A method for direct somatic embryo induction of betel nut and its suspension culture to obtain regenerated plants
A technique of regenerating plants and suspension culture, which is applied in the biological field, can solve the problems such as the research report on direct somatic embryo induction suspension culture of betel nut, and achieve the effects of shortening the breeding cycle of varieties, convenient operation, and easy control of growth conditions
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Embodiment 1
[0026] 1. Explant acquisition: select a high-yielding healthy betel nut plant, take the uncracked young inflorescence of spathe as an explant, wash the surface of the inflorescence with a saturated washing powder solution and rinse it clean, and wash it with 75% alcohol in an ultra-clean workbench. After wiping the entire surface of the inflorescence evenly several times, use 0.1% HgCl 2 Soak for 5-12min, wash 5 times with sterile water, and place on sterile filter paper to blot the surface moisture.
[0027] 2. Direct somatic embryo induction: Take the above-mentioned sterilized explants and cut them into 1-3mm thick small pieces, inoculate them into somatic embryo induction medium, and culture them in dark at 25-28°C to obtain somatic embryos ( See figure 1).
[0028] Somatic embryo induction medium: NH 4 Cl 1000mg / L+H 3 BO 3 10mg / L+KNO 3 2400mg / L+Ca(NO 3 ) 2 620mg / L+NaH 2 PO 4 300mg / L+MgSO 4 ·7H 2 O 350mg / L+CuSO 4 ·5H 2 O 0.25mg / L+Na 2 -EDTA37.3mg / L+FeSO ...
Embodiment 2
[0039] 1. Explant acquisition: select a high-yielding healthy betel nut plant, take the uncracked young inflorescence of spathe as an explant, wash the surface of the inflorescence with a saturated washing powder solution and rinse it clean, and wash it with 75% alcohol in an ultra-clean workbench. After wiping the entire surface of the inflorescence evenly several times, use 0.1% HgCl 2 Soak for 5-12min, wash 3 times with sterile water, and place on sterile filter paper to blot the surface moisture.
[0040] 2. Direct somatic embryo induction: take the above-mentioned sterilized explants and cut them into 1-3mm thick small pieces, inoculate them into somatic embryo induction medium, and culture them in dark at 25-28°C to obtain somatic embryos.
[0041] Somatic embryo induction medium: NH 4 Cl 1100mg / L+H 3 BO 3 6mg / L+KNO 3 2500mg / L+Ca(NO 3 ) 2 560mg / L+NaH 2 PO 4 325mg / L+MgSO 4 ·7H 2 O 370mg / L+CuSO 4 ·5H 2 O 0.25mg / L+Na 2 -EDTA37.3mg / L+FeSO 4 ·7H 2 O 27.8mg / L...
Embodiment 3
[0052] 1. Explant acquisition: select a high-yielding healthy betel nut plant, take the uncracked young inflorescence of spathe as an explant, wash the surface of the inflorescence with a saturated washing powder solution and rinse it clean, and wash it with 75% alcohol in an ultra-clean workbench. After wiping the entire surface of the inflorescence evenly several times, use 0.1% HgCl 2 Soak for 5-12min, wash 5 times with sterile water, and place on sterile filter paper to blot the surface moisture.
[0053] 2. Direct somatic embryo induction: take the above-mentioned sterilized explants and cut them into 1-3mm thick small pieces, inoculate them into somatic embryo induction medium, and culture them in dark at 25-28°C to obtain somatic embryos.
[0054] Somatic embryo induction medium: NH 4 Cl 1300mg / L+H 3 BO 3 5mg / L+KNO 3 2600mg / L+Ca(NO 3 ) 2 480mg / L+NaH 2 PO 4 350mg / L+MgSO 4 ·7H 2 O 390mg / L+CuSO 4 ·5H 2 O 0.25mg / L+Na 2 -EDTA37.3mg / L+FeSO 4 ·7H 2 O 27.8mg / L...
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