A kind of Phalaenopsis somatic embryogenesis synchronization regulation method
A technology of somatic embryos and Phalaenopsis, applied in the biological field of horticultural plant seedling propagation, can solve the problems of high pollution rate, large cell damage, cumbersome operation of mechanical sieving method, etc., and achieve the effect of simple operation and low cost
Inactive Publication Date: 2016-03-30
SUBTROPICAL CROPS INST OF FUJIAN PROVINCE
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[0004] The above-mentioned synchronous generation control method of somatic cells, the mechanical sieving method is cumbersome to operate and has a high pollution rate. It is suitable for liquid suspension culture of somatic embryos. Cell nutrient starvation is relatively harmful to cells. From the data point of view, optimizing the proliferation medium or improving the culture conditions is not a good idea. Main methods of synchronization control of somatic embryogenesis
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[0019] 1. Initial induction of cut protocorm explants of Phalaenopsis: The medium is 1 / 2MS (macroelement)+5mg / L6-BA+12g / L mannitol+20% coconut milk+2% sucrose, pH5. 8. Routine autoclaving. Mature protocorms of Phalaenopsis were cut into 3-5 pieces on a clean table, and inserted into the culture medium. The culture temperature was 24-26°C, the light intensity was about 1500lx, and the light was 12h·d-1.
[0020] 2. On the 11th-18th day of culture, a large number of embryogenic mother cells can be seen in the culture by microscopic observation, and they are transferred on the 18th day, and the medium is 1 / 2MS (macroelement) + 5.0mg / L6-BA + 12g / L mannitol + 20% coconut milk + 2% sucrose, autoclaved, add 100mg of vitamin C and 100mg of cysteine per liter of medium to make an aqueous solution, filter and sterilize on the ultra-clean workbench, put in the medium Add before condensation, adjust pH to 5.6. The culture is inoculated, the culture temperature is 24-26°C, the light i...
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The invention discloses a regulating and controlling method for synchronization of a somatic embryo of butterfly orchid applying an antioxidant. The method, taking a cultured protocorm slice of butterfly orchid as an explant, comprises the following steps: (1) in initial induction, adding cytokinin and high-concentration hypertonic matters into a conventional culture medium; (2) in a stage when the explant has embryo auxocytes, converting to a culture medium containing cytokinin, high-concentration hypertonic matters and the antioxidant; (3) when an embryonic callus is cultured, converting to a culture medium without cytokinin, with lower-concentration hypertonic matters and added with the antioxidant; (4) when tender protocorm is cultured, converting to a culture medium without cytokinin, without hypertonic matters and added with the antioxidant; and (5) continuing to culture to obtain protocorm like bodies which are relatively consistent in size and maturity, wherein the synchronization is about 80%. The method disclosed by the invention has the advantages of simplicity and convenience in operation, low cost and no side effects to plant materials.
Description
technical field [0001] The invention belongs to the biological technology of horticultural plant seedling propagation in the field of agriculture and forestry, and in particular relates to a method for regulating and controlling the synchronous development of Phalaenopsis somatic embryos. Background technique [0002] Plant somatic embryogenesis is an important system for studying plant cell pluripotency and embryogenesis mechanism, and a synchronized somatic embryogenesis system is necessary for studying its physiology, biochemistry, and molecular biology. In production practice, in order to use somatic embryogenesis, an effective in vitro reproduction method, to truly realize industrialization and standardized production, it is also necessary to realize the synchronization of somatic embryogenesis. In the development of artificial seeds, obtain morphological, mature Synchronized somatic embryos with similar degrees are an important prerequisite for automatic production of ...
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IPC IPC(8): A01H4/00
Inventor 刘福平
Owner SUBTROPICAL CROPS INST OF FUJIAN PROVINCE