基于GIS的马铃薯甲虫扩散与河流关系研究——以新疆沙湾县为例

马铃薯甲虫是一种重要的外来入侵害虫,自1993年传入我国新疆北部地区以来,迅速扩散传播,对我国马铃薯等茄科作物生产造成严重威胁。为明确马铃薯甲虫传播扩散与河流等生态因子之间的关系,结合马铃薯甲虫虫情调查,利用地理信息系统(GIS)的空间分析功能,对新疆沙湾县马铃薯甲虫发生与河流的关系进行研究。结果表明:在沙湾县内,马铃薯甲虫扩散方向与河流流向基本一致,均呈由南向北方向;不同缓冲区内,马铃薯甲虫危害级别差异极显著,缓冲区危害级别从大到小依次:缓冲区2(1-2 km)>缓冲区4(3-4 km)>缓冲区5(4-5 km)>缓冲区1(0-1 km)>缓冲区3(2-3 km);不同河流流域内马铃薯甲虫危害级别也差异极显著,危害级别排序依次是:金沟河>安集海河>玛纳斯河>西岸大渠。     

Northward range expansion requires synchronization of both overwintering behaviour and physiology with photoperiod in the invasive Colorado potato beetle (Leptinotarsa decemlineata)

Photoperiodic phenological adaptations are prevalent in many organisms living in seasonal environments. As both photoperiod and growth season length change with latitude, species undergoing latitudinal range expansion often need to synchronize their life cycle with a changing photoperiod and growth season length. Since adaptive synchronization often involves a large number of time-consuming genetic changes, behavioural plasticity might be a faster way to adjust to novel conditions. We compared behavioural and physiological traits in overwintering (diapause) preparation in three latitudinally different European Colorado potato beetle (Leptinotarsa decemlineata) populations reared under two photoperiods. Our aim was to study whether behavioural plasticity could play a role in rapid range expansion into seasonal environments. Our results show that while burrowing into the soil occurred in the southernmost studied population also under a non-diapause-inducing long photoperiod, the storage lipid content of these beetles was very low compared to the northern populations. However, similar behavioural plasticity was not found in the northern populations. Furthermore, the strongest suppression of energy metabolism was seen in pre-diapause beetles from the northernmost population. These results could indicate accelerated diapause preparation and possibly energetic adjustments due to temporal constraints imposed by a shorter, northern, growth season. Our results indicate that behavioural plasticity in burrowing may have facilitated initial range expansion of L. decemlineata in Europe. However, long-term persistence at high latitudes has required synchronization of burrowing behaviour with physiological traits. The results underline that eco-physiological life-history traits of insects, such as diapause, should be included in studies on range expansion.     

Hormonal regulation of proline synthesis and glucose release in the fat body of the Colorado potato beetle, Leptinotarsa decemlineata

In the adult males of the Colorado potato beetle, Leptinotarsa decemlineata, corpora cardiaca extract injected in vivo gives rise to an increase in glucose and a decrease in alanine concentration of the haemolymph. The regulation of proline synthesis and glucose release in the fat body of the Colorado potato beetle was investigated further in vitro. Proline regulates its own synthesis by a feedback inhibition. Moreover, a factor present in extracts from the corpora cardiaca stimulates synthesis in the fat body in vitro. This effect was demonstrated with corpora cardiaca extracts from beetles, locusts and cokroaches. Also, synthetic adipokinetic hormone stimulates proline synthesis in the fat body of the Colorado beetle. In addition, a release of glucose from the fat body of the beetle could be evoked by the addition of locust and beetle corpora cardiaca extracts or synthetic adipokinetic hormone. The physiological significance of both effects is discussed.     

Potato field colonization by low-density populations of Colorado potato beetle as a function of crop rotation distance

Monitoring of 10 and 12 commercial potato, Solanum tuberosum L., fields in 2004 and 2005, respectively, confirmed for a low-density population of Colorado potato beetle, Leptinotarsa decemlineata (Say), that potato fields nearest to the previous year's potato fields are significantly more colonized by this beetle than more distant fields. This pattern is partially explained by the presence of a reservoir of colonizers estimated at 35% of the season-long colonizing population in 2004 and 2005. These beetles, which emerged before potato plants broke the ground, were ready to establish themselves on nearby potato plants. The colonizing Colorado potato beetles dispersed within the maximum range of 1.5 km over a season, and the colonization risk for the new crop decreased with distance from the previous year's crop. There was no evidence that rotation distance delayed colonization. In terms of pest management, although the findings confirm that only long 1.5-km rotations can prevent Colorado potato beetle colonization, they also demonstrate that short rotations of 100 m or more can make substantial contributions to pest management programs for low-density beetle populations.     

Estimation of the role of predatory epigeic beetles (Coleoptera: Carabidae, Staphylinidae) in regulation of pest population density in agroecosystems

Epigeic polyphagous predatory beetles can regulate the pest abundance. The range of population densities at which regulation is possible is specific to each predator-prey subsystem and can be determined experimentally. In the subsystem of ground and rove beetles (Carabidae and Staphylinidae) and the cabbage maggot Delia brassicae Bouché and in that of Carabus hampei Küst. and the Colorado potato beetle Leptinotarsa decemlineata Say, regulation occurs at low pest densities: in the former subsystem, at oviposition rates not exceeding 3 eggs per day per plant, and in the latter, at the pest density varying from 1 to 36 eggs per potato plant. Within these density ranges, both the absolute and relative number of pest individuals eliminated by the predatory beetles increase. The maximum fraction of the pests destroyed by these entomophages is observed at medium prey population densities, which corresponds to functional response of type III (Holling, 1965).     

A freeze-dried diet to test pathogens of Colorado potato beetle

The Colorado potato beetle is an important pest on potato, eggplant, and tomato. Because Colorado potato beetles develop resistance to insecticides quickly, new methods are needed for control. Bacillus thuringiensis is the only bacterium to successfully control Colorado potato beetle. Until recently, one of the drawbacks to testing bacteria against the Colorado potato beetle has been the lack of an artificial diet for screening. Previous artificial diets will only be consumed by Colorado potato beetle larvae when fresh. To improve storage, we developed a freeze-dried diet, based on a 96-well plate, suitable to feed larvae for the duration of a bioassay. Individual diet components were tested both for their effect on insect growth and on pathogen toxicity. When the preservatives, methylparaben and sorbic acid, were removed from the diet, the average weight of second instar larvae increased from 7.9 mg to greater than 9.8 mg. The preservatives inhibited the growth of two of the bacteria tested, Photorhabdus luminescens HM and Chromobacterium sp. PRAA. The removal of these preservatives also allowed for fungal growth and reduced survival from 94 to 38%. Removing diet preservatives, that inhibited the growth of Chromobacterium sp. PRAA, increased the total mortality of the larvae as well as reducing the time needed to kill 50% of the larvae. Compared to incorporation of bacteria into molten diet, the total mortality of Colorado potato beetle fed either P. luminescens HM or Chromobacterium sp. PRAA on freeze-dried diet doubled. Preparation of freeze-dried diet need not be synchronized with the insect or the pathogen. The freeze-dried diet gave consistent results as measured by low control mortality and pathogen toxicity over time.     

Effect of crop rotation distance on populations of Colorado potato beetle (Coleoptera: Chrysomelidae): development of areawide Colorado potato beetle pest management strategies

The Colorado potato beetle, Leptinotarsa decemlineata (Say), overwinters adjacent to field edges and infests nearby fields in the spring, primarily by walking. Crop rotation is known to be an effective cultural control against Colorado potato beetle populations limiting spring infestations. Spatial separation is an important consideration in optimizing the rotational effect because long-distance rotations have been shown to reduce Colorado potato beetle infestations. To determine the effect of long-distance rotations on Colorado potato beetle populations in commercial Wisconsin potato production, a geographic area of >18,200 ha (>45,000) acres in southern Portage County, Wisconsin, was selected as an experimental area for areawide pest management of the Colorado potato beetle. From 1997 to 1999, beetle populations at edges within each potato field in this region were determined by spring and fall field sampling. The rotational distance between current and previous potato fields was measured and analysis was run between the distance and Colorado potato beetle populations. Long-distance rotations of >400 m were an effective cultural control management strategy to limit adult beetle infestations in the spring. This strategy can be optimized when collaborating growers are able to maximize their rotational distances by coordinating their rotational schemes within large areawide, geographic locations. Deploying long-distance rotations within a geographic area over many years would limit Colorado potato beetle populations and could result in a significantly reduced Colorado potato beetle populations entering fields in the spring.     

Flight take-off performance of Colorado potato beetle in relation to potato phenology

The flight take-off frequency of adult Colorado potato beetles, Leptinotarsa decemlineata (Say), from potato plants, Solanum tuberosum L. 'Red Pontiac' at the bloom stage of development was 2.2-2.5-fold that of Colorado potato beetle from plants at the vegetative stage. Tests were conducted in a flight chamber over a period of 3 h. Prefeeding Colorado potato beetles for 48 h on potato plants at the bloom or at the vegetative stage before placing them into the flight chamber resulted in the same significantly higher flight take-off frequency from potato plants at the bloom stage than from plants at the vegetative stage. These results demonstrate that the factor in potato plants in bloom that stimulates the flight take-off of the Colorado potato beetle is independent of the feeding history of the beetles and begins acting only when the beetles are in the presence of the plant. According to these results, the dispersal of adult Colorado potato beetles from potato fields in bloom to younger potato fields with plants at the vegetative stage, previously reported in the literature, is at least partly explained by the effect of plant phenology on the frequency of flight take-off. Results confirm the value of planting potato fields of similar phenology over as wide an area as possible to reduce Colorado potato beetle dispersal between fields. Results also imply that staggering the planting dates of conventional potato refuge areas near Colorado potato beetle transgenic or conventionally resistant potato fields is a sound management practice, because it promotes the movement of wild beetles over to the adjacent younger resistant crops.     

Pymetrozine causes a nontarget pest, the Colorado potato beetle (Coleoptera: Chrysomelidae), to leave potato plants

Pymetrozine is a selective insecticide that targets aphids. Published assessments of the effects of pymetrozine on nontarget organisms focus mainly on predatory insects, and they rarely indicate toxicity. In a laboratory bioassay, survival of Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), larvae was not affected by pymetrozine exposure. We subsequently used pymetrozine to implement low-aphid-density treatments in a field experiment that involved separate manipulations of Colorado potato beetle density. Unexpectedly, the addition of Colorado potato beetle adults and eggs did not increase the densities of Colorado potato beetle larvae in plots that were sprayed with pymetrozine (applied with water and an adjuvant). In control plots sprayed with water and adjuvant (without pymetrozine), addition of Colorado potato beetles increased densities of their larvae. Data collected on a smaller scale suggest that a behavioral mechanism underlies the population-level pattern: Colorado potato beetle larvae become more active and are less likely to remain on a host plant after exposure to pymetrozine. Thus, potato, Solanum tuberosum L., growers who use pymetrozine against aphids also might benefit in terms of Colorado potato beetle control.     

Predation on Colorado Potato Beetle Eggs by Generalist Predators in Research and Commercial Potato Plantings

Field studies quantified predation on Colorado potato beetle [Leptinotarsa decemlineata(Say)] eggs and determined the relationship between predation and egg mass abundance in research and commercial potato plantings in eastern North Carolina. Predator exclusion experiments were conducted weekly in research plantings. In addition, egg mass density and predation on egg masses were monitored throughout the season in research plots and commercial potato fields. Predation was an important source of mortality for Colorado potato beetle eggs. Survivorship of eggs exposed to predators was consistently, significantly lower than survivorship of eggs protected from predation. Averaged over 2 years, the mean survivorship of eggs protected from predation was 69%, compared with 26% survivorship of eggs exposed to predation. Regression analysis failed to detect any relationship between egg mortality due to predation and egg abundance. These results imply that efforts to reduce Colorado potato beetle populations selectively will not be offset by an according decline in abundance of natural enemies and therefore should be fully compatible with naturally occurring biological control.     

Comparative analysis of immune responses in Colorado potato beetle larvae during development of mycoses caused by <Emphasis Type="Italic">Metarhizium robertsii</Emphasis>, <Emphasis Type="Italic">M. brunneum</Emphasis>, and <Emphasis Type="Italic">M. pemphigi</Emphasis>

Development of mycoses and progress of humoral and cellular immune responses were compared in larvae of the Colorado potato beetle Leptinotarsa decemlineata infected with entomopathogenic fungi Metarhizium robertsii, M. brunneum and M. pemphigi. The larvae were found to be highly susceptible to the strains of M. robertsii and M. brunneum but weakly responsive to M. pemphigi. The extent of susceptibility to the pathogens was not related to the stimulating effect of epicuticular extracts on fungal growth. Metarhizium pemphigi, which is non-specific to the Colorado potato beetle, did not cause any significant changes in the immune response and did not colonize the hemocoel. When infected with M. robertsii and M. brunneum, the larvae exhibited an increase in hemocyte count during the early stage of mycosis (day 2) followed by a drastic decrease on day 3. The immunocompetent cells, plasmatocytes and granulocytes, exhibited the greatest decrease. Elevated phenoloxidase activity was recorded in the hemolymph and cuticle on days 2 and 3 post-infection. These changes in the immune responses correlated with strain-specific virulence. Thus, the immune response in Colorado potato beetle larvae is an important factor, which determines differences in the development of mycoses caused by different Metarhizium species.     

Cold tolerance during larval development: effects on the thermal distribution limits of Leptinotarsa decemlineata

Insects' cold tolerance during their development is a surprisingly understudied subject in ecology, despite the fact that subzero temperatures during the growing season are common at high altitudes and latitudes. Subzero temperatures can have detrimental effects on organisms, restricting a species' range. This study addresses the question whether night frosts during the growing season have an instant or delayed negative impact on larval mortality of the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae). We also tested whether populations from the centre (Poland) and margins (Russia) of the distribution range of L. decemlineata differ in their responses to subzero exposure and a low rearing temperature. Larvae of three ages were subjected to a subzero temperature (−4 °C for 3 h simulating night frost) twice, after which they were reared on a fluctuating temperature regime of 10–15 °C. These rearing conditions imitated cool summer temperatures beyond the beetles' current range, such as in Finland. Individuals of both populations were highly cold tolerant, as only 3.1% of larvae died immediately following the subzero treatment. Nonetheless, the low rearing temperature was harmful to beetles of both populations. It caused high larval (ca. 90%) and overwintering (ca. 80%) mortality. As beetle performance was affected solely by rearing temperature, low temperatures during the growing season rather than night frosts apparently retard the beetle's northern expansion.     

Influence of fertilization on the Colorado potato beetle, Leptinotarsa decemlineata, in organic potato production

The abundance of the Colorado potato beetle, Leptinotarsa decemlineata (Say), in organically grown potato did not change significantly in response to increasing rates of dehydrated poultry manure. However, peaks of abundance of larvae were shifted forward in time in response to the high rate of organic fertilizer. Tests using excised foliage showed that the shift was not caused by differential larval mortality or longer developmental times. Time allocation to resting, walking, and feeding by adults was similar regardless of fertilizer rate. Adult foliage consumption was unaffected by organic fertilizer rates in no choice tests and significantly affected in few choice tests. A 22% longer larval development time on plants treated with low fertilizer rate than on plants with high rate was the most significant effect. Even though maximum plant height, canopy, biomass, and yield were significantly smaller in the organic than in conventional plots, the suitability of the plants was not affected except for reduced feeding by summer beetles. Summer adults spent less time feeding and consumed two to five times less foliage on organic potato than on inorganically fertilized and conventionally produced plants. The overall influence of fertilizer on Colorado potato beetle populations was limited and therefore can only play a secondary role in management strategies for organic potato. Avoidance of excessive organic fertilizer that promotes short larval development time and extension of the period over which large Colorado potato beetle larvae are present should be recommended.     

Effects of temperature on survival and preimaginal development rates of Colorado potato beetle on potato and horse-nettle: potential role in host range expansion

The effect of temperature on the ability of Colorado potato beetles (Leptinotarsa decemlineata, Say) to use horse-nettle (Solanum carolinense L.) as a host plant was determined for larvae from colonies originating from two geographically separated populations, one adapted to horse-nettle (NC) and the other unadapted to horse-nettle (MA). Survival and developmental rate on horse-nettle and potato were measured for larvae from both colonies over a range of constant temperatures (12–30 °C) and one fluctuating temperature regime (22 °C to 30 °C). The ability of Colorado potato beetles to use horse-nettle as a larval host was strongly influenced by temperature, but the effects of temperature differed greatly between beetles from the two colonies. Survival of adapted larvae on horse-nettle was highest and comparable to that on potato at the constant 30 °C and the fluctuating temperature regime. Below 30 °C, survival of adapted larvae decreased drastically but some larvae survived at all temperatures except the lowest (12 °C). In contrast, survival of unadapted larvae to adult occurred only at 30 °C, and was low (10%). At lower temperatures, all larvae died. On potato, the effect of temperature was less dramatic, and consistent across colonies. At 12 °C, survival to adult was poor (ca. 10%), but at higher temperatures, survival increased sharply and larvae from both colonies survived equally well. On potato, small but statistically significant differences in developmental rates between beetle colonies were detected at the constant but not at the fluctuating temperature regimes. Also, the developmental day degree requirements (DD) and the low temperature development threshold (T0) values for the various developmental stages did not differ between colonies on potato. On horse-nettle, development times for both colonies were always significantly longer and DD requirements were greater than on potato. At 30 °C, the only constant temperature at which larvae from the unadapted colony completed development, the development rate to adult emergence was similar to that of beetles from the adapted colony. Differences between colonies in performance on horse-nettle were not a result of host-independent, genetically based differences in the thermal requirements of the two populations. Our findings are consistent with the hypothesis that adaptation to horse-nettle by Colorado potato beetle may be facilitated by a genotype × environment interaction involving temperature. These findings have important implications for host plant utilization, host range expansion and selection of pest biotypes adapted to plant resistance traits used in crop protection.     

Resistance potential of colorado potato beetle (Coleoptera: Chrysomelidae) to novaluron

Novaluron (Rimon 10 EC), a novel insect growth regulator, could play an important role in future management programs for Colorado potato beetle, Leptinotarsa decemlineata (Say). Studies were conducted to determine the potential of Colorado potato beetle to develop resistance to novaluron before its widespread use in Colorado potato beetle management. Second instars of an imidacloprid-resistant Colorado potato beetle strain exhibited reduced susceptibility (2.5-fold) to novaluron. The toxicity of novaluron to this strain was synergized by S,S,S-tributyl phosphorotrithioate (DEF) but not by piperonyl butoxide (PBO), suggesting that esterase-based detoxification mechanisms were responsible for novaluron resistance. Bioassays with treated potato foliage found that a single low- or medium-rate novaluron application was highly persistent under field conditions, resulting in up to 85% mortality of second instars 5 wk after treatment. Thus, intense selection pressure for novaluron-resistant Colorado potato beetle may continue long after population densities have been reduced below an economic threshold level. In a national survey, the susceptibility of second instars to a novaluron diagnostic dose was determined for 27 different field populations collected from six Canadian provinces in summer 2003. Despite no previous exposure to novaluron, mortalities at the diagnostic dose ranged from 55 to 100%. Although novaluron has several characteristics that should delay resistance development in insect pests, these results highlight the need for judicious use of the compound in management of Colorado potato beetle.     

Diverse enzymatic specificities of digestive proteases, 'intestains', enable Colorado potato beetle larvae to counteract the potato defence mechanism

In response to insect attack, high levels of proteinase inhibitors are synthesised in potato leaves. This can cause inefficient protein digestion in insects, leading to reduced growth, delayed development and lower fecundity. It has been suggested that Colorado potato beetle overcomes this defence mechanism by inducing the production of a set of cysteine proteases that are resistant to potato proteinase inhibitors. Experiments with gut extracts showed that these proteases have unusual inhibition profiles as they are not inhibited by most of the cystatins but are strongly inhibited by thyropins. In this study we have isolated three cysteine proteases from adapted guts of Colorado potato beetle larvae, named intestains 1, 2 and 3, the first cysteine proteases known to be involved in extracellular protein digestion. The N-terminal sequences suggest their classification into the papain family. Intestains differ in substrate specificities and inhibitory profiles. Their substrate specificities suggest that intestains 1 and 2 are general digestive enzymes, while intestain 3 has a more specific function. The inhibitory profile of intestain 1 is similar to that of proteases of the papain family. However, the Ki values for the interaction of intestain 2 with the same set of inhibitors are several hundred fold higher, which would enable the enzyme to circumvent the potato defence mechanism characterised by high concentrations of protease inhibitors in attacked potato leaves. A further, different strategy of the Colorado potato beetle to avoid potato defence is exhibited by intestain 3, which is able to cleave off the N-terminus of model cystatin and thus inactivate the inhibitor. These results suggest that the Colorado potato beetle combines different strategies to counteract plant defence mechanisms.     

Monitoring diapause development in the Colorado potato beetle, Leptinotarsa decemlineata, under field conditions using molecular biomarkers

A multiplex PCR protocol was developed using five diapause-regulated genes to monitor diapause development of the Colorado potato beetle under field conditions. A total of 870 beetles from the Red River valley of North Dakota and Minnesota, USA, were screened for three consecutive years. Out of the 32 possible expression profiles, eight could be arranged in chronological order of diapause development. These eight profiles account for over 92% of the beetles surveyed. Intra-population variation in diapause phenotypes was observed in the field. Some beetles were already in the diapause initiation phase in June when the day length was greater than 17 h. Inter-seasonal variation in the timing of diapause development was also noted. The greatest differences were before the day length decreased to less than 15 h. Anomalies in the results, e.g., the presence of the diapause maintenance phase profiles in beetles collected on the potato plants, argue that laboratory results are not always equivalent with what is observed under field conditions.     

不同寄主植物上马铃薯甲虫种群生长发育的比较研究

马铃薯甲虫Leptinotarsa decemlineata (Say)是马铃薯Solanum tuberosum等茄科作物上的一种毁灭性害虫。为了探明寄主植物对新疆马铃薯甲虫种群生长发育的影响, 本研究通过非选择性试验测定了马铃薯、 茄子Solanum melongena和番茄Lycopersicon esculentum 3种栽培寄主及野生寄主中亚天仙子Hyoscyamus pusillus对马铃薯甲虫种群生长发育、 存活、 繁殖及生命表参数的影响。结果表明： 马铃薯甲虫幼虫虽然能够取食番茄, 但幼虫的发育历期长、 存活率低且蛹不能羽化, 表明番茄不是新疆马铃薯甲虫种群的适宜寄主。马铃薯、 茄子、 中亚天仙子3种寄主植物对该虫卵、 幼虫、 蛹的存活率和发育历期及成虫产卵前期没有显著影响, 但对蛹重和繁殖力影响显著。取食不同寄主植物后, 该虫蛹重和繁殖力从大到小的次序为： 马铃薯＞茄子＞中亚天仙子; 种群的净增殖率(R₀)、 内禀增长率(r_m)和种群趋势指数(I)从大到小依次为： 马铃薯＞茄子＞中亚天仙子。因此马铃薯为新疆马铃薯甲虫种群的最适宜寄主, 其次为茄子和中亚天仙子, 而番茄为不适宜寄主。     

The relationship between predator density, community composition, and field predation of Colorado potato beetle eggs

Conservation biological control programs seek to increase natural enemy densities through the adoption of more benign farming practices, under the assumption that higher predator densities will lead to more effective pest suppression. However, predator–predator interference may lead to diminishing returns in improved pest control as predator densities increase. We examined the relationship between predator density and predation rates on Colorado potato beetle eggs in production potato fields. These potato fields naturally spanned a 10-fold range in predator density, due to differences in management practices. Periodically through the growing season we simultaneously measured predator densities and subjected sentinel eggs masses to predation, allowing us to correlate predator density and egg predation for each field on each sample date. Egg predation rates were significantly positively correlated with total predator densities, a correlation that was not improved when predator densities were scaled to reflect differences in feeding rates on potato beetle eggs of the constituent predator taxa. There was no correlation between per-capita egg predation rates and predator density, and so no evidence that predator interference increased with increasing predator density. We divided predators into six dominant taxa—dwarf spiders, crab spiders, minute pirate bugs, big-eyed bugs, damsel bugs, and Lygus bugs (together constituting 93% of all predators collected), and a seventh group, “other predators” that included all other, less common, taxa—and examined correlations between all predator combinations and egg predation rates. The highest correlation was between combined densities of the six most common predator taxa, excluding only the “other predators” grouping. This suggests that predators may be largely equivalent in their impact on Colorado potato beetle eggs, and that field scouts might be able to ignore uncommon predator taxa when sampling for natural enemies.     

Partial purification of a novel insect antidiuretic factor from the Colorado potato beetle, Leptinotarsa decemlineata (Say) (Coleoptera: Chrysomelidae), which acts on Malpighian tubules

In this study, we report for the first time the presence of an antidiuretic factor in the head of the Colorado potato beetle (Leptinotarsa decemlineata (Say)) which acts directly on Malpighian tubules. Biologically active fractions were isolated from the head and separated using molecular weight filtration and high-performance liquid chromatography (HPLC). The resulting fractions were tested for their antidiuretic activity on single isolated Malpighian tubules. Antidiuretic activity was found in the 25% acetonitrile Sep-Pak fraction and the Cn-2 (3000-10,000 MW) and Cn-3 (<3000 MW) fractions, suggesting that the antidiuretic factor was probably a peptide of 25 to 50 amino acids. The antidiuretic factor was very potent, since after five successive fractionations on two different HPLC columns, a high level of inhibition (63%) of fluid secretion by Malpighian tubules could be observed at low dose (0.14 head-equivalent/microl). The antidiuretic factor isolated from the head of the Colorado potato beetle was not affected by repeated freezing and thawing but was sensitive to heat. The differences observed between the Colorado potato beetle antidiuretic factor and other insect diuretic and antidiuretic factors may indicate the possibility of a novel family of water regulation hormones in insects.