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Observation and density measurement method for glandular hairs on Artemisia annua leaf surfaces

A technology for measuring the density of Artemisia annua leaves, applied in the field of life sciences, can solve the problems of difficult observation with glandular hairs, damage to the structure of glandular hairs, and distortion of the number and density of glandular hairs, so as to reduce the cost and time of measurement and achieve good repeatability , reagent and non-toxic effect

Inactive Publication Date: 2011-02-16
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method of observing the leaf surface can only be used for stomatal tissues that are not easily deformed. If it is applied to observe the microscopic structure of glandular hairs, it will not only destroy the structure of glandular hairs, but also lead to distortion of the number and density of the overall glandular hairs, which is difficult to use. Observation of glandular hairs on the leaf surface of Artemisia annua

Method used

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  • Observation and density measurement method for glandular hairs on Artemisia annua leaf surfaces
  • Observation and density measurement method for glandular hairs on Artemisia annua leaf surfaces
  • Observation and density measurement method for glandular hairs on Artemisia annua leaf surfaces

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Randomly select 96 plants of Artemisia annua that grow healthy and have a growth age of 3 months, and take the first leaf that has just fully expanded from the top of the main stem to be complete and vigorously growing (mainly referring to the leaves that have just fully expanded and have no pests and diseases, etc.) Artemisia annua leaves were washed once with distilled water if necessary, and 1 square centimeter each was placed in the small wells of a 96-plate in turn, and 198 μL of petroleum ether, 1.7 μL of absolute ethanol and 0.3 μL of glacial acetic acid reagent were added to each Well, incubate at room temperature for 7 min.

[0036] Take out the leaves on the glass slide, add a drop of double distilled water, gently cover the cover glass, and place it on the stage of the Motic B1 optical microscope. The part to be observed should be accurately moved to the bottom of the objective lens, and then press it with a press clamp. tight. The cover glass must be wiped cl...

Embodiment 2

[0041] Randomly select 10 Artemisia annua plants with healthy growth and a growth age of 6 months, and take the third fully expanded Artemisia annua leaf with complete leaves and vigorous growth from the top of the main stem, wash it once with distilled water if necessary, and take each 1 square centimeter was placed in a 1.5ml centrifuge tube in turn, and 980 μL of petroleum ether, 15 μL of absolute ethanol, and 5 μL of glacial acetic acid reagent were sequentially added to each centrifuge tube, and incubated at room temperature for 15 minutes.

[0042] According to the method of Example 1, take out the leaves on the glass slide, drop a drop of double distilled water, cover the cover glass, place it on the stage of the Motic B1 optical microscope, and use a 4× or 10× objective lens under the 10× eyepiece Observe, image, count the number of glandular trichomes, and finally calculate the density of glandular trichomes. The glandular hair density measured in this embodiment is 3...

Embodiment 3

[0045] Randomly select 10 Artemisia annua plants with healthy growth and a growth age of 9 months, and take the fifth Artemisia annua leaf with complete leaves and vigorous growth from the top of the main stem, wash it once with distilled water if necessary, and take each 1 square centimeter was placed in a 2ml centrifuge tube in turn, and 1900 μL of petroleum ether, 90 μL of absolute ethanol, and 10 μL of glacial acetic acid reagent were sequentially added to each centrifuge tube, and incubated at room temperature for 20 minutes.

[0046] According to the method of Example 1, take out the leaves on the glass slide, drop a drop of double distilled water, cover the cover glass, place it on the stage of the Motic B1 optical microscope, and use a 4× or 10× objective lens under the 10× eyepiece Observe, image, count the number of glandular trichomes, and finally calculate the density of glandular trichomes. The glandular trichome density measured in this embodiment is 45.9 per squ...

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Abstract

The invention discloses an observation and density measurement method for glandular hairs on Artemisia annua leaf surfaces, comprising the following steps: taking a eugonic Artemisia annua leaf, and feeding the leaf in a centrifuge tube or a microtiter plate; successively adding petroleum ether or normal hexane, absolute ethyl alcohol or carbinol and glacial acetic acid, evenly mixing, and incubating for 5-20min at room temperature; and taking out the leaf, placing the left on a glass slide, adding a drop of double distilled water, covering a cover slip slightly, compacting by a tablet clip, and observing on an optical microscope or calculating the number of the glandular hairs so as to calculate the density of the glandular hairs. The method is simple and convenient, has low requirement on equipment, greatly simplifies the preprocessing procedure of experimental materials, can complete the whole operation within half a hour, uses a little of reagent, is non-toxic, safe and environment-friendly, lowers the measurement cost and time substantially, and can be used for high pass analysis.

Description

Technical field: [0001] The invention relates to the technical field of life sciences, in particular to a method for optical microscope observation and density measurement of glandular hairs on the surface of Artemisia annua leaves. Background technique: [0002] Artemisia annua L., also known as Artemisia annua L., is a plant of the genus Artemisia in the family Asteraceae. There are two kinds of trichomes on the leaf surface of Artemisia annua, which are non-secretory trichomes and glandular trichomes. Glandular trichomes are very important for the biosynthesis of artemisinin and are considered to be the place of biosynthesis and storage of artemisinin. Artemisinin is a sesquiterpene lactone peroxide isolated from Artemisia annua for the first time by Chinese scholars. It is a special drug for treating malaria and has a great demand in the international market. Further pharmacological studies have proved that artemisinin and its derivatives have the effect of killing a va...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G02B21/00G01N21/84
CPCG01N2021/8466
Inventor 刘硕谦刘仲华田娜黄建安
Owner HUNAN AGRICULTURAL UNIV
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