Green cucumber high-frequency regeneration method

Abstract

The invention discloses a cucumber high frequency plant generative method, composed of exophytic preparation of Flamingo-bill, inducement and increasement of indefinite buds, radication and transplantation and the like. According to the invention, only one phytohormone is used during the process of tissue culture, density is lower, clone generation of somatic cells are not easy to be induced, technical cost is saved; technical link is few, operation is simple, producing efficiency is increased; only 45 d are required from inoculability to formation of the full plant, thereby greatly shortening generative time of the plant.

Description

technical field [0001] The invention belongs to the field of plant biotechnology, in particular to a cucumber high-frequency plant regeneration method. Background technique [0002] The research on cucumber tissue culture technology started in the 1970s (Cutts RHA. Plant Sci Lett, 1975, 4: 189-193), but it was in recent years that the really useful results were obtained (Chen Jifeng, Shanghai Agricultural Journal, 2007, 23:114-118). There are many types of explants used for cucumber tissue culture, such as cotyledons, true leaves, hypocotyls, petioles, shoot tips, roots, zygotic embryos, etc. Most of the studies can obtain tissue culture seedlings. Although there are many successful experiences in cucumber tissue culture, there are still many problems in some aspects. First of all, cucumber tissue culture has strong genotype specificity, the plant regeneration frequency is low and unstable, and the repeatability of the technique is poor (Hou Aiju, Zhu Yanming, Yang Aifu, e...

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Problems solved by technology

First of all, cucumber tissue culture has strong genotype specificity, the plant regeneration frequency is low and unstable, and the repeatability of the technique is poor (Hou Aiju, Zhu Yanming, Yang Aifu, etc. Acta Horticultural Science, 2003, 30: 101-103); Secondly, since cucumber regeneration plants generally go through several processes of callus formation, adventitious bud differentiation, stem bud elongation and rooting, the existing cucumber plant regeneration system has complex operation techniques and long plant regeneration cycle (Kuijpers AM, Bouman H, Klerk GJ.Plant Cell Tiss Org Cult, 1996, 1:81-83); Third, most researchers tend to obtain cucumber regenerated plants through somatic embryos, but the frequency of somatic embryos transforming into normal plants is low, And the variation of clones is large, which is not conducive to the cultivation of genetically stable and consistent regenerated plants (Lou H, Kako S. HortiScience, 29: 906-909)

These unfavorable factors limit the progress of cucumber bioengineering breeding to a certain extent

With the rapid development of bioengineering technology, rapid plant cloning technology and genetic transformation technology will be widely used in cucumber biotechnology breeding, which requires the establishment of an efficient plant regeneration system that is easy to operate, short in cycle, and stable in technology. The existing cucumber in vitro regeneration technology system cannot fully meet the needs of current cucumber biotechnology research.

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