Method for obtaining high-efficiency regeneration plant of ephemeral plant lachnoloma growing in early spring
A plant and short-lived technology, applied in the field of plant tissue culture and plant biology, can solve the problems of low natural reproduction rate, extinction threat, and shortened growth and development cycle of A. big effect
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Embodiment 1
[0027] Embodiment one: standby of aseptic material
[0028] Culture medium: MS (Murashige and Skoog, 1962) medium was prepared by selecting 10×macroelement mother liquor, 100×iron salt mother liquor, 100×trace element mother liquor, and 100×organic matter (vitamin) mother liquor, without adding any growth regulator The culture medium contains 3% sucrose and 0.6% agar powder, and the pH value is adjusted to 6.0. It is divided into 150mL Erlenmeyer flasks, and each bottle is about 50mL. Put it into a sterilizing pot after subpackaging, at 1.2×104kg / m 2 (121°C) for 20 minutes of sterilization, take it out, put it flat, and cool it for later use.
[0029]Cultivation of sterile seedlings: The mature Mianguo mustard fruit was collected in Cainan, Fukang, Xinjiang. After the fruit is picked, store it at room temperature for later use. Take the seeds that are large and plump and free from diseases and insect pests, wash them with tap water for 2-3 minutes, then soak them in 70% eth...
Embodiment 2
[0030] Embodiment two: the induction of callus
[0031] This test is for the purpose of establishing a best system for callus induction of A. sativa. The hypocotyl and cotyledon of A. sativa are used as explants, and different plant growth regulators (IAA, 2, 4-D, NAA, IBA, BAP) in different concentrations and combinations of its callus induction and growth were compared.
[0032] 1. Screening of Different Explants and Different Growth Regulators in Callus Induction
[0033] 14-15 days after the germination of mustard seeds, the hypocotyls and cotyledons of the aseptic seedlings were cut into 0.5 cm long, 0.5 × 0.3 cm 2 area, and inoculate the MS medium of adding different plant growth regulators (IAA, 2,4-D, NAA, IBA, BAP) in the prior art and the MS basic medium without adding any phytohormone as a control (CK), See Table 1. All media contain 3% sucrose and 0.6% agar powder, the pH value is adjusted to 6.0, at 1.2×104kg / m 2 (121°C) for 20 minutes (Shirin et al., 2007). ...
Embodiment 3
[0050] Embodiment three: the proliferation of callus
[0051] This test is for the purpose of establishing a best system for the callus proliferation of A. sativa, and the hypocotyl and cotyledon of A. sativa are used as explants, and different plant growth regulators (IAA, 2, 4-D, NAA, IBA, BAP) in different concentrations and combinations of its effects on callus proliferation were compared.
[0052] 1. Method for Callus Proliferation
[0053] The hypocotyl and cotyledon callus induced in the above examples were cut into 0.5×0.5cm 2 Subculture in the subculture medium supplemented with different phytohormones, see Table 2. The concentration of sucrose and agar powder added to the medium and the culture conditions are the same as those in 2.1.1.
[0054] Table 2. Different concentrations of auxin and cytokinin and their combinations
[0055]
[0056]
[0057] The above experiments show that the callus induced by cotyledons has a better proliferation efficiency than ...
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