Direct polygerm-generation regeneration plant culture method by isolated culture of Xinjiang snow locus epicotyl

A technology of in vitro culture and culture method, which is applied in the field of cotton epicotyl in vitro culture and direct multi-bud regeneration plants, which can solve the problems of low rooting efficiency, browning of explants, high deformity rate, etc., and achieve regenerated buds Robust, easy to survive after transplanting, and slow down the effect of browning

Inactive Publication Date: 2009-09-02
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] 1. At present, plants are mainly regenerated by inducing callus to differentiate into somatic embryos, which is limited by genotype, and has a high deformity rate and a long culture period;
[0016] 2. Among the many methods of using explants to directly induce bud generation to regenerate plants, it is either limited by genotype, or the bud generation efficiency is not high, or the explants are severely browned, or the rooting efficiency is low and depends on genotype, or Regenerated plants take a relatively long time to obtain

Method used

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  • Direct polygerm-generation regeneration plant culture method by isolated culture of Xinjiang snow locus epicotyl
  • Direct polygerm-generation regeneration plant culture method by isolated culture of Xinjiang snow locus epicotyl
  • Direct polygerm-generation regeneration plant culture method by isolated culture of Xinjiang snow locus epicotyl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Method for in vitro culture of sea island cotton epicotyls for direct multi-budding regenerated plants and decapitated epicotyl explants

[0049] The seeds of sea island cotton variety Xinhai No. 19 were obtained by dehulling treatment with sulfuric acid. The seeds were first surface-sterilized with 70% ethanol for 30 sec, followed by 15% hydrogen peroxide (H 2 O 2 ) The aqueous solution was sterilized for 2hr., then rinsed with sterilized water for 6 times and soaked in sterilized water. After 12 to 36hr., the water was drained and the seed coat was extruded. From 1 to 3 days, the cotyledons and the hypocotyls and radicles 4 to 6 mm below the cotyledon nodes are cut off and discarded, and the remaining part is the epicotyl explants. Make a slit of 0.3-0.7 mm longitudinally at the top of the epicotyl explant, do not split the hypocotyl, and then insert the multi-bud induction medium (MS salt + B 5 In organic+6-BA0.0~0.8mg / l+NAA0~0.06mg / l+glucose 30g / l), mu...

Embodiment 2

[0050] Example 2: Method for direct multi-budding regenerated plants by in vitro culture of upland cotton epicotyls and decapitated epicotyl explants

[0051] The seeds of the early-maturing upland cotton variety Xinluzao No. 17 were obtained by defibrating with sulfuric acid. The seeds were first surface-sterilized with 70% ethanol for 30 sec, followed by 15% hydrogen peroxide (H 2 O 2 ) The aqueous solution was sterilized for 2 hr., then rinsed with sterilized water for 5 times and soaked in sterilized water. After 36 to 48 hr., the water was drained and the seed coat was extruded. On 2-5 days, the cotyledons and the hypocotyls and radicles 6-10mm below the cotyledon nodes were cut off and discarded, and the remaining part was the epicotyl explants. Make a slit of 0.7-1.3 mm longitudinally at the top of the epicotyl explant, do not split the hypocotyl, and then insert the multi-bud induction medium (MS salt + B 5 Organic+6-BA0.8~1.3mg / l+NAA

[0052] 0.07~1.0mg / l+glucose ...

Embodiment 3

[0053] Example 3: Method for in vitro culture of epicotyl of Xintianzhi No. 1 and direct multi-budding regeneration plant and decapitated epicotyl explants

[0054] The seeds of Xintianzhi No. 1 were smashed with force and the seed coat was gently smashed to remove the seed kernels. The surface was first disinfected with 70% ethanol for 30 sec, then sterilized with 0.1% mercuric solution for 10 minutes, and then rinsed with sterilized water for 6 times. The kernels were inoculated on 1 / 2MS solid medium and cultivated in a lighted place, and the cotyledons, the hypocotyls and radicles 6-10mm below the cotyledon nodes were cut from the sterile seedlings cultivated for 5-7 days and discarded. Epicotyl explants, on which the embryos were cut with a small gap, were transferred to inducing multi-budding medium (MS salt + B5 In organic+6-BA0.1~0.5mg / l+NAA0.5~1.5mg / l+glucose 30g / l), many buds were seen after culturing for 20 days. After culturing the epicotyl explants that did not pro...

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Abstract

The invention discloses a direct polygerm-generation regeneration plant culture method by the isolated culture of Xinjiang snow locus epicotyl. The invention also discloses a decapitated epicotyl exophyte. The invention utilizes an epicotyl exophyte and the decapitated epicotyl exophyte to establish a gene-independent, simple and rapid Xinjiang snow locus isolated culture regeneration plant culture technology which directly induces polygerm generation, elongation and rootage; by the formulation of a culture medium and the optimizing screen of carbon sources, the browning degree of the exophyte is lowered to the lightest degree, an exophyte can have the maximum generation bud number more than or equal to 3 and the regeneration bud rootage rate higher than 90 percent, and a regeneration plant with a full root, a full stalk and full leaves can be obtained by only 3 to 4 months. The invention has low malformed seedling rate and lays a foundation for the hereditary improvement of Xinjiang snow locus breeds applied in production during gene engineering.

Description

[0001] The application of the present invention is a divisional application for the invention and creation titled "a method for in vitro culture of cotton epicotyls for direct multi-bud regeneration" filed by the applicant on November 13, 2006. The application date of the original application is On November 13, 2006, the application number was 200610149143.4, and the name of the invention was "a method for culturing cotton epicotyls in vitro with direct multi-bud regeneration". Field of Invention [0002] The present invention relates to the field of plant tissue culture and biotechnology. Specifically, the present invention relates to a method for culturing cotton epicotyls in vitro to directly produce regenerated plants with multiple buds. Background technique [0003] The methods of regenerating plants in vitro which have been reported on cotton are mainly used to regenerate plants by inducing the differentiation of somatic embryos through callus, which can easily lead to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 郝秀英孙立军欧阳立恒段肖霞代兴荣王新勇李雪源谢迪佳毛鸿才闫建庆
Owner THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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