Endosymbionts Differentially Alter Exploratory Probing Behavior of a Nonpersistent Plant Virus Vector.

Insect endosymbionts (hereafter, symbionts) can modify plant virus epidemiology by changing the physiology or behavior of vectors, but their role in nonpersistent virus pathosystems remains uninvestigated. Unlike propagative and circulative viruses, nonpersistent plant virus transmission occurs via transient contamination of mouthparts, making direct interaction between symbiont and virus unlikely. Nonpersistent virus transmission occurs during exploratory intracellular punctures with styletiform mouthparts when vectors assess potential host-plant quality prior to phloem feeding. Therefore, we used an electrical penetration graph (EPG) to evaluate plant probing of the cowpea aphid, Aphis craccivora Koch, an important vector of cucurbit viruses, in the presence and absence of two facultative, intracellular symbionts. We tested four isolines of A. craccivora: two isolines were from a clone from black locust (Robinia pseudoacacia L.), one infected with Arsenophonus sp. and one cured, and two derived from a clone from alfalfa (Medicago sativa L.), one infected with Hamiltonella defensa and one cured. We quantified exploratory intracellular punctures, indicated by a waveform potential drop recorded by the EPG, initiation speed and frequency within the initial 15 min on healthy and watermelon mosaic virus-infected pumpkins. Symbiont associations differentially modified exploratory intracellular puncture frequency by aphids, with H. defensa-infected aphids exhibiting depressed probing, and Arsenophonus-infected aphids an increased frequency of probing. Further, there was greater overall aphid probing on virus-infected plants, suggesting that viruses manipulate their vectors to enhance acquisition-transmission rates, independent of symbiont infection. These results suggest facultative symbionts differentially affect plant-host exploration behaviors and potentially nonpersistent virus transmission by vectors.

Location and regulation of octameric mitochondrial creatine kinase in the contact sites.

Differential extraction of creatine kinase activity (CK, EC 2.7.3.2) from rat brain mitochondria by graded concentrations of digitonin all yielded supernates varying in CK activity. As analyzed by isozyme electrophoresis and gel permeation chromatography the extracts contained different species of creatine kinase: (i) one third of the total CK activity consisting of contaminating cytosolic brain-type CK (B-CK) was liberated by 100 micrograms digitonin/mg of mitochondrial protein, (ii) approx. 20% more CK activity consisting of B-CK, as above, plus dimeric and octameric mitochondrial CK (Mi-CK), was extracted by 300 micrograms/mg digitonin, whereas (iii) all CK activity, consisting of B-CK and mainly octameric Mi-CK, were liberated by 700 micrograms/mg digitonin. In contrast to Mi-CK, B-CK associated with contaminating synaptic vesicles was readily extracted even by low concentrations of digitonin, but on the other hand octameric Mi-CK was significantly more resistant to digitonin extraction than the dimeric enzyme species. It appeared that the Mi-CK resistant to treatment with 300 micrograms/mg digitonin consisted to a large percentage of octamers and was organized as a complex between the two envelope membranes, for its activity was latent and still remained regulated by the outer membrane pore, that is: (i) the Mi-CK activity in such mitoplasts could be inhibited reversibly by cessation of the adenine nucleotide transport through the outer membrane pore with a polyanion, (ii) the ADP produced by Mi-CK in mitoplasts was not available to external pyruvate kinase, (iii) approx. 50% of total CK activity was not susceptible to inhibition by iodo acetate and phosphocreatine. In agreement with these findings a preferential association of octameric Mi-CK was also found in isolated contact site fractions indicating a physiological role of Mi-CK in energy transfer and a structure-function relationship of Mi-CK octamers at these sites. In addition some evidence for an interaction of Mi-CK with the adenylate translocator is presented.