Thomas P. Freeman

Virulent Hessian Fly (Diptera: Cecidomyiidae) Larvae Induce a Nutritive Tissue During Compatible Interactions with Wheat

Abstract The compatible interaction between virulent Hessian fly, Mayetiola destructor (Say), larvae and susceptible wheat, Triticum aestivum L., plants was investigated at the light microscope and ultrastructural levels. During the first day of larval attack at the base of the sheath of the third leaf of a wheat seedling, small punctures of the appropriate size (0.1 μm in diameter) and spacing of the paired larval mandibles were found in the outer wall of epidermal cells. Inside epidermal cells, nuclei and cytoplasmic organelles seemed to be breaking down, and the number and size of cytoplasmic vacuoles had increased. Two to 3 d later, epidermal and mesophyll cells at the base of the third leaf showed signs of becoming nutritive. Nutritive cells were identified by an increase in cytoplasmic staining, increased numbers of cellular organelles (mitochondria, proplastids, Golgi, and rough endoplasmic reticulum), numerous small vacuoles, and an irregularly shaped nucleus. The Hessian fly nutritive tissue probably acts as a sink tissue within the wheat seedling, benefiting the growth of larvae by importing photoassimilates. Breakdown of nutritive cells began soon after they were first observed, indicated by a change in the shape and density of the cell nucleus. Contents of nutritive cells moved through compromised cell walls into adjacent cells that had a more complete breakdown and loss of cytoplasm. Structural changes were not restricted to the third leaf. The sixth leaf, a leaf more recently initiated by the shoot apical meristem that was not directly fed upon by larvae, was found to consist primarily of well-developed epidermal layers, with poorly developed mesophyll cells. The implications of these findings for understanding incompatible interactions between avirulent Hessian fly larvae and R gene-defended plants are briefly discussed.

Stylet Penetration by Bemisia argentifolii (Homoptera: Aleyrodidae) into Host Leaf Tissue

Abstract Silverleaf whitefly, Bemisia argentifolii Bellows & Perring, adults and nymphs feed on the phloem tissue of host plant leaves. Infested leaves were rapidly fixed to prevent the insects from withdrawing their mouthparts. The adult stylet bundle enters the labial groove of the labium between the first and second segments and is completely contained within the labium except during feeding. Stylet length is equal to the combined length of labial segments 2, 3, and 4. Stylet penetration can be determined by examining the position that the stylet enters the labial groove. The physical force necessary for adult stylet penetration is derived from changes in the position of the whitefly head during feeding. The head is bent over the labium, which is attached to the leaf surface, forcing the stylet bundle down the labial groove and into the host tissue. Nymphal stylet lengths were found to be longer than previously reported, and of sufficient length to reach the phloem tissue in cotton and hibiscus leaves from essentially any position on the abaxial leaf surface.

Mouthpart morphology and stylet penetration of host plants by the glassy-winged sharpshooter, Homalodisca coagulata, (Homoptera: Cicadellidae)☆

The ultrastructural morphology of the mouthparts of the glassy-winged sharpshooter, Homalodisca coagulata, and method of plant penetration was examined using light microscopy, scanning electron microscopy, and transmission electron microscopy methods. The gross morphology of the labrum, labium, and stylet fascicle was consistent with what has been described for other plant-sucking homopterans. The ultrastructural examination of the mouthparts revealed unique details that have previously gone unreported. Several types of sensilla-like structures having the form of pegs and multi-lobed objects were identified on the outer surfaces of the labrum and within the labial groove. Dendritic canals terminated in an extensive network of smaller canals at the distal tip of the maxillary stylets below a series of surface denticles suggesting that this area may have a sensory function associated with locating xylem elements of host plants. Examination of salivary sheath pathways established that 65% of the plant penetrations by this insect terminated in the xylem vessels of the host plant. Probing by the insect was largely intracellular and terminal branching of a single probe site was common. Plant surface feeding sites varied with the stage of development which correlates with the depth of the xylem vessels and the length of the maxillary stylets of the various instars.

Susceptibility of Upland Cotton Cultivars to Bemisia tabaci Biotype B (Homoptera: Aleyrodidae) in Relation to Leaf Age and Trichome Density

Abstract The relationships between leaf trichome densities, leaf age, and sweetpotato whitefly, Bemisia tabaci (Gennadius) biotype B, infestations of 13 upland cotton, Gossypium hirsutum L., cultivars were investigated in 1990 and 2000. Stoneville 474 supported higher numbers of B. tabaci biotype B eggs, nymphs and adults, and also had higher numbers of stellate trichomes on abaxial leaf surfaces compared with other cotton cultivars. Siokra L-23, in general, had fewer stellate trichomes and also fewer whiteflies. However, the positive trichome–whitefly density relationships were affected by the ages of leaves from different main stem cotton nodes. The youngest leaves on main stem node 1 below the terminal for all cultivars had higher numbers of stellate trichomes but fewer whiteflies compared with older leaves.

The effect of imposed watet stress on the development and ultrastructure of wheat chloroplasts

SummaryEtiolated 6-day old wheat (Triticum aestivum L. cv. ‘Chris’) seedlings were subjected to osmotic stress by the application of polyethylene glycol 12 hours prior to exposure to continuous illumination for a 48 hours period.Stress impaired seedling growth and altered plastid development. The number of grana per plastid and the number of thylakoids per grana were significantly different in plastids from stressed and non-stressed leaves after 48 hours of development in the light. Chlorophyll production was similarly decreased in stressed leaves. After 12 hours of greening a swelling or dilation of thylakoid membranes became common. The dilation continued during the remainder of the experimental period and frequently reduced the grana and stroma thylakoid systems to a series of vesicles. There was no significant increase in the number and size of plastoglobuli as a result of the thylakoid dilation. Extensions containing crystalline-like bodies commonly developed from stressed plastids after 24 hours of greening. A reduction in both chloroplast and cytoplasmic ribosomes was noted in stressed leaves.

Waxes and lipids associated with the external waxy structures of nymphs and pupae of the giant whitefly, Aleurodicus dugesii

The nymphs and pupae of the giant whitefly, Aleurodicus dugesii, produce large quantities of external lipids, both as waxy particles and as waxy filaments. The nymphs and pupae extrude filaments from two dorsal rows of five pores each. Filaments can attain lengths of 5-8 cm. The external lipids of nymphs and pupae consist largely of long-chain aldehydes, alcohols, acetate esters and wax esters. Hydrocarbons are minor components. Soon after hatching, the nymph produced an unidentified waxy fringe extruded laterally from its margin. After molting to the second instar, long, hollow, waxy filaments were produced by the immature stages. The major lipid class associated with the filaments was saturated wax esters (89%), mainly C44, C46 and C60. Associated with formation of the filaments were waxy particles in the shape of curls, which peeled off of the extruding filaments. Similar but more tubular-shaped curls were also produced by numerous lateral pores so that, eventually, the curls completely camouflaged the nymph. The major lipid class of the curls was wax esters (50%), mainly C44 and C46. The cuticular surface lipids of the nymphs were mainly long-chain aldehydes (43%) and wax esters (27%). Unsaturated fatty acid moieties constituted 2 and 19% of the wax esters of curls and nymph cuticular surface lipids, respectively. The major lipid classes of pupae and of their palisade were long-chain aldehydes and alcohols. No unsaturated wax esters were detected in the filaments, but 30% of pupal and 21% of palisade surface wax esters were unsaturated in their fatty acid moieties, 16:1, 18:1 and 20:1.

External lipids of adults of the giant whitefly, Aleurodicus dugesii

Abstract The external lipids of male and female adults of the giant whitefly, Aleurodicus dugesii , were found in three forms; as cuticular lipids, as waxy particles, and only on the females, as waxy filaments. Collectively, the external lipids consisted of long-chain aldehydes, long-chain alcohols and wax esters. Cuticular lipids consisted mainly of wax esters. The major wax esters were C44 (tetracosanyl icosanoate and docosanyl docosanoate) and C46 (mainly triacontanyl hexadecanoate and tetracosanyl docosanoate). Hydrocarbons, largely n -alkanes, were minor components of the cuticular surface lipids. Both sexes produced waxy particles soon after eclosion by breaking off extruding strands (or ribbons) from anterior wax plates on their abdomens; three pair on males and two pair on females. These particles coated the adults and their surroundings. The waxy particles produced by the anterior wax plates of the adults were a mixture of aldehydes and alcohols, predominantly C30. The wax esters were not part of the waxy particles despite being the major lipid class on the cuticular surface. In addition, the adult female has two pair of posterior abdominal wax plates which produce waxy filaments that are broken off during oviposition and form spiral trails on the leaf. These trails appear to function to camouflage the eggs which are laid horizontally in the waxy trail. The lipid obtained from the waxy filaments from the posterior abdominal wax plates of the female was largely wax esters. The predominant wax ester was C46 (mainly triacontanyl hexadecanoate).

Cryopreservation of Embryos of the New World Screwworm Cochliomyia hominivorax (Diptera: Calliphoridae)

Abstract A method for cryopreserving embryos of the screwworm, Cochliomyia hominivorax (Coquerel), was developed for the long-term storage of strains used in research projects and for colonies maintained as back-up to production strains that are mass-reared for sterile insect release programs. This protocol, when combined with a previous procedure that was designed to render screwworm embryos permeable to water loss and the influx of cryoprotectants, yields rates of ≈53% hatching, 22% pupation, and 75% adult emergence. These yields are all lower than control levels, but by the next generation the progeny of the cryopreserved flies are near or equal to control levels and are available in sufficient numbers to reestablish laboratory colonies. Important to the success of this study was identifying the optimum embryonic stage for treatment, formulating an incubation regime, which allowed consistent retrieval of embryos at the optimum stage, establishing the correct dehydration time for the previtrification step, and developing a recovery system after liquid nitrogen storage of the embryos.

Electron transport and chloroplast ultrastructure of a chlorophyll deficient mutant of wheat

A non-lethal chlorophyll deficient mutation was induced by use of the chemical mutagen ethyl methanesulfonate. Chloroplasts from the control and mutant plants were found to be very similar ultrastructurally. Thylakoid membrane volume was only slightly greater in plastids from the control as compared with plastids from the mutant. The chlorophyll content of the mutant was reduced by over 60%. This decrease in chlorophyll was not accompanied by a similar decrease in electron transport. Uncoupled electron transport rate based on a unit chlorophyll basis was nearly twice as great for mutant chloroplasts as for control plastids. However, electron transport rate based on a unit membrane volume was similar in mutant and control plants. At high irradiance the relative quantum requirement of the control and mutant was similar when expressed on membrane volume.

The calcified puparium of the face fly, Musca autumnalis (Diptera: Muscidae)

Abstract The general composition of the calcified puparium of Musca autumnalis was determined. Ash-weight analyses show that 62% of the puparium is inorganic material. The major components of the puparium are calcium, magnesium, phosphate and carbonate. Calcium and magnesium phosphate are the predominant salts in the puparium, with a lesser contribution by their respective carbonates. Scanning electron microscopy revealed that the exocuticle was the site of calcification in the puparium. Inorganic analyses of the insoluble salt isolated from face fly larvae showed it to be compositionally identical with the puparial salt. Major organic components of the puparium are lipid, chitin and protein. The presence of carbonic anhydrase and alkaline phosphatase activity in post-feeding larvae was confirmed.