KR20110005356A - A short-term storage technique of an egg parasitoid, trichogramma sp. using developmental quiescence induced by low temperature - Google Patents

Abstract

PURPOSE: A short-term storage technique of an egg parasitoid, trichogramma sp. is provided, which can store an egg parasitoid, trichogramma sp. for a specified period without degeneration of parasitic ability. CONSTITUTION: A short-term storage technique of an egg parasitoid, trichogramma sp. comprises a step storing an egg which is expired in six days after parasitizing of an egg parasitoid, trichogramma sp. at 5-10deg.C within five weeks. An egg expired for 3-5 days after parasitizing is stored at 9-10deg.C. An egg expired for 4 days after parasitizing is stored at 10deg.C.

Description

A Short-term Storage Technique for Egg Parasitoid, Trichogramma sp. Using Developmental Quiescence Induced by Low Temperature}

The present invention relates to a storage method of egg parasitic rods of the genus Trichogramma , and more particularly to a short-term storage method at low temperatures.

Trigmatidae is a small taxa of 0.2-1.5 mm in size, belonging to the genus Chalcidoidea, which includes 620 species of 80 genera. Of these, two genera, Trichogramma and Oligosita , contain 145 and 110 species, respectively, accounting for about 40% of the bees and the whole (Pinto and Sthouthamer, 1994). Trichogramma is a globally distributed genus with a wide parasitic range, including moths, beetles, flies, stink bugs, extinct species, lumber and grass dragons (Pinto and Sthouthamer, 1994).

Trichogramma , an egg parasite, is an important biological control factor and is processed in 32 million ha annually in agricultural ecosystems worldwide (Li, 1994). The technique of radiating egg parasites in this genus has been applied to agricultural sites as one of biological control strategies to manage pests by achieving natural equilibrium between predators and predators. In addition, this method can be used in combination with various pest control techniques, and is currently widely used for comprehensive pest control. Among the parasitic rods belonging to this genus, Trichogramma sp. Nabis101 and the like. Trichogramma sp. Nabis101 is released in agroecological systems due to its wide range of target pest hosts, and it is expected that its use will be further expanded with the development of a hybrid technology with microbial pesticides.

Trichogramma barley moth (Sitotroga cerealella) is relatively easy Ju Hu-breeding pests, alrak people nabangryu (Anagasta kuehniella), people nabangryu (Corcyra cephalonica ) has been raised, and now mass farming techniques have been developed with reduced costs in alternative host or incubation conditions (Flanders, 1930; Li, 1994; Smith, 1996). However, since the demand for these egg parasites is irregular in time, there is a need for a technique capable of effectively storing them while reducing the parasitic deterioration of the mass bred live egg parasites.

As an effective storage method of conventional parasitic rods, the parasitic rod's dormant physiology is used for long term preservation (Stinner, 1977; Boivin, 1994; Chang et. al . , 1996). Some species belonging to the genus Trichogramma develop dormancy during the transitional period by work and temperature organic factors (Pizzol, 1978; Zaslavski and Umarova, 1990). This dormant development allows long term (about 6 months) preservation of egg parasites (Ventura Garcia et al. al ., 2002). However, there is a problem that many egg parasites do not show a clear dormant development.

In addition, since relatively short-term (about 1 month) storage is possible by simple low temperature treatment, such low temperature storage method has been studied in species belonging to Trichogramma (Pitchet et. al ., 2002; Ventura Garcia et al ., 2002; Rundle et al ., 2004). However, no reliable short-term cold storage method has yet been proposed that can be maintained for a period of time without a substantial change in the parasitic capacity of parasitic rods.

[Citations]

Boivin, G. 1994. Overwintering strategies of egg parasitoids. pp. 219-244. In Biological control with egg parasitoids, eds. by E. Wajnberg and SA Hassan. CAB International, Wallingford, England.

Chang, YF, MJ Tauber and CA Tauber. 1996. Reproduction and quality of F1 offspring in Chrysoperla carnea : differential influence of quiescence, artificially-induced diapause and natural diapause. J. Insect Physiol. 42: 521-528.

Curl, GD and PP Burbutis. 1977.The mode of overwintering of Trichogramma nubilale Estle and Davis. Environ. Entomol. 6: 629-632.

Dutton, A. and F. Bigler. 1993. Flight activity assessment of the egg parasitoid Trichogramma brassicae (Hym .: Trichogrammatidae) in laboratory and field conditions. Entomophaga 40: 223-233.

Flanders, SE 1930. Mass production of egg parasites of the genus Trichogramma . Hilgardia 4: 145-167.

Hutchinson, LA and JS Bale. 1994.Effects of sublethal cold stress on the aphid Rhopalosiphum padi . J. Appl. Ecol. 31: 102-108.

Jalali, S.K. and S.P. Singh. 1992. Differential response of four Trichogramma species to low temperatures for short term storage. Entomophaga 37: 159-165.

Jung, JK, JH Park, DJ Im and TM Han. 2005. Parasitism of Trichogramma evanescens and T. ostriniae (Hymenoptera: Trichogrammatidae) to eggs of the asian corn borer, Ostrinia furacalis (Lepidoptera: Pyralidae). Kor. J. Appl. Entomol. 44: 43-50.

Kim, G., H. Heo, J. Park, Y. Yu, E. Hahm, S. Kang, K. Kwon, K. Lee and Y. Kim. Efficacy of an integrated biological control of an egg parasitoid, Trichogramma evanescens Westwood, and microbial insecticide against the oriental tobacco budworm, Helicoverpa assulta (Guene) infesting hot pepper. Kor. J. Appl. Entomol. 47: 435-445.

Kumar, G.A., S.K. Jalali, T. Venkatesan, R. Stouthamer, P. Niranjana and Y. Lalitha. 2009. Internal transcribed spacer-2 restriction fragment length polymorphism (ITS-2-RFLP) tool to differentiate some exotic and indigenous trichogrammatid egg parasitoids in India. Biol. Control 49: 207-213.

Li, LY 1994. Worlwide use of Trichogramma for biological control on different crops: a survey, pp. 37-53. In Biological control with egg parasitoids, eds. by E. Wajnberg and SA Hassan. CAB international, Wallingford, England.

Pinto, JD and R. Stouthamer. 1994. Systematics of the Trichogrammatidae with emphasis on Trichogramma . pp. 1-36. In Biological control with egg parasitoids, eds. by E. Wajnberg and SA Hassan. CAB International, Wallingford, England.

Pintureau, B. and J. Daumal. 1995. Effects of diapause and host species on some morphometric characters in Trichogramma (Hym .: Trichogrammatidae). Experientia 51: 68-72.

Pitcher, SA, MP Hoffmann, J. Gardner, MG Wright and TP Kuhar. 2002. Cold storage of Trichogramma ostriniae reared on Sitotroga cerealella eggs. Biocontrol 47: 525-535.

Quinn, P. J. 1985. A lipid-phase separation model of low-temperature damage to biological membranes. Cryobiology 22: 128-146.

Rundle, BJ and AA Hoffmann. 2003. Overwintering of Trichogramma funiculatum Carver (Hymenoptera: Trichogrammatidae) under semi-natural conditions. Environ. Entomol. 32: 290-298.

Rundle, BJ, LJ Thomson and AA Hoffmann. 2004.Effects of cold storage on field and laboratory performance of Trichogramma carverae (Hymenoptera: Trichogrammatidae) and the response of three Trichogramma spp. ( T. carverae , T. nr. Brassicae , and T. funiculatum ) to cold. J. Econ. Entomol. 97: 213-221.

SAS Institute, 1988. SAS / STAT user's guide, Release 6.03, Ed. Cary, N.C.

Smith, SM 1996. Biological control with Trichogramma : advances, successes, and potential of their use. Annu. Rev. Entomol. 41: 375-406.

Stinner, R.E. 1977. Efficacy of inundative releases. Annu. Rev. Entomol. 22: 515-531.

Ventura Garcia, P., E. Wajnberg, J. Pizzol and MLM Oliveira. 2002.Diapause in the egg parasitoid Trichogramma cordubensis role of temperature. J. Insect Physiol. 48: 349-355.

Zaslavski, VA and TY Umarova. 1990. Environmental and endogenous control of diapause in Trichogramma species. Entomophaga 35: 23-29.

It is an object of the present invention to provide an effective low temperature storage technique for egg parasites in the genus Trichogramma . In particular, in the present invention, Trichogramma sp. It is intended to provide a short-term cold storage method that can be stored for a certain period of time without substantial deterioration of parasitic capacity for egg parasitic rods, which are currently being commercialized and distributed as biological control factors in Korea such as Nabis101.

Other objects and advantages of the present invention will be described below and will be better understood by practice of the present invention.

In the present invention, in view of the importance of the development of the parasitic rods to be exposed to such low temperatures with the effective low temperature selection in order to store without loss of parasitic capacity using the physiological resting state, it is confirmed through experiments and exposed to low temperatures Parasitic rod development was determined. In the present invention, it is confirmed that short-term storage is possible without profound change in parasitic ability by prolonging the emergence of adults through cold storage of egg parasites under such determined conditions.

Based on the experimental results, in the present invention,

There is provided a short-term storage method of the egg parasites in the genus Trichogramma ( Trichogramma ) comprising the storage of parasitic eggs that have elapsed within 6 days after parasitization of the egg parasites at 5 ° C to 10 ° C. Preferably, the parasitic eggs 3 to 5 days after the parasites are stored at 9 ° C to 10 ° C, and more preferably the parasitic eggs 4 days after the parasites are stored at 10 ° C.

Short-term low temperature storage method of the present invention tricot the Lamma (Trichogramma) in Al and wasps can be applied to both surfaces, especially tricot the Lamma in the nabiseu 101 (Trichogramma sp. Nabis101) and its oil genetically, for the most likely seen wasps, for It can be effectively applied to Trichogramma brasiliensis, etc. which is closest to the whole.

Another aspect of the present invention is the storage method of the present invention is currently commercialized, commercialized Trichogramma sp. It is to clarify the genetic properties of this egg parasite so that it can be used for Nabis101 and its most similar egg parasite. To this end, in the present invention, sequencing was performed based on the internal transcribed sequence (ITS) present in the ribosomal RNA gene region to determine the exact biological species characteristics of the target parasite.

Egg parasitic rods in the storage method of the present invention could be maintained for 5 weeks without significant degradation of viability, sex ratio, lifespan and parasitic capacity. The short-term low temperature storage method according to the present invention can be utilized as an effective storage method that can control the supply timing of a large number of eggs parasitic breeding without substantial deterioration of parasitic capacity.

In the present invention, it is confirmed through experiments that short-term storage is possible by prolonging the appearance of an adult through cold storage of egg parasites. Low temperature conditions were more effective at 10 ° C. than 4 ° C. to cause temporary developmental arrest. This may be due to the effect of delaying development while reducing cold damage (Quinn, 1985) such as cell membrane failure at low temperatures. 10 ° C also corresponds to the developmental zero temperature of many species of the genus Trichogramma (Curl and Burbutis, 1977; Rundle and Hoffmann, 2003). Therefore, at this temperature, it is difficult to induce growth arrest effect. In addition, lower temperatures will consume energy to maintain viability, resulting in reduced viability. Therefore, in the present invention, when stored within five weeks, the preferred temperature range is set to 5 ℃ to 10 ℃. At relatively low temperatures of 5-7 ° C., shorter storage of 1-3 weeks would be desirable. More preferably stored at 9 ℃ to 10 ℃, most preferably at 10 ℃ when stored within 5 weeks.

The development of parasitic rods to be exposed to low temperatures is also important in order to store egg parasites without significant change in parasitic capacity. It takes about 11 days for the emergence of parasites from parasitic rods at room temperature. After 4 and 7 days of parasitic storage, the parasitic embryos that were parasitic on day 4 at 10 ° C. were parasitic on day 7. The viability was shown to be higher for 5 weeks of treatment compared to fetuses. Therefore, in the present invention, the parasitic eggs 3 to 5 days after the parasite are set to the development of the parasitic rods to be exposed to the low temperature conditions, and more preferably the parasitic eggs 4 days after the parasitic are exposed to the low temperature conditions. Set the parasitic rod to the development state.

In general, even if survival is maintained after cold storage, damage can occur in several parasitic features. These damages result in decreased fertility (Jalali and Singh, 1992; Chang et. al . , 1996), reduced flight ability (Dutton and Bigler, 1995), decreased adult lifespan (Jalali and Singh, 1992; Hutchinson and Bale, 1994), and male tactile rescue (Pintureau and Daumal, 1995). . In the present invention, it was confirmed whether the sex ratio, lifespan, and parasitic decay of the adult females after storage at 10 ° C. As a result, there was no difference in other features, but the parasitic power decreased slightly after 2 weeks of storage. It is estimated that after two weeks there may be energy consumption even at 10 ° C, and this energy consumption may have affected fertility. Regardless of the storage period, however, the age-related parasitic rhythms of female adults remained unchanged. Taken together, it was found that the initial survival of host eggs at low temperature for short-term storage at low temperature was no difference in survival rate up to 5 weeks and had parasite capacity of over 80%. Therefore, it can be concluded that the egg parasites can be substantially maintained for 5 weeks without significant degradation in viability, sex ratio, life span and parasitic capacity when stored according to the conditions set forth in the present invention.

The low temperature short-term storage method of the present invention is tricogram (TrichogrammaAll the parasitic rods of the genus can be applied.Trichogramma sp. Egg parasitic rods most similar to Nabis101, for example genetically closestTrichogramma brasiliensis It can be applied primarily to the back. To this end, the present invention is currently commercializedTrichogramma sp. Genetic characteristics of Nabis101 (rice beetle bees, Nabisu, Yecheon, Korea) were first analyzed based on ITS sequences. As a result, this paperT. evanescensAlthough known as (Kimet al, 2008) The sequencing data show a big difference from this species, but ratherT. brasiliensisShowed the highest sequence homology with. Kumaret al. (2009) divided the Trichogrammatid egg parasites into three classes based on the diversity of ITS-2 lengths.Trichogrammatoidea armigeraWowTr . bactraeGroup Ⅱ, which includes 570-620 bpT. achaeae,T. embryophagum AndT. japonicumGroup III of 500-550 bpT. chilonis,T. dendrolimi,T. evanescens,T. mwanzai,T. brassicae AndT. pretiosumIncluded. PCR primers used to comply with this classification have been identified in the present invention.Trichogramma sp. Incorporating the Nabis101 sequence results in amplification up to 800-1370, resulting in an amplification length of 571 bp. In other words,Trichogramma sp. Nabis101 belongs to Group II, meaning Nabis101 belongs to Group II.T. evanescens orT. chilonisIt is backing that it is not. On the other hand, the closest similarity to phylogeneticsT. brasiliensisITS-2 analysis yields 525 bp, based on this classification.Trichogramma sp. The difference with Nabis101 is shown. Putting together the above sequence resultsTrichogramma sp. Nabis101 is one of the parasitic rods in this taxonT. brasiliensisClosest to, but showed inconsistent differences as shown in the ITS-2 analysis.

Hereinafter, the present invention will be described in more detail with reference to specific examples and experimental examples. However, these examples are only for illustrating the present invention in more detail, the scope of the present invention is not limited by these examples.

Materials and methods

1. Test insect

Egg parasitic rod Trichogramma sp. The Nabis101 (ssaljom albeol, Ltd. nabiseu, Yecheon, South Korea), give people alrak moth (Cadra cautella ) eggs were bred in indoor conditions (26 ± 2 ℃, 74 ± 5% relative humidity, 18L: 6D). Forage, 40% sugar water was fed to adult parasites.

2. Cold storage treatment

The host eggs were exposed to egg parasites for 24 hours, and then 4 eggs or 7 days old at 25 ° C. were treated at 200 ° C. and 4 ° C. and 10 ° C., respectively. The treatment period was up to 5 weeks, and the surviving rate was shifted to room temperature weekly and calculated as the number of allelic adults for 10 days.

3. Biological characterization after cold storage

Parasitic rods parasitic rods (35 per petri dish) were stored at 10 ℃ for 1, 2, 3, 4, 5 weeks and then parasitic rod development conditions (26 ± 2 ℃, 74 ± 5% relative humidity, 18L) : 6D), the allegory rate, female to female ratio, and longevity of the adult adult were investigated. In addition, after mating female adult larvae with male adult for 24 hours according to each storage period, one female adult was supplied with 100 Jouleac wild moth eggs every 24 hours, and parasitic rate and life span were examined every 24 hours. It was.

4. Statistical Analysis

The mean ± standard deviation values are plotted using the SigmaPlot (Systat Software, Inc., Point Richmond, CA, USA) program. Experimental observations were performed two-way ANOVA analysis by PROC GLM using SAS program (SAS Institute, 1988) to determine treatment effect. All treatment means were compared by least significant difference test at Type I error = 0.05.

5. Ribosomal DNA of ITS  Region Sequencing

Using 10 parasitic rods Kang et al . Genomic DNA was extracted by the method of (2005). Target DNA region amplification was performed by Vrain et. al . (1992) presented primers were used. The forward reaction primer sequence was 5'- TTGATTACGTCCCTGCCCTTT-3 'and the reverse reaction primer sequence was 5'-TTTCACTCGCCGTTACTAAGG-3'. The PCR reaction solution consisted of 10X PCR buffer (5 μl), dNTP mixture (2.5 mM each (5 μl), 25 pmol forward primer (2 μl), 25 pmol reverse primer (2 μl), Taq DNA polymerase ( 1 μl) and deionized distilled water (34 μl). Machine oil (20 μl) was added thereto, followed by reaction using a PTC-100 (MJ Research, Minnesota, USA) instrument. PCR reaction conditions were 35 repeated amplification reactions, each reaction was composed of 1 minute at 94 ℃, 1 minute at 46 ℃ and 72 minutes 2 minutes, then carried out a residual synthesis reaction at 72 ℃ 10 minutes. The PCR amplification thus obtained was recombined with TA vectors (pGEM, Promega, Madison, USA), and sequenced in both directions using Sp6 and T7 primers. The determined nucleotide sequences were tracked by similar sequences using the BlastN search program of GenBank (NCBI, http://www.ncbi.nlm.nih.gov ), which in turn was reconstructed by the DNAstar program (Version 5.01, DNAstar Inc., Madison, USA) was used to determine homology between sequences.

result

One. Trichogramma sp . Nabis101 Cold storage conditions

Two different low temperatures were treated to eggs of different parasitic states in order to develop a storage technique for egg parasites (FIG. 1). It takes about 11 days for the emergence of adult parasites from parasitic conditions at room temperature (Jung et al ., 2005), 4 and 7 days post-parasitic parasites were defined and selected as early and late parasitic rod development for storage, respectively. Storage efficiency was temperature sensitive and showed high viability at 10 ° C rather than 4 ° C. After 3 weeks of low temperature treatment, 4 ° C. showed a sharp drop in survival rate. In addition, at 10 ° C., the initial parasitic growth was higher than the late parasitic development for 5 weeks of treatment. The late parasitic developmental status showed low viability with only 50% survival after 5 weeks of storage. Based on the experimental results, subsequent experiments were performed based on eggs 4 days after parasitics at 10 ° C.

2. After cold storage Parasitic  Change in biological properties

The above results showed that the parasitic rods of early developmental condition can survive about 5 weeks under the condition of 10 ℃. In order to determine whether the surviving parasitic rods show normal adult living ability, the daily allegorical rhythm, female ratio, longevity, and parasitic rate were analyzed according to storage period (FIGS. 2A to 2D). The allegorical rhythm began to appear 4 days after transfer to room temperature, showed a maximum peak on day 5, and the remaining number of appearances on the next day (FIG. 2A). This allegorical rhythm showed the same pattern regardless of the storage period at 10 ° C (Χ ² = 3.46; df = 10; P = 0.9685). The proportion of females of the fabled adults according to the cold storage period (Fig. 2b) showed a difference of about 55-65% according to the treatment, but it was not statistically significant (Χ ² = 0.20; df = 5; P = 0.991). Adult lifespan (FIG. 2C) was about 6 days and lifespan was reduced in 5 parking storage treatments. Parasiticity (FIG. 2D) spawned approximately 52 per female for untreated treatment, while decreasing as the storage period continued, representing less than 40 adult adults appearing in stored eggs for 5 weeks.

Parasitic rhythm of egg parasitic rods was analyzed according to cold storage (FIG. 3). Regardless of the storage period (F = 0.11; df = 5, 695; P = 0.9902), all of them became parasitic and declined rapidly after 24 hours of fable (F = 981.75; df = 4, 695; P <0.0001 ) Trends.

3. Commercial Parasitic  Genetic characteristics

Currently available Trichogramma sp. The ITS sequence of Nabis101 was analyzed (SEQ ID NO: 1). To identify both military ITS, DNA amplification was performed based on the conserved sequences of 18S rRNA and 28S rRNA. The size of the amplification was 1,447 bp and included all 5.8S rRNAs as well as some 18S rRNAs and 28S rRNAs as well as both ITS regions.

This sequence was compared with the DNA sequence data registered in NCBI to analyze the similarity (Fig. 4). High agreement was found in T. minutum , T. ostrinae , T. chilonis , T. exiguum and T. brasiliersis centered on E value, but the highest similarity was found to be T. minutum as 97%. The results are shown in Table 1 below. The results were analyzed systematically using Trichogramma sp. Nabis101 is T. chilonis And T. brasiliersis were close, and T. minutum with high similarity in Blast search showed systematic differences (FIG. 4). Taken together, these parasites have the highest genetic relationship with T. brasiliensis .

1 shows Trichogramma sp. The parasitic eggs of day 4 ('4 pp') and day 7 ('7 pp') after parasitic storage of Nabis101 were stored at two low temperature conditions of 4 ° C and 10 ° C, respectively.

Figure 2 shows the change in biological properties of the parasitic rods after cold storage, Figure 2a is a daily allegorical rhythm, Figure 2b is a female ratio, Figure 2c is a life span, and Figure 2d is a result of analyzing the parasitic rate. Parasitic eggs were stored at 10 ° C for different weeks ('wk') and used for analysis.

3 is a result of analyzing the parasitic rhythm of egg parasitic rods under cold storage. Parasitic eggs were stored at 10 ° C for different weeks ('wk') and used for analysis.

4 is Trichogramma sp. This is the result of analyzing the similarity by comparing the base sequence of Nabis101 with the DNA sequence data registered in NCBI.

<110> Andong National University Industry-Academic Cooperation Foundation <120> A Short-term Storage Technique of an Egg Parasitoid, Trichogramma          sp. Using Developmental Quiescence Induced by Low Temperature <160> 1 <170> KopatentIn 1.71 <210> 1 <211> 1447 <212> DNA <213> Trichogramma sp. Nabis101 <400> 1 tgattacgtc cctgcccttt gtacacaccg cccgtcgcta ctaccgattg aatgatttag 60 tgaggtcttc ggactgatgc gcggcaatgt gtttactacg ttgccgattg tgccgggaag 120 atgaccaaac ttgatcattt agaggaagta aaagtcgtaa caaggtttcc gtaggtgaac 180 ctgcggaagg atcattaacg ttcttgctcc cgctgtgcag ctttgtatag agaggcacac 240 gcggagagat aaacaagcac cgacgtgtat cttttgtcga tgacgacgac gacgacgacg 300 acgacgaatc gttgttgcct gcgtctcgtc tcgacatttg cgtcttcgcg tgaaactgct 360 gcgcgcactt tcttacgctg tttttggttt tttttctctg cgcgtgtgtg cgtgtcacgc 420 acacacacac gcacagttga attttttttg gttttctgct ttctttggtt gtgctcggtt 480 tcctcgtgcg atcatcgaga tcgagagaga aatttacaat gaacacgaaa gaaaggacaa 540 tagaaaacga caataaaaaa ctttgcgctt ttttttttct tctcggtgtg cttgccgcgc 600 gccgagagcg agaaaaaaaa aagaaacaaa catgtgattt tctttcgctc tcgactcctc 660 gcgcgcgcgc gctacgtgcg tgtgtgctgc gagtagttcg ggggtgttgg caaaattaac 720 aaactttttt atgattaccc tgaacggtgg atcacttggc tcgtggatcg atgaagaacg 780 cagctaattg cgcgtcaact tgtgaactgc aggacacatg aacatcgaca tttcgaacgc 840 acattgcggt ccacggatct cgttcccgga ccacgcctgg ctgagggtcg tttataaaaa 900 cgaacccgac tgctctctcg caagagagag cgttgatctg ggcgctcgtc tctaactcta 960 cgcgctcttt ttcttccgag aggagggtag tgtagcagtg tgatacgtcg cctcaaacga 1020 gacgcaagaa aaaagatgaa ttcgttcgtc tagctggcgc gcgcgcgctt accgcttgga 1080 gagtacgcga gtacttccga tcgttctgcg tcgagtcccg gagctttctc gactactcgt 1140 cgagcagcgg accgacgtct agcacacgat caggctcgtc catgcatcgg tcattgaacg 1200 cgcgcgcgct cgtgcccttt tacgcgctca cacacacaca cacacacacg cactgtgtgc 1260 tgtgctctgt gctgctgctg ctgcgtatca gagtcaaagt ggctagctcg aagaagcttt 1320 ttgctgaacg agtctttttt ctcgatcgac gacctcagag caggcgaggc cacccgctga 1380 atttaagcat attattaagc ggaggaaaag aaactaacga ggatttcctt agtaacggcg 1440 agtgaaa 1447  

Claims (4)

A method of short-term storage of egg parasites in the genus Trichogramma ( Trichogramma ) comprising storing the parasitic eggs that have elapsed within 6 days after the parasites of egg parasites within 5 weeks at 5 ℃ to 10 ℃. The method of claim 1, wherein the parasitic eggs 3 to 5 days after the parasites are stored at 9 ° C to 10 ° C. The method of claim 1, wherein the parasitic eggs stored 4 days after the parasites are stored at 10 ° C. The method of claim 1, wherein the egg parasitic rod is Trichogramma sp. Nabis 101 genus Trichogramma short-term storage method of egg parasitic rods.

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