Method for detecting starch degrading microorganisms in tobacco leaf

A starch degradation and microbial technology, applied in the field of microbial detection, can solve the problems of inability to form a complete and unified tobacco leaf fermentation technology, lack of in-situ information of starch degrading bacteria groups, and inability of fermentation conditions to be suitable for action conditions, and achieve accurate experimental results and cost. Low, the effect of improving the quality of taste absorption

Active Publication Date: 2016-12-07
KUNMING UNIV
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AI Technical Summary

Problems solved by technology

However, the artificial amylase preparation obtained by pure culture has the following disadvantages: (1) No matter whether it is improving the technical conditions of tobacco leaf fermentation or adding external enzyme preparations, due to the different internal quality of tobacco leaves in various places, a complete and unified tobacco leaf fermentation technology cannot be formed; (2) Since the activity of the enzyme is affected by factors such as temperature, pH, and inhibitors, and the chemical composition of different types of tobacco leaves is not completely the same, the fermentation conditions cannot be suitable for the action conditions of each enzyme; Commercialized commercial enzymes, because the catalytic activity of enzymes is regulated and controlled by many aspects (such as temperature, pH, concentration of enzymes and substrates, and the presence of inhibitors, etc.), and the tobacco leaf fe

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  • Method for detecting starch degrading microorganisms in tobacco leaf
  • Method for detecting starch degrading microorganisms in tobacco leaf

Examples

Experimental program
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Example Embodiment

[0037] Example 1: A method for detecting starch-degrading microorganisms in tobacco leaves, the steps are as follows:

[0038] Sample collection: Wear sterile gloves and use a disposable sterile collection bag to randomly collect 1kg tobacco leaf samples during baking or aging, and store them at 37°C under constant temperature conditions and send them to the laboratory for sample processing within 20 minutes;

[0039] Sample processing: In the laboratory, first use 70% alcohol to sterilize scissors. Under aseptic conditions, cut the tobacco leaf sample into 1-4cm with sterilized scissors 2 Small pieces, and carefully remove useless tissues such as stems and veins of tobacco leaves;

[0040] Dilute the sample: under aseptic conditions, use a sterile 50ml beaker to weigh 30g of fresh broken leaf sample, add 300ml sample treatment reagent A, and mix with the broken leaf sample evenly;

[0041] Cell separation: Pour the mixed liquid into a Philips (PHILIPS) household mixer for 3 minutes ea...

Example Embodiment

[0051] Embodiment 2: A method for detecting starch-degrading microorganisms in tobacco leaves, the steps are as follows:

[0052] Sample collection: Wear sterile gloves and use a disposable sterile collection bag to randomly collect 1 kg of tobacco leaf samples during baking or aging, and store them at a constant temperature of 30°C and send them to the laboratory for sample processing within 20 minutes;

[0053] Sample processing: In the laboratory, first use 70% alcohol to sterilize scissors. Under aseptic conditions, cut the tobacco leaf sample into 1-4cm with sterilized scissors 2 Small pieces, and carefully remove useless tissues such as stems and veins of tobacco leaves;

[0054] Dilute the sample: under aseptic conditions, use a sterile 50ml beaker to weigh 30g of fresh broken leaf sample, add 600ml sample treatment reagent A, and mix with the broken leaf sample evenly;

[0055] Cell separation: Pour the mixed liquid into a PHILIPS household mixer for 4 minutes each time, and th...

Example Embodiment

[0065] Embodiment 3: A method for detecting starch-degrading microorganisms in tobacco leaves, the steps are as follows:

[0066] Sample collection: Wear sterile gloves and use a disposable sterile collection bag to randomly collect 1 kg of tobacco leaf samples during baking or aging, and store them at a constant temperature of 25°C and send them to the laboratory for sample processing within 20 minutes;

[0067] Sample processing: In the laboratory, first use 70% alcohol to sterilize scissors. Under aseptic conditions, cut the tobacco leaf sample into 1-4cm with sterilized scissors 2 Small pieces, and carefully remove useless tissues such as stems and veins of tobacco leaves;

[0068] Dilute the sample: under aseptic conditions, use a sterile 50ml beaker to weigh out 50g of fresh broken leaf sample, add 1000ml sample treatment reagent A, and mix with the broken leaf sample evenly;

[0069] Cell separation: Pour the mixed liquid into a PHILIPS household mixer for 5 minutes each time, a...

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Abstract

The invention relates to a method for detecting starch degrading microorganisms in a tobacco leaf. The method for detecting the starch degrading microorganisms in the tobacco leaf comprises the specific steps of sample collection, sample treatment, sample dilution, cell separation, ultrasonic treatment, calibration and tracing of the starch degrading microorganisms, microscopic observation and the like. The method can be applied to tobacco leaves of different varieties, different positions and different maturity levels for flue-cured tobacco; the information of strain degrading flora in the tea leaf in an in-situ state is obtained; the labor intensity is decreased more greatly; the cost is lower; the operation process is simple and convenient; an operator does not need to be specially trained; a special requirement on a sampling position does not exist; the experimental result is accurate and errorless; relevant data can be quickly provided for a technician. Meanwhile, the method is also helpful to the technician to ascertain the specific fermentation and aroma enhancement mechanisms of the tobacco leaf, so as to better guide practices, explore and perfect the treatment technical measure and condition of the tobacco leaf, broaden and innovate the search thoughts of the fermentation technique of the tobacco leaf and positively carry out the search of a new field relevant to the method for detecting the starch degrading microorganisms in the tobacco leaf.

Description

technical field [0001] The invention relates to a method for detecting microorganisms, in particular to a method for detecting starch-degrading microorganisms in tobacco leaves. Background technique [0002] Flue-cured tobacco is one of the important pillar industries in our country and occupies an important position in the development of tobacco in our country. Due to the reasons during the cultivation, baking and aging process of tobacco leaves, there are generally problems such as high starch residues in domestic flue-cured tobacco. Starch is the main product of photosynthesis in tobacco leaves. Starch is a storage form of tobacco leaf matter and energy. In chloroplasts, in addition to meeting the matter and energy required by the cell's own metabolic activities, the excess part of photosynthetic products is basically converted into starch. When stored, starch can be degraded and converted into other forms of sugars to supply various needs when the life activities of the...

Claims

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Application Information

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IPC IPC(8): C12Q1/40C12Q1/04
Inventor 孔云虹夏云韩雁冰黄鹤平王定康夏体渊
Owner KUNMING UNIV
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