Method capable of effectively inhibiting browning for multiplication culture of buckwheat callus
A technology for callus, proliferation and culture, applied in horticultural methods, botany equipment and methods, plant regeneration, etc., can solve the problems of browning of culture materials, browning of culture materials, death, etc., to reduce browning rate, improve The effect of multiplier
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Embodiment 1
[0016] (1) Obtaining buckwheat callus: select buckwheat petioles that are sown in spring and grow healthy, and they are disinfected with 0.1% mercury liters for 4 minutes, then washed with sterile water 3 times, and then cut into lengths of 1.5 cm. Access to callus induction medium MS+6-BA 0.5mg / L+2,4-D 4mg / L+IAA 0.1mg / L, under the conditions of 25℃, 10 hours of light per day, and 2000 lx light intensity Cultivation is carried out under the condition of pH 6.0, sucrose 30g / L and agar 6.8 g / L;
[0017] (2) Pre-culture of callus: select buckwheat callus with 20 days of induction culture, no browning and fast growth rate, and put it in an incubator at a temperature of 8°C and no light for 10 days;
[0018] (3) Multiplication and culture of callus: Cut the pre-cultured callus along the transverse direction (to reduce the wound area of the callus), and then quickly insert the cut end into the multiplication medium 1 / 2 MS+6-BA 1.0 In mg / L+IAA 2 mg / L + citric acid 10 mg / L+ L-cysteine ...
Embodiment 2
[0020] (1) Obtainment of buckwheat callus: select buckwheat petioles that are sown in spring and grow healthy, disinfected with 0.1% mercury liters for 5 minutes, then washed with sterile water 4 times, and then cut into 1.5cm length. Connected to callus induction medium MS+6-BA 1.5mg / L+2,4-D 2mg / L+IAA 0.5mg / L, at 25℃, 14 hours of light per day, light intensity of 2000 lx Cultivation is carried out under the condition of pH 6.0, sucrose 30g / L and agar 6.8 g / L;
[0021] (2) Pre-culture of callus: select buckwheat callus with 25 days of induction culture, no browning and fast growth rate, and put it in an incubator at a temperature of 12°C and no light for 8 days;
[0022] (3) Multiplication culture of callus: After cutting the pre-cultured callus, quickly insert the cut end into the multiplication medium 1 / 2 MS+6-BA 1.5 mg / L+IAA 1 mg / L + citric acid In 7 mg / L+ L-cysteine 140 mg / L, cultured for 3 days at a temperature of 8°C and no light, and then quickly performed at 22°C, 10 hou...
Embodiment 3
[0024] (1) Obtaining buckwheat callus: select buckwheat petioles that are sown in spring and grow healthy. First, sterilize with 0.1% mercury liters for 4 minutes, then wash with sterile water 4 times, and then cut into 2cm lengths. Into the callus induction medium MS+6-BA 1.0mg / L+2,4-D 5mg / L, cultivate under the conditions of 25℃, 12 hours light per day, and 2000 lx light intensity. pH value is 6.0, sucrose 30g / L, agar 6.8 g / L;
[0025] (2) Pre-culture of callus: select buckwheat callus that has been induced and cultured for 20 days without browning, and put it in an incubator at a temperature of 6°C and no light for 12 days;
[0026] (3) Multiplication and culture of callus: After cutting the pre-cultured callus, quickly insert the cut end into the multiplication medium 1 / 2 MS+6-BA 0.6mg / L+IAA 3mg / L + citric acid 12 In mg / L+ L-cysteine 60 mg / L, cultured for 5 days at a temperature of 6°C and no light, and then fasted at 27°C, 7 hours of light per day, and light intensity of 18...
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