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Technique for epicotyl of cotton culturing in vitro regenerated plant, and decapitated epicotyl explant

A technology for in vitro culture and regeneration of plants, applied in plant regeneration, plant cells, botany equipment and methods, etc., can solve the problems of browning of explants, low rooting efficiency, high deformity rate, etc., and achieve easy survival after transplanting, The regenerated buds are strong and the effect of slowing down browning

Inactive Publication Date: 2007-07-18
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

[0014] 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;
[0015] 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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  • Technique for epicotyl of cotton culturing in vitro regenerated plant, and decapitated epicotyl explant
  • Technique for epicotyl of cotton culturing in vitro regenerated plant, and decapitated epicotyl explant
  • Technique for epicotyl of cotton culturing in vitro regenerated plant, and decapitated epicotyl explant

Examples

Experimental program
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Effect test

Embodiment 1

[0047] 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

[0048] 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

[0049] Example 2: The method of in vitro culture of upland cotton epicotyls for direct multi-bud regeneration and decapitated epicotyl explants

[0050] 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

[0051] Example 3: The method of in vitro culture of Xintiansi No. 1 epicotyl and direct multi-bud regeneration plant and decapitated epicotyl explant

[0052] 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

A technique for directly polygemmic regenerative plant of cotton by in vitro culture of its epicotyl explant and a decapitated epicotyl explant are disclosed.

Description

field of invention [0001] The invention relates to the fields of plant tissue culture and biotechnology. Specifically, the present invention relates to a technique for in vitro culture of cotton epicotyls and direct multiple shoots to regenerate plants and decapitated epicotyl explants. Background technique [0002] 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 is invalid for the main cultivated cotton varieties. Despite the commercial success of genetically modified cotton, genetic transformation and regeneration of cotton remains challenging compared to other crops. These issues are mainly genotype-dependent regeneration, efficiency of somatic embryogenesis, and somatic variation. An important a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C12N5/04
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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