CN1555707A - Application of fungus activated protein in biological pesticide - Google Patents
Application of fungus activated protein in biological pesticide Download PDFInfo
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- CN1555707A CN1555707A CNA2004100228211A CN200410022821A CN1555707A CN 1555707 A CN1555707 A CN 1555707A CN A2004100228211 A CNA2004100228211 A CN A2004100228211A CN 200410022821 A CN200410022821 A CN 200410022821A CN 1555707 A CN1555707 A CN 1555707A
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Abstract
An application of activated mycoprotein in bioinsecticide is disclosed. After the activated mycoprotein in the form of wettable powder and the Bacillus thuringienis are proportionally mixed, the resultant mixture is added to feed or used to immerse leaves or pests for killing the lepidopterous pests.
Description
Technical field:
The albumen that the present invention relates to extract from biology is specifically related to a kind of mycoprotein.
Background technology:
The microprotein agricultural chemicals that research and development form from protokaryon bacterium Harpin (allergen protein) is the representative pest-resistant disease-preventing microbial albumen novel pesticide of generally acknowledging in the world at present, about the frontier of plant molecular immune drug and the rise of promotion microprotein novel pesticide have been started in the research of Harpin and the successful exploitation of Messenger.In recent years, the Qiu Dewen doctor isolates the fungi activator protein of new construction first from eukaryotic microorganisms fungi (Alternaria) (the Chinese patent application publication number is: CN1344727A), except having tangible enhancing via plant immunity, the significantly short increasing crop yield effect (more than 10%) of giving birth to, can increase substantially beyond agricultural product quality and the international competitiveness, the control efficiency of multiple virus diseases such as plant tobacco mosaic virus, cucumber mosaic viral disease and aphid also more than 75%, has been shown outstanding DEVELOPMENT PROSPECT.And whether this activator protein has control efficiency to lepidoptera pest, particularly whether the biological insecticides bacillus thuringiensis,Bt (Bacillusthuringiensis Bt) with very high occupation rate of market being had the insecticidal synergistic effect, is the problem that is worth exploration in the pesticide research.
Summary of the invention:
The present invention is intended to study the effect of mycoprotein in biological insecticides, to improve the insecticidal effect of biological insecticides.
The present invention is achieved through the following technical solutions the foregoing invention purpose.
After fungi activator protein wetting powder with 3.0% and the former powder of 50000IU/ μ l bacillus thuringiensis,Bt mix in 0.03～1.5: 1 ratio of Bt consumption with activator protein, contaminate, soak leaf with feed and soak worm or leaf dipping method processing lepidoptera pest, make the insecticidal synergistic index reached 1.5～18.5 of bacillus thuringiensis,Bt.
Be described in further detail the present invention below.
The present invention adopts feed contamination method, leaf dipping method, soak multiple bioassay method such as leaf dip method, as testing insect, measured the synergistic effect of fungi activator protein to Bt with important pest cotton bollworm, diamond-back moth and beet armyworm.The result is conforming to be shown, this activator protein has good synergistic effect to Bt, and with the increase of activator protein additional proportion, obvious more to the synergistic effect of Bt.
Test method and result of the test are as follows:
1 materials and methods
1.1 reagent agent
The former powder of 50000IU/ μ l Bt (Bt research center, Wuhan City product)
3% fungi activator protein wetting powder (protein drug engineering experiment chamber, the Chinese Academy of Agricultural Sciences biological control research institute provides)
1.2 for examination worm kind
Cotton bollworm: adult was picked up from cotton field, 4 different regions, Hebei in 1996, and indoor mixing breeding is raised so far with artificial feed, and raising method is seen the virtuous woods 1998 of model
[1]In this test for examination larva worm be age: the feed bulk method adopts 2 instar larvaes, soak the leaf dip method and leaf dipping method adopts 3 instar larvaes.
Diamond-back moth: pick up from Beijing suburb vegetables field, raise many generations at indoor cabbage leaves, 25 ± 1.0 ℃ of raising temperatures, periodicity of illumination 16L: 8D adopts 3 instar larvaes for examination.
Beet armyworm: same diamond-back moth.
1.3 test method
1.3.1 feed contamination method: establish activator protein, Bt pulvis, Bt and mix 3 kinds of processing by a certain percentage with activator protein.Required each medicament is configured to series concentration respectively with after artificial feed evenly mix by measuring, adds in the 24 hole test boxs.Heavy uniform 2 instar larvaes of selective body move into respectively in the hole, and add a cover after adding preservative film in 1 in every hole, tightens with rubber band, place 25 ± 1.0 ℃, the incubator of photoperiod L: D=16h: 8h, and every processing repeats 3 times, 72 of every repetitions.Check result after 5 days.
1.3.2 infusion process: establish activator protein, Bt pulvis, Bt and mix 3 kinds of processing by a certain percentage with activator protein.Each handles 6-8 concentration of dilute with water, and is stand-by.
Cotton bollworm is with two kinds of methods.Leaf dipping method is, fresh rape leaf disk was immersed in the soup 10 seconds, takes out the back and dries naturally on paper, adds in the 24 hole test boxs 1 of every hole adding cotton bollworm 3 instar larvae respectively.Soaking the leaf dip method is, respectively rape leave disk and larva is soaked for 10 seconds in the soup of same concentrations, treat blade dry with the unnecessary soup of polypide inhale go after, move in the 24 hole boxes 1 on every hole 1 leaf.24 hole boxes are tightened with rubber band, and each concentration repeats 3 times, and 72 of larvas are established blank.Examination worm after the processing places 25 ± 1.0 ℃, the incubator of photoperiod L: D=16h: 8h to cultivate check result after 48 hours and 72 hours.
Diamond-back moth is used leaf dipping method.Cabbage leaves was flooded in soup 10 seconds, dry, put into 10 centimetres of culture dishes of diameter, add larva, other same cotton bollworm.
Beet armyworm is used leaf dipping method, uses leaf dipping method with diamond-back moth.
The dead criterion of the examination worm of above-mentioned test is: touch polypide with dialling pin, the complete motionless person of examination worm is dead.Go out the LC of former powder+activator protein of Bt and the former powder of Bt with the calculating computer
50Value, 95% confidence limit, LC
90Value, and use LC
50Value is calculated the synergy index.
2 result of the tests
2.1 activator protein adds among the Bt synergistic effect to cotton bollworm
2.1.1 the measurement result of feed contamination method
Feed contamination method was handled cotton bollworm after 5 days, and the Bt+ activator protein is with the LC of 1: 0.015,1: 0.03 ratio mixed determining
50Value and Bt use comparison separately, and the synergy index is respectively 1.1 times and 2.1 times, and the result shows that adding activator protein with 1: 0.03 ratio in Bt has good synergistic effect (seeing Table 1) to Bt.
Measured the biologically active of 3% activator protein wetting powder itself with the method, at set activator protein is that 15.6 μ g/ml, 31.25 μ g/ml, 62.5 μ g/ml, 125 μ g/ml, 250 μ g/ml, 500 μ g/ml and 1000 μ g/ml are in totally 7 concentration, the lethality of cotton bollworm does not have significant difference with contrast lethality 8.3% after 5 days between 8.8%-18.8%.
Table 1 feed contamination method is measured the synergism test (cotton bollworm, 5 day) of activator protein to Bt
| Medicament | Toxicity regression formula (Y=) | LC 50Value (95% confidence limit) (μ g/ml) | ???LC 90Value (μ g/ml) | The synergy index |
| Former powder+the activator protein of Bt 1: 0.03 | ????3.76+1.59X | ??6.01(4.49-8.05) | ????38.67 | ????2.1 |
| Former powder+the activator protein of Bt 1: 0.015 | ????2.25+2.63X | ??11.13(7.56-16.39) | ????34.24 | ????1.1 |
| The former powder of Bt | ????2.99+1.82X | ??12.63(9.92-16.08) | ????64.25 | ????1.0 |
Annotate: the LC of the former powder of synergy index=Bt
50The LC of the former powder+activator protein of value/Bt
50Value, down together.
2.1.2 leaf dipping method measurement result
Measure the activity of different medicaments to cotton bollworm with leaf dipping method, the Bt+ activator protein adds with the consumption of 1: 0.095 ratio, can reach 2.74 times to the synergy index of Bt, compares with feed contamination method, increase the consumption of activator protein after, synergistic effect more obvious (seeing Table 2).
Table 2 leaf dipping method is measured the synergism test (cotton bollworm, 72 hour) of activator protein to Bt
Toxicity regression formula LC
50Value (95% confidence limit) LC
90Value
Medicament
The synergy index
(Y＝)????????????(μg/ml)???????????(μg/ml)
The former powder of Bt+activation egg
2.54+0.92X?????484.7(223.70-1050.25)??12176.41????2.74
White 1: 0.095
1328.19(592.759-2976.
The former powder 1.75+1.04X 22667.61 1.0 of Bt
0893)
2.1.3 soak the measurement result of leaf dip method
With soaking the leaf dip method cotton bollworm is measured, the Bt+ activator protein adds with the consumption of 1: 1.5 ratio, and more up to 18.5 times, it is not only remarkable to the synergistic effect of Bt to further illustrate this novel activator protein to the synergy index of Bt.And, with the increase of activator protein additional proportion, to the synergistic effect good more (seeing Table 3) of Bt.
Table 3 soaks the leaf dip method and measures the synergism test (cotton bollworm, 72 hour) of activator protein to Bt
Toxicity regression formula LC
50Value (95% confidence limit) LC
90Value
Medicament synergy index
(Y＝)????????????(μg/ml)????????(μg/ml)
The former powder of Bt+activation egg
4.51+0.81X?????4.08(0.66-25.23)????158.1402????18.5
White 1: 1.5
75.50
The former powder 3.85+0.61X 9326.626 1.0 of Bt
(20.51-277.92)
Activator protein＞11000
2.2 activator protein adds among the Bt synergistic effect to diamond-back moth
Show with the measurement result of leaf dipping method, handle after 60 hours that add activator protein by 60% of Bt albumen consumption and also Bt is had significant effect, the synergy index (sees Table 4) up to 5.5 times to diamond-back moth.
Table 4 leaf dipping method is measured the activator protein synergistic effect (diamond-back moth, 60 hour) right to Bt
| Medicament | Toxicity regression formula (Y=) | LC 50Value (95% confidence limit) | ???LC 90Value (μ g/ml) | The synergy index |
| Former powder+the activator protein of Bt 1: 0.6 | ??3.29+2.39X | ?5.20(3.57-7.57) | ???17.93 | ????5.5 |
| The former powder of Bt | ??2.86+1.47X | ?28.67(10.29-79.87) | ???215.21 | ????1.0 |
2.3 activator protein adds among the Bt synergistic effect to beet armyworm
Show with the measurement result of leaf dipping method beet armyworm 3 instar larvaes; Under the situation of 1000 times of Bt dilutions, add a spot of activator protein Ap, make the lethality and the effect identical (table 5) of using 500 times of Bt of dilution of beet armyworm
Table 5 activator protein is to the synergism test (beet armyworm) of Bt
| Processing time (my god) | Lethality (%) | |||||
| ??CK | ???Bt ??500X | ???Bt+Ap ?1000X+10％ | ????Bt+Ap ??1000X+30％ | |||
| ????2 | ??4.2 | ??88.9 | ????63.9 | ????59.7 | ||
| ????3 | ??8.3 | ??100.0 | ????97.2 | ????91.7 | ||
| ??8.3 | ??96.6 | ????100 | ????100 | |||
| On average | ??98.3 | ????98.6 | ????95.9 | |||
Annotate: drug concentration is Bt (extension rate)+activator protein (%)
Method: leaf dipping method;
Worm kind: beet armyworm 3 instar larvaes
Because activator protein itself is nontoxic, do not change the DNA of plant, in plant corpus and in the soil, easily decompose, noresidue is promoting plant growth, and the effect of prevention and elimination of disease and pests is remarkable, can obviously improve the insecticidal effect of Bt again, by systematic research more, might develop the novel synergist of Bt, promote with Bt to be the great development of the biopesticide of representative.Therefore be hopeful to be developed to a kind of Multifunction albumen agricultural chemicals in the agriculture healthy production technology system.
Claims (1)
1. the application of fungi activator protein in biological insecticides, it is characterized in that the former powder of bacillus thuringiensis,Bt with 3.0% fungi activator protein wetting powder and 50000IU/ μ l, mix the post processing lepidoptera pest with activator protein in 0.03～1.5: 1 ratio of Bt consumption, make the insecticidal synergistic index reached 1.5～18.5 of bacillus thuringiensis,Bt.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2004100228211A CN1555707A (en) | 2004-01-08 | 2004-01-08 | Application of fungus activated protein in biological pesticide |
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|---|---|---|---|
| CNA2004100228211A CN1555707A (en) | 2004-01-08 | 2004-01-08 | Application of fungus activated protein in biological pesticide |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102648713A (en) * | 2011-02-23 | 2012-08-29 | 天津市植物保护研究所 | Technology for improving biological disease-prevention activity of trichoderma preparation by utilizing fungi activator protein |
| CN102687730A (en) * | 2011-03-25 | 2012-09-26 | 中国农业科学院植物保护研究所 | Biological compound preparation for controlling plant virus disease and application thereof |
| CN102823626A (en) * | 2012-09-20 | 2012-12-19 | 联保作物科技有限公司 | Fungus biological protein water dispersible granules |
-
2004
- 2004-01-08 CN CNA2004100228211A patent/CN1555707A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102648713A (en) * | 2011-02-23 | 2012-08-29 | 天津市植物保护研究所 | Technology for improving biological disease-prevention activity of trichoderma preparation by utilizing fungi activator protein |
| CN102687730A (en) * | 2011-03-25 | 2012-09-26 | 中国农业科学院植物保护研究所 | Biological compound preparation for controlling plant virus disease and application thereof |
| CN102823626A (en) * | 2012-09-20 | 2012-12-19 | 联保作物科技有限公司 | Fungus biological protein water dispersible granules |
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