An Efficient Method for Isolating and Determining Endogenous Cytokinins in Turfgrass
A cytokinin and turfgrass technology is applied in the field of determination of plant endogenous cytokinin, which can solve the problems of few detection types, difficult purification, low recovery rate, etc. The effect of high recovery rate
Inactive Publication Date: 2015-08-19
WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI
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[0009] The purpose of the present invention is to provide a method for efficiently separating and measuring turfgrass endogenous cytokinins for the detection of endogenous cytokinins in plants, such as complicated operation, long time consumption, difficult purification, few detection types and low recovery rate. method
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[0025] Below in conjunction with embodiment describe in detail:
[0026] 1. Test material: Bermudagrass material treated with salt stress (treatment concentration: 200mmol / L NaCl) for 35 days (C43 bermudagrass from the Germplasm Resource Garden of Wuhan Botanical Garden, Chinese Academy of Sciences).
[0027] 2. Detection steps
[0028] 1) Extraction of endogenous cytokinin
[0029] A. Extraction: Take 0.5g of fresh sample, grind it with liquid nitrogen, then add 5ml of extraction solution, and extract overnight at -20°C to obtain the extraction solution;
[0030] The extract is methanol: formic acid: water = 15:1:4, according to v / v, plus 40mg / ml 2,6-di-tert-butyl-4-methylphenol as an antioxidant;
[0031] B. Centrifugation: Centrifuge the extract at 15,000 rpm and 4°C for 10 minutes, collect the supernatant and store it in a new tube at -20°C, extract the residue with 5ml of extract at -20°C for 1 hour, and centrifuge. Collect the supernatant and mix it with the previous ...
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Abstract
The invention discloses a method for effectively separating and measuring endogenesis cytokinins of turfgrass, and relates to the measuring field of plant endogenesis cytokinins. The method disclosed by the invention comprises the following steps of (1) extraction of the endogenesis cytokinin: leaching by adding an extracting solution, centrifuging, filtering, decoloring, and carrying out vacuum concentration on the endogenesis cytokinins, and thus finishing the extraction of the endogenesis cytokinins; (2) solid-phase extraction elution: dissolving the endogenesis cytokinins to obtain a dissolution liquid, and carrying out vacuum concentration on the eluate so as to obtain a solid-phase extraction eluate; (3) high performance liquid chromatography-mass spectrometry. The method disclosed by the invention has the advantages that the pretreatment is simple, the dosage of a chemical reagent is less, the cost is effectively saved, and the impurity interference is small; the detection sensitivity is high, the selectivity is good, and the derivation is not needed; multiple endogenesis cytokinins can be directly detected in a crude extract solution of a plant sample, a separation and extraction method, a chromatographic separation condition and a mass spectrometric detection method are suitable for the endogenesis cytokinins of the turfgrass, and the recovery rate is high and can achieve 93-96%.
Description
technical field [0001] The invention relates to the field of determination of plant endogenous cytokinins, in particular to a method for efficiently separating and measuring lawn grass endogenous cytokinins. Background technique [0002] Based on its unique ecological, social, landscape, aesthetic and economic functions, the lawn industry has been paid more and more attention and has shown great development momentum and potential. As an important part of the urban ecosystem, lawns not only have strong ornamental and entertaining properties, but also play a huge role in preventing pollution and improving the ecological environment. In view of this, the demand for turfgrass in society continues to increase. The development of the lawn industry also ushered in a good opportunity. In today's frequent smog, how to improve our living environment is a long way to go, and the role of turfgrass in improving the environment is also very important. [0003] Plant endogenous cytokinin...
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IPC IPC(8): G01N30/88G01N30/06
Inventor 傅金民胡龙兴娄燕宏
Owner WUHAN BOTANICAL GARDEN CHINESE ACAD OF SCI