A method for inducing and cultivating hybrid liquidambar tetraploid
A tetraploid and hybrid technology, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve problems such as induced breeding and reporting of allotetraploid of Liquidambar genus, and achieve improvement of tetraploid Induction rate, high ratio, good effect of plant growth
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Embodiment 1
[0086] 1. Test materials
[0087] 1. Obtained from hybrid sweetgum seeds
[0088] The present invention uses the sweetgum tree and the sweetgum tree of North America that are well-grown, mature, healthy, and free from insect damage in Shanghai Chenshan Botanical Garden (Songjiang District, Shanghai, longitude: 121.22, latitude: 31.03).
[0089] In mid-March 2016 (10‐15), unopened flower branches of liquidambar formosana were collected, and then the flower branches were hydroponically cultured. After 3‐5 days, the water content of the stamen flower flower dropped and cracked, releasing gold. Yellow pollen, collect the pollen of the liquidambar tree, and then store it in a test tube containing a desiccant at low temperature;
[0090] At the end of March, 2016 (20-30), the female inflorescence and the male inflorescence can be completely separated, and the bracts on the male inflorescence have not fallen off. 3-5 days before the male flower looses powder, remove the male inflore...
Embodiment 2
[0117] Embodiment 2 prepares liquidambar aseptic seedling
[0118] 1. Sterilization of hybrid sweetgum seeds
[0119] Select plump and healthy seeds that are stored dry at 4°C, and disinfect them under an ultra-clean workbench, that is, first put the seeds into 75% alcohol (v / v) for 30-60s (preferably 45s (seconds) ), wash 1-2 times with sterile water, then put the seeds into 2% sodium hypochlorite solution (w / w) and soak them for 6-10min (preferably 8min), then clean them several times with sterile water (preferably 3 -5 times); then the hybrid sweetgum seeds are placed on sterile dry filter paper to dry the seed surface moisture to obtain sterilized hybrid sweetgum seeds (350);
[0120] 2. Seed germination treatment
[0121]Place the sterilized hybrid sweetgum seeds in the seed germination medium (WPM basic medium+sucrose 30g / L+agar 2g / L+Bilicon 4g / L, pH5.8-5.9), carry out the germination culture of seeds, 4 After -8 days, the seeds began to germinate. After 15 days, the ...
Embodiment 3
[0122] Embodiment 3 prepares aseptic explant
[0123] 1. After the seeds are germinated and cultivated for about 2 months (60-65 days), select 3 vigorously grown hybrid liquidambar aseptic seedlings (i.e. 3 genotype hybrid liquidambar aseptic seedlings named as Z1, Z2, Z3 genotype hybrid sweetgum sweet aseptic seedling), and 3 strains of hybrid sweetgum sweet aseptic seedlings are pruned to leave 2 blades at the top respectively, and the 2 blades at the top are cut in half, long 1.5-2.5cm (preferably 2cm ), and then inoculate them in the rooting medium (WPM medium + IBA 2.0mg / L + NAA 0.1mg / L + sucrose 30g / L + agar 2g / L + beilicon 4g / L, pH 5.8‐5.9) , carry out rooting culture, wherein, the culture condition of rooting culture of sweetgum seedling section is as follows: culture temperature is 25 ± 2 ℃, and light is 2000lx, and photoperiod is 16h light / 8h dark; After -15 days), the plant begins to take root, obtains respectively the rooted sweetgum seedling section (ie Z1, Z2, Z...
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