CN103076217A - Preparation method for wheat leaf stomata guard cell observation samples - Google Patents
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Abstract
The invention discloses a preparation method for wheat leaf stomata guard cell observation samples, and relates to the plant research technology. The method comprises the steps of preparation of a material to be treated, fixation of the material to be treated, dehydration and whole transparency. The method is characterized in that a transparent reagent adopted by the whole transparency is obtained by mixing chloral hydrate, distilled water and glycerol according to the weight ratio of 4 g: 1 ml: (0-0.5) ml (the ratio can be rationally widened to a range). For preparing the wheat leaf stomata guard cell observation samples, clear microscopic observation images are obtained.
Description
Technical field
The present invention relates to the plant research technology, particularly a kind of wheat leaf blade Stomacal guard cell is observed the preparation method of sample.
Background technology
The wheat leaf blade pore is comprised of the guard cell of two dumbbell shapeds, studies show that, the length of Stomacal guard cell can be used as an index (Wang Pei etc. that identify the wheat plant ploidy, 1989), and can tentatively judge genotypic drought tolerance (model light-year etc. according to boot leaf point Stomacal guard cell length difference under irrigated land and the nonirrigated farmland condition, 1990,1991).Because the observation of Stomacal guard cell has simply, quick, cost is low, plays an important role in the indirect evaluation of ploidy and genotype drought tolerance.
The general employing of preparation that the wheat leaf blade Stomacal guard cell is observed sample directly tears and follows the example of (Liu Pei and Gu Peiwen, 1993), scrape method for making (Wang Pei etc., 1989), sticking (Chen Baihong etc., 2004 of following the example of of cellophane method; Duan Yunfeng etc., 2008) and segregation process (envelope great waves and Hu Dong, 2008; Teacher sea etc., 2010).Because wheat leaf blade is elongated, directly tear and often follow the example of under the mesophyll band, can't accurately observe; The sticking problem that also has epidermis adhesion mesophyll of following the example of of adhesive tape; The chromic acid segregation process destroys the mesophyll in the blade, peels off epidermis again, easily epidermis is torn get incoherent in tearing the process of getting; Above several method all needs the step of getting the blade epidermis through tearing, and the problem that all exists epidermis to separate with the mesophyll difficulty is difficult for tearing and gets successfully, thereby affects the efficient that sample is observed.Striking dynamics and the angle of scraping method for making blade when striking are difficult for grasping, and mesophyll can occur and strike off halfway phenomenon, thereby affect the sharpness that sample is observed.These methods shortcomings such as all in various degree existence is not easy to operate in actual applications, efficient is low, the preparation that is not suitable for a large amount of samples is observed.
The whole clearing technology is a kind of technology commonly used in the botany research, often is used to observation of plant pollen granule, embryo and ovary (Zhao Shixu etc., 1993; Van baarlen etc., 2002; Yang Lu gives birth to and Li Guoping, and 2004; Chen Qingliang etc., 2005; Noyes etc., 2005), its principle is after plant tissue or organ are processed through clarifier, because the thickness of different tissues and refractive index is different, therefore present different states, need be through dyeing after the transparent processing, utilize the microscope such as bright-field can carry out Observation of Histological Structure.Clarifier commonly used has highly basic, wintergreen, glycerine, " 4
1/ 2 " composite transparent agent, chloral hydrate, dimethylbenzene, chloroform, cloves wet goods (Herr, 1971; Hao Jianhua and prosperity, 2007).Hao Jianhua and prosperous and powerful wait [2007] utilize glycerine to study the growth of tomato lateral-root primordia as clarifier, have studied feverfew Radix Conyzae sumatrensis during sexual reproduction take wintergreen as clarifier; Li Yankun etc. (2011) have studied embryo's growth in the reproductive process take outstanding bell leaf ramie female flower as test material, have compared the combination of different immobile liquids and transparent liquid to the impact of transparent effect; Zhao Zhijing etc. (2010) utilize highly basic potassium hydroxide to be clarifier, have observed tuber of pinellia leaf epidermis form, have obtained preferably effect.But it is bad to be used for wheat leaf blade sample effect with these at present known schemes.
Summary of the invention
The present invention is based on the blank in above-mentioned field, be provided for the whole clearing technology that wheat leaf blade is observed sample, technical scheme is as follows:
A kind of wheat leaf blade Stomacal guard cell is observed the preparation method of sample, comprise and prepare pending material, pending material is fixed, dehydration, three steps of whole clearing is characterized in that: the clarifier that described whole clearing adopts is: chloral hydrate, distilled water and glycerine can rationally be expanded into scope with this ratio of weight ratio 4g:1ml:0～0.5ml() ratio mix and get.
To be chloral hydrate, distilled water and glycerine with the ratio of 4g:1ml:0.5ml mix described clarifier gets.
Described solid adopts absolute ethyl alcohol and glacial acetic acid to press the mixed liquor of 6:1 volume ratio.
The transparent time is 6 hours.
Described dehydration refers to: the pending material after fixing with 70% ethanol soaking at room temperature 30 minutes, then use the absolute ethyl alcohol soaking at room temperature 2 times, each 30 minutes.
The present invention is applied to the whole clearing technology preparation that the wheat leaf blade Stomacal guard cell is observed sample first.And the clarifier prescription that can obtain better effects is provided, after saturated trichloroacetaldehyde, chloral hydrate and 2 kinds of clarifiers of glycerine mixed liquor are processed, all obtained clearly microexamination image; After adding glycerine sample is had the preservation effect, can prolong the holding time of leaf sample, observe for sample and provide convenience.Compared with the control, Zhao Zhijing etc. observe the 10% potassium hydroxide clarifier that tuber of pinellia leaf epidermis form adopts, effect is bad in the wheat leaf blade sample preparation, after utilizing 10% potassium hydroxide treatment wheat leaf blade, blade construction is damaged, very easily make blade broken during with the tweezers gripping, guard cell's obscure boundary is clear during microexamination, is unfavorable for the Measurement accuracy Guard Cell Length.
This sample preparation technology only need to blade be fixed, dehydration, transparent three steps can observe clearly leaf epidermis tissue and stomatal apparatus observation image under simple microscope, having removed epidermis from tears and gets or this step of mesophyll striking, easy to operation, draw materials few, cost is low, the compressing tablet sharpness is high, the pore authenticity is good, be applicable to the stomatic observation of the tender and mature leaf of wheat children, observe for Stomacal guard cell a large amount of, in batch in the Ploidy of Wheat Pollen Plants identification research, a kind of convenient, fast, efficient technical method is provided.
Description of drawings
Wheat leaf blade under Fig. 1 OlympusSZX16 Stereo microscope after the different disposal
A: fresh blade; B: the fixing rear blade of processing; C: processed rear blade; D:10% potassium hydroxide treatment rear blade; E: glycerite is processed rear blade; F: saturated trichloroacetaldehyde solution-treated rear blade; G: trichloroacetaldehyde and glycerine mixed liquor are processed rear blade;
The different clarifiers of Fig. 2 OlympusBX51 microscopically are processed the observations of blade
A:10% potassium hydroxide treatment rear blade, 10 times of object lens; B:10% potassium hydroxide treatment rear blade, 40 times of object lens; C: glycerite is processed rear blade, 10 times of object lens; D: saturated trichloroacetaldehyde solution-treated rear blade, 10 times of object lens; E: saturated trichloroacetaldehyde solution-treated rear blade, 40 times of object lens; F: trichloroacetaldehyde and glycerine mixed liquor are processed rear blade, 10 times of object lens; G: trichloroacetaldehyde and glycerine mixed liquor are processed rear blade, 40 times of object lens
Embodiment
Materials and methods
1. experiment material
Test material is the Wheat Pollen that utilizes the vitro anther culture technology to obtain, and doubles in the Anther Culture process to process, and the existing monoploid of pollen plant also has dliploid.All take a sample and listing mark to pollen plant before the plant jointing in pollen plant kind garden factory experimental plot in the Chinese Academy of Agricultural Sciences.
2 experimental techniques
2.1 solution preparation
Fixing agent:
Ethanol/acetic acid mixed liquor: 6 parts of absolute ethyl alcohols mix with 1 part of glacial acetic acid, are used for the fixing processing of sample.
Clarifier:
10% potassium hydroxide solution: 1 gram potassium hydroxide is dissolved in 10 ml distilled waters.
Glycerite: 1 milliliter of glycerine is dissolved in 2 ml distilled waters.
Saturated chloral hydrate solution: 4 gram chloral hydrates are dissolved in 1 ml distilled water.
Trichloroacetaldehyde and glycerine mixed liquor: 8 gram chloral hydrates are dissolved in 1 milliliter of glycerine and 2 ml distilled waters.
Agents useful for same is all available from Beijing chemical reagents corporation
2.2 blade disposal route
(1) draws materials and fixing: fall about 1.0 centimetres of two leaf blade tip parts with what scissors taked to list Wheat Pollen, put into the centrifuge tube room temperature that ethanol/acetic acid mixed liquor is housed and fix 24 hours.
(2) flushing and dehydration: remove the alcohol, acetic acid mixed liquor, use first 70% ethanol soaking at room temperature 30 minutes, the absolute ethyl alcohol soaking at room temperature is 2 times afterwards, each 30 minutes.
(3) transparent: as to remove ethanolic solution, with knife blade blade is cut into long and the wide fritter that is about 0.5 centimetre, use 4 kinds of transparent liquid instead and processed respectively sample 6 hours.
2.3 film-making is observed
The wheat leaf blade sample utilizes the OlympusSZX16 Stereo microscope to observe, and the background of taking pictures is black.The leaf sample that transparent processing is crossed places on the microslide, with knife blade blade is cut into 1 millimeter size, and covered after transparent liquid is dripped at the sample place is observed at the OlympusBX51 microscopically.
2.4 Guard Cell Length is measured and pollen plant fertility statistics
Utilize DP2-BSW software that the pollen plant blade is carried out the Stomacal guard cell measurement of length, each blade is measured 10 pores, with the mean value of 10 Stomacal guard cell length as this sample value.
Fertility is added up in maturity stage investigation, and take setting percentage 10% as the criteria for classifying, pollen plant respectively tillers setting percentage all less than 10% the sterile strain that is, pollen plant respectively tillers setting percentage all greater than 10% the fertile plant that is.
3. experimental result
3.1 the preparation of wheat leaf blade sample
Three steps have been passed through in the wheat leaf blade sample preparation of this experiment, at first utilize ethanol and acetic acid mixed liquor that the leaf sample of clip is fixed, so that leaf sample stops division and keeps condition of living organism, can be found out by Figure 1B, the visible blade of naked eyes faded fully after the Wheat Pollen vanes was crossed fixing the processing, blade yellowing and becoming fragile.
Utilize afterwards absolute ethyl alcohol and 70% ethanol to carry out respectively processed, the transparent processing that is beneficial to next step reaches better effects, can be found out by Fig. 1 C, still be yellow after leaf abscission is processed, but the blade deliquescing is easy to utilize knife blade cutting.
4 kinds of clarifiers of mixed liquor of 10% potassium hydroxide solution, glycerite, saturated chloral hydrate and chloral hydrate and glycerine are selected in this experiment, to having carried out transparent processing through the leaf sample after fixing and the processed, the result shows, wheat leaf blade is after 10%KOH solution transparent processing, it is light yellow that blade is, blade construction is damaged, and very easily makes blade broken (Fig. 1 D) during with the tweezers gripping; After 3 kinds of clarifiers of the mixed liquor of glycerite, saturated chloral hydrate and chloral hydrate and glycerine are processed wheat leaf blade, to blade construction all without destroying, it is complete that blade construction keeps, wherein glycerite processing rear blade is light yellow (Fig. 1 E), the full trichloroacetaldehyde that closes carries out transparent processing rear blade be white in color (Fig. 1 F), after the mixed liquor of chloral hydrate and glycerine was processed wheat leaf blade, blade became fully transparent (Fig. 1 G).
3.2 the microexamination of wheat leaf blade sample
Utilize the OlympusBX51 microscope that the Wheat Pollen leaf sample after processing through 4 kinds of clarifiers is observed, the result shows, after the 10% potassium hydroxide solution transparent processing, can be observed clearly wheat leaf epidermis image (Fig. 2 A) under 10 times of object lens, can be observed the stomatal apparatus structure under 40 times of object lens, but guard cell's obscure boundary is clear, is unfavorable for Measurement accuracy Guard Cell Length (Fig. 2 B).Glycerite transparent processing effect is bad, 10 times of object lens hypographs unintelligible (Fig. 2 C).After saturated trichloroacetaldehyde, chloral hydrate and 2 kinds of clarifiers of glycerine mixed liquor are processed, microexamination has all obtained better effects, when being 10 times, the micro objective enlargement factor can clearly observe the long cell of wheat leaf blade epidermis and the short cell between the long cell, vascular bundle, stomatal apparatus and epidermis fluff structures (Fig. 2 D, 2F), be beneficial to and carry out blade epidermis long cell, short cell, the observation that vascular bundle, stomatal apparatus and epidermis fluff structures distribute; When being 40 times, the micro objective enlargement factor can be observed clearly pore, guard cell, accessory cell and air hole structure (Fig. 2 E, 2G), for the Measurement accuracy of wheat Stomacal guard cell is laid a good foundation.Consider behind the adding glycerine sample is had the preservation effect, can prolong the holding time of leaf sample, observe for sample and to provide convenience, therefore in carrying out wheat leaf blade sample preparation research work suggestion with the mixed liquor of chloral hydrate and glycerine as clarifier.
3.3 this sample preparation methods is applied to the effect that Ploidy of Wheat Pollen Plants is identified
Utilize this sample preparation methods that the blade of 231 strain Wheat Pollens is processed, by utilizing the OlympusBX51 microscope observation of sample is found, the leaf sample of all Wheat Pollens has all obtained clearly stomatal apparatus image, illustrate and utilize this sample preparation methods to process wheat leaf blade, not only clearly sample image can be obtained, the stability of leaf sample treatment effect can be guaranteed simultaneously.
Utilize DP2-BSW software that the Guard Cell Length of Wheat Pollen leaf sample is measured, according to [1] results of study such as Wang Pei the Guard Cell Length measurement result is analyzed, Guard Cell Length is classified as dliploid greater than the Wheat Pollen of 65 μ m, and its maturity stage plant performance can be educated; Guard Cell Length is classified as monoploid less than the Wheat Pollen of 65 μ m, and its maturity stage plant performance is sterile.And according to the field fertility investigation result that obtains in the Wheat Pollen maturity stage, the Ploidy Identification result who utilizes Stomacal guard cell length to carry out is verified, the result shows that the Ploidy of Wheat Pollen Plants evaluation Average Accuracy that utilizes this sample preparation methods to carry out can reach 98.16%(table 1), proved that the wheat leaf blade sample preparation methods in this experiment is used for the Guard Cell Length measurement, and then carried out the feasibility of pollen plant Ploidy Identification.
This sample preparation methods of table 1 is applied to the accuracy rate that Ploidy of Wheat Pollen Plants is identified
List of references
Wang Pei, Chen Yurong, Fang Ren, Wu Lanpei, Zhu Zhiqing (1989). identify the research of pollen plant ploidy method with the guard cell. Beijing Agricultural University's journal, 15 (2): 141-145.
Model light-year (1991). the guard cell identifies the genotypic index of wheat drought tolerance. biology magazine, 2:13-15.
The model light-year, Wang Pei, Fang Ren (1990). genotype and irrigation are on the impact of Guard Cell Length In The Flag Leaf of Triticum Aestivum. North China agronomy newspaper, 5 (4): 24-26.
Liu Pei, Gu Peiwen (1993). utilize Guard Cell Length that the ploidy of Wheat Pollen is identified. Ningxia agricultural college journal, 14 (3): 49-55.
Chen Baihong, Li is newborn, Cao Ziyi, Yao Qingrong (2004). and a kind of with the sticking method of getting blade epidermis observation pore of adhesive tape. Plant Physiology Communications .40 (2): 215-218.
Duan Yunfeng, Wang Youning, Li Xia (2008). a kind of simple and easy method and application thereof that obtains blade epidermis observation pore. North China agronomy newspaper, 23 (supplementary issue): 73-76.
The envelope great waves, Hu Dong (2008). the research of monocotyledon Stoma of Leaves observational technique improvement. plant research, 28 (1): 82-84.
The teacher sea, Li Yuxin, Dong Baodi, tall even week, Liu Mengyu (2010). the sample preparation methods improvement of Leaf-Blade of Poaceae epidermis stomatic observation. Plant Physiology Communications, 46 (4): 395-398.
Zhao Shixu, among Du, Ling Zuming (1993). with the embryonic development of ovary whole clearing method and Differential interference contrast microscope Study On Rice. heredity, 15 (4): 33.
Chen Qingliang, Zhai Zhixi, Yang Chongjun (2005). the whole clearing Study on Technology of Dyeing of cistanche deserticola seed and embryo. Chinese herbal medicine .36 (10): 1559-1561.
Yang Lu gives birth to, Li Guoping (2004). the observational technique of angiosperm blastular structure. and laboratory study and exploration, 23 (2): 9-11.
Van?Baarlen?P,de?Jong?JH,VanDijk?PJ(2002).Comparative?cyto-embryologicalinvestigations?of?sexual?and?apomictic?dandelions?(Taraxacum)?and?their?apomictic?hybrids.Sex?Plant?Reprod?15,31-38.
Noyes?RD,Allison?JR?(2005).Cytology,ovule?development,and?pollen?qualityin?sexual?Erigeron?strigosus?(Asteraceae).Int?J?Plant?Sci?166,49-59.
Herr?JMJ(1971).A?new?clearing-squash?technique?for?the?study?of?ovule?development?in?angiosperms.Am?J?Bot?58,785-790.
Hao Jianhua, prosperous and powerful (2007). application example and the analysis thereof of whole clearing technology in phytobiology. BULLETIN OF BOTANY Vol., 24 (4): 490-497.
Li Yankun hides and consolidates, Zhao Lining, Li Yujun, the dragonfly Cheng Chao of Tang China (2011). and the whole clearing technology is observed the method research of outstanding bell leaf ramie embryonic development. China's fiber crops industry science, 33 (3): 142-146.
Zhao Zhijing, Zheng Daoguang, strict teacher's joint, Chen Jishuan (2010). observe the epidermal shape of three kinds of blade profile tuber of pinellia leaves with transillumination. science and technology circular, 26 (3): 413-416.
Claims (5)
1. a wheat leaf blade Stomacal guard cell is observed the preparation method of sample, comprise and prepare pending material, pending material is fixed, dehydration, three steps of whole clearing is characterized in that: the clarifier that described whole clearing adopts is: chloral hydrate, distilled water and glycerine can rationally be expanded into scope with this ratio of weight ratio 4g:1ml:0～0.5ml() ratio mix and get.
2. preparation method according to claim 1, described clarifier is that chloral hydrate, distilled water and glycerine mix with the ratio of 4g:1ml:0～0.5ml and gets.
3. preparation method according to claim 1, described solid adopts absolute ethyl alcohol and glacial acetic acid to press the mixed liquor of 6:1 volume ratio.
4. preparation method according to claim 1, the transparent time is 6 hours.
5. preparation method according to claim 1, described dehydration refers to: the pending material after fixing with 70% ethanol soaking at room temperature 30 minutes, then use the absolute ethyl alcohol soaking at room temperature 2 times, each 30 minutes.
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105067620A (en) * | 2015-07-30 | 2015-11-18 | 广西南亚热带农业科学研究所 | Hevea brasiliensis leaf pore observation method |
| CN106011240A (en) * | 2016-05-19 | 2016-10-12 | 合肥艾迪康临床检验所有限公司 | Fusion gene indoor quality control product, manufacturing method and recipe of fixing solution of fusion gene indoor quality control product |
| 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 |
| CN108427005A (en) * | 2017-12-01 | 2018-08-21 | 北京大学 | A method of whole detection and localization being carried out to hormone in plant organ or tissue |
| CN108624547A (en) * | 2018-05-22 | 2018-10-09 | 天津农学院 | A method of separation monocotyledon guard cell |
| CN109187144A (en) * | 2018-07-30 | 2019-01-11 | 贵州医科大学 | The method of fly blow whole stain-clearing technique |
| CN109443883A (en) * | 2018-12-28 | 2019-03-08 | 齐鲁师范学院 | It is a kind of handle plant root tip clarifier and its application |
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-
2012
- 2012-12-28 CN CN2012105899240A patent/CN103076217A/en active Pending
Non-Patent Citations (5)
| Title |
|---|
| CHEN-FANG WANG 等: "Histochemical studies on the accumulation of reactive oxygen species (O2 and H2O2) in the incompatible and compatible interaction of wheat—Puccinia striiformis f. sp. tritici", 《PHYSIOLOGICAL AND MOLECULAR PLANT PATHOLOGY》 * |
| ELEONORA COMINELLI 等: "Over-expression of the Arabidopsis AtMYB41 gene alters cell expansion and leaf surface permeability", 《THE PLANT JOURNAL》 * |
| GOROU HORIGUCHI 等: "Large-scale histological analysis of leaf mutants using two simple leaf observation methods: identification of novel genetic pathways governing the size and shape of leaves", 《THE PLANT JOURNAL》 * |
| PATRYCJA NIEWIADOMSKI 等: "The Arabidopsis Plastidic Glucose 6-Phosphate/Phosphate Translocator GPT1 Is Essential for Pollen Maturation and Embryo Sac Development", 《THE PLANT CELL》 * |
| 梁卫红 等: "水稻光敏色素A和B的突变对气孔发育和OsAQP基因表达的影响", 《生物技术通报》 * |
Cited By (9)
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| CN105067620A (en) * | 2015-07-30 | 2015-11-18 | 广西南亚热带农业科学研究所 | Hevea brasiliensis leaf pore observation method |
| CN106011240A (en) * | 2016-05-19 | 2016-10-12 | 合肥艾迪康临床检验所有限公司 | Fusion gene indoor quality control product, manufacturing method and recipe of fixing solution of fusion gene indoor quality control product |
| 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 |
| CN108427005A (en) * | 2017-12-01 | 2018-08-21 | 北京大学 | A method of whole detection and localization being carried out to hormone in plant organ or tissue |
| CN108624547A (en) * | 2018-05-22 | 2018-10-09 | 天津农学院 | A method of separation monocotyledon guard cell |
| CN108624547B (en) * | 2018-05-22 | 2021-08-13 | 天津农学院 | Method for separating monocotyledon guard cells |
| CN109187144A (en) * | 2018-07-30 | 2019-01-11 | 贵州医科大学 | The method of fly blow whole stain-clearing technique |
| CN109443883A (en) * | 2018-12-28 | 2019-03-08 | 齐鲁师范学院 | It is a kind of handle plant root tip clarifier and its application |
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Application publication date: 20130501 |