Histological investigation on gall development induced by a worldwide invasive pest,Dryocosmus kuriphilus,on Castanea sativa

The Asian chestnut gall wasp Dryocosmus kuriphilus (Hymenoptera: Cynipidae) is one of the most serious pests of chestnut trees worldwide. Wasps lay eggs into chestnut buds from mid Jane to late July, depending on latitude, and galls develop the following spring on new vegetation, reducing photosynthesis and nut production. We observed that modification of tissues surrounding D. kuriphilus eggs, which differentiate to form the larval chamber, started approximately 1 month after oviposition, shortly after eggs hatch. The larval chambers continued to increase slightly in size throughout the autumn months until January. After that, a period of stagnation, which corresponds to the plant's dormancy, occurred, followed by rapid growth from March to May, during the period from bud swelling to bud break. Galls continued to grow during the leaf expansion after bud break and stopped when plant organs achieved their final size. Our results have implications for the management of the pest, providing a better understanding of the critical time periods for the effective control.     

Transformation of five grape rootstocks with plant virus genes and a virE2 gene from Agrobacterium tumefaciens

Summary To facilitate the development of transgenic grapevines that are resistant to grapevine fanleaf virus (GFLV), grapevine leafroll-associated closterovirus (GLRaV-3) and crown gall diseases, we developed a rapid system for regenerating root-stocks: Couderc 3309, Vitis riparia ‘Gloire de Montpellier’, Teleki 5C, Millardet et De Grasset 101-14, and 110 Richter via somatic embryogenesis. Embryo culture and grape regeneration were accomplished with four media. Embryogenic calluses from anthers were induced in the initiation medium [MS basic medium containing 20 g sucrose per L, 1.1 mg 2,4-dichlorophenoxyacetic acid (2,4-D) per L, 0.2 mg N⁶-benzyladenine (BA) per L, and 0.8% Noble agar). The percentage of anthers that developed into embryogenic calli ranged from 2 to 16.3% depending on the rootstock. Calluses with early globular stage embryos were cocultivated with Agrobacterium tumefaciens strain C58Z707 containing the gene constructs of interest. The genes were sense-oriented translatable and antisense coat protein genes from GFLV and GLRaV-3, a truncated HSP90-related gene of GLRaV-3 (43K), and a virE2 del B gene from A. tumefaciens strain C58. Twenty independent transformation experiments were performed on five rootstocks. After 3–4 mo. under kanamycin selection, secondary embryos were recovered on differentiation medium (1/2 MS salts with 10 g sucrose per L, 4.6 g glycerol per L, and 0.8% Noble agar). Embryos that were transformed were regenerated on a medium containing MS salts with 20 g sucrose per L, 4.6 g glycerol per L, 1 g casein hydrolysate per L, and 0.8% Noble agar. Elongated embryos were then transferred to a rooting medium supplemented with 0.1 mg BA per L, 3 g activated charcoal per L, 1.5% sucrose, and 0.65% Bacto agar. A total of 928 independent putative transgenic plants were propagated in the greenhouse. All plants were tested for neomycin phosphotransferase II expression by enzyme-linked immunosorbent assay (ELISA). The presence of transgenes was assessed by polymerase chain reaction and Southern analysis. ELISA revealed various levels of expression of GFLV coat protein in transgenic plants of Couderc 3309. The transgenic rootstocks that have been generated are being screened to determine whether transgenes have conferred resistance to the virus and crown gall diseases.     

Structural modification of Quercus petraea leaf caused by Cynips quercusfolii – histological study of galls

Galls formed by the interaction of insects on plant tissues are an example of the unusual transformation and use of plants by insects. The aim of this study was to characterize the structure of galls formed by Cynips quercusfolii L. on sessile oak leaves. In the structure of galls, we distinguished the following: (1) the protective ‘first contact zone’ created by epidermal and sub-epidermal sclerenchyma rings, (2) the wide parenchymatous ring, (3) the internal protective zone created by the sclerenchyma ring, and (4) the nutritional zone consisting of cells filled with amyloplasts containing starch. A characteristic for galls in the development stage is the centripetal starch gradient in which starch accumulates in a ‘ring of amyloplasts’ in the larval chamber.     

Ultrastructure of Enteromyxum leei (Diamant, Lom, & Dyková, 1994) (Myxozoa), an Enteric Parasite Infecting Gilthead Sea Bream (Sparus aurata) and Sharpsnout Sea Bream (Diplodus puntazzo)

ABSTRACT. The ultrastructure of the developmental stages of the myxozoan Enteromyxum leei parasitizing gilthead seabream ( Sparus aurata ) intestine and sharpsnout sea bream ( Diplodus puntazzo ) intestine and gallbladder are described. The earliest stage observed was a small dense trophozoite located among enterocytes. Proliferative stages, observed intercellularly in the epithelium of the intestine and gallbladder as well as in the lumen, possessed the typical cell-in-cell configuration throughout their development. Secondary cells were seen undergoing division within a common vacuolar membrane that also encompassed pairs of tertiary cells. Cytochemical studies showed that primary cells stored mainly lipids whereas secondary cells stored abundant β-glycogen granules. Sporogonic development resembled that described for other disporous myxozoans. Within sporogonic stages, nonsporogonic secondary cells were observed accompanying two developing spores. Mature spores had a binucleated sporoplasm in which glycogen stores were abundant and no sporoplasmosomes were found. Our observations are discussed in relation to our knowledge on other myxozoans of the genus Enteromyxum .     

Linking Avicennia germinans (Acanthaceae) architecture to gall richness and abundance in Brazilian Amazon mangroves

The diversity and abundance of gall‐inducing organisms are directly proportional to the structural complexity of the host plant. This hypothesis is controversial for forest environments, such as mangroves. Avicennia germinans (L.), a principal mangrove tree species found in the Neotropical region, is considered to be a superhost for gall‐inducing insects. Using a generalized linear mixed model (GLMM) based on the analysis of 1000 apical branches from 50 A. germinans trees, we examined the diversity and abundance of gall morphotypes (GM), together with the structural attributes of replanted 5‐ to 9‐year‐old mangroves, in the Amazon coast of Brazil. A total of 7602 galls were registered, averaging 1.3 ± 0.4 galls per leaf. Sixteen of the 22 morphotypes identified were found at all study sites. Two gall morphotypes (GM7 and GM4) were the most abundant, representing approximately 40 percent of the total. The structural complexity of the plant (mainly based on the number of leaves) directly affected the abundance and diversity of these organisms. While A. germinans is a superhost, this type of parasitism did not affect plant development or survival. The ample distribution of A. germinans, the formation of monospecific forests, and the high palatability of this plant make it an essential resource for the survival of the gall‐inducing guild in the mangroves of the Neotropics.     

Three-dimensional analysis of the association of viral particles with mitochondria during the replication of Rice gall dwarf virus

Examination of cultured insect vector cells that had been infected with Rice gall dwarf virus (RGDV), using transmission electron microscopy and confocal microscopy, revealed the presence of clusters of virus-coated mitochondria around viroplasms in which replication and assembly of RGDV occurred, suggesting a role for mitochondria in supplying the energy required for viral morphogenetic processes. Electron tomography revealed that RGDV particles on the surface of mitochondria are arrayed in an orderly but loose manner, unlike tightly packaged particles in vesicular compartments, suggesting the presence of counterpart molecules on the surface of mitochondria. The viral particles in close proximity to mitochondria were aligned along intermediate filaments, which might serve as scaffolds for the anchorage of these particles. RGDV has a putative mitochondrion-targeting sequence on the outer surface of the outer-capsid protein P8. The arrangement of RGDV particles around mitochondria suggests that the region of the P8 protein containing the mitochondrion-targeting sequence might attach to a molecule like a receptor on the outer mitochondrial membrane. Our analysis demonstrates the three-dimensional arrangement and molecular basis for the mitochondrial proximity of RGDV particles during viral replication.     

Evolutionary relationships of Pemphigus and allied genera (Hemiptera: Aphididae: Eriosomatinae) and their primary endosymbiont,Buchnera aphidicola

Aphids harbor primary endosymbionts, Buchnera aphidicola, in specialized cells within their body cavities. Aphids and Buchnera have strict mutualistic relationships in nutrition exchange. This ancient association has received much attention from researchers who are interested in endosymbiotic evolution. Previous studies have found parallel phylogenetic relationships between non‐galling aphids and Buchnera at lower taxonomic levels (genus, species). To understand whether relatively isolated habitats such as galls have effect on the parallel relationships between aphids and Buchnera, the present paper investigated the phylogenetic relationships of gall aphids from Pemphigus and allied genera, which induce pseudo‐galls or galls on Populus spp. (poplar) and Buchnera. The molecular phylogenies inferred from three aphid genes (COI, COII and EF‐1α) and two Buchnera genes (gnd, 16S rRNA gene) indicated significant congruence between aphids and Buchnera at generic as well as interspecific levels. Interestingly, both aphid and Buchnera phylogenies supported three main clades corresponding to the galling locations of aphids, namely leaf, the joint of leaf blade and petiole, and branch of the host plant. The results suggest phylogenetic conservatism of gall characters, which indicates gall characters are more strongly affected by aphid phylogeny, rather than host plants.