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Method of screening mutants being insensitive to strigolactone

A strigolactone and mutant technology, applied in the field of agricultural biology, can solve the problems of lack of mutant materials, lack of creation of mutants, etc., and achieve the effect of inhibiting root growth

Inactive Publication Date: 2017-05-31
INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Although strigolactone is the current research hotspot, there are two problems: First, compared with the five major hormones of auxin, gibberellin, abscisic acid, cytokinin, and ethephon, strigolactone The research on this technology has been a research hotspot in recent years, mainly due to the lack of mutant materials; secondly, through literature search, there is no patent technology or non-patent literature similar to this technology patent at home and abroad. technology

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  • Method of screening mutants being insensitive to strigolactone
  • Method of screening mutants being insensitive to strigolactone
  • Method of screening mutants being insensitive to strigolactone

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Embodiment 1

[0033] A method for screening strigolactone-insensitive mutants, the steps of which are:

[0034] (1) EMS treatment of wild-type Arabidopsis seeds:

[0035] A, select the Arabidopsis seeds harvested within one year, weigh 0.2g, put into a 50mL centrifuge tube, mix evenly with 0.1% (weight ratio) Tween 15 (0.1g Tween 15 is put into 100mL distilled water), process Rinse with distilled water after 15 minutes until there are no bubbles;

[0036] B. Add 15-45mL EMS (0.1% or 0.2% or 0.3%), usually 0.2% (weight ratio), must be operated in a fume hood, shake well, mix well for 8 or 10 or 12 or 14 or 16 hours ;

[0037] C. Pour the treated EMS into freshly prepared 0.5M sodium hydroxide (NaOH) overnight and pour it out, put the seeds in 50mL distilled water, mix and shake well and wash for 2 or 3 or 4 hours;

[0038]D, the seeds after rinsing are moved into 100mL 0.1% (weight ratio) agar (Agar), and are moved into the soil with a pipette gun to grow, every small bowl (area 100cm 2 ...

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Abstract

The invention discloses to a method of screening mutants being insensitive to strigolactone. The method includes the steps of: 1) processing of wile-type arabidopsis thaliana seeds: A) weighing seeds and washing the seeds in distilled water; B) adding ethylmethane sulfonate and uniformly mixing the components in a fume cupboard; C) pouring processed ethylmethane sulfonate in NaOH and cleaning the seeds; D) placing the cleaned seeds in an agar solution and moving the seeds into soil by means of a pipette gun for growth; 2) screening of the mutants being insensitive to strigolactone: A) when the seeds are matured, harvesting the seeds and placing the seeds in a centrifugal tube, and disinfecting the seeds with a bleaching agent solution and placing the seeds in a refrigerator; B) sowing the seeds on an AT culture medium containing the strigolactone by means of the pipette gun; C) irradiating the culture dish, and placing the culture dish in a dark place so that the seeds are germinated; and D) selecting a long-root-system arabidopsis thaliana mutant single plant and transplanting the single plant in soil. The method has simple operations and short screening period, and can quickly and high-effectively screen the arabidopsis thaliana mutants being insensitive to the strigolactone.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and more specifically relates to a method for screening strigolactone-insensitive mutants. The wild-type Arabidopsis seeds are treated with ethyl methanesulfonate, and the offspring seeds are treated with strigolactone at a certain concentration. Lactone acts as a selective pressure, screening for root length-insensitive mutants. The invention provides a technical reference for the screening of plant hormone-sensitive differential materials. Background technique [0002] EMS is ethyl methanesulfonate, which is the most commonly used type of alkylating agent for mutagenesis, and has a high mutagenesis rate. More than 99% of the mutations are base C to T mutations, so CG pairings are changed to TA pairings. EMS mutant materials can be used in forward genetics and reverse genetics research. The frequency of mutations depends on the position of the gene in the genome and the treatment con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/04
CPCA01H1/04
Inventor 张丰琪马霓张春雷胡超万林
Owner INST OF OIL CROPS RES CHINESE ACAD OF AGRI SCI
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