HPLC determination method for gibberellin content in sugarcane leaves

A technology of sugarcane leaves and detection methods, which is applied in the directions of measuring devices, instruments, scientific instruments, etc., to achieve the effects of short extraction time, high purity, and stable sample retention time

Inactive Publication Date: 2017-02-22
SUGARCANE RES INST GUANGXI ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is still no efficient and accurate detection method for gibberellin content of sugarcane tip rot leaves. In view of the increasing trend of sugarcane tip rot in my country, a rapid and effective detection method for gibberellin content of sugarcane tip rot leaves has been established. The method is of great significance for the study of the biochemical mechanism that mediates the stress response of the sugarcane tip rot pathogen

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  • HPLC determination method for gibberellin content in sugarcane leaves
  • HPLC determination method for gibberellin content in sugarcane leaves
  • HPLC determination method for gibberellin content in sugarcane leaves

Examples

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Embodiment 1

[0025] Example 1 HPLC detection method of gibberellin content in sugarcane leaves.

[0026] S1. Select sugarcane leaves infected with the pathogen of sugarcane tip rot as the sample to be tested. Take 2 g of the sample to be tested and add liquid nitrogen to grind it. Add 10 mL of 80% methanol aqueous solution pre-cooled to 4 ° C. Extract for 16 hours; then centrifuge at 8000 r / min for 10 minutes at 4°C to obtain the supernatant, and extract the residue with 5 mL of 80% methanol aqueous solution pre-cooled to 4°C for 2 hours, and take out the supernatant after centrifuging for 10 minutes. The two supernatants were combined and evaporated under reduced pressure at 40°C until methanol-free (about 0.3 mL of aqueous solution remained), added 15 mL of petroleum ether for extraction and decolorization three times, discarded the upper organic phase, and evaporated the lower aqueous phase to dryness at 40°C. Add 0.5mL of the mobile phase used for HPLC detection to dissolve, dilute to ...

Embodiment 2

[0040] Example 2 Study on the difference of susceptibility and resistance of sugarcane varieties infected with tip rot.

[0041](1) Preparation of inoculum: Inoculate the pathogenic bacteria of sugarcane tip rot on PDA medium for 3 days to activate, pick the mycelia from the edge of PDA medium and inoculate them in sterilized potato glucose water medium for 3 days, centrifuge, Use sterilized absorbent cotton to filter and collect the obtained bacteria, and use sterile water and bacteria to prepare a concentration of 1 × 10 6 CFU / ml sugarcane tip rot pathogen spore suspension.

[0042] (2) Treatment of inoculation materials: sugarcane variety YT94-128 and sugarcane variety GT37 were selected as inoculation materials, and 30 barrels of each variety were planted under greenhouse conditions, with 4 buds per barrel, and the seeds were soaked with 0.3% carbendazim for 20 minutes before planting. Manage normally after emergence, and inoculate when 5 to 6 complete leaves grow.

[00...

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Abstract

The invention discloses an HPLC determination method for the gibberellin content in sugarcane leaves. The HPLC detection method comprises the following steps: (S1) adding liquid nitrogen into to-be-detected sugarcane leaves, grinding, carrying out methanol extraction and centrifugation, extracting supernate, extracting residues with methanol, extracting supernate after the centrifugation, combining the types of supernate, carrying out reduced pressure vaporization, carrying out petroleum ether extraction and decoloration, carrying out reduced pressure steaming on a low-layer water phase, adding a flow phase to dissolve, carrying out volume fixation, and filtering to obtain liquid as a sample solution for later use; and (S2) determining the gibberellin content in the sample solution by virtue of an HPLC. By virtue of the HPLC determination method, the gibberellin content in the sugarcane leaves can be determined, and the scientific basis is provided for the research of a biochemical mechanism of the gibberellin in the mediation of the stress response of pathogenic bacteria of the top rot of the sugarcanes.

Description

technical field [0001] The invention relates to the technical field of sugarcane tip rot detection, in particular to an HPLC detection method for gibberellin content in sugarcane leaves. Background technique [0002] sugar cane( Saccharum officinarum ) is the most important sugar crop in my country, and sucrose accounts for 92% of my country's total sugar production. Because tip rot gradually shows the characteristics of non-regional and irregular, the harm of the disease to the sugarcane industry is becoming more and more serious. Symptoms of various types of tip rot emerge in endlessly in sugarcane areas such as Brazil, India, Iran, Malaysia and my country, especially in high temperature and high humidity seasons. According to the 2012-2014 Guangxi sugarcane variety regional test and the 2013-2014 national sugarcane variety regional test survey, the incidence rate of the tested varieties reached 100%. This disease can lead to a significant reduction in sugarcane plant h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/062
Inventor 王泽平刘璐李毅杰段维兴梁强张革民罗霆韦金菊高轶静林波周珊杨翠芳林善海张保青
Owner SUGARCANE RES INST GUANGXI ACADEMY OF AGRI SCI
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