Sieve element occlusion: Interactions with phloem sap-feeding insects. A review.

Phloem sieve element (SE) occlusion has been hypothesized for decades to be a mechanism of resistance against phloem sap-feeding insects. Few studies have tested this hypothesis although it is likely a widespread phenomenon. This review focuses on SE occlusion by callose and P-proteins. Both are reversible, which would allow the plant to defend itself against phloem sap-feeders when SEs are penetrated and resume normal function when the insects give up and withdraw their stylets. Callose (ß-1,3 glucans with some ß-1,6 branches) serves many roles in plant physiology in many different tissues, each being under the control of different callose synthase genes; only callose deposited in SE sieve pores is relevant to SE occlusion. The amount of callose in sieve pores (and consequently how much it impedes sap flow) is determined by the balance in activity between callose synthase and ß-1,3 glucanase. Sieve pore callose deposition has been shown to provide resistance to some phloem sap-feeders in a few studies, and in one, the difference in resistance between a susceptible and resistant rice variety was due to the ability or inability of the insect to upregulate the plants' ß-1,3 glucanase that degrades the callose deposition. P-proteins occur only in dicotyledons and include a variety of proteins, not all of which are involved in SE occlusion. In some plants, P-proteins form distinct bodies in mature functional SEs. In papilionid legumes, these discrete bodies, called forisomes can expand and contract. In their expanded state, they effectively plug SEs and stop the flow of sap while in their contracted state, they provide negligible resistance to sap flow. Expansion of forisomes is triggered by an influx of Ca2+ into the SE. Penetration of a legume (Vicia faba) SE by a generalist aphid not adapted to legumes triggers forisome expansion which occludes the SE and prevents the aphid from ingesting sap. In contrast, a legume specialist aphid, Acyrthosiphon pisum, does not trigger forisome expansion and readily ingests sap from V. faba. P-protein bodies in SEs of non-legumes do not appear to be involved in SE occlusion. In most dicotyledons, P-proteins do not form discrete bodies, but rather occur as filamentous aggregations adhering to the parietal margins of the SE and in response to damage, are released into the lumen where they are carried by the flow of sap to the downstream sieve plate where they back up and clog the sieve pores. Their effectiveness at actually stopping the flow of sap is controversial. In one study, they seemed to provide little resistance to the flow of sap while in other studies, they provided considerable resistance. In response to injury in melon, they completely stop the flow of sap, and in an aphid-resistant melon, penetration of SEs by the melon aphid, Aphis gossypii, triggers P-protein occlusion which prevents the aphids from ingesting sap. The first P-protein described, PP1, occurs only in the genus Cucurbita, and although it has been often cited to function as a SE occlusion protein, experimental evidence suggests it does not play a significant role in SE occlusion. The most common strategy for phloem sap-feeders to mitigate P-protein occlusion seems to be avoid triggering it. A widely cited in vitro study suggested that aphid saliva can reverse P-protein occlusion, but a subsequent study demonstrated that saliva was ineffective at reversing P-protein occlusion in vivo. Lastly, SE callose deposition in wheat triggered by Russian wheat aphid has been hypothesized to create an artificial sink that benefits the aphid, but additional studies are needed to test that hypothesis.

Does aphid salivation affect phloem sieve element occlusion in vivo?

To protect against loss of photo-assimilate-rich phloem sap, plants have evolved several mechanisms to plug phloem sieve tubes in response to damage. In many Fabaceae, each sieve element contains a discrete proteinaceous body called a forisome, which, in response to damage, rapidly transforms from a condensed configuration that does not impede the flow of sap to a dispersed configuration that plugs the sieve element. Aphids and other specialized phloem sap feeders can ingest phloem sap from a single sieve element for hours or days, and to do this, they must be able to suppress or reverse phloem plugging. A recent study provided in vitro evidence that aphid saliva can reverse forisome plugs. The present study tested this hypothesis in vivo by inducing forisome plugs which triggered aphids to switch behaviour from phloem sap ingestion to salivation into the sieve element. After salivating into the sieve element for various periods of time, the aphids were instantaneously cryofixed (freeze fixed) in situ on their leaf. The state of the forisome was then determined in the penetrated sieve element and in nearby non-penetrated sieve elements which served as controls for sieve elements not subjected to direct aphid salivation. Forisomes were almost always in close contact with the stylet tips and thus came into direct contact with the saliva. Nonetheless, forisome plugs in the penetrated sieve element did not revert back to a non-plugging state any faster than those in neighbouring sieve elements that were not subjected to direct aphid salivation.

Larval morphology of Metaphycus flavus and its role in host attachment and larval cannibalism.

Metaphycus flavus (Howard) (Hymenoptera: Encyrtidae) is a facultatively gregarious endoparasitoid of soft scales (Hemiptera: Coccidae). When it develops in superparasitised hosts, the larvae often attack and consume brood mates six or more days post oviposition. Under our laboratory conditions (25±1°C and 14 hours of light followed by 18±1°C and ten hours of darkness in 50-70% R.H.), M. flavus eggs hatched three days after oviposition. Measurements of the mandibles and tentorium indicate there are four larval instars, and M. flavus reaches the fourth instar by day six post oviposition, and pupates on day eight. Thus, cannibalism among M. flavus larvae occurs during the fourth instar. During this instar, M. flavus larvae separate from their attachment to the scale cuticle, to which they were tethered by a respiratory structure during the previous three larval instars. Once detached, they are free to move within the scale, which increases the probability of larval encounters and aggressive behaviours. Moreover, the mandibles of the fourth instar are better adapted for fighting than are those of the first three larval instars, since they are larger and more sclerotized. The cranium and mouthparts of M. flavus have four different types of sensory organs, some of which are almost certainly olfactory, an unexpected function for a larva that presumably is surrounded by an aqueous medium where gustatory sensilla would seem to be more appropriate. The cranium also bears two pairs of what appear to be secretory pores.

Selenium levels in cows fed pasture and concentrates or a total mixed ration and supplemented with selenized yeast to produce milk with supra-nutritional selenium concentrations.

Seventy multiparous Holstein-Friesian cows were fed different amounts of pasture and concentrates, or a total mixed ration (TMR), for 42 d in mid-lactation to test the hypothesis that the concentration of Se in milk would depend on the amount of Se consumed, when the Se is primarily organic in nature, regardless of the diet of the cows. Of the 70 cows, 60 grazed irrigated perennial pasture at daily allowances of either 20 or 40 kg of dry matter (DM)/cow. These cows received 1 of 3 amounts of concentrates, either 1, 3, or 6 kg of DM/cow per day of pellets, and at each level of concentrate feeding, the pellets were formulated to provide 1 of 2 quantities of Se from Se yeast, either about 16 or 32 mg of Se/d. The other 10 cows were included in 2 additional treatments where a TMR diet was supplemented with 1 kg of DM/d of pellets formulated to include 1 of the 2 quantities of supplemental Se. Total Se intakes ranged from 14.5 to 35.9 mg/d, and of this, the Se-enriched pellets provided 93, 91, and 72% of the Se for cows allocated 20 and 40 kg of pasture DM/d or the TMR, respectively. No effects of the amount of Se consumed on any milk production variable, or on somatic cell count, body weight, and body condition score, for either the pasture-fed or TMR-fed cows were found. Milk Se concentrations responded quickly to the commencement of Se supplementation, reaching 89% of steady state levels at d 5. When milk Se concentrations were at steady state (d 12 to 40), each 1mg of Se eaten increased the Se concentration of milk by 5.0 µg/kg (R(2)=0.97), and this response did not seem to be affected by the diet of the cows or their milk production. The concentration of Se in whole blood was more variable than that in milk, and took much longer to respond to change in Se status, but it was not affected by diet at any time either. For the on-farm production of Se-enriched milk, it is important to be able to predict milk Se concentration from Se input. In our study, type of diet did not affect this relationship.

Output of selenium in milk, urine, and feces is proportional to selenium intake in dairy cows fed a total mixed ration supplemented with selenium yeast.

Fifteen rumen fistulated Holstein cows in late lactation and fed a total mixed ration offered ad libitum were supplemented with Se yeast to provide 0, 11, 20, 30, or 42 mg of supplemental Se/day to test the hypothesis that amounts of Se secreted in milk, excreted in urine and feces, and apparently retained in tissues would increase in direct proportion to Se intake. One-half of the yeast supplement was placed directly into the rumen through the fistula of each cow just before milking in the morning and again in the evening, and estimates of average daily excretion of Se were made using total collections of urine and feces from 25 to 31 d after treatments commenced. Amounts of Se secreted daily in milk and apparently retained in tissues increased linearly with average daily intake of Se. The amount of Se excreted in feces and total excretion of Se in urine plus feces increased curvilinearly with Se intake, such that proportionately less Se was excreted as the amount of Se fed increased. On average, total Se excretion accounted for 66%, Se secretion in milk accounted for 17%, and Se apparently retained in tissues accounted for 17% of total Se intake by cows. Thus, in herds fed large amounts of Se yeast, most of the Se will be excreted and retained on-farm. High concentrations of Se will be found where urine and feces accumulate (e.g., yards and effluent ponds), and effluent management practices must be tailored to avoid environmental issues.

Phoretic dispersal of armored scale crawlers (Hemiptera: Diaspididae).

Dispersal and colonization of new areas by armored scale insects (Hemiptera: Diaspididae) is achieved by mobile first-instar nymphs, called crawlers. Few studies have considered the actual mechanisms by which crawlers disperse, and although crawlers are capable of actively wandering over short distances (generally < 1 m), their dispersal over longer distances has been thought to be wind-mediated. Here, we present evidence of a potentially more important means of dispersal over longer distances (> 1 m). We first confirmed that crawlers of four species of Diaspididae [Abgrallaspis aguacatae Evans, Watson & Miller; Hemiberlesia lataniae (Signoret); Aspidiotus nerii Bouché; and Diaspidiotus perniciosus (Comstock)] have four hairs on the end of each of their legs and that each of these hairs ends in a suction cup-like structure, reminiscent of the attachment structures possessed by phoretic mites. In a controlled environment, using crawlers of A. nerii, we then showed that the crawlers use these structures to attach themselves to three different insect species [Musca domestica L., Cryptolaemus montrouzieri Mulsant and Linepithema humile (Mayr)] and can effectively be moved phoretically by these insects. Crawlers can remain attached to flying insects for considerable periods of time, suggesting that this may be an important means of dispersal for armored scale insects. The importance of phoresy for diaspidid dispersal in the field remains to be determined.

Discovery of localized regions of excess 10-TeV cosmic rays.

The 7 year data set of the Milagro TeV observatory contains 2.2 x 10(11) events of which most are due to hadronic cosmic rays. These data are searched for evidence of intermediate scale structure. Excess emission on angular scales of approximately 10 degrees has been found in two localized regions of unknown origin with greater than 12sigma significance. Both regions are inconsistent with pure gamma-ray emission with high confidence. One of the regions has a different energy spectrum than the isotropic cosmic-ray flux at a level of 4.6sigma, and it is consistent with hard spectrum protons with an exponential cutoff, with the most significant excess at approximately 10 TeV. Potential causes of these excesses are explored, but no compelling explanations are found.

Increasing selenium concentration in milk: effects of amount of selenium from yeast and cereal grain supplements.

Two experiments were conducted to establish responses in milk Se concentrations in grazing dairy cows to different amounts of dietary Se yeast, and to determine the effects of the Se concentration of the basal diet. The hypothesis tested was that the response in milk, blood, and tissue Se concentrations to supplemental Se would not be affected by whether the Se was from the basal diet or from Se yeast. In addition, by conducting a similar experiment in either early (spring; experiment 1) or late (autumn; experiment 2) lactation, we hypothesized that different Se input-output relationships would result. Both 6-wk experiments involved 60 multiparous Holstein-Friesian cows, all of which had calved in spring. They were allocated to 1 of 10 dietary Se treatments that included 2 types of crushed triticale grain (low Se, approximately 165 microg of Se/kg of DM; or high Se, approximately 580 microg/kg of DM) fed at 4 kg of DM/d, and 1 kg of DM/d of pellets formulated to carry 5 quantities of Se yeast (0, 4, 8, 12, or 16 mg of Se). Daily total Se intakes ranged from <2 to >18 mg/cow in both experiments. Milk Se concentrations plateaued after 15 and 7 d of supplementation in experiments 1 and 2, respectively, and then remained at plateau concentrations. Average milk Se concentrations for the plateau period increased as the amount of Se yeast increased, and low- and high-Se grain treatments were different at all quantities of Se yeast, although there was a tendency for this difference to diminish at the greatest concentrations of yeast. There were significant positive, linear relationships between Se intake and the concentrations of Se in milk, which were not affected by the source of Se, and the relationships were similar for both experiments. Therefore, the output of Se in milk in experiment 1 was greater than that in experiment 2 because the milk yield of the cows in early lactation was greater. The estimated proportions of Se partitioned to destinations other than milk and feces increased with the amount of Se in the diet and were greater in experiment 2 than in experiment 1, a result that was supported by Se concentrations in whole blood and plasma and in semitendinosus muscle tissue. If high-Se products are to be produced for human nutrition, it is important to be able to develop feeding systems that produce milk with consistent and predictable Se concentrations so that products can consistently meet specifications. The results indicate that this objective is achievable.

Development of economic thresholds and monitoring systems for Helicoverpa armigera (Lepidoptera: Noctuidae) in tomatoes.

Field sampling methods and economic thresholds were developed to provide management recommendations for Helicoverpa armigera (Hübner) on processing tomatoes, based on a commercially acceptable damage level of 5% fruit damage. Population estimates from destructive sampling and a rapid 1-min plant scouting method were related to fruit damage, and a nominal economic threshold of one larva per plant was derived. The economic threshold was confirmed in a designed trial where it resulted in acceptable levels of fruit damage. The 1-min scouting method and economic threshold was validated in 17 commercial crops in the Gisborne, Poverty Bay region of New Zealand. Scouting in these fields was based on 10 plants in each of four quadrants proportionally representing the topography of each field. In unsprayed areas, egg and larval populations were usually below the economic threshold in early-planted crops but often exceeded thresholds in late-planted crops. In commercial demonstration trials where standard calendar spraying practice was compared with no spraying, calendar-based applications maintained fruit damage below 3.4%, but insecticide was applied unnecessarily to more than half the crops. Larval populations were a significant predictor of damage in these commercial crops. In 12 implementation trials, where spraying recommendations were based on the 1-min scouting threshold of one larva per plant, the worst fruit damage observed was 2.3%. The definition of an economic threshold, scouting methods, and establishment of parasitoids have reduced spray applications and contributed to the implementation of an integrated pest management program for processing tomatoes in New Zealand.