A kind of broccoli isolated cell rapid proliferation method
A broccoli, in vitro technology, applied in the field of plant biology, can solve the problems of low content, high cost, dependence, etc., and achieve the effects of reducing production costs, improving capabilities, and simple methods
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Embodiment 1
[0025] Example 1 A method for the rapid proliferation of broccoli isolated cells, comprising the following steps:
[0026] ⑴Preparation of MS basic medium:
[0027] First, in a 1000mL beaker, add 4.5g~5.5g of agar into distilled water at a temperature of 85~95°C, and stir until the agar is completely dissolved to obtain an aqueous agar solution; Trace element mother liquor 20mL, iron salt mother liquor 20mL, calcium salt mother liquor 20mL, magnesium salt mother liquor 20mL, organic mother liquor 20mL, inositol mother liquor 20mL, stir well and add 30g of sucrose, stir until the sucrose is completely dissolved, then dilute to 1000mL scale with distilled water After stirring well, add 0.1mol / L NaOH or 0.1mol / L HCl to adjust the pH value to 5.8~6.0; finally, divide each 40~60mL into a 100mL Erlenmeyer flask and sterilize at 121℃ for 20min Afterwards, place it horizontally at room temperature and cool for 4 hours to obtain the MS basal medium.
Embodiment 2
[0032] Example 2 A method for the rapid proliferation of broccoli isolated cells, comprising the following steps:
[0033] ⑴ Preparation of MS basic medium is the same as Example 1 .
[0034] ⑵ Broccoli in vitro cell acquisition and inoculation method:
[0035] After the broccoli seeds of the current year were rinsed with water for 40 minutes, they were first sterilized with 75% alcohol for 0.75 minutes, and then sterilized with 1% NaClO for 6 minutes to obtain sterile seeds; the sterile seeds were rinsed with sterile water for 4 minutes. ~5 times and inoculated on the MS basic medium, cultured for 14 days under the conditions of light intensity of 6000LX and temperature of 26°C to obtain sterile seedlings, in which 8 seeds were inoculated in every 60mL of MS basic medium; the cutting length was 5mm The embryos were inoculated on the MS induction medium with the face up, and 6 embryos were inoculated in each 60mL MS induction medium, and dedifferentiated and cultured for ...
Embodiment 3
[0038] Example 3 A method for the rapid proliferation of broccoli isolated cells, comprising the following steps:
[0039] ⑴ Preparation of MS basic medium is the same as Example 1 .
[0040] ⑵ Broccoli in vitro cell acquisition and inoculation method:
[0041] After washing the broccoli seeds of that year with clean water for 30 minutes, first sterilize them with 75% alcohol for 0.50 minutes, and then sterilize them with 1% NaClO for 4 minutes to obtain sterile seeds; rinse the sterile seeds with sterile water for 4 minutes. ~5 times and inoculated on the MS basic medium, cultured for 14 days under the condition of light intensity of 5000LX and temperature of 25°C to obtain sterile seedlings, in which 6 seeds were inoculated in every 50mL of MS basic medium; the cutting length was 5mm The embryos were inoculated face-up on the MS induction medium, wherein 5 embryos were inoculated per 50 mL of MS induction medium, and dedifferentiated cultured for 30 days in the dark at ...
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