Chinese sweetgum tissue culture and quick propagation method
A technology of tissue culture and sweetgum, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve problems that have not yet been published with a complete set of technologies
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Embodiment 1
[0095] Embodiment 1: adopt Chinese liquidambar 1-year-old seedling tip to carry out tissue culture
[0096] In the last ten days of March, select actively growing branches from the vigorously growing 1-year-old seedlings of Liquidambar chinensis in the field, take the 5cm tops, and disinfect them with 75% ethanol for 30 seconds, 0.1% mercury liter for 2 minutes, and rinse them with sterile water for 4 to 5 times. Finally, 1.0 to 1.5 cm shoot tips were cut as explants.
[0097] The basic medium used for shoot tip growth and bud differentiation and proliferation is WPM and MS medium, and the additional plant hormone combinations are BA 0, 0.1, 0.5, 1.0, 2.0mg / l, NAA 0, 0.05, 0.2mg / l, Remove the phytohormone combination whose BA concentration is lower than NAA, and add TDZ 0.1, 0.01mg / l for WPM culture, the medium contains 3% sucrose, 0.65% agar, the pH before sterilization is adjusted to pH5.8, 121°C high temperature and high pressure Sterilize for 16 minutes, cultivate at a te...
Embodiment 2
[0109] Embodiment 2: adopt the leaf and the petiole that the shoot tip tissue culture of Chinese sweetgum 1-year-old seedling is cultured to carry out tissue culture
[0110]On the differentiated buds of Chinese sweetgum cultivated from the shoot tip, select the young leaves that are unfolding or just unfolding, and cut them off together with the petioles. Under sterile conditions, use a scalpel to separate the leaf from the petiole, leaving 1-2 mm of the end of the petiole on the leaf, then cut the leaf transversely into thin strips of 3-5 mm wide with a scalpel, and place the adaxial side up on the organogenesis On the culture medium, the petioles were cut into small pieces of about 5 mm and placed flat on the organogenesis medium.
[0111] The minimal medium used was WPM and the additional phytohormone combinations were NAA 0, 0.1 mg / l, BA 0.5, 1.0, 2.5, 5.0 mg / l. In addition, TDZ 0.01 mg / l was added alone. The medium contains 3% sucrose and 0.65% agar, and the medium is ...
Embodiment 3
[0115] Embodiment 3: Cultivate Chinese sweetgum test-tube plantlet with seed hypocotyl
[0116] Liquidambar chinensis seeds were washed with running water for 3 hours, sterilized with 75% ethanol for 30 seconds and 0.1% mercuric chloride for 10 minutes, rinsed with sterile water for 4 to 5 times, and inoculated on hormone-free MS medium. After 1 week, the seeds began to germinate, and when the germinated seedlings grew to 1-2 cm, the hypocotyls were removed and cut into 3-5 mm segments as explants. The minimal media for callus induction were MS and MS / 2. The hormones used in the medium are 0.5-3.0 mg / l 2,4-D, 0-2.0 mg / l BA. The hormones used for adventitious bud induction are 0.5-1.0 mg / l ZT, 0.5-1.0 mg / l BA and 0.1 mg / l NAA.
[0117] When the callus was transferred from the callus proliferation medium to the adventitious bud differentiation medium, three pretreatment methods were used:
[0118] 1. Direct transfer without pretreatment, this method is used as a control;
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