CN105067620A - Hevea brasiliensis leaf pore observation method - Google Patents
Hevea brasiliensis leaf pore observation method Download PDFInfo
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- CN105067620A CN105067620A CN201510457298.3A CN201510457298A CN105067620A CN 105067620 A CN105067620 A CN 105067620A CN 201510457298 A CN201510457298 A CN 201510457298A CN 105067620 A CN105067620 A CN 105067620A
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Abstract
The invention discloses a hevea brasiliensis leaf pore observation method. A good observation sample can be obtained through steps of sampling, alkali treatment, and observation. The area of the processed leaf is identical with the area of the obtained cuticle, and thus the clearest part can be observed. At the same time, the method is not limited by the time or whether, and can be performed in clear day, rainy day, morning and afternoon. Furthermore, the method can be carried out continuously, thus the work efficiency is improved, and the operation is simple and easy to master. The method can be applied to the research and practices in the fields such as bioecology, phytophysiology, plant germplasm resource science, thremmatology, cultivation science, and the like.
Description
Technical field
The present invention relates to plant physioecology studying technological domain, the method for particularly Stoma of Leaves observation.
Background technology
Blade is the major organs that plant carries out photosynthesis and transpiration, CO required for leaf photosynthesis
2from air, people's blade interior is entered by pore.Meanwhile, the moisture in plant is lost to air by the transpiration of blade from pore, and therefore, the physiological function of the form of leaf epidermis pore, quantity and distribution and blade is closely related.In addition, also closely related with the growing environment of plant.
In the evaluation of germ plasm resource, pore, as one of important Agronomic trait, can be used as and observes record project.Pore is the vitals remained under natural selection and artificial selection, has larger stability, and thus it can be used for studying the sibship of plant, evolutionary process, the ecotype, chromosomal characteristics and geographic origin.The change belonging to general stomatal frequency and pore length between plant not of the same race together is relatively little, and the plant do not belonged to together then changes greatly.Stomatal frequency, gas accumulation belts number are relevant to ploidy.As can be seen here, the research of stomatic observation method is an important content in botany research.
In plant research, often carry out the observation of Stoma of Leaves.And the method for observing has multiple, one is impression method, is namely coated onto leaf back with collodion, makes it form thin film, is torn after film drying with tweezers, observes as under microscope.Another kind method is directly torn by the lower epidermis tissue at its back side with tweezers, can observe after certain process.In addition, also have pyridine solution method, blade is immersed in pyridine solution, then take out and observe.Above several method, when carrying out Hevea rubber Stoma of Leaves and observing, although collodion impression method can see pore, film easily broken and rainy day can not carry out, and the epidermis of rubber leaf often takes a long time to take off, the waste of manpower time.Pyridine solution method, although also can see pore, image is clear not, can not take again.
Summary of the invention
For problems of the prior art and defect, the invention provides a kind of under field test condition of living body, the technical method of not damaged, a large amount of research rubber tree stomata characteristics sampled can be realized.
To achieve these goals, present invention employs following technical scheme:
The method that Hevea rubber Stoma of Leaves is observed, comprises the following steps:
1) sample; Take off sampling blade, and put into water;
2) base extraction; Be 1:1 ~ 2 respectively weighing sodium hydroxide and natrium carbonicum calcinatums according to mass ratio, again they are incorporated in water the alkali lye making 5% ~ 7.5%, thing to be mixed dissolves complete, obtain mixed solution, after mixed solution being heated to boiling, sampling blade is put into, continue heating, treat that epidermis starts to float off, sampling blade is picked up and puts into clear water;
3) observe; Swing back and forth in water with the lower epidermis that tweezers are clamped through the sampling blade of base extraction or with tap water, obtain pellicle film, pellicle film being placed on to carry on glass after dyeing process and shakeouing, then examine under a microscope pore.
Further illustrate of the present invention, continue in the process of heating in described step (2), the leaf look of sampling blade from green to yellow, in leaf, heave, and after 10 ~ 15min, epidermis starts to float off by bubble.
Further illustrate of the present invention, described pellicle film is through following steps dyeing process:
1) dye 8 ~ 12min in aceto-camine;
2) in 70% alcohol, 0.5 ~ 2min is soaked;
3) in 90% alcohol, 0.5 ~ 2min is soaked;
4) in anhydrous alcohol, 0.5 ~ 2min is soaked;
5) at 1:1(V/V) alcohol and normal butyl alcohol mixed solution in leaching 1 ~ 2min;
6) in normal butyl alcohol, 0.5 ~ 2min is soaked.
Further illustrate of the present invention, before base extraction, sampling blade is cut into 1 × 2 centimetre of size.
Further illustrate of the present invention, described sampling blade is on the sunny side, grows the intermediate blade of normal fluffy leaf second from the bottom.
In the heating process of base extraction, firepower can not be too violent, otherwise blade rolls up and down to exaggerate and causes excoriation, is hard to tell epicuticle or lower epidermis, has influence on work efficiency.
Described pellicle film makes image more clear through dyeing process, is convenient to shooting.
To make permanent sheet in the present invention, then epidermal tissue should be cut into the fritter of about 0.5 × 0.5 centimetre, then use normal butyl alcohol natural gum mounting.Mounting is noted not making bubble enter, in order to avoid affect the quality of sheet.
Advantage of the present invention:
1. have much by the leaf block of the inventive method process, the epidermis of acquisition just has much, is convenient to select position the most clearly to observe.
2. adopt the inventive method not to be subject to the restriction of time and weather, no matter all can carry out fine day, rainy day and morning and afternoon, more convenient and flexible.
3. adopt the inventive method to work continuously, increase work efficiency, simple to operate, be easy to grasp.
4. the inventive method may be used for the Researching and practicing in the fields such as bioecology, plant physiology, plant germplasm resource, thremmatology, cultivation, is widely used flexibly.
Accompanying drawing explanation
Fig. 1 is the embodiment of the present invention 1 para ruber stomatic observation figure.
Fig. 2 is the embodiment of the present invention 2 para ruber (dyeing transparent process) stomatic observation figure.
Embodiment
Below in conjunction with drawings and Examples, the present invention is further detailed.
example 1
Choose the normal photosynthetic functional leaf of sunny slope (intermediate blade of the second fluffy leaf), after first taking off functional leaf with knife, insert immediately in water.Blade is cut into 1 × 2 centimetre of size, then the alkali lye that seethed with excitement is put into, they for being 1:1 respectively weighing sodium hydroxide and natrium carbonicum calcinatum according to mass ratio, then are incorporated in water the alkali lye making 5% by described alkali lye, continue heating 10min, after starting to float off Deng epidermis, blade is picked up and puts into clear water, clamp lower epidermis with tweezers and swing back and forth in water, obtain one piece of pellicle film, be placed on microslide and shakeout, 200 power microscopes are observed.Observations as shown in Figure 1.
example 2
Difference from Example 1 is, described before pellicle film is observed priority dye in aceto-camine 10min, in 70% alcohol, soak 1min, in 90% alcohol, soak 1min, in anhydrous alcohol, soak 1min, at 1:1(V/V) alcohol and normal butyl alcohol mixed solution in soak 1min and soak 1min in normal butyl alcohol.Observations as shown in Figure 2.
example 3
Difference from Example 1 is, they for being 1:1 respectively weighing sodium hydroxide and natrium carbonicum calcinatum according to mass ratio, then are incorporated in water the alkali lye making 6% by described alkali lye.
example 4
Difference from Example 2 is, described alkali lye is for according to mass ratio being 1:1.5 weighing sodium hydroxide and natrium carbonicum calcinatum respectively, again they are incorporated in water the alkali lye making 7%, continue heating 12min, after starting to float off Deng epidermis, blade is picked up and puts into clear water, clamp lower epidermis with tweezers to swing back and forth in water, obtain one piece of pellicle film, then successively dye pellicle film 8min in aceto-camine, 1.5min is soaked in 70% alcohol, 1.5min is soaked in 90% alcohol, 1.5min is soaked in anhydrous alcohol, at 1:1(V/V) alcohol and normal butyl alcohol mixed solution in soak 1.5min and soak 1.5min in normal butyl alcohol after, be placed on microslide and shakeout, 200 power microscopes are observed.
example 5
Difference from Example 2 is, they for being 1:2 respectively weighing sodium hydroxide and natrium carbonicum calcinatum according to mass ratio, then are incorporated in water the alkali lye making 7.5% by described alkali lye.Continue heating 14min, after starting to float off Deng epidermis, blade is picked up and puts into clear water, clamp lower epidermis with tweezers to swing back and forth in water, obtain one piece of pellicle film, then pellicle film is successively dyeed in aceto-camine 12min, in 70% alcohol, soak 2min, in 90% alcohol, soak 2min, in anhydrous alcohol, soak 2min, at 1:1(V/V) alcohol and normal butyl alcohol mixed solution in soak 2min and soak 2min in normal butyl alcohol after, be placed on microslide and shakeout, 200 power microscopes are observed.
example 6
Difference from Example 2 is, described pellicle film successively dye in aceto-camine before observing 12min, in 70% alcohol, soak 2min, in 90% alcohol, soak 2min, in anhydrous alcohol, soak 2min, at 1:1(V/V) alcohol and normal butyl alcohol mixed solution in soak 2min and soak 2min in normal butyl alcohol.
Claims (5)
1. a method for Hevea rubber Stoma of Leaves observation, is characterized in that: comprise the following steps:
(1) sample; Take off sampling blade, and put into water;
(2) base extraction; Be 1:1 ~ 2 respectively weighing sodium hydroxide and natrium carbonicum calcinatums according to mass ratio, again they are incorporated in water the alkali lye making 5% ~ 7.5%, thing to be mixed dissolves complete, obtain mixed solution, after mixed solution being heated to boiling, sampling blade is put into, continue heating, treat that epidermis starts to float off, sampling blade is picked up and puts into clear water;
(3) observe; Swing back and forth in water with the lower epidermis that tweezers are clamped through the sampling blade of base extraction or with tap water, obtain pellicle film, pellicle film being placed on to carry on glass after dyeing process and shakeouing, then examine under a microscope pore.
2. the method for Hevea rubber Stoma of Leaves observation according to claim 1, is characterized in that: continue in the process of heating in described step (2), the leaf look of sampling blade from green to yellow, in leaf, heave, and after 10 ~ 15min, epidermis starts to float off by bubble.
3. the method for Hevea rubber Stoma of Leaves observation according to claim 1, is characterized in that: described pellicle film is through following steps dyeing process:
(1) dye 8 ~ 12min in aceto-camine;
(2) in 70% alcohol, 0.5 ~ 2min is soaked;
(3) in 90% alcohol, 0.5 ~ 2min is soaked;
(4) in anhydrous alcohol, 0.5 ~ 2min is soaked;
(5) at 1:1(V/V) alcohol and normal butyl alcohol mixed solution in leaching 1 ~ 2min;
(6) in normal butyl alcohol, 0.5 ~ 2min is soaked.
4. the method for Hevea rubber Stoma of Leaves observation according to claim 1, is characterized in that: before base extraction, sampling blade is cut into 1 × 2 centimetre of size.
5. according to claim 1-4
arbitrarythe method that described Hevea rubber Stoma of Leaves is observed, is characterized in that: described sampling blade is on the sunny side, grows the intermediate blade of normal fluffy leaf second from the bottom.
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| CN201510457298.3A CN105067620A (en) | 2015-07-30 | 2015-07-30 | Hevea brasiliensis leaf pore observation method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106909906A (en) * | 2017-03-03 | 2017-06-30 | 河南科技学院 | A kind of wheat leaf blade stomatal frequency measuring method based on micro-image |
| CN106969939A (en) * | 2017-05-24 | 2017-07-21 | 刘小楠 | One kind makes onion slide and instrument and its application method is quickly taken with onion epidermis |
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| CN101556215A (en) * | 2009-05-13 | 2009-10-14 | 西北农林科技大学 | Sampling method for non-damage fixed position observation of plant stomata in field experiment |
| CN103076217A (en) * | 2012-12-28 | 2013-05-01 | 中国农业科学院作物科学研究所 | Preparation method for wheat leaf stomata guard cell observation samples |
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2015
- 2015-07-30 CN CN201510457298.3A patent/CN105067620A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101556215A (en) * | 2009-05-13 | 2009-10-14 | 西北农林科技大学 | Sampling method for non-damage fixed position observation of plant stomata in field experiment |
| CN103076217A (en) * | 2012-12-28 | 2013-05-01 | 中国农业科学院作物科学研究所 | Preparation method for wheat leaf stomata guard cell observation samples |
Non-Patent Citations (5)
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| 姜孝成，莫湘涛，彭贤锦: "《生物学实验教程》", 31 July 2007 * |
| 王建书: "《植物学技术 第二版》", 30 June 2013 * |
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| 黄镜浩 等: "观察柑橘叶片表皮细胞与气孔结构的简易制片方法", 《中国园艺学会热带南亚热带果树分会第二届学术研讨会暨全国龙眼产业发展论坛论文集》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106909906A (en) * | 2017-03-03 | 2017-06-30 | 河南科技学院 | A kind of wheat leaf blade stomatal frequency measuring method based on micro-image |
| CN106969939A (en) * | 2017-05-24 | 2017-07-21 | 刘小楠 | One kind makes onion slide and instrument and its application method is quickly taken with onion epidermis |
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Application publication date: 20151118 |