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Rapid and efficient cymbopogon citratus protoplast preparation method

A technology for protoplasts and citronella, which is applied in the field of rapid and efficient preparation of citronella protoplasts, can solve problems such as undiscovered, and achieve the effects of simple operation, high activity, and short enzymatic hydrolysis time.

Active Publication Date: 2021-06-18
SHANGHAI CHENSHAN BOTANICAL GARDEN
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Problems solved by technology

No research report on the preparation of citronella protoplasts has been found. Therefore, a fast and efficient method for preparing citronella protoplasts is urgently needed for the subsequent use of protoplasts for gene function identification, subcellular localization, protein interaction, and cell fusion. Lay a solid foundation for research on single-cell sequencing and other aspects

Method used

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  • Rapid and efficient cymbopogon citratus protoplast preparation method
  • Rapid and efficient cymbopogon citratus protoplast preparation method
  • Rapid and efficient cymbopogon citratus protoplast preparation method

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Embodiment 1

[0049] This embodiment relates to a fast and efficient preparation method of citronella protoplasts, specifically comprising the following steps: culturing citronella tissue culture seedlings; enzymolysis of citronella protoplasts; separation and purification of citronella protoplasts; collection of citronella protoplasts; Mao protoplast observation, the preparation process is as follows figure 1 As shown, it specifically includes the following steps:

[0050] (1) Cultivation of citronella tissue culture seedlings: clip the strong young stems of citronella, rinse with running water after soaking in sterile water, then soak 10min with 75% alcohol in an ultra-clean bench, rinse with sterile water for 3 times, and then use Soak in 10% sodium hypochlorite solution for 5 minutes, and finally rinse with sterile water for 5 times, then move to sterilized 1 / 2 MS medium and cultivate for about 3 weeks until leaves grow out.

[0051] (2) Enzymatic hydrolysis of citronella protoplasts: ...

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Abstract

The invention discloses a rapid and efficient cymbopogon citratus protoplast preparation method, and belongs to the field of plant cell engineering. Healthy-growing cymbopogon citratus tissue culture seedlings are adopted as experimental materials, a CPW solution is used for preparing enzymatic hydrolysate, a low-speed oscillation enzymolysis method is adopted for carrying out enzymolysis digestion on cymbopogon citratus leaves, and the protoplast is efficiently prepared. And finally, purifying and collecting of the high-quality cymbopogon citratus protoplast is conducted by using a sucrose density gradient centrifugation method. The method is simple, convenient and easy to operate, and the separated protoplast is high in yield, complete in cell, spherical, rich in chloroplast and relatively high in activity. The invention provides an important basis for research and application in the aspects of gene function identification, subcellular localization, protein interaction, citronella cell fusion, single cell sequencing and the like by utilizing a cymbopogon citratus leaf instantaneous transformation system.

Description

technical field [0001] The invention belongs to the field of plant cell engineering and relates to a fast and efficient method for preparing citronella protoplasts. Background technique [0002] Citronella (Cymbopogon citratus) is a perennial herbaceous plant of the genus Citronella. It originates in tropical regions, such as India and Sri Lanka, and is widely distributed in Guangdong, Hainan, Yunnan and other places in my country. Ancient books record that lemongrass is sweet, pungent, and warm in nature, and has the effects of dredging wind and dredging collaterals, warming the middle, relieving pain, refreshing the brain, and stopping diarrhea. Lemongrass has a certain health care effect when used as tea, and its pharmacological effects are mainly antibacterial, anti-inflammatory, analgesic, anti-oxidant, anti-tumor, anti-anxiety, antihypertensive, hypoglycemic, etc. The essential oil extracted from its stems and leaves has a wide range of applications in food, antisepsi...

Claims

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Application Information

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IPC IPC(8): C12N5/04
CPCC12N5/04
Inventor 曹建美储昭庆张凯蒋敏宋昌梅
Owner SHANGHAI CHENSHAN BOTANICAL GARDEN
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