Method for cultivating meloidogyne hapla by adopting peanut rooting
A technology for root-knot nematodes and peanut roots, which is applied in animal husbandry and other directions to achieve the effects of high reproduction rate, simple operation, and beneficial to infection and reproduction
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Embodiment 1
[0032] a Preparation of peanut hairy roots
[0033] Select the seeds of Huayu No. 45 with large and plump seeds, wash them with tap water for 20 minutes, then soak them in alcohol with a mass percentage concentration of 75% for 1 minute under sterile conditions, and then soak them in 0.1% mercury liter for 15 minutes to carry out surface disinfection. Finally, wash with sterile water 3 times, 3min each time. After disinfection, use a sterile knife to peel off the seed coat on sterile filter paper, and inoculate it on the culture medium.
[0034] Under sterile conditions, use an inoculation loop to pick up the Agrobacterium rhizogenes A4 strain frozen in a low-temperature refrigerator at -20°C, inoculate it on a solid plate of LB+50mg / L Kan, and then culture it at a constant temperature of 28°C Cultivate in the dark in the box. After a single colony grows, pick a single colony and transfer it to 10ml LB liquid medium (with 50mg / L Kan added), and culture and recover in the dark...
Embodiment 2
[0040] a Preparation of peanut hairy roots
[0041] Select the large and plump Huayu No. 45 seeds, wash them with tap water for 30 minutes, then soak them in alcohol with a mass percentage concentration of 75% for 0.5 minutes under sterile conditions, and then soak them in 0.1% mercury liter for 15 minutes to carry out surface disinfection. , and finally washed 4 times with sterile water, 3 min each time. After disinfection, the seed coat was peeled off on sterile filter paper with a sterile knife, and inoculated on hormone-free MSB5 medium for culture. MSB5 medium is MS inorganic salt + B5 organic component (referred to as MSB5).
[0042] Under sterile conditions, use an inoculation loop to pick up the Agrobacterium rhizogenes A4 strain frozen in a low-temperature refrigerator at -20°C, inoculate it on a solid plate of LB+50mg / L Kan, and then culture it at a constant temperature of 28°C Cultivate in the dark in the box. After a single colony grows, pick a single colony and ...
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