A method for rapidly detecting the activity of Gexianmi seeds
A technology of kudzu rice and seeds, applied in the field of agricultural biology, can solve the problems such as the entry of tribendan blue and hinder the entry of dyes into cells, etc., and achieve the effect of convenient operation.
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Embodiment 1
[0026] Pectinase solution is configured as follows: 0.5g pectinase is dissolved in pH=3.6NaH 2 PO 4 (0.2mol / L)-citric acid (0.1mol / L) buffer solution 100ml, store in refrigerator at 4°C.
[0027] Physiological saline: dissolve 0.9g NaCl in 100ml distilled water and store at room temperature.
[0028] Quliben blue working solution: Weigh 0.2g of Quliben blue powder, put it in a mortar, add 5ml of double distilled water and grind until no particles appear and fine dark blue solution, filter with filter paper to get 4% Quliben blue solution, put into The narrow-mouth reagent bottle should be stored at 4°C. When in use, it should be diluted ten times with normal saline to obtain a 0.4% Triben blue solution.
Embodiment 2
[0030] Ge Xianmi seed pretreatment. The seeds of Ge Xianmi were disinfected with hydrogen peroxide diluted 100 times for 2 to 3 times, each time for 30 to 40 seconds, and then washed with pure water for 2 to 3 times, and then transferred to BG-110 culture solution diluted 10 times to make Pueraria The mass-to-volume ratio of the mass of Xianmi to the 10-fold diluted culture solution is 1:10 g / mL.
Embodiment 3
[0032] The parameters of pectinase treatment were determined. Take 50ml of the homogenized algae filament prepared in Example 2, kill the Gexianmi cells by high temperature and high pressure sterilization at 115° C. / 10min, and observe under a microscope. The cell morphology is complete and the same as the active Gexianmi cells. It was found that the optimum pH of pectinase was 3.0-6.0, and the optimum temperature was 40-65°C. Set a temperature gradient of 10°C, 30°C, 50°C and an enzyme concentration gradient of 0ul / ml, 50ul / ml, 150ul / ml, 300ul / ml to act on the active and inactivated Ge Xianmi homogenate respectively.
[0033] Table 1 Different treatment conditions of active and inactivated Gexianmi homogenate
[0034]
[0035]
[0036] A total of 24 treatments were mixed in a 1.5ml centrifuge tube and reacted in a water bath for 4 minutes, then centrifuged at 7000r / min for 1 minute, and the supernatant was discarded to terminate the reaction. The mass volume ratio of t...
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