KR20130063209A - New entomopathogenic fungi isaria javanica pf04 having controlling of bemisia tabaci q type and microbial insecticide for controlling bemisia tabaci q type containing the same - Google Patents

Abstract

PURPOSE: An insecticide containing entomopathogenic fungi, Isaria javanica Pf04, for controlling Bemisia tabaci Q-type is provided to effectively prevent drug-resistant pest and viral diseases, and to enhance economic value of crops which are cultivated in a greenhouse. CONSTITUTION: A new entomopathogenic fungi, Isaria javanica Pf04(KACC 93122P), prevents Bemisia tabaci Q-type and is denoted by sequence number 1. The strain is isolated from white fly pest. An insecticide for controlling Bemisia tabaci contains Isaria javanica Pf04(KACC 93122P). The insecticide contains the fungi and/or a culture medium thereof.

Description

New Insect Pathogenic Fungus Isaria Javanica P04 with Control Effect against Tobacco Powder Type and Microbial Insecticide for Tobacco Powder Type { FOR CONTROLLING BEMISIA TABACI Q TYPE CONTAINING THE SAME}

The present invention relates to a novel insectogenic pathogenic fungus Isaria Javanica Pf04 and a microbial insecticide for tobacco powder Q type control comprising the same, and particularly to tobacco powder Q biotype worms The present invention relates to a novel insecticidal fungus Isaria Javanica Pf04 exhibiting excellent insecticidal properties and a microbial insecticide for tobacco powder Q type control comprising the same.

Tobacco powder ( Bemisia tabaci ) has been distributed throughout the country since it was first discovered in a rose greenhouse in 1998 in Korea, and is known as a harmful pest due to direct absorption of vegetative and plant virus diseases in ornamental plants and crops.

Tobacco powder has been reported to kill more than 600 plants, including tomatoes, peppers, beans, eggplants, cucumbers, as well as other vegetables.

Tobacco powder is a species complex of which 24 biotypes are known, and biotypes are classified according to host range, extent of host infestation, insecticide resistance and viral transmission capacity.

Among them, the biotype Q nymph is known as a pest that is difficult to control due to its rapid development of resistance to drugs, and recently, the global interest is focused on the rapid expansion of the distribution area. In Mediterranean countries, the main cause of the conversion of biotype B nymph to Q-type nymph is pointed out the increase of insecticide resistance.

For the biological control of tobacco powder, methods using natural enemies such as predators and parasites and methods using insect pathogens are widely used.

As a pest control means using insect pathogenic fungi, Beauveria bassiana ), Aschersonia aleyrodis ), Isaria Farinosus farinosus ) and others have been reported.

On the other hand, in many countries, microbial insecticides for tobacco powder control have been registered, but most of these microorganisms are the result of insects of unknown biotype or unknown biotype of tobacco powder B biotype. In 2007, only the drug resistance of tobacco powder Q biotype was reported, and the research on the selection of microorganisms to be used for the control of insect powder of tobacco powder Q biotype for environmentally friendly control was conducted. Not reported yet.

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel insecticidal fungus Isaria Javanica Pf04 and a microbial insecticide for controlling tobacco powder Q type, which exhibits excellent insecticidal activity against tobacco powder Q type.

Technical problems to be achieved by the present invention are not limited to the above-mentioned technical problems, and other technical problems not mentioned above may be clearly understood by those skilled in the art from the following description. There will be.

In order to achieve the above object, the novel insect pathogenic fungus isaria javanica ( Isaria) of the present invention javanica ) Pf04 strain (KACC93122P) has a control effect against tobacco powder Q type, and in particular, has a control effect against tobacco powder Q type nymphs.

The strain is represented by SEQ ID NO: 1, it is characterized in that it is isolated from the powdered insect pests.

The microbial insecticide for controlling tobacco dust of the present invention is a novel insect pathogenic fungus isaria javanica ( Isaria). javanica ) Pf04 strain (KACC93122P), and in particular, tobacco powder is characterized by controlling Q type nymphs.

The microbial insecticide is characterized in that it comprises any one or more of the cells or culture of the strain.

The microbial insecticide is characterized in that it consists of granules, powders, liquid hydrating agents, hydrating agents, or encapsulated preparations.

Tobacco powder Q-type insect control of the present invention is characterized by applying the above-described microbial insecticide to crops or soil.

Insect pathogenic fungi Isaria exhibiting excellent insecticidal activity against tobacco powder Q type nymphs by the present invention. By providing a microbial insecticide for Q-type control of tobacco powder containing javanica Pf04 microorganisms, it is expected to increase the economic value of plant-grown crops by effectively controlling drug-resistant pests and preventing viral diseases caused by virus-mediated insect control. .

In addition, the stable production and supply of eco-friendly microbial insecticides can increase the production of safe agricultural products in Korea and activate microbial insecticide development industries, thereby promoting domestic job creation and related industries.

1 is a graph showing the insecticidal rate for tobacco powder Q type nymph according to the spore suspension single concentration treatment for each strain (30 strains).
Figure 2 is a graph showing the cumulative insecticidal rate for tobacco powder Q type nymph according to the spore suspension concentration of the strain.
(A) Beauveriabassiana Bb06
(B) B. bassiana Bb13
(C) Isaria javanica Pf04
(D) Lecanicillium lecanii L14
(E) Aschersonia aleyrodes AS1260
(F) B. bassiana GHA
3 is Isaria selected by relative humidity A graph showing the insecticidal rate against tobacco powder Q type insects of the javanica Pf04 strain.
ck: control
Pf04: Selected Isaria javanica Pf04 strain
4 isaria selected by temperature A graph showing the insecticidal rate against tobacco powder Q type insects of the javanica Pf04 strain.
5 is a detailed Isaria A graph showing the insecticidal rate according to the development stage of tobacco powder of javanica Pf04 strain.
Figure 6 isaria selected Figure showing the tobacco powder Q type adult and nymph infected with the javanica Pf04 strain.

The present invention is for environmentally friendly control of tobacco powder Q type which is difficult to control, and selects the highly pathogenic microorganisms of tobacco powder Q type among the insect pathogenic fungi well adapted to the domestic environment, and the proper pesticide concentration of the selected microorganisms. In this regard, the insecticides by environmental conditions and the developmental stages that can be effectively controlled were selected, and thus new strains having excellent control effects on the tobacco powder Q type were selected.

The thus obtained from the starting strain 16S Lai bojom Ria bar, moving a comparative analysis of the homology of the RNA Java NIKA (Isaria A novel strain of javanica has been identified, which is called ' Isaria Javanica'. javanica) Pf04 strain, and was deposited with the National Agricultural Science Institute Agricultural Genetic Support Center strain. 2011.11. On day 14, accession number KACC93122P was assigned.

The new insect pathogenic fungus isaria javanica javanica ) Pf04 strain consists of all or part of the nucleotide sequence set forth in SEQ ID NO: 1, and consists of an open reading frame (ORF) of 579bp.

The microbial insecticide of the present invention is the novel insect pathogenic fungus isaria javanica ( Isaria) javanica ) Pf04 strain (KACC93122P) is provided as a microbial insecticide for tobacco powder Q type control.

At this time, the novel entomopathogenic fungus Isaria javanica Pf04 strain (KACC93122P) is suitable for 10 to 20 days at 24 to 26 ° C, preferably at 25 ° C for 15 days in a suitable medium, for example, a conventional PDA medium. After culturing to produce a spore suspension, the spore suspension is centrifuged to harvest the cells, and the cells (10 ⁶ to 10 ⁸ conidia ml ^-1 ) or the culture solution of the cells are mixed with a conventionally used additive to kill microorganisms. It may be prepared, and at this time, it may be used by adding any one or more of a surfactant, extender, or nutrient as an additive.

As the surfactant, polycarboxylate (polycarboxylate), sodium lignosulfonate, sodium lignosulfonate sodium dialkyl sulfosuccinate, sodium alkyl sulfonate, poly Oxyethylene alkyl phenyl ether, sodium tripolyphosphate (sodium concentrate), sodium alkyl aryl sulfonate, polyoxyethylene alkyl phenyl ether, polyoxyethylene alkyl aryl phosphoric ester (polyoxyethylene aryl phosphoric ester), polyoxyethylene alkyl aryl ether, polyoxyethylene alkyl aryl polymer, polyoxyethylene alkyl aryl polymer special, polyoxyalkylone alkyl phenyl ether alkyl phenyl ether), polyoxyethylene nonyl phenyl One or two or more selected from the group consisting of ether (polyoxyethylene nonyl phenyl ether), sodium sulfonate naphthalene formaldehyde, triton 100 and tween 80, and as extenders and nutrients, soy flour, rice, wheat One or two or more selected from the group consisting of ocher, diatomaceous earth, dextrin, glucose and starch.

The microbial insecticide may be formulated in the form of granules, powders, liquid hydrating agents, hydrating agents, or encapsulating agents, but is not limited thereto.

Formulated insecticides can be diluted 500-2000 times, preferably 1000 times, with water before use, and can be treated on the surface of crops or on the soil in which they are growing, or on the surface of crops in transit or storage. Can be processed.

As a result of confirming the insecticidal effect on the tobacco powder Q type nymph using the microbial insecticide for tobacco powder Q type control of the present invention prepared as described above, it showed a high insecticidal effect of more than 98%, in particular even at a relative humidity of 75% or more It was confirmed that 70% of tobacco powder can be controlled.

In addition, regardless of the growth stage of the tobacco powder Q type nymphs showed an insecticidal effect at all stages, especially in the second and third age, it was confirmed that the best insecticidal effect, over time the tobacco powder Q type nymphs The insect pathogenic fungus Isaria Javanica javanica ) confirmed that the probability of infection with Pf04 microorganisms increased.

Hereinafter, the content of the present invention will be described in more detail by examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1 : The cigarette is Louis  Q type On nymph  Screening and Spore Separation of Microorganisms with Insecticidal Effects

To select microorganisms with insecticidal effects against tobacco powder Q type nymphs, various fungi isolated from insect species or soil were selected.

The selected fungi were cultured in PDA (Potato-Dextrose Agar) medium at 25 ° C. for 15 days to produce spore suspension.

Into the spore suspension thus produced, the surfactant Tween 80 was added and the spores of each fungus were harvested, and insecticide experiments were conducted on 30 kinds of tobacco powder Q type insects.

That is, the insecticidal rate of the tobacco powder Q type nymphs after the single concentration treatment of the spore suspension of the selected microorganisms was confirmed.

As a result, as shown in Figure 1, the treated tobacco powder Q type nymphs were different depending on the microorganism (F = 35.65; P <0.0001).

That is, after 6 days of treatment, Isaria javanica Pf04 microorganism showed the highest insecticidal rate and suppression rate of adult development among the bioassay microorganisms with the cumulative insecticide rate of 98% and adult development rate of 1.8%.

Also. The killing time for the tobacco powder Q type nymphs of the selected microorganisms was confirmed, and the results are shown in Table 1 below.

Species 50% lethal time
LT ₅₀ (days) 90% fatality
LT ₉₀ (days) Beauveria bassiana Bb01 4.3 (4.1 4.5) ^bcd 6.3 (6.0 6.6) ^abc B. bassiana Bb02 4.6 (4.3 4.8) ^bcde 6.9 (6.6 7.2) ^abc B. bassiana Bb03 4.0 (3.8 4.2) ^cdef 5.9 (5.6 6.2) ^abc Isaria javanica Pf04 3.3 (3.1 3.4) ^f 4.8 (4.6 5.0) ^c B. bassiana Bb05 4.1 (3.9 4.2) ^def 6.0 (5.8 6.3) ^bc B. bassiana Bb06 2.9 (2.8 3.1) ^f 4.8 (4.5 5.0) ^c B. bassiana Bb07 3.3 (3.1 3.5) ^ef 5.2 (5.0 5.4) ^c B. bassiana Bb08 3.9 (3.7 4.1) ^def 6.2 (5.9 6.5) ^bc B. bassiana Bb09 4.6 (4.4 4.7) ^def 6.1 (6.0 6.3) ^bc B. bassiana Bb10 4.3 (4.2 4.5) ^abc 6.0 (5.8 6.2) ^a B. bassiana Bb11 4.2 (4.1 4.4) ^def 5.7 (5.6 5.9) ^bc B. bassiana Bb12 3.2 (3.1 3.4) ^f 4.8 (4.7 5.0) ^c B. bassiana Bb13 3.7 (3.6 3.8) ^def 4.9 (4.8 5.0) ^c B. bassiana Bb14 4.3 (4.2 4.4) ^def 5.4 (5.3 5.6) ^bc B. bassiana Bb15 4.6 (4.5 4.8) ^cdef 6.4 (6.2 6.6) ^abc B. bassiana Bb16 4.5 (4.3 4.6) ^def 6.2 (6.0 6.4) ^bc B. bassiana Bb17 5.3 (5.2 5.4) ^cdef 6.4 (6.3 6.5) ^bc B. bassiana Bb18 3.5 (3.4 3.7) ^def 5.1 (4.9 5.2) ^c B. bassiana Bb19 4.1 (3.9 4.2) ^def 5.5 (5.3 5.7) ^bc B. bassiana Bb20 3.8 (3.6 3.9) ^def 5.3 (5.1 5.5) ^bc I. fumosorosea Pf59 4.3 (4.1 4.5) ^cdef 6.1 (5.8 6.3) ^abc B. bassiana Bb1058 3.5 (3.4 3.7) ^def 5.3 (5.1 5.5) ^bc Aschersonia aleyrodes AS1260 4.7 (4.2 5.1) ^def 6.9 (6.2 7.6) ^bc Lecanicillium lecanii L18 4.6 ( ^4.3-4.8 ) ^bcde 6.9 (6.5 7.3) ^abc B. bassiana Bb11218 4.1 (4.0 4.3) ^def 5.6 (5.5 5.8) ^bc L. lecanii L22 4.4 (4.3 4.6) ^def 5.8 (5.6 5.9) ^bc L. lecanii L14 3.4 (3.3 3.6) ^def 5.1 (4.9 5.4) ^c L. attenuatum L625 3.8 (3.6 4.0) ^cdef 5.9 (5.6 6.3) ^abc I. fumosorosea Pf109 5.6 (5.3 6.0) ^ab 8.0 (7.4 8.6) ^ab I. cateni Pc366 5.8 (5.4 6.3) ^a 9.6 (8.5 10.7) ^a

That is, as shown in Table 1, Isaria even at 50% and 90% insecticidal time of tobacco powder Q type insects The microorganisms of javanica Pf04 were 3.3 days and 4.8 days, leading to faster insecticides than other microorganisms.

Example  2: above Example  By concentration of microorganism selected from 1 The cigarette is Louis  Q type Nymph  Insecticide Effect

Among the 30 strains shown in Table 1, six candidate strains having excellent insecticidal effects were selected, and half the lethal times of the tobacco powder Q type II nymphs by concentration of these microorganisms were confirmed.

As a result, it was shown in Table 2 and FIG.

Species LT ₅₀ (days)
(Confidenceintervals) Conidia concentration (X conidia / ml) 10 ⁶ 10 ⁷ 10 ⁸ A. aleyrodes AS1260 5.4 (5.2 5.5) 5.1 (5.0 5.1) 4.4 (4.3 4.5) c B. bassiana Bb06 5.4 (5.1 5.7) 4.6 (4.4 4.7) 3.7 (3.6 3.8) B. bassiana Bb13 5.1 (4.8 5.3) 4.2 (4.1 4.4) 3.8 (3.6 3.9) L. lecanii L14 4.5 (4.3 4.7) 4.0 (3.8 4.1) 3.5 (3.4 3.6) I. javanica Pf 04 5.3 (5.1 5.6) 4.4 (4.3 4.5) 3.3 (3.2 3.4) B. bassiana GHA 4.9 (4.7 5.1) 4.1 (4.0 4.3) 3.9 (3.8 4.1)

As shown in Table 2 and Figure 2, the insecticidal rate for tobacco powder Q type nymphs increased as the treatment concentration increased.

In addition, the half-life lethality time (LT ₅₀ ) of the Isaria javanica Pf04 strain was the lowest as 3.3 days, but the half-life lethality was 269 conidia / mm ² , similar to other strains.

Considering the above results and the degree of spore formation and convenience of collection, Isaria It is determined that the javanica Pf04 strain is suitable for the development of tobacco powder Q biotype control agent in the present invention provides a microbial insecticide comprising the selected Isaria javanica Pf04 strain as described above.

Example  3: The cigarette is Louis  Q type On nymph  Sequence analysis of novel strains showing insecticidal effect against

Isaria finally selected from Table 2 for tobacco powder Q type nymphs based on the above results Sequencing was performed on the javanica Pf04 strain.

For the analyzed sequences, homology analysis with previously known genes was performed.

As a result, the selected strain was found to be a novel strain of Isaria javanica , as described in the attached depositary certificate, which is referred to as ' Isaria Javanica'. javanica) Pf04 strain, and was deposited with the National Agricultural Science Institute Agricultural Genetic Support Center strain and received accession number KACC93122P on November 14, 2011.

Example  4: by relative humidity The cigarette is Louis  Q type On nymph  Insecticide Effect

New strain Isaria identified in Example 3 above The insecticidal effect of the javanica Pf04 strain (KACC93122P) on tobacco powder Q type nymph by relative humidity was confirmed.

1. Experimental Method

After spraying the spore suspension of Isaria javanica Pf04 on the leaves inoculated with tobacco powder nymphs, the leaves were kept at the relative humidity of 53, 75, 85, and 97% to protect the insecticidal effects of tobacco type Q nymphs. Confirmed.

2. Experimental results

As a result of the experiment, Isaria selected in Example 2 as shown in FIG. The javanica Pf04 strain exhibited a pesticidal rate at 53% relative humidity and 70% tobacco dust at a relative humidity of 75% or higher. .

Example  5: The cigarette is Louis  Q type On nymph  Insecticide effect by temperature

New strain Isaria identified in Example 3 above The insecticidal effect on the tobacco powder Q type nymph of the javanica Pf04 strain (KACC93122P) was confirmed.

1. Experimental Method

After spraying the spore suspension of Isaria javanica Pf04 on the leaves inoculated with tobacco powder, the plant leaves were kept at 15, 20, 25, 30, and 35 ℃ to control the insecticidal effect of tobacco powder Q type insects. Confirmed.

2. Experimental results

As a result of the experiment, as shown in FIG. 4, the insecticidal rate was shown at 15 ° C. or higher, and it was confirmed that the highest insecticidal rate was shown at 30 ° C. in particular.

Example  6: The cigarette is Louis  Q type Nymph  Confirmation of pesticidal effect by developmental stage

New strain Isaria identified in Example 3 above The insecticidal effect on the tobacco powder Q type nymph by the developmental stage of the javanica Pf04 strain (KACC93122P) was confirmed.

1. Experimental Method

Tobacco powder adult insects were infected with plant leaves, and after 12 hours, the adult insects were removed, and tobacco powder was bred to reach eggs, 1, 2, 3, 4 year old insects and adults of Q type.

After spraying the spore suspension of Isaria javanica Pf04 on the tobacco powder of each developmental stage, the plant leaves were stored at 25 ℃ and the relative humidity of 97% to confirm the insecticidal effect of the tobacco powder Q type nymph It was.

2. Experimental results

As a result of the experiment, as shown in Figure 5 showed the insecticidal effect in all stages of development, in particular it was observed to show a high pesticide rate in the second stage (second instar) and third stage (third instar) stage.

In other words, the growth rate from adult to nymph was less than 5%, which is lower than other microorganisms, and the next generation density inhibitory effect was also expected. As shown in FIG. 6, tobacco powder Q type nymph is infected with the Isaria javanica Pf04 strain as time passes It could be confirmed.

Thus, the present invention is an insect pathogenic fungi Isaria exhibiting excellent insecticidal properties against tobacco powder Q type nymphs By providing microbial insecticides for controlling Q-type insecticides containing javanica Pf04 microorganisms, it is expected to increase the economic value of plant-grown crops by effectively controlling drug-resistant pests and preventing viral diseases caused by virus-mediated insect control. In addition, the stable production and supply of eco-friendly microbial insecticides can increase the production of safe agricultural products in Korea and activate microbial insecticide development industries, thereby promoting domestic job creation and related industries.

The present invention has been described with reference to the preferred embodiments, and those skilled in the art to which the present invention pertains to the detailed description of the present invention and other forms of embodiments within the essential technical scope of the present invention. Could be. Here, the essential technical scope of the present invention is shown in the claims, and all differences within the equivalent range will be construed as being included in the present invention.

National Agricultural Science Institute Agricultural Genetic Support Center KACC93122P 20111114

<110> REPUBLIC OF KOREA (MANAGEMENT: RERAL DEVELOPMENT ADMINISTRATION) <120> NEW ENTOMOPATHOGENIC FUNGI ISARIA JAVANICA Pf04 HAVING          CONTROLLING OF BEMISIA TABACI Q TYPE AND MICROBIAL INSECTICIDE          FOR CONTROLLING BEMISIA TABACI Q TYPE CONTAINING THE SAME <130> P2011-2165 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 579 <212> DNA <213> ISARIA JAVANICA Pf04 <400> 1 ctgggaggga tcattaacga gttttttcaa actccctaac cctttgtgaa catacctatc 60 gttgcttcgg cggactcgcc ccggcgtccg gacggccctg cgccgcccgc gacccggacc 120 caggcggccg ccggagaccc acaaattctg tttctatcag tctttctgaa tccgccgcaa 180 ggcaaaacaa atgaatcaaa actttcaaca acggatctct tggttctggc atcgatgaag 240 aacgcagcga aatgcgataa gtaatgtgaa ttgcagaatt cagtgaatca tcgaatcttt 300 gaacgcacat tgcgcccgcc agcattctgg cgggcatgcc tgttcgagcg tcatttcaac 360 cctcgacacc ccttcggggg agtcggcgtt ggggaccggc agcataccgc cggccccgaa 420 atacagtggc ggcccgtccg cggcgacctc tgcgtagtac tccaacgcgc accgggaacc 480 cgacgcggcc acgccgtaaa acacccaact tctgaacgtt gacctcggat caaggtagga 540 ctacccgctg aacttaagca tatataaaac ccggaagaa 579

Claims (8)

Tobacco powder has a control effect on Q type and is a new insect pathogenic fungus Isaria Javanica represented by SEQ ID NO: 1. javanica ) Pf04 strain (KACC93122P). The method of claim 1,
Tobacco powder has a control effect against Q type nymphs,
New Insect Pathogenic Fungus Isaria Jabnica javanica ) Pf04 strain (KACC93122P). The method according to claim 1 or 2,
The strain is isolated from arachnid infected pest,
New Insect Pathogenic Fungus Isaria Jabnica javanica ) Pf04 strain (KACC93122P). The novel insectogenic pathogenic fungus Isaria Javanica of claim 1. javanica ) containing Pf04 strain (KACC93122P),
Microbial insecticide for tobacco powder control. 5. The method of claim 4,
The microbial insecticide includes any one or more of the cells or culture of the strain,
Microbial insecticide for tobacco powder control. The microbial insecticide of any one of claims 4 or 5,
To control tobacco powder Q type nymph,
Microbial insecticide for tobacco powder control. The microbial insecticide of any one of claims 4 or 5,
In the form of granules, powders, liquid hydrating agents, hydrating agents, or encapsulating agents,
Microbial insecticide for tobacco powder control. A method for controlling tobacco powder Q type nymphs by applying the microbial insecticide of claim 4 to crops or soil.

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