Photophosphorylation Associated with Photosystem II: IV. KINETIC ANALYSES OF PHOTOSYSTEM II CYCLIC PHOTOPHOSPHORYLATION ACTIVITY: EVIDENCE FOR TWO CYCLIC REACTIONS

Photosystem II-dependent cyclic photophosphorylation activity produced by addition of p-phenylenediamines to KCN-Hg-NH₂OH-inhibited chloroplasts is the product of two separate reactions when a proton/electron donor is the catalyst. The activity observed with an electron donor as catalyst consists of a single reaction. One of the cyclic reactions, evoked by low (≤40 micromolar) concentrations of a proton/electron donor is sensitive to dibromothymoquinone and to perturbation of membrane organization by sonication. The second reaction, requiring higher catalyst concentrations, is less sensitive to either dibromothymoquinone or membrane perturbation. These results indicate that at low concentrations, proton/electron or electron donor catalysts act to produce a photosystem II cyclic reaction which is dependent on membrane-bound electron carriers. High concentrations of proton/electron donors, on the other hand, can produce a phosphorylation reaction in which the catalyst itself is largely responsible for cyclic activity.     

GENETIC EVIDENCE FOR DIPLOIDY IN EQUISETUM

Sporophytes and gametophytes of Equisetum arvense, E. laevigatum, and E. telmateia were analyzed using enzyme electrophoresis to estimate isozyme number. Despite their uniformly high chromosome numbers (2n = 216), these three species exhibited isozyme numbers typical of diploid seed plants for the enzymes AAT, ADH, ALD, GDH, [NADP]IDH, LAP, MDH, [NADP]ME, PGI, PGM, SkDH, and 6PGDH. All three species exhibited an additional isozyme for TPI. There is, therefore, no genetic evidence for low base numbers such as x = 9 and x = 12 suggested for Equisetum. Intact chloroplasts were isolated from E. arvense and the chloroplast extract compared electrophoretically to whole plant extracts. The single enzymes observed for LAP, GDH, [NADP]IDH, and [NADP]ME were absent from the chloroplast extract. Isozymes AAT-1, ALD-2, MDH-3, PGI-1, PGM-2, SkDH-2, 6PGDH-2, TPI-2, and TPI-3 were active in the chloroplast fraction; 6PGDH-1, PGI-2, PGM-1, and TPI-1 were lacking from the chloroplast fraction and were considered cytosolic. Isozymes AAT-2, MDH-1, MDH-2, MDH-4, and SkDH-1 were also lacking from the chloroplast fraction but because AAT, MDH, and SkDH have been reported from several subcellular compartments, their localization is unknown. These findings indicate that isozymes in Equisetum species are subcellularly compartmentalized as has also been demonstrated for homosporous ferns, gymnosperms, and angiosperms.     

MICROTUBULES: EVIDENCE FOR 13 PROTOFILAMENTS

When microtubules are fixed in glutaraldehyde in the presence of tannic acid and thin sections cut, the subunit structure of the microtubule is readily observed without the need of image reinforcement. Seven types of microtubules were analyzed: those in the heliozoan axoneme, the mitotic apparatus, the contractile axostyle, repolymerized microtubules derived from the chick brain, the central pair in flagella, and the A tubules of flagella and the basal body. In all cases microtubules were composed of 13 equally spaced protofilaments. The B tubules in flagella and the basal body appear to be composed of 11 subunits. The connections of the B to the A and the C to the B are described. A model of a microtubule is presented.     

EVIDENCE FOR GENETIC HETEROZYGOSITY IN A HOMOSPOROUS FERN

Homosporous fern sporophytes from natural populations exhibited heterozygous electrophoretic patterns for several enzyme systems. Genetic tests utilizing individual gametophytes demonstrate that the observed heterozygosity is coded by alleles at single loci. This simple procedure makes it possible to distinguish segregating from fixed or phenotypic heterozygosity, previously a problem in homosporous vascular plants.     

EVIDENCE FOR BATESIAN MIMICRY IN A POLYMORPHIC HOVERFLY

Palatable Batesian mimics are avoided by predators because they resemble noxious or defended species. The striking resemblance of many hoverflies to noxious Hymenoptera is a “textbook” example of Batesian mimicry, but evidence that selection by predators has shaped the evolution of hoverfly patterns is weak. We looked for geographical and temporal trends in frequencies of morphs of the polymorphic hoverfly Volucella bombylans that would support the hypothesis that these morphs are Batesian mimics of different bumblebee species. The frequency of the black and yellow hoverfly morph was significantly positively related to the frequency of black and yellow bumblebees across 52 sites. Similarly, the frequency of the red‐tailed hoverfly morph was positively related to the frequency of red‐tailed bumblebees. However, the frequencies of hoverfly morphs were positively spatially autocorrelated, and after controlling for this, only one of the two common hoverfly morphs showed a significant positive relationship with its putative model. We conclude that the distribution of V. bombylans morphs probably reflects geographical variation in selection by predators resulting from differences in the frequencies of noxious bumblebee species.     

ELECTROPHORETIC EVIDENCE FOR GENETIC DIPLOIDY IN PSILOTUM NUDUM

Psilotum nudum (2n = 104) has been considered an ancient polyploid, having resulted from repeated cycles of hybridization and allopolyploidy. However, electrophoretic analysis indicates that this species is genetically diploid despite its high chromosome number. Sixteen enzymes, encoded by 28 loci, revealed in P. nudum the number of isozymes typical of diploid seed plants. There is, therefore, no evidence of polyploid gene expression for the enzymes analyzed. These results for Psilotophyta are similar to those obtained for other lineages of homosporous pteridophytes, i.e., Arthrophyta and homosporous Microphyllophyta and Pteridophyta, all of which should be considered genetically diploid. Several hypotheses have been proposed to explain these results, most notably 1) cycles of allopolyploidy followed by massive gene silencing, and 2) initiation of these lineages with high chromosome numbers, possibly via chromosomal fission. Discrimination between these hypotheses awaits testing with molecular genetic techniques.     

GENETIC EVIDENCE FOR ZYGOTIC MEIOSIS IN PHYTOPHTHORA CAPSICI

Variation of several traits among single-oospore isolates of the heterothallic oomycete, Phytophthora capsici, was studied to determine genetically the place of meiosis in the life cycle. Low percentages of met^– offspring were recovered from two crosses of met^– X prototrophic isolates. Arg^– cultures were also recovered from the crosses indicating the possible presence of a suppressed arg^– gene in one of the parental cultures. Three arg^– X met^– crosses yielded all four of the possible classes of offspring. Thus meiosis was either zygotic or, if gametangial, the original parental cultures must have been heterozygous at both the arg and met loci. However, when these parental cultures were crossed, only prototrophic offspring were recovered. A ratio of 3 prototrophs: 1 met^– was obtained from cross R in which two prototrophic offspring from a previous arg^– X met^– cross were mated. This indicated the presence of a suppressor of met^– and may explain the low recovery of met^– offspring from earlier crosses. From backcrosses of met^– offspring from this cross to the A² prototrophic parent, ratios close to the expected 1:1 were obtained. When the A¹ prototrophic parent of cross R (K-10) was crossed to an A² prototrophic isolate, met^– offspring were again recovered, which indicated that K-10 carried a suppressed met deficiency. In two crosses of streptomycin-resistant (S^r) X sensitive (S^s) isolates, ratio of 108 S^s: 14 S^r and 84 S^s:14 S^r were obtained. Although 1:1 ratios would have been expected if meiosis were zygotic, segregation of S^s and S^r in the F₁ generation indicated that gametangial meiosis was unlikely. Segregation of the A¹ and A² mating types in all crosses also indicated that meiosis was zygotic in P. capsici. Possible control of mating type by a single pair of alleles, by plasmids, and by more complex mechanisms is discussed in relation to the mating type ratios obtained.     

BEHAVIORAL EVIDENCE FOR HOST-RACE FORMATION IN EUROSTA SOLIDAGINIS

We report behavioral evidence that Eurosta solidaginis, a stem-galling tephritid fly, has formed host races on its two goldenrod hosts, Solidago altissima and S. gigantea. Previous work has shown that flies from each host plant differ electrophoretically at the level of host races. The two host-associated populations were truly sympatric and were frequently found on host plants of the two species growing interdigitated with each other. Each host-associated population demonstrated a strong preference for ovipuncturing its own host. The S. gigantea–associated population emerged 10 to 14 d earlier than the S. altissima–associated population, contributing to the reproductive isolation between populations. Partial reproductive isolation is also maintained by a preference for mating on the host from which the fly emerged. The populations meet the criteria established for host races, suggesting that they may be in an intermediate stage of sympatric speciation.     

THE EGG WHITE PROTEIN EVIDENCE FOR RATITE AFFINITIES

The egg white proteins of the large ratites ( Struthio, Casuarius, Dromaius, Rhea ), the kiwis ( Apteryx ) and several tinamous (Tinamidae) were compared with one another and representatives of several other groups of birds using the technique of isoelectric focusing in acrylamide gel. The tryptic peptides of the ovalbumins of the same groups were compared by thin-layer electrophoresis. The results indicate that the large ratites are more closely related to one another than any one of them is to any other living bird; that the kiwis are not closely related to any of the other groups with which they were compared; and that the tinamous are not closely related to any of the large ratites but may be distantly related to the Galliformes.