HPLC detection method of content of salicylic acid in sugarcane leaves

A technology of sugarcane leaves and detection methods, applied in the direction of measuring devices, instruments, scientific instruments, etc., to achieve the effects of stable sample retention time, high purity, and complete active ingredients

Inactive Publication Date: 2017-03-22
SUGARCANE RES INST GUANGXI ACADEMY OF AGRI SCI
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  • Application Information

AI Technical Summary

Problems solved by technology

However, no efficient and accurate detection method for salicylic acid content in leaves has been established so far. In view of this, a set of rapid and effective detection methods for salicylic acid content in sugarcane leaves has been established. The biochemical mechanism of acid endogenous hormone mediating the stress response of sugarcane tip rot pathogen is of great significance

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  • HPLC detection method of content of salicylic acid in sugarcane leaves
  • HPLC detection method of content of salicylic acid in sugarcane leaves
  • HPLC detection method of content of salicylic acid in sugarcane leaves

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Embodiment 1

[0027] The HPLC detection method of salicylic acid content in the sugarcane leaf of embodiment 1

[0028] 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, add 10 mL of 80% methanol aqueous solution pre-cooled to 4 ° C, and extract at 4 ° C for 16 hours, Centrifuge at 8000 r / min for 10min, take the supernatant, add 5mL of the residue to pre-cool to 4°C with a volume fraction of 80% methanol aqueous solution for 2h at 4°C, take out the supernatant after centrifugation, combine the supernatant, 40°C Evaporate under reduced pressure to no methanol, add 15mL of petroleum ether for extraction and decolorization 3 times, discard the upper organic phase, and evaporate the lower aqueous phase to dryness at 40°C, add 0.5mL of mobile phase used for HPLC detection to dissolve, and dilute to 2mL , filtered through a 0.45 μm microporous membrane into a sample bottle w...

Embodiment 2

[0043] Example 2 Study on the difference of susceptibility and resistance of sugarcane varieties infected with tip rot.

[0044] (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.

[0045] (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.

[0...

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Abstract

The invention discloses an HPLC detection method of the content of salicylic acid in sugarcane leaves. The method comprises the following steps: S1, adding the sugarcane leaves to be detected into liquid nitrogen, grinding the leaves, carrying out methanol extraction, centrifuging the obtained extract, taking obtained supernatant, carrying out methanol extraction on obtained residues, centrifuging the residue extract, taking obtained supernatant, mixing the supernatants, carrying out reduced pressure evaporation, carrying out extraction decolorizing, carrying out reduced pressure evaporation on obtained lower layer water phase, adding a mobile phase to dissolve the evaporated water phase, allowing the volume of the obtained solution to reach a constant value, and filtering the solution to obtain a sample solution for later use; and 2, detecting the content of salicylic acid in the sample solution through adopting HPLC. The method allows the content of salicylic acid in sugarcane leaves to be accurately detected, and provides a scientific and powerful detection technology for the top rot resistance difference research of sugarcane varieties and the stress response biochemical mechanism of salicylic acid mediated sugarcane top rot pathogens.

Description

technical field [0001] The invention relates to the technical field of sugarcane endogenous hormone detection, in particular to an HPLC detection method for salicylic acid 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. The phytohormones contained in sugarcane are involved in the physiological regulation of almost all processes of its growth and development, from cell growth, division and differentiation, to seed dormancy, fruit development, sex differentiation and aging processes. For sugarcane, it has two very important indicators that directly affect the economic value of sugarcane, namely, sugar content and yield. However, if sugarcane is affected by diseases and insect pests and cannot be detected in time, it will greatly affect the sugar content and yield of sugarcane. If it is possible to determine the di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88G01N30/89
CPCG01N30/06G01N30/88G01N30/89G01N2030/884
Inventor 王泽平李毅杰刘璐蒋洪涛李长宁张保青高轶静罗霆张革民韦金菊段维兴周珊杨翠芳林善海
Owner SUGARCANE RES INST GUANGXI ACADEMY OF AGRI SCI
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