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Method for inducing haploid endosperm callus tissues by using ripe taxales chinensis var.mairei endosperms

A technology of callus and mature embryo, which is applied in the field of haploid endosperm callus induced by mature endosperm of Taxus chinensis, and achieves the effects of saving water resources, low pollution rate and simple pretreatment

Active Publication Date: 2012-09-26
HUNAN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] According to the current research reports, there has been no successful report on the haploid callus induced by the endosperm callus of Taxus chinensis

Method used

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  • Method for inducing haploid endosperm callus tissues by using ripe taxales chinensis var.mairei endosperms
  • Method for inducing haploid endosperm callus tissues by using ripe taxales chinensis var.mairei endosperms

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Embodiment 1

[0029] (1) Stripping of seed embryos: Collect the seeds of the southern yew in the current year, use mechanical and manual methods to break the shell, and peel the seeds from the outer testa;

[0030] (2) Disinfection of explants: first put the seeds with the outer testa removed into a sterile container, disinfect them with 70% alcohol for 90 seconds and 0.1% mercury chloride for 12 minutes in an ultra-clean workbench, and rinse them with sterile water for 5 times Reserve later, soak the seeds for about 5 minutes each time when cleaning.

[0031] (3) Pretreatment of seeds: after the surface disinfection treatment of step (2), the seeds with the outer testa removed are sealed and soaked in sterile distilled water at room temperature for 1-3 days.

[0032] (4) Separation of endosperm tissue: the endosperm tissue of the seeds is stripped under sterile conditions, and the other parts are discarded;

[0033] (5) Induction of endosperm callus: Inoculate the endosperm tissue onto th...

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Abstract

The invention discloses a method for inducing haploid endosperm callus tissues by using ripe taxales chinensis var.mairei endosperms, and the method comprises the following steps of: (1) seed exotesta peeling: mechanically breaking a shell, and manually peeling exotesta; (2) explants sterilization: sequentially sterilizing by using 70% alcohol for 90 seconds and 0.1% mercuric chloride for 12 minutes, and washing five times with sterile water for later use; (3) seed pretreatment: sealing with the sterile water, and soaking seeds for 1-5 days at normal temperature; (4) separation of endosperm tissues; and (5) in vitro induction of endosperm callus tissues: inoculating seed endosperms to a callus tissue-inducing culture medium, so as to obtain the 97.92% induction rate after 30 days. By utilizing the method, ripe taxales chinensis var.mairei endosperm callus tissues can be obtained conveniently and quickly, and the ripe taxales chinensis var.mairei endosperm callus tissues lay a foundation for simplifying genetic analysis and chromosome operation and are used as transgenic materials and genomics research materials.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a method for inducing haploid endosperm callus from yew mature endosperm. Background technique [0002] Paclitaxel (trade name Taxol) is a natural tetracyclic diterpene alkaloid isolated from the bark of Taxus L. in the 1960s and 1970s. It has remarkable curative effect in the treatment of cervical cancer, breast cancer, small cell lung cancer, melanoma and colon cancer. Paclitaxel was first extracted from the bark and cambium of the Pacific yew. According to the current purification technology, about 10 tons of yew bark are needed to produce 1 kg of paclitaxel. This needs to be obtained from about 2000 to 4000 yew trees that are 60-70 years old. According to relevant information, the world needs to consume about 250kg of pure paclitaxel every year. This needs to be extracted from 2.5 million kilograms of bark or 750,000 yew trees to meet the needs of this mark...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 熊兴耀李炎林张家银石梦蝶秦宇
Owner HUNAN AGRICULTURAL UNIV
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