Tissue culture propagation and in vitro preservation method of a critically endangered plant Rhododendron grandiflora Guizhou

A technique for in vitro preservation and rhododendron flowers, applied in the biological field, to achieve the effects of increasing the reproduction coefficient, prolonging the transfer and preservation time, and protecting endangered species

Active Publication Date: 2021-12-21
贵州思源农旅综合开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there are no reports on the tissue culture propagation and in vitro preservation methods for this species.

Method used

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  • Tissue culture propagation and in vitro preservation method of a critically endangered plant Rhododendron grandiflora Guizhou
  • Tissue culture propagation and in vitro preservation method of a critically endangered plant Rhododendron grandiflora Guizhou

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Example 1 Tissue culture and in vitro preservation steps of Rhododendron grandiflora in Guizhou:

[0018] Step (1): Collect explants and sterilize them: Collect new stem segments that have just grown after flowering from the mother plant of Rhododendron grandiflorum Guizhou, with 3-5 latent buds, take the explants and wash them in tap water for 12 hours, On the ultra-clean workbench, disinfect with 75% alcohol for 30 seconds, rinse with sterile water for 3-5 times, disinfect with 0.1% mercury chloride for 8-9 minutes, and rinse with sterile water for 3-5 times;

[0019] Step (2): Cumulative bud induction culture: cut the sterilized explants into small sections on the ultra-clean workbench, each section has an axillary bud, and insert obliquely into the induction medium modified WPM + 2.5mg / L 2-iP + 0.8 mg / L IBA, sucrose 28g / L, agar 7g / L, pH 5.6, dark culture for 5 days, light culture for 30 days, culture for 35-50 days, light and dark time 12h / 12h, light intensity 1500 ...

Embodiment 2

[0023] Example 2 Media screening experiment for tissue culture propagation of Rhododendron grandiflora in Guizhou:

[0024] 1) Basal medium screening experiment. The media Read, WPM, 1 / 4MS, and improved WPM commonly used by rhododendrons were selected for comparative experiments, and the hormone-free stem segment survival experiment was carried out. Each group of experiments was inoculated with 10 bottles, and each bottle was inoculated with 1 segment of explant stem segment after disinfection. Compare:

[0025]

[0026] The results show that the species grows well on the medium of the improved WPM, and the growth is relatively vigorous, and no death or non-germination occurs. Therefore, the basic medium is selected as the improved WPM.

[0027] 2) In vitro preservation medium screening experiment. The design temperature is two gradients: low temperature 13-18°C and normal temperature 20-23°C. The designed medium filling volume is two gradients of 45ml and 65ml. Each grou...

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Abstract

The invention discloses a method for tissue culture propagation and in vitro preservation of an extremely endangered plant Rhododendron guizhouensis. The method is suitable for tissue culture rapid propagation and in vitro preservation of Rhododendron guizhouensis in the family Rhododendronaceae, and belongs to the field of plant biotechnology. The present invention uses freshly drawn tender stems after flowering as explants, and through a series of steps of explant disinfection, cluster bud induction, strong seedlings, in vitro preservation, and rooting, effectively solves the problem of the distribution area of ​​Rhododendron grandiflora in Guizhou. Narrow and difficult to introduce and domesticate, the number of plants is small, and it is adjacent to the status quo of endangered species, so as to achieve the purpose of species protection. The tissue culture rapid propagation method provided by the present invention has a multiplication coefficient of 8-10 within one month, a rooting rate of 95%, and a transplanting survival rate of 95%-98%, which greatly improves the reproduction coefficient of Rhododendron grandiflora in Guizhou , The in vitro preservation can reach more than 240d, which provides technical support for the introduction and domestication of this species, in vitro preservation, and seedling expansion and return to planting.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for tissue culture propagation and in vitro preservation of an extremely endangered plant Rhododendron grandiflorum. Background technique [0002] Guizhou Rhododendron (Rhododendron magniflorum ) is a species of the subgenus Rhododendron subgenus Rhododendron in the family Rhododendronaceae, which was published in 1985 and is the rhododendron with the largest flowers in the genus Rhododendron. It is listed as critically endangered in the latest edition of The Red List of Rhododendron. This species is distributed in Longtou Mountain, Anlong County, Guizhou Province. The diameter of a single flower corolla is about 10cm, which has high ornamental value. In the latest survey, only one population of this species has been found so far, and there are only 3 strains left. Due to the scarcity of populations and poor natural regeneration, effective tissue culture propa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 田晓玲马永鹏乔琴路登宇何娇朱强
Owner 贵州思源农旅综合开发有限公司
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