Plant regeneration method of humid euphoriba somatic embryogenesis path and culture medium for said method

A culture medium and Dijin technology, applied in the field of plant tissue and cell culture, can solve the problems of difficult long-term subculture and differentiation ability of callus, low reproduction efficiency, and difficulty in realizing factory production, etc., and achieve callus induction rate and somatic embryo differentiation rate, and the effect of good proliferation effect

Inactive Publication Date: 2006-03-22
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is difficult to subculture and maintain the differentiation ability for long-term subculture and maintain the differentiation ability of the callus produced by the Dijin tissue or cell culture under the above-mentioned in vitro conditions, so it is difficult to be the recipient of genetic transformation
At the same time, the prior art reports that the tissue culture of broccoli has low reproductive efficiency and is difficult to realize factory production.

Method used

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  • Plant regeneration method of humid euphoriba somatic embryogenesis path and culture medium for said method
  • Plant regeneration method of humid euphoriba somatic embryogenesis path and culture medium for said method
  • Plant regeneration method of humid euphoriba somatic embryogenesis path and culture medium for said method

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Experimental program
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Effect test

Embodiment 1

[0040] Test material selection, culture medium design and inoculation culture

[0041]1. Sources of test materials and their processing:

[0042] The test material of the present invention is selected from the bud and stem section of Dijindai, collected from the flower test base of Huazhong Agricultural University, Wuhan City, Hubei Province, and each Dijina is a genotype. The six plants collected by the applicant were numbered as PTC, PT1, PT2, PT3, PT4, and PT5, respectively. The stem sections of Dijina chinensis collected in the field were sterilized by conventional methods (refer to: compiled by Li Mingjun, Plant Tissue Culture, China Agricultural Press, 1992 edition), and the sterilized Dijina chinensis stem sections were inoculated on MS basic medium + 6-BA 1.0 mg Axillary buds were induced to germinate on / L medium to obtain sterile seedlings. After the sterile seedlings were obtained, they were inoculated on MS basic medium+6-BA1.5mg / L+IAA0.01mg / L medium for prolifera...

Embodiment 2

[0059] Effects of Different Concentrations of 2,4-D on Induction of Petiole Callus of Digum chinensis

[0060] In plant tissue culture, 2,4-D is a widely used auxin growth regulator, which is recognized as one of the most effective auxins to initiate cell dedifferentiation and form callus or organogenesis .

[0061] After cultivating the petiole of Dijina for about 1 week, a small amount of white callus can be seen at the swollen incision. About 1 month, two types of callus can be observed, one is white and translucent, more compact, with glossy with few adventitious roots (see figure 1 A), most of the callus can produce somatic embryos in subsequent passages. The other is white, loose and vigorous growth or water-soaked or browned. 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 produce callus on the medium of four different c...

Embodiment 3

[0065] Effects of Different Concentrations of BA and NAA on Induction of Somatic Embryos from Petiole Callus of Dijina chinensis

[0066] Plant hormones are important for somatic embryo differentiation and seedling formation, and can stimulate somatic cells to form embryos by changing the level of external auxin or the ratio of auxin / cytokinin. In order to investigate the effects of BA and NAA on somatic embryo induction, callus derived from genotype numbered PTC (after 2 passages) was inoculated on different somatic embryo induction media (see Table 3).

[0067] 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 emb...

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Abstract

The present invention relates to plant tissue and cell culture technology, and is especially the plant regeneration process via humid euphorbia somatic embryogenesis path. The process includes the following steps: A. culturing humid euphorbia explant in culture medicine with inducing agent and inducing callus from the explant; B. inoculating the callus to proliferation culture medium for proliferation; C. inoculating well developed callus to somatic embryo inducing culture medium to induce somatic embryo differentiation or generate fragile embryonic callus, and inoculating the fragile embryonic callus to other culture medium for secondary culture or differentiation; D. inoculating the un-mature somatic embryo to somatic embryo proliferating culture medium for proliferation and further differentiation; and E. inoculating the mature somatic embryo to seedling culture medium to culture complete plant. The present invention also relates to the different culture mediums for the process.

Description

technical field [0001] The invention belongs to the technical field of plant tissue and cell culture, is related to the rapid propagation of garden plants, and in particular relates to a method for regenerating a chinensis plant through somatic cell generation by using a petiole and a stem segment as explants, and a method for the method. 's medium. Background technique [0002] Parthenocissus tricuspidata Planch, also known as wall-crawling tiger, belongs to the genus Parthenocissus tricuspidata Planch. The plant grows rapidly, has strong climbing ability, dark green in summer, purple-red in autumn, wide adaptability and good sand-fixing ability. of cover vegetation. [0003] In the prior art, the method for regeneration of plants through the generation of broccoli somatic cells is usually as follows, inoculating immature embryos or cotyledon leaves and petioles on an induction medium to induce callus, and sub-culturing the induced callus, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 包满珠阳永学刘国锋张俊卫高丽萍胡惠蓉叶要妹
Owner HUAZHONG AGRI UNIV
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