Phyllostachys edulis protoplast culture method

A technology of protoplasts and moso bamboo, applied in the field of tree biotechnology and cell engineering breeding

Active Publication Date: 2015-12-16
RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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  • Summary
  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

As one of the most common and important bamboo species in my country and the world, there is no report on the cultivation of moso bamboo protoplasts

Method used

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Embodiment Construction

[0031] The present invention is described in detail below in conjunction with Phyllostachysedulis example of Phyllostachysedulis.

[0032] (1) Select a number of dense light yellow embryogenic calli of Phyllostachys pubescens, place them on the basis of MS medium, add 3mg / L of 2,4-D, 500mg / L of proline, and 500mg / L of glutamine Amide, 300mg / L of hydrolyzed casein, 30g / L of glucose, 8g / L of agar powder culture medium, 26 ℃ dark culture for 10d, white loose embryogenic callus grows on the edge of callus; The white and loose embryogenic callus was separated and put into the cellulase containing 2% cellulase, 1.5% macerozyme, 0.1% MES, 0.7M manitol, 0.1% CaCl 2 .2H 2 O, 1% BSA, pH 5.80 in the culture dish of the enzymolysis solution, placed on a shaker at 70rpm to carry out enzymolysis culture in the dark for 3h;

[0033] (2) Use a 40μm cell sieve to filter the enzymolysis solution, collect the filtered liquid, centrifuge at 200g for 10min at 4°C, pour off the supernatant carefu...

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Abstract

The invention discloses a phyllostachys edulis protoplast culture method. The method comprises the steps of performing enzymolysis to embryonic calluses, performing filtration, centrifugal separation and purification to obtained protoplasts after the enzymolysis, then culturing the purified protoplasts on a proper culture medium to enable the protoplasts to regenerate cell walls to form embryonic calluses which germinate to form regenerated plants, finally performing seedling strengthening and hardening to the regenerated plants and then transplanting seedlings to survive. The phyllostachys edulis protoplast culture method can solve the technical problem of phyllostachys edulis protoplast culture, provides a technical platform for the cell engineering breeding of phyllostachys edulis, and also provides a reference for related researches on other bamboo plants.

Description

technical field [0001] The invention relates to the fields of tree biotechnology and cell engineering breeding, in particular to the cultivation of moso bamboo protoplasts. Background technique [0002] As an important part of plant cell engineering, protoplast culture technology emerged in the 1960s and 1970s and has been widely used in the field of plant genetics and breeding research for decades. With the development of genetic engineering, the protoplast system has also become an important tool for gene expression and intracellular positioning, and plays an important role in the field of biotechnology. [0003] In 1960, Cocking succeeded in separating protoplasts by enzymatic method, provided a method for preparing a large number of viable protoplasts, and initiated the research field of plant protoplast culture and cell hybridization (Reference 1). [0004] In 1971, Nagata and Takebe obtained intact regenerative cells from cultured tobacco mesophyll protoplasts, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 袁金玲顾小平岳晋军吴晓丽
Owner RES INST OF SUBTROPICAL FORESTRY CHINESE ACAD OF FORESTRY
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