:
Take 0.5mL of each bacterial liquid, dilute it with normal saline, take 10-5, 10-6, 10-7 (two plates each time), pour it into complete culture medium, and count it after 24 hours at 36℃. This is the total number of bacteria that have not been treated with enzymes. ?
4. Away from the wall:
Take 5mL of bacterial suspension from each of the two parent strains, add 5mL of lysozyme solution with lysozyme concentration of 100μ g/ml, mix well, and then keep the temperature in a water bath at 36℃ for 30min. Samples were taken regularly and the formation of protoplasts was observed under a microscope. When more than 95% cells become spherical protoplasts, centrifuge at 4000r/min for 65438±00min, discard the supernatant and use hypertonic buffer. Immediately determine the number of remaining bacteria.
5. Determination of residual bacteria:
Take 0.5mL of the above protoplast suspension, dilute it with sterile water, and make the protoplast crack and die. Take 0. 1-2, 0. 1-3, 0. 1-4 dilutions respectively, spread them on a complete medium plate, and cultivate them at 36℃ for 24 ~ 48h. The colonies grown should be intact. The number of remaining cells after enzyme treatment was calculated, and the protoplast formation rate of the two parents was calculated respectively. Protoplast formation rate = the total number of bacteria without enzyme treatment-the number of remaining cells after enzyme treatment.
References:
/baike.baidu.com/item/%E6% 9e% AF% E8% 8d% 89% E8% 8a% BD% E5% AD% A2% E6% 9d% 86% E8% 8f% 8c/4980560 "target =" _ blank "title.