阅读说明：本技术 一种测定衣藻低温荧光光谱的生物稳定溶剂 (Biostable solvent for measuring low-temperature fluorescence spectrum of chlamydomonas ) 是由 于倩倩 于 2020-03-31 设计创作，主要内容包括：本发明涉及一种测定衣藻荧光光谱生物缓冲试剂。由以下重量份的原料制成：N-(羟甲基)甲基甘氨酸1-2份,三羟甲基氨基甲烷0.5-1.5份,丙三醇40-60份,氢氧化钠0.5-1份,水40-50份。本发明可以保持衣藻细胞形态,维持衣藻生理状态不变,避免衣藻培养液因低温冷冻固化膨胀造成的样品通透性差后果,荧光信号强。(The invention relates to a biological buffer reagent for measuring fluorescence spectrum of chlamydomonas. The feed is prepared from the following raw materials in parts by weight: 1-2 parts of N- (hydroxymethyl) methylglycine, 0.5-1.5 parts of tris (hydroxymethyl) aminomethane, 40-60 parts of glycerol, 0.5-1 part of sodium hydroxide and 40-50 parts of water. The invention can keep the cell shape of the chlamydomonas, keep the physiological state of the chlamydomonas unchanged, avoid the result of poor sample permeability caused by low-temperature freezing solidification expansion of the chlamydomonas culture solution and have strong fluorescence signals.)

1. A biological buffer reagent for measuring a low-temperature fluorescence spectrum of Chlamydomonas is characterized by being prepared from the following raw materials in parts by weight:

1-2 parts of N- (hydroxymethyl) methylglycine, 0.5-1.5 parts of tris (hydroxymethyl) aminomethane, 40-60 parts of glycerol, 0.5-1 part of sodium hydroxide and 40-50 parts of water.

2. Preferably, the Chlamydomonas measuring low-temperature fluorescence biological buffer reagent is prepared from the following raw materials in parts by weight:

1.4 parts of N- (hydroxymethyl) methylglycine, 1 part of tris (hydroxymethyl) aminomethane, 50 parts of glycerol, 0.6 part of sodium hydroxide and 45 parts of water.

3. A process for the preparation of a formulation according to claim 1 or 2, characterized in that it comprises the following steps:

(1) weighing appropriate amount of N- (hydroxymethyl) methylglycine and tris (hydroxymethyl) aminomethane, adding distilled water, stirring and dissolving

(2) And (2) adding glycerol into the mixed solution prepared in the step (1) and uniformly stirring.

(3) And (3) adding sodium hydroxide into the mixed liquor prepared in the step (2), and adjusting the pH value to 7.0.

4. Use of the formulation of claim 1 or 2 for the determination of chlamydomonas low temperature fluorescence spectroscopy.

5. The method for providing the chlamydomonas low-temperature fluorescence spectrum comprises the following steps:

the preparation is prepared according to the sample ratio of 1: 5-15.

6. Preferably, in the above method, the dosage sample ratio of the preparation is 1: 10.

Technical Field

The invention relates to a biostable solvent for measuring a low-temperature fluorescence spectrum of chlamydomonas.

Background

Chloroplasts are important sites for photosynthesis in green plants, where light energy is absorbed, transmitted, and finally converted to chemical energy, a process that requires photosystem II (PSII), cytochrome b6f, and,

Photosystem i (psi), ATP synthase, four supramolecular complexes act synergistically. The smooth proceeding of the photosynthesis linear electron transfer process requires the electron transfer rates of the two optical systems to be coordinated. However, due to differences in the absorption characteristics of PSI and PSII and natural variations in the light quality of sunlight reaching the photosynthetic mechanism at different times of the day, the absorption and excitation of light energy by the two photosystems tends to be unbalanced. To adapt to complex light environments, plants need to balance the distribution of excitation energy between two photosystems to avoid damage to photosystems due to over-excitation, and state transition is an important short-term regulatory mechanism for photosynthetic mechanisms to maintain the balanced distribution of light energy between two photosystems and the balanced excitation of chlorophyll molecules in the reaction center. The state conversion is one of the rapid regulation modes of plant light energy capture, can be completed within a few minutes to tens of minutes, and only relates to the change of the relative position of the light capturing antenna and not to the change of the abundance of the light capturing antenna.

77K low temperature fluorescence refers to chlorophyll fluorescence observed under the temperature condition of 196 ℃ below zero, and the low temperature condition avoids the influence of temperature-dependent enzyme reaction and electron transfer on fluorescence measurement, so that only the change of the original photochemical reaction can be reflected, and the method is generally used for the state conversion research between two optical states. Chlamydomonas (Chlamydomonas) is a unicellular flagellated eukaryotic aquatic organism belonging to Chlamydomonas of the order Volvocaridae and contains a large goblet chloroplast in the body which accounts for approximately 70% of the total cell volume. The cell structure is simple and is a model organism for researching a state conversion mechanism, but the cell stability is poor, water is easy to volatilize and shrink, and the cell form is changed, so that the real-time physiological and biochemical state of the chlamydomonas cannot be truly reflected.

The biological buffer plays an important role in determining a sample, can keep the form of the chlamydomonas cells stable, maintain the physiological and biochemical states of the chlamydomonas cells, avoid the situations of poor permeability, low fluorescence signal intensity and the like of the sample after solidification and expansion in a low-temperature environment, can better research how the chlamydomonas is dispatched in a short time, and arrange different light-capturing antennas to be combined at proper positions so as to achieve the balance of excitation energy between two optical systems.

Disclosure of Invention

In view of the prior art, the invention aims to provide a biological buffer reagent for measuring a low-temperature fluorescence spectrum of chlamydomonas. The invention can avoid the situation that the permeability of the chlamydomonas sample is deteriorated after solidification and expansion in low-temperature liquid nitrogen, maintain the cell morphology and keep the real-time physiological and biochemical state of the chlamydomonas.

The preparation method of the biological buffer reagent for measuring the low-temperature fluorescence of the chlamydomonas comprises the following steps:

1-2 parts of N- (hydroxymethyl) methylglycine, 0.5-1.5 parts of tris (hydroxymethyl) aminomethane, 40-60 parts of glycerol, 0.5-1 part of sodium hydroxide and 40-50 parts of water.

Preferably, the chlamydomonas low-temperature fluorescence biological buffering agent is prepared from the following raw materials in parts by weight:

1.4 parts of N- (hydroxymethyl) methylglycine, 1 part of tris (hydroxymethyl) aminomethane, 50 parts of glycerol, 0.6 part of sodium hydroxide and 45 parts of water.

In a second aspect of the present invention, there is provided a method for preparing the above formulation, comprising the steps of:

(1) weighing appropriate amount of N- (hydroxymethyl) methylglycine and tris (hydroxymethyl) aminomethane, adding distilled water, stirring and dissolving;

(2) adding glycerol into the mixed solution prepared in the step (1) and uniformly stirring;

(3) and (3) adding sodium hydroxide into the mixed liquor prepared in the step (2), and adjusting the pH value to 7.0.

In a third aspect of the invention, there is provided the use of the above formulation in a low temperature fluorescence spectroscopy assay for Chlamydomonas.

In a fourth aspect of the present invention, there is provided a method for chlamydomonas low-temperature fluorescence spectroscopy, comprising the steps of:

the preparation is prepared according to the sample ratio of 1: 5-15.

Preferably, in the above method, the dosage sample ratio of the preparation is 1: 10.

The invention has the beneficial effects that:

the invention can keep the cell shape of the chlamydomonas, maintain the physiological state of the chlamydomonas unchanged, avoid the result of poor sample permeability caused by solidification and expansion of the chlamydomonas culture solution and have strong fluorescence signals.