植物病害标本的采集制作与保存Collectionandpreservationofplantdiseasespecimens

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1、植物病害标本的采集制作与保存（Collection and preservation of plant disease specimens）Collection and preservation of plant disease specimensPlant disease is an indispensable material for the undergraduate teaching of plant pathology in higher agricultural colleges. It has the characteristics of intuitionistic, unse

2、asonal and regional restriction in teaching. Therefore, the collection, production and preservation of plant disease specimens are essential knowledge and skills in the experimental teaching of plant pathology.Collection of plant disease specimensBecause of the clear season in the north, the crop va

3、riety is abundant, the plant disease is frequent, most of the plant disease samples can be collected from the fields. However, the occurrence of disease has a certain rule, it is necessary to master the seasonal law of the disease and the growth rule of the host to obtain the typical specimens.1.1 t

4、ools for collection of disease specimensPlant disease occurs mostly in the root, stem, leaf and fruit, so the tools used for different parts, the commonly used tools have knives, scissors, hoe, saw, etc., and specimen holder, small specimens of specimen box, paper, glass bottle, paper bags, tags, an

5、d pads, etc.1.2 specimen collectionThe field collection is mainly divided into season and host collection. Seasonal collection is mainly aimed at the diseases that occur at certain times of the year. The wheat powdery mildew can be collected from may to June every year, such as powdery mildew. The p

6、lants grey mould is prone to disease in cold and rainy conditions and is easiest to pick up from march to may. According to host collection, mainly for the wide range of diseases, such as downy mildew, can be found on the leaves of many plant diseases, common lettuce, grape, cabbage, etc. Diseases s

7、uch as tomato, potato, pepper and so on.1.3 record of specimen collectionIn order to distinguish different specimens collected, records should be recorded. It mainly includes the host name, time and place, the name of the collection person, the main occurrence situation and the necessary ecological

8、environment factor.The preparation of plant disease samplesAfter the specimen has been collected, it is usually preserved by drying or impregnation, and the original character of the specimen can be preserved as far as possible. Minor can make glass specimens, more difficult to save and special of s

9、pecimens by multimedia scanning system or microscopic system in the computer, made the corresponding style graphics, animation, video, images, etc.2.1 specimen drying method2.1.1 suppression. The collected plant disease specimens, if dry and easy to retract (such as rice leaves), preferably with the

10、 pressure. The other can be put in the collection box to take back pressure on the water absorbent paper, with the specimen clamping, sun drying. In the meantime, change the specimen paper frequently. Summer temperature and humidity are high, easy to make the specimen mouldy and discolored, specimen

11、 paper to be replaced frequently. Usually the first 3 4 d is 1 2 / d, and 2 3d-3d-3d-1, until the specimen is completely dry.2.1.2 hot dry or silica gel drying method. These two methods can be used for specimens that are not prepared for separation. Specimens in order to speed up the drying speed, b

12、etter keep specimens from the original color, just the samples can be rapidly back and forth on absorbent paper or cloth with electric iron very hot dry or paper and loads it dry specimen holder of silicon powder were bound tightly,In 50 oven for 2 3 d, make the dry quickly.2.1.3 microwave oven dryi

13、ng and preserving color method. In order to make up for the deficiency of hot dry method, liu chunyuan et al. (2003) reported on the quality of the samples of plant diseases, and fumbled a set of microwave drying and colorizing method. This method is dry and fast, the color preserving effect is bett

14、er, but it can be kept for a long time and will not fade. If the leaf of leguminous plant disease 1 min, the effect is optimal, the symptom is preserved intact.2.1.4 freeze-drying method. Its a good way to do it. The specimen is dried in freezing conditions and almost completely retains its original

15、 color and shape.2.1.5 natural drying method. Water few specimens such as the branch of stem disease, do not need to take any dry means, in the air naturally dry can.The method of preparation of the dipping specimenThe impregnation can be preserved by impregnation of the fruit disease and the diseas

16、e of the juice and the granule of the fleshy and basidiomycetes. However, this method occupies a large area and has limited storage time. It can only be used to save a small number of specimens, which are mostly used for teaching and demonstration. The preparation of the impregnated specimen require

17、s various types of impregnation fluid, mainly including various preservative and preservative preservatives that keep the original color.The preparation of the liquid of the percolate. According to the color of the host plant of the plant fruit, the impregnating liquid can be divided into the preser

18、vation of green impregnation liquid, the preservation of yellow and orange color, and the preservation of the red liquid. Preserve the green immersion fluid. It is necessary to select the appropriate method according to different plant disease samples to achieve good results. Such as: copper acetate

19、 pickling liquid will copper acetate crystal gradually add in 50% acetic acid solution, until no longer dissolve, concentrate according to the different color samples will be diluted 3 4 times, and then use. Specimens will be added to the boiling of diluent, continue to heat, specimens of green bega

20、n to fade, after 3 4 min, make green after recovery, to remove and rinse with clear water, preserved in 5% formalin solution, or pressed into dry specimens. This method is good for color protection, but slightly different from the original color. Uncooked samples of green apples, pears, green grapes

21、 can be soaked directly in 1% subsulfuric acid 100Ml, 95% alcohol 100 ml, water 800 ml of the solution soaked 10 20d, then remove, rinse clean, then preserved in 5 % of formaldehyde solution. In addition, there are also sulfuric sulfuric acid impregnation, Vacha impregnating fluid, etc., which are a

22、lso commonly used in preserving green specimens. Preserve the red impregnation liquid. For red fruit diseases such as strawberries, chili, potato and other plant red group, choose disease symptom of typical specimen, can look up red impregnation liquid immersed specimens tile (15 g, cobalt nitrate a

23、nd formalin 25 ml, tin chloride 10 g, water 000 ml) 1, 2 weeks after rinse clean out, soak in sulfite 30 50 ml, formalin 10 ml, alcohol (95%), 10 ml, 000 ml water with a solution to save them. The impregnation of yellow and orange is preserved. The fruits such as apricot, pear, persimmon and citrus

24、fruits, such as apricot, pear, citrus, etc., can be preserved with sulfuric acid solution.The method is to match the sulfuric acid (including SO2, 5% 6%) with water solution of 4% 10% (including SO2 0.2% 0.5%). In conclusion, the impregnating liquid of various kinds of color, except some of the impr

25、egnated liquid that preserves green, the impregnation of other colors, the effect is not ideal. In order to better display the hazards of plant diseases, it is better to use the multimedia system to scan or photograph them, and make corresponding pictures and video to save them.2.2.2 anti-corrosive

26、impregnation liquid. Corrosion is the most important part of the preserved specimen. Therefore, it is necessary to preserve the impregnated liquid. The main ingredients are as follows: formalin 50 ml, alcohol (95%) 300 ml, water 2 000ml, or formalin and alcohol.The preservation of plant disease spec

27、imens3.1 preservation of the specimen of the wax leaf3.1.1 bulk specimens. Put dry good exsiccate in sterilizing room or disinfection cabinet, the traditional way is to use of alcohol, mercuric chloride liquid disinfection, or dichlorvos or carbon tetrachloride and carbon disulfide in the mixture in

28、 a glass dish, using gas fume kill the insect or worm eggs on the specimen, take out after about 3 d. Conditional - 40 low temperature disinfection. To identify and organize disinfected wax specimens, put them in the bag of specimens and put them on the label and use them as a physical specimen for

29、students to use in class.3.1.2 make boxed specimens. The typical specimen of the plant disease and the specimen of dried fruit can be preserved by using the method of glass box. According to the size of the specimen, choosing the appropriate box (generally for 28 cm long and 20 cm wide, 1.5 3.0 cm h

30、igh), 1 layer of cotton in the carton, samples and labels are put on cotton, indicate the name of the host plant, fungus, acquisition time and so on, then add the glass cover. A small amount of camphor or insect repellent may be added to the cotton. This method can be preserved for long time and rec

31、ycled.3.1.3 plastic seal specimen. Suitable for preserved leaf specimens and smaller stalks (e.g. wheat, rice) specimens. Way is: according to the size of the specimens of plant disease choose proper protection film, will be 1 smaller than protecting card membrane blank sheet of paper into the card

32、in the film, write the label for the specimens on white paper at the bottom right, and then put the samples in white paper, put the film flat again, into the preheated to 160 in the molding machine seal model. If the specimen is smaller and narrower, a few specimens can be placed on one piece of pap

33、er, which can be classified and bound as required. Specimens of this kind of method to make more than makes up for the box method has many disadvantages, such as specimen box of large volume, carry inconvenience, high cost, only with the naked eye and specimens of local armed with a magnifying glass

34、 observation, difficult for small organization and structure observation, specimen is easy to change color, mildewy, bug eat by moth, not resistant to storage, etc.3.2 preservation of impregnated specimensAfter proper handling of the identified specimens, place them in a specimen bottle containing t

35、he preserved fluid. After sealing with wax or colloidal glue, the label will be placed on the specimen bottle for preservation. Because of the pickling liquid medicines used by mostly volatile, or easy to oxidation, so impregnated specimens in the dark, best to reduce the oxidation of liquid, or bot

36、tle rupture caused by temperature change.4 considerationsPlant disease specimen collection by the plant growth season, the climate conditions, the sampling location, time and the influence of many factors, such as crop varieties layout should grasp the opportunity to adopt to the need of symptoms ty

37、pical of the specimens. In addition, different parts are made and preserved in different ways. Exsiccate in the preservation process, keep dry specimen can prevent mildew, but vulnerable to dry specimen bug eat by moth, so its best to wait for drug fumigation with methyl bromide each year, to ensure

38、 that the samples in good condition. The preserved fluid should also be replaced regularly to ensure that the specimen does not decompose. In this way, gradually accumulating and supplementing various diseases and diseases can make the experiment course go smoothly and achieve the desired good resul

39、ts.Method of making insect specimenThe dry method of insect specimenUse a needle to dry. The insect needle is pierced by the insects chest and inserted into the soft wood. The pins should be vertical, and the height of the inserted person should be fully fixed with the stinger.2. Sticky adhesive pap

40、er. Smaller insects, such as fleas and mosquitoes, stick them on sticky paper, cut off pieces of paper, and stick them on paper to dry.3. Spread the wings to dry. The winged insects, often two wings after acupuncture, should use the spread fins to spread their wings and dry them.Remove internal orga

41、ns. For larger insects, the viscera is highly susceptible to corruption, and it should remove the internal organs. In the abdomen or dorsal side openings take out the internal organs, then fill with the non-fat cotton, and put a small amount of phenol, and then use the insect needle to burn in the c

42、hest.5. Wax filling. For larger moth larvae, after a gut (viscera from the anus with a syringe stabbing people all the internal gettering), the dissolution of paraffin wax from the anus into the cavity with a syringe, rendering the original full status. Put in cold water to solidify.Go dirty. After

43、the guts of the insect are pulled out, the air is slowly pumped through the air with a two-ball ball, plus an infrared lamp to dry.7. Burial laws. After the insect has been fixed and dehydrated, it is slowly watered with melted alcohol and the insects are buried in the rosin. The other method is to

44、dehydrate the insect and use the gelatine liquid to cover it. The gelatin solution is mixed with water and carbonic acid, and then the water bath method is melted and then planted. In addition, you can also use the universal glue to hide, the effect is better. Put the dehydrated material on the whit

45、e paper and bury the whole body with glue, then drop it again, and then cover the dust. This kind of method effect is better, the bag is buried extremely thin layer of glue, do not affect observation, increase materials solid property, can moistureproof the mildew.2. Insect specimen impregnation met

46、hodFormalin immersion method. Soak it in 5 % formalin. After 3-5d, change liquid into 5 % formaldehyde forest preservation.Alcohol immersion method. Use 50% alcohol to fix 12-24h and transfer to 70% alcohol.3. Alcohol glycerol, formaldehyde glycerin leaching method. It is better to add 20% to 40% gl

47、ycerine respectively. It can increase the softness of insect limb joint and prevent the weakness of alcohol dehydration.The specimen collection and preservation of rodentsLiu shu, institute of economics of Shanxi Province, Shanxi ProvinceIn rodent fauna, ecological investigation, and was used in the

48、 prevention and control work, must want to do one thing, is to collect the specimen material, good preservation and the production of various specimens of rats and mice, as evidence of the research of biological sciences, specimen feature is one of the most important recording materials, collecting

49、specimens, rats mousetrap is the most commonly used tools, kinds of models a lot, according to the activity of rat Habitat and mice size selection, respectively. In order not to damage the captive animals, also can choose various traps to catch alive. In addition, there are also shotguns and small c

50、aliber rifles to hunt animals. In the case of the drug kill, the treatment can be collected properly. Especially in the area of animal epidemics or areas where there is no doubt that there is no safe and reliable sterilization measures and methods, do not randomly collect. It must be pointed out tha

51、t in carrying out the work, anti-epidemic clothing, shoes, socks and so on should be worn to avoid the infection of animal epidemics by flea and other bites.(1) the items needed for making the specimen1. Disinfection products: chloroform, laisu, alcohol, soap, detergent, etc.2, preservatives: arseni

52、c vitriol powder (formula: the arsenic acid evenly developed into a fine powder, alum) or arsenic soap cream (formula of arsenic acid, camphor, two 5, one 5 two 5 mix minced soap water boil cream) camphor or naphthalene, etc.3, tools and goods: smoked barrels, scalpel, small scissors, pliers, tweeze

53、rs, balance, or small said, steel tape, lead wire according to the specimen (thickness), board, towel, cotton, needles, thread, tag, pen, pencil, ink, directives, pins, etc. When making an ecological specimen, prepare the false eye, etc.(2) handling before strippingFor the specimen obtained, be sure

54、 to place the bag in the rat bag and tighten the bag. At the same time, use the label to indicate the location, time and principle of a rat bag. Will be back to the rat body, first put inside fume barrels, pour into a certain amount of ethyl ether, chloroform or calcium cyanide, the external parasit

55、es, fleas, mites, such as smoked die and temporarily anesthesia, and then, with insects tweezers or close comb qing jian insects. The captive live mice can destroy the medullary, crush, and suffocate in situ. Or placed in a sealed container for the death of CO2. For rare species of mice, even the pu

56、ps and blood stains, or a slightly decomposed corpse, should be scrubbed clean with cloth or talcum powder and managed to make a specimen. After the dead animals treated above, the specimen can be measured and identified.(3) external measurement and identificationThe dead animals that have been kill

57、ed in vitro have been selected for the specimens of fur, skulls, limbs and the intact tail. There may be damage, but it may be rare, precious or suspicious. The selected specimens should be recorded as weight, in grams, and then external measurements.The external measure is measured in millimeters a

58、nd the integer can be taken. Measuring instruments can be used with two feet, steel tape, caliper, etc. Before measuring, put the rat body on the table or enamel plate, take its natural posture, if the stiff and crouching specimen, should gently knead the upright, it will restore the natural positio

59、n.Determination and recorded in the project include: specimen number, mills, date, gender, body length, long hind legs, long tail, ears (see figure 1) animal sex discrimination, main structure is decided by the external genital organs of animals, both male and female have developed on both sides of

60、the mammary gland is located in the chest and mouse mouse department; The male testicles descend from the abdominal cavity into the scrotum of the mouse tram.These characteristics are not easy to be easily identified and are not easy to identify. Generally speaking, the distance between the genitour

61、inary nipple and the anus at the back end of the midline of the abdomen can be determined. The male is longer than the female. Neither of these methods can identify the gender or the unfamiliar type, can open the abdomen, through the internal genital observation. The male can see the penis and the t

62、esticles, and the female can see the fallopian tubes and the uterine belt.(4) stripping methodPlace the mouse on the tablet, gently gently along the midline of the abdomen, using a small scissors or a scalpel, to the front of the genitals. (not too hard, or you will cut abs to reveal internal organs

63、), at the blade carefully separate the skin and muscles on both sides, until the knee joint, knee joint and the hind leg cut with scissors to the knee joints. (see figure 2) to make this work more convenient, push the hind legs a little bit toward the inside, then turn the knee up and the joints go

64、up. Clear the thigh muscles and turn the skin over to restore the original state. The other side is treated the same way.By cutting off the genitals and the rectum with the skin joint, the tail can be removed. Then trim the connective tissue fibers around the tail, and then use a hands index finger

65、and thumb to tighten the base of the base of the base of the base, gently pull the tail and pull it out (see figure 3). Remember not to overexert, or the tail will break.After peeling the tail, continue to remove the sock forward, and then remove to the forelimb and cut off from the scapula.The skin is more complicated than the skin and should be especially careful not to touch the skin or skull. When you turn to the upper part of your neck you will encounter a translucent soft periosteum, which i