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A HPLC detection method of malic acid 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 rapid extraction, good impurity removal effect and high purity

Inactive Publication Date: 2019-06-07
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, no efficient and accurate detection method for malic acid content in sugarcane leaves has been established so far. In view of this, the establishment of a rapid and effective detection method for malic acid content in sugarcane leaves is helpful for timely judging the difference in susceptibility and resistance of sugarcane varieties infected with pathogens and for studying the content of malic acid. The biochemical mechanism of source hormone mediating the stress response of sugarcane tip rot pathogen is of great significance

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  • A HPLC detection method of malic acid content in sugarcane leaves
  • A HPLC detection method of malic acid content in sugarcane leaves
  • A HPLC detection method of malic acid content in sugarcane leaves

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Embodiment 1

[0026] The HPLC detection method of malic acid content in the sugarcane leaf of embodiment 1

[0027] 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, add 4 mL of methanol with a volume fraction of 80% pre-cooled to 4 ° C, add liquid nitrogen to grind into a slurry, and set the sample with ultra-clean water. Make up to 10ml, extract at 4°C for 1 hour, ultrasonically extract for 2 hours, centrifuge at 10,000r / min for 10min, take the supernatant, add 2mL of the residue to 80% methanol pre-cooled to 4°C, extract ultrasonically for 30min, and centrifuge at 10,000r / min for 10min , take out the supernatant after centrifugation, combine the supernatant, and dilute to 8mL with 80% methanol by volume fraction, take an appropriate amount of the dilute solution and filter it into a sample bottle with a liner with a syringe filter, as The sample solution is ready for use;

[0028] S2. Using HPLC t...

Embodiment 2

[0042] Example 2 Judgment of the difference in susceptibility and resistance of sugarcane varieties infected with tip rot

[0043] (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.

[0044] (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.

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Abstract

The invention discloses a HPLC detection method for the malic acid content of a sugarcane leaf. The method comprises the following steps: S1, adding a to-be-detected sugarcane leaf into liquid nitrogen for trituration, then adding an aqueous methanol solution with a concentration of 70 to 90%, carrying out grinding to obtain slurry, carrying out extraction at 2 to 5 DEG C, then carrying out ultrasonic extraction and centrifugation, taking a supernatant, adding the aqueous methanol solution with a concentration of 70 to 90% into residues for ultrasonic extraction, carrying out centrifugation, taking another supernatant, repeating extraction and centrifugation of the residues once to three times, combining all the supernatants, fixing the combined supernatants to a certain volume with methanol with a concentration of 70 to 90% and carrying out filtering; and S2, detecting the content of malic acid in a sample solution by using HPLC. The method can accurately detect the malic acid content of the sugarcane leaf and provides scientific and powerful detection technology for research on difference of sugarcane varieties in inductive reactance to top rot and biochemical mechanism of stress response to pathogens of malic acid-mediated top rot of sugarcane.

Description

technical field [0001] The invention relates to the technical field of sugarcane endogenous hormone detection, in particular to an HPLC detection method for malic 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 differ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/06G01N30/89
CPCG01N30/06G01N30/89
Inventor 王泽平梁强林善海周珊罗霆韦金菊刘璐张革民高轶静杨翠芳张保青林波蒋洪涛段维兴
Owner SUGARCANE RES INST GUANGXI ACADEMY OF AGRI SCI
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