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

An in vitro culture and culture method technology, applied in plant tissue culture and biological fields, can solve the problems of low rooting efficiency, browning of explants, high deformity rate, etc., and achieve robust regeneration buds, easy survival after transplanting, and slow down browning Effect

Inactive Publication Date: 2011-10-19
THE INST OF MICROBIOLOGY XINJIANG ACADEMY OF AGRI SCI
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  • Description
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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

[0044] Example 1: The method of in vitro culture of sea-island cotton epicotyls for direct multi-bud generation and regenerated plants and decapitated epicotyl explants

[0045] The seeds of sea-island cotton variety Xinhai No. 19 were treated with sulfuric acid to obtain light seeds. The seeds were first surface sterilized with 70% ethanol for 30 sec, followed by 15% hydrogen peroxide (H 2 o 2 ) aqueous solution for 2hr., then rinsed with sterilized water for 6 times and soaked in sterilized water, drained after 12-36hr., squeezed out the seed coat, inoculated the seed kernel on 1 / 2MS solid medium and cultured in a dark place On the 1st to 3rd day, the cotyledon and the hypocotyl and radicle 4-6 mm below the cotyledon node were cut off and discarded, and the remaining part was the epicotyl explant. Cut a gap of 0.3-0.7 mm longitudinally at the top of the epicotyl explant, avoid splitting the hypocotyl, and then insert the multi-bud induction medium (MS salt+B) with the 4-6 ...

Embodiment 2

[0046] Example 2: The method of in vitro culture of upland cotton epicotyls for direct multi-bud regeneration and decapitated epicotyl explants

[0047] The seeds of Xinluzao 17, an early-maturing upland cotton variety, were treated with sulfuric acid to obtain light seeds. The seeds were first surface sterilized with 70% ethanol for 30 sec, followed by 15% hydrogen peroxide (H 2 o 2 ) aqueous solution for 2hr., then rinse with sterilized water for 5 times and soak in sterilized water. After 36-48hr., drain the water and squeeze out the seed coat, inoculate the seed kernel on 1 / 2MS solid medium and culture in a dark place On 2-5 days, cut off and discard the cotyledons and the hypocotyls and radicles 6-10 mm below the nodes of the cotyledons, and the remaining part is the epicotyl explant. Cut a gap of 0.7-1.3 mm longitudinally at the top of the epicotyl explant, avoid splitting the hypocotyl, and then insert the multi-bud induction medium (MS salt+B) with the 6-10 mm hypoco...

Embodiment 3

[0048] Example 3: The method of in vitro culture of Xintiansi No. 1 epicotyl and direct multi-bud regeneration plant and decapitated epicotyl explant

[0049] Gently crush the seed coat of Xintiansi No. 1 seed to remove the kernel, first sterilize the surface with 70% ethanol for 30 sec, then sterilize with 0.1% mercuric chloride aqueous solution for 10 min., and then rinse with sterilized water for 6 times. Kernels were inoculated on 1 / 2MS solid medium and cultured under light. Cut off the cotyledons and hypocotyls and radicles 6-10mm below the cotyledon nodes from the aseptic seedlings that had been cultured for 5-7 days. For the epicotyl explants, the upper embryo was treated with a small gap, and then transferred to the medium for inducing multiple shoots (MS salt+B 5 Organic +6-BA 0.1~0.5mg / l+NAA 0.5~1.5mg / l+glucose 30g / l), after 20 days of cultivation, multiple buds can be seen. After culturing the epicotyl explants that did not produce multiple buds for 15 days, the bu...

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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 exopThe 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 culturhyte. 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 exophytee 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 exophyteis 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 plan 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 st 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.now locus breeds applied in production during gene engineering.

Description

[0001] The application of the present invention is a divisional application of the invention-creation titled "a method for cultivating cotton epicotyls in vitro and direct multi-bud generation and regeneration of plants" filed by the applicant on November 13, 2006. The filing 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 in vitro culture of cotton epicotyls for direct multi-bud generation and regeneration of plants". field of invention [0002] The invention relates to the fields of plant tissue culture and biotechnology. Background technique [0003] Tracing back to the methods of in vitro culture and regeneration of plants reported on cotton, the main method is to induce differentiation of somatic embryos through callus to regenerate plants, which can easily cause high deformity rate and long culture period, and only It is suitable for Kezi cotton or wild cotton, and it i...

Claims

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Application Information

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Patent Type & Authority Patents(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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