Plant regeneration method of humid euphoriba somatic embryogenesis path and culture medium for said method
A technology of culture medium and proliferation medium, which is applied in plant regeneration, botanical equipment and methods, horticultural methods, etc., can solve the problem that callus has difficulty in long-term subculture differentiation ability, low reproduction efficiency, and difficulty in realizing industrial production, etc. To achieve the effect of improving callus induction rate and somatic embryo differentiation rate and good proliferation effect
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Embodiment 1
[0040] Test material selection, medium design and inoculation culture
[0041] 1. Source of test materials and their treatment:
[0042]The test materials of the present invention are selected from the buds and stems of the brocade band, collected from the flower test base of Huazhong Agricultural University, Wuhan City, Hubei Province, and each plant of the brocade is a genotype. According to the difference in differentiation rate, the applicant will collect six brocade plants and number them PTC, PT1, PT2, PT3, PT4, PT5 respectively. The stems of Radix chinensis collected in the field are sterilized according to conventional methods (refer to: Compiled by Li Mingjun, Plant Tissue Culture, China Agricultural Publishing House, 1992 edition), inoculate the stems of Radix radiata after disinfection in MS basic medium+6-BA1.0mg / L medium to induce axillary bud germination to obtain aseptic seedlings, and inoculate them on MS basic medium + 6-BA1.5mg / L + IAA0.01mg / L medium to pro...
Embodiment 2
[0060] Effects of Different Concentrations of 2,4-D on Callus Induction of Petiole of Radix Digitata
[0061] In plant tissue culture, 2,4-D is a widely used auxin growth regulator, and it is recognized as the most effective auxin for initiating cell dedifferentiation, forming callus or organogenesis .
[0062] After cultured for about 1 week, a small amount of white callus can be seen at the enlarged incision. After about 1 month, two types of callus can be observed, one is white and translucent, firmer, with Glossy with few adventitious roots (see figure 1 A), most of the callus can produce somatic embryos in subsequent subcultures. The other is white, loose and vigorous growth or water-stained or brown. The state of this callus is difficult to adjust, and somatic embryos cannot be produced during differentiation. In the method of the present invention, the petioles of the six genotypes tested can all produce callus on the medium of four different concentrations of 2,4-D,...
Embodiment 3
[0067] Effects of Different Concentrations of BA and NAA on the Induction of Somatic Embryos from Petiole Callus
[0068] Phytohormones play an important role in somatic embryo differentiation and seedling formation, and can stimulate somatic cells to form embryos by changing the level of auxin or the ratio of auxin / cytokinin. In order to explore the effects of BA and NAA on somatic embryo induction, the callus derived from genotype coded PTC (after subculture twice) was inoculated on different somatic embryo induction media (see Table 3).
[0069] Different concentrations of 6-BA and NAA and their interaction had significant effects on somatic embryo induction. LSD analysis showed that there were significant differences between BA1.5mg / L and BA1.0mg / L and BA2.0mg / L, but there was no significant difference between BA1.0mg / L and BA2.0mg / L. When the NAA concentration is within a certain range (0.01-0.05mg / L), with the increase of BA concentration, the somatic embryo differentia...
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