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Immunofluorescent method for detecting plant gibberellin

An immunofluorescence and gibberellin technology is applied in the field of immunofluorescence detection of plant gibberellins, which can solve the problems of high cost, expensive instruments, cumbersome operations, inability to locate gibberellins in situ, etc. simple effect

Inactive Publication Date: 2019-08-02
南京烁朴生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Therefore, understanding the distribution of gibberellin in plants is of great significance for understanding the physiological and biochemical indicators of plants. At present, gibberellin detection is mainly through ELISA, HPLC, GC-MS and other detection methods. In situ localization of gibberellins on tissues or cells

Method used

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  • Immunofluorescent method for detecting plant gibberellin
  • Immunofluorescent method for detecting plant gibberellin
  • Immunofluorescent method for detecting plant gibberellin

Examples

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Embodiment 1

[0033] An immunofluorescence method for detecting phytogibberellins, comprising the following steps:

[0034] S1. Dewaxing and rehydration: After the plant tissue is made into paraffin sections, put 1,3-xylene with a concentration of ≥99% in sequence for 10 minutes, 1,3-xylene with a concentration of ≥99% for 10 minutes, and a concentration of ≥99% for 10 minutes. 99% 1,3-xylene for 10 minutes, absolute ethanol I for 5 minutes, absolute ethanol II for 5 minutes, 95% ethanol for 5 minutes, 85% ethanol for 5 minutes, 75% ethanol for 5 minutes, and finally washed with distilled water to obtain dewaxed and rehydrated tissues slice;

[0035] S2. Antigen retrieval: the dewaxed and rehydrated tissue sections were placed in a mixture of 1.8 mM / L citric acid monohydrate and 0.2 mM / L trisodium citrate dihydrate at a pH of 6.0, and placed in Heat in a microwave oven on medium-high heat for 10 minutes, heat on medium-high heat for 5 minutes, take it out and cool it to room temperature na...

Embodiment 2

[0045] An immunofluorescence method for detecting phytogibberellins, comprising the following steps:

[0046] S1. Dewaxing and rehydration: After the plant tissue is made into paraffin sections, put 1,3-xylene with a concentration of ≥99% in sequence for 10 minutes, 1,3-xylene with a concentration of ≥99% for 10 minutes, and a concentration of ≥99% for 10 minutes. 99% 1,3-xylene for 10 minutes, absolute ethanol I for 5 minutes, absolute ethanol II for 5 minutes, 95% ethanol for 5 minutes, 85% ethanol for 5 minutes, 75% ethanol for 5 minutes, and finally washed with distilled water to obtain dewaxed and rehydrated tissues slice;

[0047] S2. Antigen retrieval: the dewaxed and rehydrated tissue sections were placed in a mixture of 1.8 mM / L citric acid monohydrate and 0.2 mM / L trisodium citrate dihydrate at a pH of 6.0, and placed in Heat in a microwave oven on medium-high heat for 8 minutes, then take it out and cool it to room temperature naturally, then place it in 0.05mM / L d...

Embodiment 3

[0057] An immunofluorescence method for detecting phytogibberellins, comprising the following steps:

[0058] S1. Dewaxing and rehydration: After the plant tissue is made into paraffin sections, put 1,3-xylene with a concentration of ≥99% in sequence for 10 minutes, 1,3-xylene with a concentration of ≥99% for 10 minutes, and a concentration of ≥99% for 10 minutes. 99% 1,3-xylene for 10 minutes, absolute ethanol I for 5 minutes, absolute ethanol II for 5 minutes, 95% ethanol for 5 minutes, 85% ethanol for 5 minutes, 75% ethanol for 5 minutes, and finally washed with distilled water to obtain dewaxed and rehydrated tissues slice;

[0059] S2. Antigen retrieval: the dewaxed and rehydrated tissue sections were placed in a mixture of 1.8 mM / L citric acid monohydrate and 0.2 mM / L trisodium citrate dihydrate at a pH of 6.0, and placed in Heat in a microwave oven on medium-high heat for 9 minutes, heat on medium-high heat for 4 minutes, take it out and cool it to room temperature nat...

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Abstract

The invention discloses an immunofluorescent method for detecting plant gibberellin, and belongs to the field of plant biotechnology. The immunofluorescent method for detecting plant gibberellin comprises the following steps: S1, dewaxing rehydration; S2, antigen retrieval; S3, autofluorescence quenching; S4, serum sealing; S5, addition of one antibody; S6, addition of two antibodies; S7, DAPI cell nucleus redyeing; S8, mounting; and S9, microscopic examination. The method is simple, rapid, safe and non-toxic, the detection cost is reduced effectively, histocytes are positioned by using DAPI redyeing cell nucleuses, and the position of antigen distribution can be observed and determined preferably; the distribution condition of gibberellin inside plant tissues can be observed through a fluorescence microscope after dyeing; the original states of plant tissues and cells are maintained to the maximum extent in the experimental process; and the biological activity and immunocompetence canbe maintained after marking.

Description

technical field [0001] The invention belongs to the technical field of plant biology and relates to an immunofluorescence method for detecting plant gibberellins. Background technique [0002] Gibberellin is a widely existing plant hormone, and its chemical structure belongs to diterpenoid acid, which is derived from a tetracyclic skeleton. Gibberellins have important physiological effects on plant growth, the most prominent physiological effects are promoting stem elongation and inducing bolting and flowering of long-day plants under short-day conditions. Genetically dwarf plants such as dwarf corn and pea are The plant type after gibberellin treatment was similar to that of non-dwarf plants; non-dwarf plants had only slight responses. [0003] Gibberellin plays a regulatory role in seed germination. The starch in the seeds of many cereal plants such as barley is rapidly hydrolyzed during germination; if the embryo is removed, the starch will not be hydrolyzed, and the emb...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531
CPCG01N33/53G01N33/531
Inventor 白文霞黄双徐明智严鹏
Owner 南京烁朴生物科技有限公司
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