CN103766146B - Inoculation device and inoculation method - Google Patents
Inoculation device and inoculation method Download PDFInfo
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- CN103766146B CN103766146B CN201410044984.3A CN201410044984A CN103766146B CN 103766146 B CN103766146 B CN 103766146B CN 201410044984 A CN201410044984 A CN 201410044984A CN 103766146 B CN103766146 B CN 103766146B
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Abstract
The invention relates to an inoculation device, provides an inoculation method realized by the aid of the device simultaneously, and belongs to the technical field of pesticide biological assay for plant protection. The device comprises a vertical cylindrical tower, wherein a lifting mesh cage is suspended in the center of the top of the tower; at least three upwards inclined pneumatic nozzles are uniformly and circumferentially distributed on the side wall of the lower part of the tower; spraying spread angles of the pneumatic nozzles form an airflow convergence zone which can contain the mesh cage; and a drawer is mounted at the bottom of the tower. According to the device, conidial powder can be blown and separated from attacked leaves once by the aid of vertical airflow, separated conidial powder is floated uniformly, rotation cannot be caused when the conidial powder is blown, and obstruction is free when the conidial powder falls, so that the conidial powder can uniformly fall and spread on a host plant sample plate with a larger area, and biological assay and assessment with high identity and repeatability are realized.
Description
Technical field
The present invention relates to a kind of classification inoculation apparatus, especially a kind of classification inoculation apparatus evenly dusted, provides the inoculation method realized by this device simultaneously, belongs to the pesticide bioassay technical field of plant protection.
Background technology
" inoculation " in pesticide bioassay field refers to and manually makes pathogen and the susceptible location contacts of host plant, thus creates conditions and pathogen is invaded and leads to host plant to fall ill.Inoculation is the important step demonstrate,proving sick process, and research parasitics occurrence regularity, mensuration variety resistance and medicament protection effect all be can't do without " inoculation ".
According to the applicant understood, conventional inoculation method has semar technique and spray-on process.The shortcoming of semar technique to accomplish every subjob uniformity, and spray-on process is for the circulation way changing conidia powder the germ that air-flow is propagated, and is difficult to accomplish correct assessment.In addition, also have direct by incidence of leaf conidia powder directly with the dynamic method people of hand shaking for being spread across on different host plant sample, the deposition spore count be difficult to like this on each sample of control is basically identical.
People devise special classification inoculation apparatus for this reason, to realize the uniform settlement of germ conidia powder, such as " Jiangsu's agriculture science " 05 phase in 1986 discloses " the simple and easy inoculation tower of powdery mildew ", electric fan towards up-draught is housed in the middle part of its tower cylinder, rotatable articles holding table is equipped with in bottom, tower mouth from top during use shakes disease leaf, thus fall on the host plant model on articles holding table after making it dispel, its shortcoming is that the air-flow that electric fan upwards blows can form the swirling eddy circumferentially moved, the conidia powder of following flow rotation has the trend along the sedimentation of tower body perisporium, cause the surrounding sedimentation conidia powder of articles holding table more, radially sedimentation distribution is very uneven, and electric fan has blocked the decline of middle part conidia powder.
And for example, " The American Phytopathological Society " Vol78 of the U.S., No11, 1988-1463 discloses " A Vacuum-Operated Settling Tower for Inoculation of Powdery Mildew Fungi. powdery mildew fungi inoculation vacuum deposition tower ", the top of this tower is the leaf loculus that pretends to be sick with seal cover, articles holding table is equipped with in bottom, after vacuumizing, open seal cover, on the host plant model that the air-flow that can cause by negative pressure makes germ conidia powder be scattering on articles holding table, its shortcoming relies on the limited cavity of sealing property to vacuumize formed negative-pressure air-flow to be difficult to blow off disposable for the conidia powder on incidence of leaf, and this negative-pressure air-flow only opens moment disposable formation at seal cover, downward air-flow is difficult to conidia powder uniform settlement on the larger model of area.
Summary of the invention
The object of the invention is to: the shortcoming existed for above-mentioned prior art, propose one and effectively can realize conidia powder to be seeded at the equally distributed classification inoculation apparatus of larger area host plant model, provide the inoculation method realized by this device simultaneously.
In order to reach above object, classification inoculation apparatus of the present invention comprises vertical tubular tower cylinder, the center of top of described tower cylinder hangs liftable cylinder mould, the lower sides of described tower cylinder is equipped with at least three uniform pneumatic energy nozzles towards surface thereof of circumference, and the spraying divergence angle of described pneumatic energy nozzle forms the air-flow confluence can containing cylinder mould; Drawer is equipped with in the bottom of described tower cylinder.
The inclination angle of described pneumatic energy nozzle is 15 °-35 °, best 30 °.
The height of described tower cylinder makes the axes extending line of pneumatic energy nozzle at least turn back for twice in the formation of tower cylinder inwall.
The inoculation method of classification inoculation apparatus of the present invention is used to comprise the steps:
The first step, open drawer, fall cylinder mould, the incidence of leaf with conidia powder to be seeded is put into cylinder mould;
Second step, cylinder mould is risen to air-flow confluence, open pneumatic energy nozzle and brush conidia powder in cylinder mould on incidence of leaf;
3rd step, after the scheduled time, host plant model is placed in drawer, closing drawers, and closes pneumatic energy nozzle;
4th step, cylinder mould is risen to tower top gradually, on the host plant sample that the conidia powder of brushing rear even suspension is dropped in drawer.
The present invention, by the device of advantages of simple, under the air-flow effect of crossing that several pneumatic energy nozzles spray, makes to be positioned at conidia powder on the cylinder mould incidence of leaf of air-flow confluence by the strength effect in side direction, leaves blade and move to barrel direction; Enter the immixture district of straight blowing gas stream and reflected gas stream afterwards, the trend to the motion of barrel direction dies down, the intensifying trend moved upward; Entering afterwards is reflected gas stream region completely again, and conidia powder is mainly moved upward airflow function, is uniformly suspended in tower cylinder top.After shower nozzle is closed, the conidia powder of suspension is evenly fallen on the host plant sample in drawer at the bottom of tower under gravity gradually.
The present invention effectively can make by three-dimensional air-flow that conidia powder is disposable from incidence of leaf dispels separation, and then make the conidia powder even suspension of separation, and can not rotation be produced when conidia powder is brushed, do not hinder during whereabouts, therefore evenly can land and be distributed on the host plant model of larger area, so as to realize homogeneity, repeatability strong raw test and appraisal estimate.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the present invention is further illustrated.
Fig. 1 is the structural representation of one embodiment of the invention.
Fig. 2 is the B-B sectional view of Fig. 1.
Fig. 3 is the A place partial enlarged drawing of Fig. 1.
Fig. 4 is the jet-stream wind confluence schematic diagram of Fig. 1 embodiment.
Fig. 5 is the top view of Fig. 4.
Fig. 6 is another using state schematic diagram of the present invention.
In figure: rotary drawers 1, drawer handle 2, culture dish 3, rotating pin 4, pneumatic energy nozzle 5, connecting bolt 6, L shape installing plate 7, nut 8,9, tower cylinder 10, control panel 11, cylinder mould door bolt 12, stainless steel cylinder mould 13, positioning rope 14, screw 15, rotational positioning wheel 16, stepper motor 17, motor fixing plate 18.
Detailed description of the invention
Embodiment one
The classification inoculation apparatus basic structure of the present embodiment as shown in Figure 1 and Figure 2, stepper motor 17 is equipped with at the top of vertical tubular tower cylinder 10, the rotational positioning wheel 16 of its end hangs liftable stainless steel cylinder mould 13 by positioning rope 14 from tower cylinder 10 center of top, and this stainless steel cylinder mould 13 is equipped with cage door able to turn on or off by cylinder mould door bolt 12.The lower sides of tower cylinder 10 is equipped with six pneumatic energy nozzle 5(towards surface thereof 30 ° (β=30 °) see Fig. 3 respectively by the L shape installing plate 7 that six circumferences are uniform), the β angle of L shape installing plate 7 can regulate as required.The spraying divergence angle of six pneumatic energy nozzles 5 forms the air-flow confluence (see Fig. 5) can containing cylinder mould 13 shown in Fig. 4 profile.The height of tower cylinder 10 ensures that pneumatic energy nozzle 5 axes extending line is formed at tower cylinder inwall and turns back (namely reflecting twice) for twice.Transposition drawer (see Fig. 2) is equipped with by vertical rotating pin 4 in the bottom of tower cylinder 10, and it is convenient to open, and closes sealing.
During use, drawer is opened in first transposition, falls cylinder mould by control panel, the incidence of leaf containing standard quantitative bacterial classification conidia powder to be seeded is put into cylinder mould, and buckles well cage door; Restart stepper motor, cylinder mould is risen to air-flow confluence, adjusting air valve pressure is to 0.2MPa, open pneumatic energy nozzle, brush the conidia powder in cylinder mould on incidence of leaf, after 8-12s, conidia powder is fully brushed, and the large granular impurity on blade also comes off, evenly be placed in drawer by put as host plant model one group of culture dish 3, reverse indexing closing drawers also closes pneumatic energy nozzle; Again start stepper motor, cylinder mould is risen to tower top gradually, make the bacterium colony conidia powder free settling of brushing rear even suspension be about 120s, drop on the culture dish in drawer.Be added with 1.5% water agar in culture dish, every culture dish is put 9 2.5cm
2circular leaf dish, uniform settlement fungal spore powder, cultivates parasitic bacterium colony.
In above process, stainless steel cylinder mould is positioned the presumptive area of classification inoculation apparatus, under six pneumatic energy nozzle direct effects, conidia powder on incidence of leaf is by the strength effect in side direction, first mainly to move to barrel direction, then after moving to the two-way function region by direct gas flow and barrel reflected gas stream, die down gradually to barrel motion, and the trend that moves upward strengthens, enter into total reflection airflow function region afterwards, conidia powder substantially only moves upward, be uniformly suspended in tower cylinder top, after closing shower nozzle, under gravity, conidia powder slowly moves downward, evenly fall.Test setting whiff pressure 0.2MPa, air blowing duration is 5 seconds, 10 seconds, 20 seconds, checks respectively to be blown on rear bacterium powder blade to be detained clump count, determines that 10 seconds air blowing durations are advisable.The determination test setting of sedimentation duration is after air blowing terminates, sedimentation 30 seconds, 60 seconds, 120 seconds, 180 seconds respectively, then read the quantity of bacterium colony in every culture dish with 40 power microscopes, result of the test 120 seconds and sedimentation in 180 seconds as broad as long, determine that sedimentation duration is 120 seconds.
Equalsettlement assessment reads random 19.6mm in every culture dish with 40 power microscopes equally
2the quantity Nx of bacterium colony in (diameter 5mm garden) region, every culture dish reads different 4 place's data, standard deviation (coefficient of variation) Cv calculating 12 culture dish 48 place's sedimentation clump counts is 7.2%, and far below external vacuum deposition tower 19.76%, uniformity is promoted greatly.
In addition to the implementation, the present invention can also have other embodiments.Such as, drawer structure is not limited to transposition drawer; For another example, as host plant model put except culture dish, also can be as shown in Figure 6, adopt 96 orifice plates: in each hole, add 100 μ l fluid nutrient mediums in advance, after uniform settlement fungal spore powder, in each hole, add the bactericide of the different doubling dilution concentration of 100 μ l more respectively, do bactericidal effect experiment.All employings are equal to the technical scheme of replacement or equivalent transformation formation, all drop on the protection domain of application claims.
Claims (6)
1. a classification inoculation apparatus, comprise vertical tubular tower cylinder, it is characterized in that: the center of top of described tower cylinder hangs liftable cylinder mould, the lower sides of described tower cylinder is equipped with at least three uniform pneumatic energy nozzles towards surface thereof of circumference, and the spraying divergence angle of described pneumatic energy nozzle forms the air-flow confluence can containing cylinder mould; Drawer is equipped with in the bottom of described tower cylinder.
2. classification inoculation apparatus according to claim 1, is characterized in that: the inclination angle of described pneumatic energy nozzle is 15 °-35 °.
3. classification inoculation apparatus according to claim 2, is characterized in that: the height of described tower cylinder makes the axes extending line of pneumatic energy nozzle at least turn back for twice in the formation of tower cylinder inwall.
4. classification inoculation apparatus according to claim 3, is characterized in that: the lower sides of described tower cylinder is equipped with six pneumatic energy nozzles towards surface thereof 30 ° respectively by the L shape installing plate that six circumferences are uniform.
5. classification inoculation apparatus according to claim 4, is characterized in that: transposition drawer is equipped with by vertical rotating pin in the bottom of described tower cylinder.
6. use the inoculation method of classification inoculation apparatus described in claim 1, it is characterized in that comprising the steps:
The first step, open drawer, fall cylinder mould, the incidence of leaf with conidia powder to be seeded is put into cylinder mould;
Second step, cylinder mould is risen to air-flow confluence, open pneumatic energy nozzle and brush conidia powder in cylinder mould on incidence of leaf;
3rd step, after the scheduled time, host plant model is placed in drawer, closing drawers, and closes pneumatic energy nozzle;
4th step, cylinder mould is risen to tower top gradually, on the host plant sample that the conidia powder of brushing rear even suspension is dropped in drawer.
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| CN201410044984.3A CN103766146B (en) | 2014-02-07 | 2014-02-07 | Inoculation device and inoculation method |
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| CN201410044984.3A CN103766146B (en) | 2014-02-07 | 2014-02-07 | Inoculation device and inoculation method |
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| CN114480090A (en) * | 2021-12-14 | 2022-05-13 | 杭州市农业科学研究院 | Powdery mildew inoculation device and powdery mildew rapid batch inoculation method |
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| BE786536A (en) * | 1971-07-23 | 1972-11-16 | Aquitaine Pour L Etude Et Le C | PREPARATION OF SELECTED MICROBIAL POPULATIONS FOR USE IN POLLUTION CONTROL |
| CN2584608Y (en) * | 2002-12-13 | 2003-11-05 | 任荣泽 | Mold starter culturing machine |
| CN101671625B (en) * | 2009-09-28 | 2012-08-15 | 厦门北大泰普科技有限公司 | Method and device for preparing trichoderma conidium by liquid state submerged fermentation |
| CN101712931A (en) * | 2009-10-20 | 2010-05-26 | 上海卉芸农业科技有限公司 | Production method and production equipment of haze mycorrhiza fungus |
| EP2505634B1 (en) * | 2011-03-31 | 2013-12-04 | Technelep srl | A solid state fermentation plant |
| CN102229893B (en) * | 2011-06-02 | 2012-09-05 | 刘方一 | High-efficiency high-yield production method of mycorrhizal fungi |
| CN203105161U (en) * | 2013-03-26 | 2013-08-07 | 陕西众兴菌业科技有限公司 | Equipment capable of shortening inoculating time of edible mushroom liquid spawns |
| CN203735134U (en) * | 2014-02-07 | 2014-07-30 | 农业部南京农业机械化研究所 | Inoculation device |
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Address after: No. 100, Liuying, Nanjing, Jiangsu Province, 210013 Patentee after: NANJING Research Institute FOR AGRICULTURAL MECHANIZATION MINISTRY OF AGRICULTURE Address before: No. 100, Liuying, Nanjing, Jiangsu Province, 210013 Patentee before: NANJING Research Institute FOR AGRICULTURAL MECHANIZATION MINISTRY OF AGRICULTURE |
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Effective date of registration: 20230314 Address after: 210019 area a, 4 / F, building A8, 8 Bailongjiang East Street, Jianye District, Nanjing City, Jiangsu Province Patentee after: Nanji Agricultural Machinery Research Institute Co.,Ltd. Address before: No. 100, Liuying, Nanjing, Jiangsu Province, 210013 Patentee before: NANJING Research Institute FOR AGRICULTURAL MECHANIZATION MINISTRY OF AGRICULTURE |
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