CN103937868A - Method for forecasting field kendir rust disease and special device therefor - Google Patents
Method for forecasting field kendir rust disease and special device therefor Download PDFInfo
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Abstract
The invention discloses a method for forecasting a kendir rust disease caused by kendir Melampsora and a special device therefor. The method mainly comprises the following steps: manufacturing of a carrier, preparation of a sampling device, sampling and trapping of spores, isolated culture and pathogenicity calculation. The occurrence of the Melampsora of the field-planted kendir can be accurately forecast and predicted, so that a control plan can be effectively made. Compared with the prior art, the invention can effectively determine the quantity of pathogenic bacteria having infectivity while the quantity of pathogenic bacteria can be determined, and especially can solve the problem that isolated kendir leaves which have been sampled in an aeroscope can be easily polluted and rotten in the culture process. Thus, the invention is suitable for forecasting and predicting the Melampsora of the field commercially-planted kendir.
Description
Technical field
The invention belongs to phytopathy prevention category, relate generally to the technology that the important disease rust of kendir is detected to forecast, particularly a kind of to the method for being predicted by the caused kendir rust of kendir grid rest fungus and the special device of the method.
Background technology
Along with kendir cultivated area and output expanding day, grid rust has become the Major Diseases in kendir plantation, kendir growth and quality are caused to important impact, particularly bring great financial loss to production, seriously hindered the development of kendir industry.
Kendir grid rest fungus uredospore is the source of infection that development occurs grid rust, therefore, measure exactly land for growing field crops air uredospore quantity, particularly virulence, can be for accurately in time for prediction kendir rust provides foundation, reduce rust and kendir is produced to the loss causing.
Mainly sample by air microorganism sampler at present, then under measuring microscope distinguishable or in substratum educable target pathogenic bacteria quantity, then according to this numerical value, Plant diseases and development thereof are carried out to prediction.
Due to the passing in time in physical environment of the pathogenic bacterias such as kendir grid rest fungus, active can remarkable reduction even completely losing.Therefore, only measure the pathogenecity that pathogenic bacteria quantity can not reflect that it is actual, thereby usually cause wrong report.
And, the current method of sampling be directly excised leaf is placed in water or moisturizing thing on, then sample by air microorganism sampler.Because needing certain humidity environment, the excised leaf of inoculation grid rest fungus just can infect, show disease, and the kendir blade of land for growing field crops sampling is easily caused blade fast rotting by other living contaminants, therefore impact is infected and is produced spore effect, finally make whole detection and reality mutually wrong very far away, also become the major reason that causes wrong report.
Therefore need to sample accurately and effectively, effectively determine the pathogenic bacteria quantity with infection ability, the generation of prediction kendir rust and development accurately and effectively, the particularly power of kendir grid rest fungus uredospore virulence, the forecast and control prediction scheme of making prediction exactly, and then avoid or reduce rust kendir is produced and caused damage.
Therefore, one can be sampled accurately and effectively, particularly can effectively determine the pathogenic bacteria quantity with infection ability, and then prediction kendir rust accurately and effectively, as the method for being predicted by the caused kendir rust of kendir grid rest fungus is just arisen at the historic moment.
Summary of the invention
The object of the present invention is to provide one to sample accurately and effectively, can effectively determine method and the special purpose device thereof of the pathogenic bacteria quantity with infection ability, and then prediction kendir rust accurately and effectively, particularly to being predicted by the caused kendir rust of kendir grid rest fungus, work up effectively preventing prediction scheme, ensure the sound development of high yield, sustainability and the kendir plantation industry of kendir plantation.
Object of the present invention mainly realizes by following steps: carrier is made, and sampling unit is prepared, and the sampling of spore catches, isolated culture, virulence measuring and calculating.Specific operation process is as follows:
Carrier is made: in gnotobasis, cultivate kendir plant and get its blade as catching carrier.
A, substratum make: select after the abundant sterilizing of commercially available vermiculite as substratum.
B, vernalization: by kendir seed, with concentration be 0.1～0.5% chlorine bleach liquor, 3～5 min that sterilize, with using again aseptic water washing after filtered through gauze 3～5 times, then by seed vernalization 12～15h in the water-bath of 38～42 DEG C, treat that showing money or valuables one carries unintentionally appears in seed bud bud, anhydrates for subsequent use.
Described sterilized water is preferably the benzimidazolyl solution that concentration is 80～100ppm.
C, field planting: by field planting, the generally field planting in 5～10 h in the inherent described substratum of 10 h of the seed after vernalization.
D, grow seedlings: when plant strain growth to 3 is taken turns with blade.
Sampling unit is prepared: in special sampling unit, lay sampling carrier, sampling carrier is slide glass and kendir blade.
The preparation of a, sampling unit: with upper opening and be provided with the box body of lid, a slide glass and a moisturizing body are set in box body, described slide glass is the plates of a rectangle the bottom that is adhered to box body by the object of non-setting adhesive or viscous silica gel one class, meanwhile, described moisturizing body and slide glass are placed in the bottom of box body side by side.
Described box body can be the one in circle or rectangle, and box body diameter or the length of side be at 9～15cm, and high 2～5cm is for well.
B, make spore and catch slide: slide glass apart from evenly smear one deck Vaseline in region between lower edge 10～15mm and catch slide as spore.
C, making spore catch blade: the healthy leaves of getting cultivated downward 3rd～5 places of wheel, kendir plant top catches blade as spore, the kendir blade blade back of choosing is upwards placed on moisturizing body, the leaf stalk place gland fritter absorbent cotton of kendir blade, along blade tip upper limb 0.5～1cm place, moisturizing body and blade are clinged with adhesive tape, with adhesive tape, the absorbent cotton of blade lower end gland is clinged equally, sampling unit is upright, to moisturizing body upper limb note sterilized water, till occurring dripping to lower edge, preservation closes the lid.
Sampling: the sampling unit of having laid sampling carrier is placed in to large Tanaka and samples.
A, sampling spot is set: select after sampling spot, before rust occurs, start sampling.
B, lay sampling unit: use constant volume type aeroscope, being fixed on apart from floor level is on 30～50cm support.
C, installation sampling unit: the sampling unit of having laid making spore seizure slide and making spore seizure blade is placed in constant volume type aeroscope, be that every day 8～10 start sampling sample time, afternoon 1～3 respectively exchange primary sample device for 7～9 of dusks, and the rainy day does not sample.
Above-mentioned sampling preferably arranges 10～12 circulations at every turn, and each sampler carries out primary sample and samples by following process: every sampling 80～120L air is a circulation, starts next circulation after the 15～20min of interval again.
Above-mentioned sampling is best according to disease situation occurred, at the each cycle sampling 100～120L of disease early period of origination air, each circulation in mid-term occurs disease is 90～100L air, and the each circulation of disease occurance peak is 80～90L air, and disease occurs latter stage is 90～100L air.
D, statistics slide glass sampling spore count: by sampling unit cover lid complete sampling, take back indoorly, slide glass is taken off, be placed in storage case and preserve after 30 days, add up under the microscope uredospore number.
Isolated culture: the spore after sampling is caught to blade and carry out isolated culture.
A, culture apparatus: as culture dish, arrange in box body bottom and be strip shape body that paliform arranges as substrate with upper opening the box body that is provided with lid.
Described strip shape body is preferably built on stilts shape apart from 0.8～1cm place, box body bottom.Can adopt the support bar longitudinal with strip shape body built on stilts, also can take the mode that strip shape body two ends is fixed on to box body side to make somebody a mere figurehead.
Described box body can be the one in circle or rectangle, and box body diameter or the length of side be at 9～15cm, and high 2～5cm is for well.
B, isolated culture: by the substrate of the placement culture apparatus of the kendir blade after sampling, direction blade is longitudinally crossing with strip shape body direction, preferably be vertical, leaf stalk portion wraps up with absorbent cotton and uses bar shaped filter paper to cover, bar shaped filter paper has at least an end, place to contact with culture apparatus bottom, then in culture apparatus, add sterilized water to concordant with substrate, the benzimidazolyl solution that is 80～100ppm to filter paper dropping by the concentration of sterilized water configuration again, till having water droplet to occur, evenly spray sterilized water to blade face 2～3 times with watering can, covering culture apparatus upper cover is placed in incubator and at 20～23 DEG C, cultivates 8～10 days, and by 10～12 h alternation of light and darkness, intensity of illumination 8000～12000LUX.
Carry out afterwards virulence measuring and calculating.
A, the pathogenic spore count of statistics: cultivate and add up uredinium number after 8～10 days, user's ruled paper or scanner calculate leaf area.
B, virulence calculate: carry out virulence calculating with following formula:
Total spore amount: S1(/ liter)=(S1 * A1/a1)/V;
Cause a disease spore amount: S2(/ liter)=(S2*A1/a2)/V;
Virulence index: S2/S1=(a1*S2)/(a2*S1).
Above-mentioned parameter is defined as: A1 sampling unit floorage; A1 slide glass is smeared Vaseline region area; A2 blade area; The uredospore number that S1 slide glass captures; S2 blade uredinium number; S1 unit volume every day air spore count in ancient name for China; The uredospore number of tool virulence in S2 unit volume every day air; V air sampling volume.
According to above-mentioned virulence index comparatively exactly prediction go out the grid rust of field planting kendir a situation arises, and then effectively work up and prevent and treat prediction scheme.
Compared with prior art, the invention solves in measuring pathogenic bacteria quantity, can effectively measure the pathogenic bacteria quantity with infection ability, particularly solve the rotten problem of vulnerable to pollution in the time that aeroscope completes the kendir leaf culture in vitro of sampling, be suitable for the grid rust prediction of land for growing field crops commercialization plantation kendir.
Brief description of the drawings
Fig. 1 is the structural representation of kendir grid rest fungus uredospore Special sampling device.
Fig. 2 is the structural representation of Figure 1A-A section
Fig. 3 is the structural representation of kendir grid rest fungus uredospore culture apparatus special.
Fig. 4 is the structural representation of Fig. 3 B-B section
In diagram: 11 is box body, 12 is moisturizing body, and 13 is slide glass, and 14 is kendir blade, and 15 is adhesive tape, and 16 is absorbent cotton, and 17 is non-setting adhesive, and 21 is culture dish, and 22 is substrate, and 23 is support bar, and 24 is filter paper.
Embodiment
Below in conjunction with embodiment, the present invention is further described.
Carrier is made: in gnotobasis, cultivate kendir plant and get its blade as catching carrier.
A, substratum make: select particle thicker, the commercially available vermiculite of good permeability, generally select particle diameter 1～3mm better, by vermiculite sterilizing 48～72 h at 120～171 DEG C, baking oven, take out and pack in seedling pan.
B, vernalization: choose then kendir seed 80～100 g that gather, with concentration be 0.1～0.5% chlorine bleach liquor, 3～5 min that sterilize, with using again aseptic water washing after filtered through gauze 3～5 times.Then seed is put into triangular flask, tie bottleneck with sealed membrane, put into the water-bath vernalization 12～15h of 40 DEG C.Treat that showing money or valuables one carries unintentionally appears in seed bud bud, removes the water in triangular flask.
C, field planting: urged the seed of bud to carry out field planting in the inherent greenhouse of 5～10 h.Select seed full, that show money or valuables one carries unintentionally, with sterilization after tweezers gripping point plant in installing the seedling pan of vermiculite, 10～15 seeds of every nest.Cover seedling pan with preservative film, be placed in plastic chassis, at chassis side wall mark gauging line, be advisable with 1/2～2/3 of chassis volume.Interval 3～5 d moisturizings, to gauging line, ensure that vermiculite moisture is in state of saturation.
D, grow seedlings: after emerging, outwell the water in chassis, water the Hoagland ˊ s nutritive medium of a time 1/2 to gauging line, moisture evaporate to dryness in all seedling pans is treated at interval after 7～10 days, water for the second time nutritive medium to gauging line.After 30～35 days, start thinning, every nest consistent seedling of 4～5 strain growing way of selecting and remain.Wrap up seedling pan with black plastic film, kendir seedling is exposed.20～25 DEG C of greenhouse day temperatures are set, 15～20 DEG C of nights, illumination 10000～12000LUX, relative air humidity 60～70%, cultivates 7～10 weeks.
Sampling unit is prepared: in Special sampling device, lay sampling carrier.
A, prepare sampling unit: select diameter be 9～15cm culture dish as box body 11(Dispoable medical culture dish or through sterilization glass culture dish all can), get the sheet glass of the length × wide 76.2 × 25.4mm of being as slide glass 13, slide glass 13 is adhered to the bottom of box body 11 by the object of non-setting adhesive 17 or viscous silica gel one class, simultaneously, settling side by side long × wide with slide glass 13 is the sampling moisturizing body 12 of 65～75 × 25～30mm, and sampling moisturizing body 12 forms according to 1～3 layer of thieving paper of actual placement.
Certainly, described slide glass 13 also can be selected plastic sheet, and sampling moisturizing body 12 also can adopt other the material that has stronger water sorption, as fabric.
B, make spore and catch slide: slide glass 11 apart from get the Vaseline of pea grain size in region between lower edge 10～15mm, evenly spread upon on slide glass 11 and catch slide as spore.
C, making spore catch blade: the healthy leaves of getting cultivated downward 3rd～5 places of wheel, kendir plant top catches blade as spore, kendir blade 14 blade backs of choosing are upwards placed on moisturizing body 12, the leaf stalk place gland fritter absorbent cotton 16 of kendir blade 14, along blade tip upper limb 0.5～1cm place, moisturizing body and blade are clinged with adhesive tape 15, with adhesive tape 15, the absorbent cotton 16 of blade lower end gland is clinged equally, sampling unit is upright, to moisturizing body upper limb note sterilized water, till occurring dripping to lower edge, preservation closes the lid.
The seizure sampling of spore:
A, sampling spot is set: before rust occurs, start sampling, select East, West, South, North and central zone large Tanaka, each orientation arranges 3～5 sampling spots.
B, sampling unit: use constant volume type aeroscope, being fixed on apart from floor level is on 30～50cm support.
C, installation sampling unit: the sampling unit of making is placed in constant volume type aeroscope.Be sample time: 9 of every days start sampling, before and after 2 pm and at dusk 8 front and back respectively exchange primary sample device for.Every sub-sampling arranges 10～12 circulations of pattern, and each cycle sampling 80～120L air, according to disease situation occurred, initial stage sampling 100～120L, be 90～100L mid-term, the Sheng phase is 80～90L, be 90～100L latter stage, intercycle 15～20min, and the rainy day does not sample.
D, add up total spore count: by sampling unit cover lid complete sampling, take back indoorly, slide glass is taken off, be placed in storage case and preserve after 30 days, add up under the microscope uredospore number.
Isolated culture: the spore after sampling is caught to blade and carry out isolated culture.
A, leaf in vitro device: with upper opening and be provided with lid, diameter or the length of side are 9～15cm, the box body of high 2～5cm is as culture dish 21, bamboo let or sticking plaster is set in box body bottom and is paliform and arranges as substrate 22, and bamboo let or sticking plaster adopt longitudinal support bar 23 to make somebody a mere figurehead.
Certainly the strip shape body of leaf in vitro device also can be maked somebody a mere figurehead by the glue mode being bonded at apart from the sidewall at 0.8～1cm place at the bottom of ware that is parallel to each other.
The isolated culture of b, sampling rear blade: will take the blade of sample place with bamboo let direction vertical position on, leaf stalk portion wraps up with an absorbent cotton, respectively cover with 3～5 layers of bar shaped filter paper 24 up and down, centre sandwich "
"" upper end of shape filter paper 24, lower end contacts with at the bottom of ware.Then to adding sterilized water at the bottom of ware to till soon approaching bamboo let, to the filter paper 24 benzimidazolyl solution that to drip by the concentration of sterilized water configuration be 80～100ppm, till having water droplet to occur.Evenly spray sterilized water to blade face 2～3 times with watering can, finally cover ware lid and be placed in incubator and cultivate, set temperature is 20～23 DEG C, alternation of light and darkness 12h, intensity of illumination 8000～12000LUX.
The box body of sampling unit and isolated culturing device also can adopt circular or square plastics, glass or other analogous material to make.
Virulence measuring and calculating
A, the pathogenic spore count of statistics: cultivate and add up uredinium number after 8～10 days, user's ruled paper or scanner calculate leaf area.
B, virulence are calculated:
Total spore amount: S1(/ liter)=(S1 * A1/a1)/V
Cause a disease spore amount: S2(/ liter)=(S2*A1/a2)/V
Virulence index: S2/S1=(a1*S2)/(a2*S1)
P, parameter-definition:
A1 sampling unit floorage; A1 slide glass is smeared Vaseline region area; A2 blade area; The uredospore number that s1 slide glass captures; S2 blade uredinium number; S1 unit volume every day air spore count in ancient name for China; The uredospore number of tool virulence in S2 unit volume every day air; V air sampling volume.
According to above-mentioned virulence index comparatively exactly prediction go out the grid rust of field planting kendir a situation arises, and then effectively work up and prevent and treat prediction scheme.
Need statement, the specific embodiment of the present invention is to further description of the present invention, can not assert that specific embodiment of the invention is confined to these explanations.For general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, can also make some simple deduction or replace, all should be considered as belonging to protection scope of the present invention.
Claims (9)
1. a method of land for growing field crops kendir rust being predicted, is characterized in that mainly realizing by following steps: carrier is made, and sampling unit is prepared, and the sampling of spore catches, isolated culture, and virulence measuring and calculating, specific operation process is as follows:
Carrier is made: in gnotobasis, cultivate kendir plant and get its blade as catching carrier;
A, substratum make: select after the abundant sterilizing of vermiculite as substratum;
B, vernalization: by kendir seed, with concentration be 0.1～0.5% chlorine bleach liquor, 3～5 min that sterilize, with using again aseptic water washing after filtered through gauze 3～5 times, then by seed vernalization 12～15h in the water-bath of 38～42 DEG C, treat that showing money or valuables one carries unintentionally appears in seed bud bud, anhydrates for subsequent use;
C, field planting: by the field planting in the inherent described substratum of 10 h of the seed after vernalization;
D, grow seedlings: when plant strain growth to 3 is taken turns with blade;
Sampling unit is prepared: in special sampling unit, lay sampling carrier;
The preparation of a, sampling unit: with upper opening and be provided with lid, diameter or the length of side are 9～15cm, the box body of high 2～5cm, a slide glass and a thieving paper are set in box body, slide glass is adhered to the bottom of box body by the object of non-setting adhesive or viscous silica gel one class, meanwhile, side by side moisturizing body is placed in to the bottom of box body with slide glass;
B, make spore and catch slide: slide glass apart from evenly smear one deck Vaseline in region between lower edge 10～15mm and catch slide as spore;
C, making spore catch blade: the healthy leaves of getting cultivated downward 3rd～5 places of wheel, kendir plant top catches blade as spore, the kendir blade blade back of choosing is upwards placed on moisturizing body, the leaf stalk place gland fritter absorbent cotton of kendir blade, along blade tip upper limb 0.5～1cm place, moisturizing body and blade are clinged with adhesive tape, with adhesive tape, the absorbent cotton of blade lower end gland is clinged equally, sampling unit is upright, to moisturizing body upper limb note sterilized water, till occurring dripping to lower edge, preservation closes the lid;
Sampling: the sampling unit of having laid sampling carrier is placed in to large Tanaka and samples;
A, sampling spot is set: select after sampling spot, before rust occurs, start sampling;
B, lay sampling unit: use constant volume type aeroscope, being fixed on apart from floor level is on 30～50cm support;
C, installation sampling unit: the sampling unit of having laid making spore seizure slide and making spore seizure blade is placed in instrument, be that every day 8～10 start sampling sample time, afternoon 1～3 respectively exchange primary sample device for 7～9 of dusks, and the rainy day does not sample;
D, statistics slide glass sampling spore count: by sampling unit cover lid complete sampling, take back indoorly, slide glass is taken off, be placed in storage case and preserve after 30 days, add up under the microscope the uredospore number on slide glass;
Isolated culture: the spore after sampling is caught to blade and carry out isolated culture;
A, culture apparatus: with upper opening and be provided with lid, diameter or the length of side are 9～15cm, the box body of high 2～5cm is as culture dish, arranges be strip shape body that paliform arranges as substrate in box body bottom;
B, isolated culture: by the substrate of the placement culture apparatus of the kendir blade after sampling, direction blade is longitudinally crossing with strip shape body direction, leaf stalk portion wraps up with absorbent cotton and uses bar shaped filter paper to cover, bar shaped filter paper has at least an end, place to contact with culture apparatus bottom, then in culture apparatus, add sterilized water to concordant with substrate, the benzimidazolyl solution that is 80～100ppm to filter paper dropping by the concentration of sterilized water configuration again, till having water droplet to occur, evenly spray sterilized water to blade face 2～3 times again, covering culture apparatus upper cover is placed in incubator and at 20～23 DEG C, cultivates 8～10 days, and by 10～12 h alternation of light and darkness, intensity of illumination 8000～12000LUX,
Carry out afterwards virulence measuring and calculating.
2. the method that land for growing field crops kendir rust is predicted according to claim 1, is characterized in that: described sterilized water is that concentration is the benzimidazolyl solution of 80～100ppm.
3. the method that land for growing field crops kendir rust is predicted according to claim 1 and 2, is characterized in that: described sampling arranges 10～12 circulations at every turn, each cycle sampling 80～120L air, intercycle 15～20min.
4. the method that land for growing field crops kendir rust is predicted according to claim 3, it is characterized in that: according to disease situation occurred, described is sampled as: the each cycle sampling 100～120L of disease early period of origination air, each circulation in mid-term occurs disease is 90～100L air, the each circulation of disease occurance peak is 80～90L air, and disease occurs latter stage is 90～100L air.
5. a kendir grid rest fungus uredospore sampling unit, comprise upper opening and be provided with the box body of lid, it is characterized in that: a slide glass and a moisturizing body are set in box body, described slide glass is the plates of a rectangle the bottom that is adhered to box body by the object of non-setting adhesive or viscous silica gel one class, meanwhile, described moisturizing body and slide glass are placed in the bottom of box body side by side.
6. kendir grid rest fungus uredospore sampling unit according to claim 5, is characterized in that: described box body is the one in circle or rectangle, and box body diameter or the length of side are 9～15cm, high 2～5cm.
7. a kendir grid rest fungus uredospore isolated culturing device, comprises upper opening and is provided with the box body of lid, it is characterized in that: arrange in box body bottom and be strip shape body that paliform arranges as substrate.
8. kendir grid rest fungus uredospore isolated culturing device according to claim 7, is characterized in that: described strip shape body is built on stilts shape apart from 0.8～1cm place, box body bottom.
9. according to the kendir grid rest fungus uredospore isolated culturing device described in claim 7 or 8, it is characterized in that: described box body is the one in circle or rectangle, box body diameter or the length of side are 9～15cm, high 2～5cm.
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| CN110036860A (en) * | 2019-04-26 | 2019-07-23 | 中国烟草总公司郑州烟草研究院 | Method for judging tobacco root Disease pressure |
| CN110468041A (en) * | 2019-09-25 | 2019-11-19 | 福建省农业科学院植物保护研究所 | A kind of phytopathogen spore monitoring device and monitoring method |
| CN110468041B (en) * | 2019-09-25 | 2024-04-30 | 福建省农业科学院植物保护研究所 | Plant germ spore monitoring device and monitoring method |
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Address after: 833000. The Xinjiang Uygur Autonomous Region, Bortala, Mongolia Autonomous Prefecture, Jinghe County, Toto Township, Ebinur Lake, Ma Bao Ma base Patentee after: Xinjiang Ebinur Lake Gaubau Hemp Co., Ltd. Patentee after: Lanzhou University Patentee after: Al tiger Bao tea Limited by Share Ltd Address before: 833000. The Xinjiang Uygur Autonomous Region, Bortala, Mongolia Autonomous Prefecture, Jinghe County, Toto Township, Ebinur Lake, Ma Bao Ma base Patentee before: Xinjiang Ebinur Lake Gaubau Hemp Co., Ltd. Patentee before: Lanzhou University Patentee before: Xinjiang Altay Gaubau Hemp Co., Ltd. |
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Effective date of registration: 20161101 Address after: 836509 the Xinjiang Uygur Autonomous Region, Aletai Saline Lake, Aletai Bao Ma Ma base Patentee after: Al tiger Bao tea Limited by Share Ltd Address before: 833000. The Xinjiang Uygur Autonomous Region, Bortala, Mongolia Autonomous Prefecture, Jinghe County, Toto Township, Ebinur Lake, Ma Bao Ma base Patentee before: Xinjiang Ebinur Lake Gaubau Hemp Co., Ltd. Patentee before: Lanzhou University Patentee before: Al tiger Bao tea Limited by Share Ltd |