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Production method of anoectochilus symbiosis seedling

A production method and a technology of tissue culture seedlings, which are applied in the field of clematis symbiotic seedlings, can solve the problems of low invasion frequency and low production efficiency, and achieve the effects of reducing production costs, shortening the growth cycle, and improving the survival rate

Inactive Publication Date: 2010-10-13
东莞市生物技术研究所 +1
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Problems solved by technology

The symbiosis between whole seedlings of tissue culture and mycorrhizal fungi, due to the natural structural barrier of cells, leads to low invasion frequency and low production efficiency

Method used

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  • Production method of anoectochilus symbiosis seedling
  • Production method of anoectochilus symbiosis seedling
  • Production method of anoectochilus symbiosis seedling

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specific Embodiment

[0018] First activate Rhizoctonia sp. JSH-1 from the slant test tube strain preserved at low temperature, transfer it to a PDA plate, and culture it in the dark at 28° C. for 3 to 5 days for later use.

[0019] Use a sterilized puncher (0.5-1 cm in diameter) or a burnt inoculation needle to make a block on the edge of the colony or pick a mycelial agar block with a side length of 0.4-0.7 cm.

[0020] Select thicker aseptic clematis twigs with a height of 5 ± 0.5 cm, remove the top, and cut into 2 to 4 stem segments with a sterilized scalpel, each stem segment must have more than one stem node, Then received in the symbiotic medium, each bottle received 4 stem segments. Finally, the agar block of Rhizoctonia sp. JSH-1 mycelium was picked and placed in the middle of the four stem segments, and a control was set at the same time, that is, no inoculation. All the above operations were performed on a sterile ultra-clean bench.

[0021] Culture conditions: temperature: 25°C; light...

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Abstract

The invention relates to a production method of anoectochilus symbiosis seedling, which comprises the following steps that: (1) endophytic fungi culturing: after being activated from a slant tube specie preserved at low temperature, rhizoctonia sp. JSH-1 with the preservation number of CGMCC No. 3074 which is separated from the root of wild anoectochilus is transferred into a PDA plate and cultured for 2 to 4 days in dark at 28DEG C; then a mycelial agar block is prepared; and (2) anoectochilus tissue culture symbiosis: the top leaf bud of the sterile anoectochilus tissue culture seedling are removed, sterile operation is adopted to cut off the leaf bud from the middle parts of the two eustipes to obtain 2 to 4 stems, and then the stems are simultaneously grafted to a symbiotic culture medium to be symbiotically cultured for 2 to 3 months together with the rhizoctonia sp. JSH-1 mycelial agar block. The stem symbiotic anoectochilus tissue culture symbiosis culture method can greatly reduce the use cost of anoectochilus tissue culture seedling, shorten the growth cycle, improve the survival rate so as to reduce the production cost and facilitate the realization of industrial production.

Description

Technical field: [0001] The invention relates to a method for obtaining clematis symbiotic seedlings through the symbiosis of tissue cultured seedlings of clematis and mycorrhizal fungi. Background technique: [0002] Clematis clematis belongs to the perennial herb of the family Orchidaceae. It is a traditional precious medicinal material widely used by the people. It is mostly used to treat tuberculosis, lung heat cough, rheumatoid arthritis, etc. In recent years, the effectiveness of clematis in the treatment of difficult diseases such as hypertension, diabetes and tumors has attracted more and more attention from the medical field. Especially in Taiwan, clematis is known as the "king of traditional Chinese medicine". [0003] However, clematis is very rare in natural resources. The reason is that it has very harsh requirements on the ecological environment and is damaged by small animals and people in nature. The more important thing is that the seeds of clematis are very...

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Application Information

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IPC IPC(8): A01H4/00C12N1/14
Inventor 方祥江绮晴张善信陈春满
Owner 东莞市生物技术研究所
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