곤충의 머리를크게 cranium(머리겉뼈)과 mouth part(입틀)로 나눌 때 머리 겉뼈 부분을 frontoclypeal area(이마둔순부), parietals(머리옆뼈), occipital arch(뒷머리 아치), postocciput(뒷머리뒷판) 및 subgeneal area(아래뺨부)로 나누던 것 중 clpeus(얼굴앞판)와 subgeneal area(아래뺨부)를 입틀쪽으로 옮겨서 입틀을 peristome(입둘레)과 4개의 부속지(appendages)로 합치는 것이 타당할 것으로 생 각된다.
따라서 epistomal sulcus(입윗판홈)와 subgeneal sulcus(아래뺨홈)는 한 개의 연 결된 홈이므로 peristomal sulcus(입둘레홈) 또는 cranio-stomal sulcus(머리와 입 틀 사이홈)로 칭하는 것을 제안한다.
The alfalfa weevil is a destructive invasive pest of Chinese milk vetch, a main source of honey products. Because apiarists and farmers disfavor the use of insecticides, a specialist larval parasitoid wasp, Bathyplectes anurus, was introduced for biological control of the weevil. Although the introduced parasitoid initially had showed very low percentages of parasitism in early 1990’s, it gradually increased its parasitism and hence effectiveness of control. Finally, the parasitoid successfully suppresses the alfalfa weevil in 2000’s. Although this is a good example of successful biological control, one major question remains unanswered: why does the parasitoid expand and increase so slowly? Field and laboratory experiments show: (1) the parasitoid prefers alfalfa weevils on common vetch rather than those on Chinese milk vetch; (2) Chinese milk vetch fields (= rice paddies) are not the source of parasitoids, but weedy areas with common vetch plants around rice paddies are the main habitat of the parasitoid where it can successfully overwinter; (3) hunger level of adult parasitoids searching in Chinese milk vetch fields is much higher than that in common vetch fields; (4) common vetch plants have extra-floral nectary glands which are found to be a main sugar source for the introduced parasitoid; and the effectiveness of the parasitoid is rather low in areas where common vetch is rather rare. These results strongly indicate that the conservation of parasitoid habitats, in this case, weedy areas with common vetch, is a key factor affecting the success of biological control with B. anurus parasitoids. Thus, great care should be taken whether we can make the agro-environment suitable for natural enemies.
In insects, the sense of smell is a complex and highly sensitive modality, governing essential decisions such as choice of food and oviposition sites. Plants emit substantial amounts of volatile organic compounds (VOCs), and the characteristic scent represents a dynamic communication channel. Understanding this odor-mediated system is critically important in the habitat management and in the largest view of the conservation biological control. Here we suggest that the fitness of the egg parasitoid ,an important biological control agent of the green vegetable bug may be improved through the understanding of chemical communication in the biological control system. Initially, the attractiveness of four flowering plant species, chosen from apanelofplants based on the longevity of the eggparasitoid on these plants, to T. basalis was assessed, which indicated significant behavioral attraction of T. basalis to the buckwheat flowers. Subsequently, and GC-MS analysis were carried out to identify the olfactory-active VOC semanated from buck wheat, demonstrating that the antennalol factory receptor neurons of T. basalis were responsive to some aliphatic acids as well as a few common plant volatiles. In behavioral bioassays using synthetic formulation based on the chemical and electrophysiological analysis, T. basalis exhibited significant behavioral attraction to the synthetic blend at optimum dose. The findings, in a wider perspective, form the basis for further improvement of the use of the companion plants that may increase the insects’ communities’ ability to persist in an environment.
In Korea, in 1970s and 1980s a few research for biological control program was conducted mostly for agricultural field crop and forest insect pests. Since 1990s when the greenhouse cultivation area increased significantly and the insect pest problem thereby increased highly, the needs for practical biological control program for greenhouse pests increased accordingly. In 1990s especially when Tetranychus urticae in strawberry, Trialeurodes vaporariorum and Liriomyza bryoniae in cucumber and tomato, and Aphis gossypii in cucumber became a major problem, demand for use of commercially produced biological control agents such as Phytoseiulus persimilis, Encasia formosa, and Aphidius colemani increase rapidly but these commercially available biological control agents in foreign countries were not allowed to be imported at that time. Therefore, temporary pilot study for application these natural enemies began in mid 1990s by the National Institute of Agricultural Science and Technology, RDA and some provincial agricultural extension center. The greenhouse biological control research and natural enemy industry began to flourish when 7 species of exotic biological control agents were officially permitted for import in 2003 and the promotion program for horticultural insect pest management using natural enemies was implemented from 2005. Unfortunately, when this promotion program was terminated in 2010 the Korean natural enemy industry began to collapse and also biological control research activity decreased significantly. In this presentation, past and current status of the biological control research activity and natural enemy industry in Korea will be addressed and the future perspective will be cautiously discussed.
Trichogramma is widely used in biological control of many Lepidoptera insect pests in the world. Although many species of host eggs, e.g. Sitotroga cerealela, Corcyra cephalonica, Ephestia kuehniella, Mamestra brassicae and even artificial host eggs are explored in mass production of Trichogramma, eggs dissected from the ovaries in female of Chinese (Oak) tussah moth (Antheraea pernyi) are still the most popular host eggs in China. Here I introduce technology processes of Trichogramma mass production that are popularly used in China, including: 1) selecting female cocoons from mixed sexes commercial cocoons, 2) cold storage of cocoons, 3) warming up the cocoons, 4) collecting female moths and dissecting ovaries, 5) separating, washing and drying host eggs, 6) multiplying Trichogramma with the host eggs, 7) producing Trichogramma products, i.e. the host egg cards, 8) storing Trichogramma under low temperatures, 9) releasing Trichogramma in the field. Nevertheless, problems are frequently encountered in these processes, e.g. high mortality of long-term storage and multiplying too many generations etc. To solve these problems and explore a method for long-term storage, we studied the diapause of T. dendrolimi in detail. We found that the optimal processes for diapause induction were exposing hosts for parasitization at 26℃ for 8 h before keeping them at 26℃ for another 40 h, and finally moving them into 10℃ for 4 weeks. The optimal processes for diapause termination were setting the diapause Trichogramma under 1℃ for two months and then exposing them at 26℃ until their emergence.
Biological control as a tactic of environmentally safer agriculture has not been widely accepted by farmers. In Japan integrated biological control with native natural enemies is rapidly developing for greenhouse vegetables, because of high prices of commercially mass-reared biological control agents. We have studied the effective use of natural enemies at farm level, focusing on generalist predators, although there are differences in using natural enemies for augmentation in greenhouses and conservation in open fields. Recently, however, insectary plants (crops) have been considered as a key component for enhancing natural enemies. We have examined the effectiveness of candidate insectary plants on natural enemies and implemented biointensive IPM with insectary plants at farm-level. Our recent study suggested that some herbs and crops are highly effective on improving the survival and reproduction of generalist predators. By growing such plants in vegetable greenhouses and open-fields, natural enemy populations could be enhanced and thus stably persist even after targeted prey populations are exploited. Moreover, insectary plants may result in a shift of IPM from level I to level II.
There are relatively few cases of classical biological control in Korea. During 1904 to 1905 the wooly apple aphid, Eriosoma langigerum, had been happen to invade into Korea from Japan together with imported seedling of apple and its damage spread to Jinampo near Pyoung-Yang City and it occurred all across the country by 1934. In 1934 a parasitic wasp, Aphelinus mali, was introduced and released to apple orchards and found established in nature. And it was the first case of classical biological control in Korea. Afterward couple of classical biological control programs against invaded pests were conducted mainly by the RDA with variable results. For example the orange scale, Ceroplastes rubens, and the Eggar which were successfully controlled by introduction Anicetus beneficus and Thecodiplosis pinico, respectively.
Nucleopolyhedrovirus (NPV), which belongs to Baculoviridae, is a rod-shaped, double-stranded circular DNA virus which infects arthropods, mostly insects. NPVs are highly species-specific, and make unique crystalline polyhedral structure made of polyhedrin protein. The NPVs do not replicate in mammalian cells, are safe to human, and can be observed the viral replication with conventional compound microscope, plus the availability of susceptible insect cell lines, therefore, the NPVs became an ideal model system to study basic virology.
Also, NPVs became popular because of its applications for baculovirus expression vector system (BEVS). A foreign gene will be cloned into a shuttle vector, and introduced to the NPV chromosome to make recombinant virus. This NPV will produce the protein in culture cells or host insects under the control of the strong polyhedrin promoter. So far, the commercially available BEVS has been widely used because of its high efficiency and eukaryotic characteristics, however, the hidden bottleneck is finding new useful genes which will maximize the capability of BEVS.
Since the human genome project, next generation sequencing technique (NGS) is getting useful in life science field along with the development of sequence analysis algorithms and increase of computational power. Especially, RNA-seq and de novo sequence assembly technics make discovery of new genes easier even in a non-model species with a proteomics approach, and these useful tools will be the key to catalyze the insect biotechnology.
The genus Diadegma is a well known parasitoid group and some are known to have symbiotic virus, PDV. A novel IV was discovered from the calyx of D. fenestrale female. This virus was named as D. fenestrale Ichnovirus (DfIV). The encapsidated DfIV genome contains 65 circular DNA segments with an aggregate size of 247,191 bp. Based on BLAST analysis, a total of 120 ORFs were predicted as follows: rep; 48, cys-motif; 11, vinnexin; 10, vankyrin; 9, PRRP; 3 and other unassigned genes (39). These viral genes were expressed in lepidopteran hosts (Phthorimaea operculella and Plutella xylostella) after parasitization which means DfIV genome segments were integrated into lepidopteran hosts. This study was focused on this result. Based on gene expression profile, candidate promoter and integration motifs were selected and then, fused with eGFP as a reporter gene. Modified DfIV genome segment was ligated to a commercial containing f1 ori and Ampr gene to propagate in E. coli. We have named this fusion vector as pIN. The construction methodology of pIN and its application would be further discussed in detail.
캐드헤린 (cadherin)은 캴슘 의존성 막관통 단백질로써 세포질영역, 막관통영역, 세포외영역의 세 부분으로 구성되어 있다. 캐드헤린은 세포외세포와 세포간 또는 세포와 조직간 밀착연접을 형성하는 역할을 수행하며, 생체내에서 주로 세포와 세 포 또는 기질간 정보 교환과 조직의 항상성을 유지한다. 최근 곤충에서 캐드헤린은 나비목과 파리목을 대상으로 한 연구에서 비티독소 (Cry toxin)의 중요한 작용점으 로 알려지기 시작하였으며, 비티독소에 대한 곤충의 감수성에 영향을 미친다.
옥수수의 뿌리를 가해하여 심각한 피해를 야기하는 딱정벌레목의 western corn rootworm (WCR, Diabrotica virgifera virgifera)에서 얻어진 캐드헤린 유전자를 토대로 캐드헤린 반복서열의 일부를 대장균을 통하여 재조합단백질 (DvCad1 CR8-10)로 만들 수 있었다. 딱정벌레목에 속하는 일부 해충들을 대상으로 DvCad1 CR8-10은 Cry toxin과의 혼합처리에서 일부 Cry toxin의 반수치사농도 감소를 나 타내었다. 또한 Cry toxin과 캐드헤린 단백질간의 결합력 조사와, 캐드헤린 유전자 에 대한 RNA interference 연구를 통해서 캐드헤린이 Cry toxin의 중요한 수용체라 는 것을 확인할 수 있었다.
비티독소를 이용한 농업해충과 위생해충의 방제는 지금까지 성공한 생물적방 제로 인식되고 있으며, 현재에는 다양한 중요작물에 비티독소가 도입되어 발현되 는 형질전환 비티작물로 까지 비티공학기술이 발전되어 왔다. 그러나 야외에서 배 추좀나방에 대한 비티독소의 저항성 문제가 처음으로 출현하며, 딱정벌레목까지 다양한 해충들이 비티에 대한 저항성을 보이고 있다. 따라서 이러한 해충들에 대한 비티의 효과적인 이용과 저항성 개체의 출현을 조절하기 위해서 곤충유래의 캐드 헤린 단백질을 이용한 생물공학적 해충관리가 도입 될 것으로 전망된다.
Insect cuticle is a complex biocomposite material consisting of three major morphologically distinct layers, the waterproofing envelope, the protein-rich epicuticle and the chitin/protein-rich procuticle. Structural cuticular proteins (CPs) and the polysaccharide chitin are the major components of the exo- and endocuticular layers that comprise the procuticle. During cuticle tanning (sclerotization and pigmentation), CPs are cross-linked by quinones derived from the oxidation of catechols, resulting in hardening of the exoskeleton. However, the factors that lead to synthesis and assembly of cuticular regions with differing mechanical properties are not well understood.
To gain a better understanding of the development and differentiation of rigid cuticle, we performed transmission electron microscopic (TEM) analysis of elytral cuticle (highly sclerotized and pigmented forewing) from 2 d-old pupae to 9 d-old adults of the red flour beetle, Tribolium castaneum. In 2-3 d-old pupae, pupal cuticle separated from the underlining epidermal cells (apolysis), and outermost envelope and protein-rich epicuticle begun to form. A numerous horizontal chitinous laminae and vertical pore canals were evident in the procuticle of 4-5 d-old pupae. By one day after adult eclosion, less-compact horizontal chitinous laminae were deposited, followed by block-type cuticular layers with no horizontal laminae were formed by 9 days. These results will lead to a) a better understanding of insect cuticle formation, structure and mechanics, b) the potential for development of novel insect control agents that target cuticle physiology, and c) the production of biomimetic materials with physical properties like those of the insect exoskeleton for use in biomedical or other technological devices.
This work was supported by NRF (NRF-2012R1A2A1A01006467).
Symbiotic bacteria are common in insects. Because symbiotic bacteria are known to intimately affect the various aspects of insect host biology, ideally insects can be controlled by manipulating their symbiont. However, the attempts to control insects through their symbiont have been very limited. The paucity of the insect pest control using their symbiont is most likely due to the poor understanding of the symbiotic interactions between host insect and symbiont, which is attributed to the difficulty in cultivation of insect symbionts. However, the recently established bean bug, Riptortus pedestris, symbiotic system provides good opportunities to study insect’s symbiont in molecular level through their cultivable symbionts. Bean bugs acquire genus Burkholderia cells from environment and harbor them as their gut symbionts in the specialized posterior midgut. The genome of the Burkholderia symbiont was sequenced, and the genomic information has been used to generate the genetically manipulated Burkholderia symbiont strains. After orally administering the mutant Burkholderia symbionts into bean bugs for symbiotic association, the bacterial colonization levels in the host gut and host phenotypes were analyzed. As a result, we have identified novel symbiotic factors necessary for establishing successful association with host. Our recent understandings on the bacterial symbiotic factors demonstrate a great possibility to control the bean bug pest using genetically modified Burkholderia symbiont.
Insects constitute the largest and most diverse group of animals on Earth. They also serve as the hosts or nutrient sources for an immense assemblage of parasites, pathogens, and predators, ranging from bacteria and fungi to plant and animals. More than 700 known fungal species from 100 genera have adopted an entomopathogenic lifestyle. These fungi are widely distributed, occurring in aquatic, forest, and agricultural habitats, and are often used as active ingredients in microbial insect pest control agents. Their mode of action against insects involves the attachment of conidia to the insect cuticle, followed by germination, cuticle penetration, and internal dissemination throughout the insect. During this process, secreted enzymes, protein toxins, and secondary metabolites can be used by the fungus to overcome the host immune system, modify host behavior, and defend host resources against competing pathogens and saprophytes. In some cases, the host insect relationship has been found to be associated with bioactive fungal metabolites. These metabolites exhibit a wide variety of insecticidal, antibacterial, antifungal, anticancer, antioxidant, and antiviral activities. Using molecular techniques and phylogenetic analyses, both the asexual (anamorphic) stages and sexual (teleomorphic) stages of entomopathogenic fungi have recently been shown as linked together. Therefore, entomopathogenic fungi, especially in complex with the host insect, might be a promising source of bioactive molecules of pharmaceutical and various industrial interests. Here, we evaluated the antimicrobial activity of entomopathogenic fungi metabolites against plant pathogenic bacteria and fungi for the use in agriculture. The radicals scavenging activity and the anticancer activity were also evaluated for pharmaceutical interests.
After firstly identified sex pheromone components of Bombxy mori, those of many insect pests were synthesized by organic chemistry methodology. These synthesized components were used for monitoring, mass trapping, and mating disruption during five decades. For identification of pheromone biosynthesis mechanisms and control to many pests bring to serious damages also were proceeded. The transcriptome analysis from pheromone glands by Next Generation Sequence (NGS) showed many genes and pathway involved on sex pheromone biosynthesis.. The two main genes involved on production of acetate and alcohol, and aldehyde from fatty acid, fatty acid desaturase and fatty acid reductase (FAR) were identified and functional characterized via gene introduction to Brewer’s yeast Saccharomyces cerevisiae. This S. cerevisiae now used as a mediator as well as cell factory for sex pheromone producing. Recently, One group was published that the plant factory for producing via genetically modified plant (tobacco, Nicotiana benthamiana) as a step of semisynthetic preparation. These trials will be suggest that firstly, the possibility of yeast as a molecular toolbox to produce pheromone components and secondly, a novel and cost-effective way of producing moderate to large quantities of pheromones with high purity and a minimum of hazardous waste.
For replacement of methyl bromide(MB) in Korea, there are several chemical options being introduced and developed in preplant and postharvest sections. In soil case, methyl-isothiocyanate(MITC) generating fumigant such as dazomet and metham sodium are currently being adapted for prevent from spreading soilborne disease and nematode, although there will be more successful application technology needed to develop in domestic circumstances. Also, ethanedinitrile(EDN), is newly developed chemical, could be one of the options to eradicate notorious soilborne diseases and nematodes near future. Ethyl formate(EF) now is showing the potential to replace MB in terms of consumer and worker safety and unwanted phytotoxic damage in routine MB fumigation. A phosphine gas(PH3) could be the last option to protect the import and export stored grains from stored grain insects and to solve the current issue such as PH3-resistant insect and consumer safety issues to replace metal phosphide. EDN also should be extended its use in not only timber and log fumigation in quarantine purpose but in use of national heritage and forest to keep wood infested pest safer. This presentation will be good opportunities to share the information and discuss with further research needs in current MB alterative program.
This study was done to evaluate the efficacy of Vapormate (ethyl formate 16.7 % + CO2) as an methyl bromide-alternative fumigant. Some insect pests and mites were found under calyx of sweet persimmon fruits, including grape-myrtle scale, two-spotted spider mite, Amblyseius eharai. Vapormate fumigation was equally toxic to miticide- susceptible and -resistant T. urticae showing 100 % mortality at higher concentration than 146.7 and 211.0 mg/L, respectively. In persimmon-filled desiccator, summer and overwintering types of T. urticae showed 100 % mortality at higher concentration than 163.7 and 265.8 mg/L, respectively. Sweet persimmon damage from vapormate fumigation was not shown by the days after treatment. Tolerance of each live stage of grape-myrtle scale was in descending order: egg < nymph < adult. EF 141.5 mg h L-1 for 6 h at 5 ℃ killed 100 % of the larvae of D. punctiferalis.
The final scaled-up fumigation confirmed the efficacy of EF to target pests and non-phytotoxicity on persimmons. In conclusion, EF may become a MB alternative fumigant for sweet persimmon, and the LC99 values of EF may be used to practical applications to all kinds of insects and mites on harvested sweet persimmon.
Methyl bromide(MB) is a fumigant for quarantine and pre-shipment(QPS) that can eradicate target pests within short fumigation periods. However, MB, an ozone-depleting substance, is scheduled to be phased out in the second decade of this century. In contrast to MB alternatives for soil fumigation, there is no ideal alternative fumigant to MB for QPS of perishable commodities. It is important that all target insect pests are killed within a short fumigation time.
The citrus mealybug(Planococcuscitri), a problematicquarantine pest from Korea, was exposed to ethyl formate and phosphine for different times and to different concentrations of EF(0.0∼33.4 mg/L) and PH3(0.25∼1.0 mg/L) in small desiccators. The eggs showed more tolerance than the nymphs and adults. To control the eggs, they were treated with a mixture of EF and PH3 for 2 and 4-hr. The egg stage showed 100% mortality with EF:PH3(25.1:0.25, 16.7:0.5, and 16.7:1.0 mg/L) at 4-hr. Treatment with a mixture of EF and PH3 for 4-hr at 8°C can control all stages of the citrus mealybug.
목재와 섬유, 지류 등의 유기질 문화재는 그 재질적 특성과 여름철 고온다습한 우리나라의 기후 특성으로 인해 충균에 의한 생물적 피해가 크게 발생한다. 우리나 라의 주요 문화재 가해곤충은 8목에 이르며, 특히 딱정벌레목과 흰개미목, 벌목의 피해가 심하게 발생한다. 이러한 문화재의 곤충 피해를 방제하기 위해서 살충살균 력과 침투력이 뛰어난 훈증제가 사용되어 왔으며, 1982년 최초로 야외 목조건축물 에 대한 훈증처리가 실시되었다. 기존에 문화재 훈증에 사용되어온 Methyl Bromide(이하 MB)는 오존층 파괴물질로서 2015년부터 사용이 금지될 예정이며, 이에 따른 대체가스선정 및 친환경 생물방제방안의 개발이 요구되는 실정이다.
문화재용 대체훈증가스 선정 및 개발을 위해 Ethylene Oxide(이하 EO)를 유효 성분으로 하는 약제 2종과 Sulfuryl Fluoride(이하 SF)의 문화재 가해충균에 대한 살충살균력을 평가하고, 각종 문화재 재질에 대한 안정성(반응성)을 평가하여 현 장적용을 위한 매뉴얼을 제작하였다. 또한 검역, 임산 등 다양한 분야에서 MB 대체 물질로 평가 중인 Ethanedinitrile(이하 EDN)의 살충살균력 및 목재침투성, 잔류성 과 함께 각종 문화재 재질에 대한 안정성 평가를 수행하였다. 이 외에도 인체와 환 경에 무해하고 문화재 재질에 영향을 미치지 않는 생물방제법으로서 야외 목조건 축물에 대한 저산소처리 및 천연약제의 적용성을 평가하였으며, 이에 그 결과를 보 고하고자 한다.
식물체정유란 식물체에 함유된 휘발성분을 증기증류를 이용하여 추출한 액상 형태의 추출물로 살충, 살균, 항세균, 살선충 등 다양한 활성들이 보고되어 있다. 특 히 최근 메틸브로마이드 등 가스형태의 훈증제들이 가지고 있는 독성 등 여러 가지 부작용 때문에 식물체정유들이 가지고 있는 살충, 살균,항세균 활성들이 관심을 받 고 있다. 식물체정유에는 다양한 화합물들이 함유되어 있는데(cynohydrins, monoterpenoids, sesquiterpenoids, sulphur compounds, thiocyanates 등) 이들 화 합물들은 단독 혹은 협력하여 다양한 활성들을 보인다. 이들 식물체 정유 혹은 정유 유래 화합물들은 발육단계별로도 다양한 활성에 차이를 보이며 농약에 저항성인 계체에 대해서도 활성을 나타낸다. 그리고 식물체정유는 이산화탄소나 ethyl formate 등과 혼합하였을 때 훨씬 좋은 활성을 나타낸다는 보고가 있다. 식물체정 유 성분들의 살충 혹은 살균 활성을 나타내는 기작에 대한 연구는 일부 수행이 되었 으나 현재까지 정확한 기작은 알려진 것이 별로 없는 실정이다. 본 발표에서는 식물 체정유 및 정유성분들의 살충, 살균, 항세균 등 다양한 활성을 알아보고 앞으로 식 물체정유를 이용한 병해충 방제제의 전망을 알아보고자 한다.
훈증제는 식물검역에 문제되는 해충을 방제하기 위해 사용되는데 특히, 목재류 검역에 있어서 많이 사용되고 있다. 그 중 phosphine(PH3)은 메틸브로마이드 (Methyl bromide)보다 해충에 대한 독성이 강하고 침투성 및 작업자의 안전성 측 면에서 우수한 것으로 알려져 저장류에 대한 검역용 훈증제로 널리 사용되고 있다. 포스핀은 상온에서 가장 효과적이라고 알려져 있으며 고온과 저온에서는 그 효과 가 떨어지는 것으로 알려져 있는데, 목재해충인 북방수염하늘소(Monochamus saltuaris)와 흰개미(Reticulitermes speratus)에 대한 PH3의 훈증활성을 온도조건 에 따라 살펴보고자 한다.
북방수염하늘소에서 포스핀의 훈증활성은 5°C보다20°C에서 더 높은 활성을 보였으나 알과 번데기 시기에서는 온도와 상관없이 포스핀의 활성이 거의 나타나 지 않는 것을 알 수 있었다. 또한, 흰개미에서 PH3의 훈증활성을 탐색한 결과 북방 수염하늘소와 마찬가지로 저온에서 활성이 감소하였는데 5°C보다 15°C 에서 8배 정도 높은 살충활성을 보였다. 그러나 5°C에서 산소(O2)의 함량을 높였을 때 훈증 효과가 증가하는 것을 알 수 있었다. 이는 포스핀이 기문을 통해 해충 체내로 들어 가 cytochrome C 산화효소를 저해함으로써 산소호흡을 억제시켜 해충을 질식시키 는데, 저온에서는 호흡량이 줄어들기 때문에 포스핀의 흡수량이 떨어졌다가 산소 포화도가 높아지면서 흡수하는 산소와 포스핀의 양 또한 증가하기 때문으로 생각 된다. 일반적으로 포스핀의 훈증활성은 상온에서 효과적이고 저온에서 감소하지 만 산소협력과 같은 방법을 사용한다면 효과적인 훈증활성을 얻을 수 있다고 판단 된다. 그러므로 목재류에 대한 훈증활성을 증가시킬 수 있는 방법에 대한 지속적인 연구가 필요할 것으로 생각된다.