The application discloses a kind of preparation and
suspension culture of blackland
callus method, belong to the field of
industrial biotechnology.It includes the following steps: after sterilization, take flower stem, access pH value is 5-7 in induction medium, under the condition of temperature is 20-30 DEG C and no light, induction culture 15-21 days, namely obtain blackland
callus, wherein, the induction medium is made of MS basic medium,
sucrose 25-35g / L,
agar 6-7g / L, 6-BA 1.0-2mg / L, NAA 0.5-1.5mg / L, 2, 4-D 0.3-1.0mg / L and coconut water 0.5-1.5g / L composition.Under aseptic conditions, clamp the growth state of good blackland embryogenic
callus 30g,
inoculation is carried out in 200mL / 500mL pH value is 6.0 in MS
liquid medium, temperature 25 DEG C, 110 rpm full light oscillation culture, subculture period 7 days, when subculture, the mixture of
cell and
culture fluid is mixed with fresh
culture fluid according to 1:1 volume ratio, after equal division, continue to culture, namely obtain blackland suspension
cell, wherein, the
liquid medium is made of MS basic medium,
sucrose 30g / L, 6-BA 1.5mg / L, NAA 0.5mg / L, 2, 4-D 0.5mg / L and coconut water 1.5g / L composition.The callus out of the rate of 88.6% is realized, and the induction rate of embryogenic callus that can be suspended culture is 68.6%-91.8%, and good callus state is provided for large-scale culture blackland
cell.