Bud Structure and Shoot Architecture of Canopy and Understorey Evergreen Broad-leaved Trees at their Northern Limit in East Asia

The morphology of winter buds, shoot growth and branching architecturewas studied in evergreen broad-leaved trees of subtropical/warm-temperaterain forests of southern and central Japan. Winter buds werecategorized into three types based on external morphology anddevelopmental processes: naked, hypsophyllary and scaled buds.Each shoot tip with intermittent growth was covered with a smallnumber of immature leaves or hypsophylls when growth ceased.Hypsophylls protect the apical meristem during its resting period,hence we termed them hypsophyllary buds. In trees with nakedbuds, immature leaves resumed their growth and developed tomature leaves the following spring; thus these trees had nospecial organs to cover shoot tips during winter. In trees withhypsophyllary buds, some hypsophylls covering the shoot tipsthrough the year were shed without further growth when new shootsstarted to grow in the spring. In trees with scaled buds, newlygrowing shoots had hypsophyllary buds at their tips in spring.After the completion of stem elongation, the buds were replacedby scaled buds (often covered with more than 30 scales) in summer.These scaled buds grew during autumn and winter until a newflush of growth the following spring. The three bud types correspondedto forest stratification in the northern-limit forest: the nakedbuds of Rubiaceae and Myrsinaceae in the ground layer; the hypsophyllarybuds of various families (e.g. Symplocaceae, Myrsinaceae) inthe understorey; and the scaled buds of Fagaceae and Lauraceaein the forest canopy. The position and activity of buds on abranch were reflected in the architectural patterns of the treesin different layers of the forest. The scaled-bud trees hadwell-protected, abundant axillary buds and are probably suitedto survive in the forest canopy (with frequent disturbances),whereas the single terminal bud of hypsophyllary-bud trees cansurvive in the less disturbed, resource-limited understoreyof the forest.Copyright 1998 Annals of Botany Company Bud structural type; bud formation; bud growth; shoot elongation; shoot-growth cycle; branching architecture; forest stratification.     

Coalescent under the evolution of coadaptation

Adaptation to novel environments arises either from new beneficial mutations or by utilizing pre‐existing genetic variation. When standing variation is used as the source of new adaptation, fitness effects of alleles may be altered through an environmental change. Alternatively, changes in epistatic genetic backgrounds may convert formerly neutral mutations into beneficial alleles in the new genetic background. By extending the coalescent theory to describe the genealogical histories of two interacting loci, I here investigated the hitchhiking effect of epistatic selection on the amount and pattern of sequence diversity at the linked neutral regions. Assuming a specific form of epistasis between two new mutations that are independently neutral, but together form a coadapted haplotype, I demonstrate that the footprints of epistatic selection differ markedly between the interacting loci depending on the order and relative timing of the two mutational events, even though both mutations are equally essential for the formation of an adaptive gene combination. Our results imply that even when neutrality tests could detect just a single instance of adaptive substitution, there may, in fact, be numerous other hidden mutations that are left undetected, but still play indispensable roles in the evolution of a new adaptation. We expect that the integration of the coalescent framework into the general theory of polygenic inheritance would clarify the connection between factors driving phenotypic evolution and their consequences on underlying DNA sequence changes, which should further illuminate the evolutionary foundation of coadapted systems.     

Development of Plasmodium berghei Ookinetes to Young Oocysts In Vitro

ABSTRACT. The mosquito stage of Plasmodium berghei was cultivated in vitro, with special attention to ookinete transformation into early oocyst. The ookinetes were obtained by in vitro culture of gametocytes taken from infected mice, purified by density gradient of metrizoic acid or a lymphocyte separation medium, and incubated either in acellular culture or in co-cultivations with mosquito cells. In acellular culture, the ookinetes were found to aggregate with each other and transformed from banana to round shapes. Their inner pellicular membranes and subpellicular microtubules partially disappeared, indicating that development to early oocyst had occurred. Co-cultivation with Aedes albopictus cells (C6/36 clone) revealed that ookinetes transformed into early oocyst in the medium, or invaded the cells and then transformed to early oocysts within the cell cytoplasm as well. However, all of these transformed cells failed to develop further, i.e. neither deposition of the oocyst capsule nor nuclear division was observed. Many ookinetes which failed to penetrate the Aedes cells were phagocytized within three days of culture. A significant difference between invaded and transformed oocysts and phagocytized ookinetes was seen in that the former lacked vacuole membrane. Co-cultivation with Toxorhynchites amboinensis cells (TRA-284-SFG clone) permitted transformation of ookinetes into early oocysts in the medium as in the acellular culture, but no ookinete invasion nor phagocytosis by the cell was observed.     

Genetic analysis and population survey of sex pheromone variation in the adzuki bean borer moth, Ostrinia scapulalis

Sexual communication in many moths occurs between females emitting a sex pheromone and males responding to it. Females of Ostrinia scapulalis (Lepidoptera: Crambidae) show a large variation in blend ratios of the two sex pheromone components (E)‐ and (Z)‐11‐tetradecenyl acetates. E type females produce a pheromone with a high percentage of (E)‐11‐tetradecenyl acetate, whereas Z type females produce the opposite blend. We established laboratory cultures of E and Z types. Females of the F₁ generation produced an intermediate blend (I type) in both reciprocal crosses of the E and Z cultures. Results of further crossing experiments suggested that the three pheromone types are primarily controlled by a single autosomal locus with two alleles. Also, analyses of the variation in pheromone blend within F₁, backcross and F₂ families suggested that other genetic factors modify the pheromone blend of the I and Z types. Investigation of the pheromone variation in natural populations at 14 localities in Japan has shown that the E type was predominant in northern Japan, whereas the pheromone was highly polymorphic in central Japan. At a locality in central Japan, the pheromone was constantly polymorphic for several years, and the pheromone type frequencies did not deviate from Hardy–Weinberg expectations, providing no evidence of selection or assortative mating between the pheromone types. Analyses of pheromone variation within families derived from feral females indicated that matings between a pair with different genotypes for pheromone production was occurring in natural populations. Overall, this study showed that the genetic basis of the pheromone variation in O. scapulalis is very similar to that in its sibling species Ostrinia nubilalis although the state of pheromone polymorphisms in natural populations appears to differ between the two species. © 2005 The Linnean Society of London, Biological Journal of the Linnean Society, 2005, 84 , 143–160.     

Differential anatomical responses to elevated CO2 in saplings of four hardwood species

To determine whether an elevated carbon dioxide concentration ([CO₂]) can induce changes in the wood structure and stem radial growth in forest trees, we investigated the anatomical features of conduit cells and cambial activity in 4‐year‐old saplings of four deciduous broadleaved tree species – two ring‐porous (Quercus mongolica and Kalopanax septemlobus) and two diffuse‐porous species (Betula maximowicziana and Acer mono) – grown for three growing seasons in a free‐air CO₂ enrichment system. Elevated [CO₂] had no effects on vessels, growth and physiological traits of Q. mongolica, whereas tree height, photosynthesis and vessel area tended to increase in K. septemlobus. No effects of [CO₂] on growth, physiological traits and vessels were seen in the two diffuse‐porous woods. Elevated [CO₂] increased larger vessels in all species, except B. maximowicziana and number of cambial cells in two ring‐porous species. Our results showed that the vessel anatomy and radial stem growth of Q. mongolica, B. maximowicziana and A. mono were not affected by elevated [CO₂], although vessel size frequency and cambial activity in Q. mongolica were altered. In contrast, changes in vessel anatomy and cambial activity were induced by elevated [CO₂] in K. septemlobus. The different responses to elevated [CO₂] suggest that the sensitivity of forest trees to CO₂ is species dependent.     

Generation of Zika virus–specific T cells from seropositive and virus-naïve donors for potential use as an autologous or “off-the-shelf” immunotherapeutic

BackgroundZika virus (ZIKV) infection can cause severe birth defects in newborns with no effective currently available treatment. Adoptive transfer of virus-specific T cells has proven to be safe and effective for the prevention or treatment of many viral infections, and could represent a novel treatment approach for patients with ZIKV infection. However, extending this strategy to the ZIKV setting has been hampered by limited data on immunogenic T-cell antigens within ZIKV. Hence, we have generated ZIKV-specific T cells and characterized the cellular immune responses against ZIKV antigens.MethodsT-cell products were generated from peripheral blood of ZIKV-exposed donors, ZIKV-naive adult donors and umbilical cord blood by stimulation with pentadecamer (15mer) overlapping peptide libraries spanning four ZIKV polyproteins (C, M, E and NS1) using a Good Manufacturing Practice–compliant protocol.ResultsWe successfully generated T cells targeting ZIKV antigens with clinically relevant numbers. The ex vivo–expanded T cells comprised both CD4⁺ and CD8⁺ T cells that were able to produce Th1-polarized effector cytokines and kill ZIKV-infected HLA-matched monocytes, confirming functionality of this unique T-cell product as a potential “off-the-shelf” therapeutic. Epitope mapping using peptide arrays identified several novel HLA class I and class II–restricted epitopes within NS1 antigen, which is essential for viral replication and immune evasion.DiscussionOur findings demonstrate that it is feasible to generate potent ZIKV-specific T cells from a variety of cell sources including virus naïve donors for future clinical use in an “off-the-shelf” setting.     

Airborne‐pollen pool and mating pattern in a hybrid zone between Pinus pumila and P. parviflora var. pentaphylla

The reproductive isolation barriers and the mating patterns among Pinus pumila, P. parviflora var. pentaphylla and their hybrids were examined by flowering phenology and genetic assays of three life stages: airborne‐pollen grains, adults and seeds, in a hybrid zone on Mount Apoi, Hokkaido, Japan. Chloroplast DNA composition of the airborne‐pollen was determined by single‐pollen polymerase chain reaction. Mating patterns were analysed by estimating the molecular hybrid index of the seed parent, their seed embryos and pollen parents. The observation of flowering phenology showed that the flowering of P. pumila precedes that of P. parviflora var. pentaphylla by about 6 to 10 days within the same altitudinal ranges. Although this prezygotic isolation barrier is effective, the genetic assay of airborne‐pollen showed that the two pine species, particularly P. pumila, still have chances to form F₁ hybrid seeds. Both parental species showed a strong assortative mating pattern; F₁ seeds were found in only 1.4% of seeds from P. pumila mother trees and not at all in P. parviflora var. pentaphylla. The assortative mating was concluded as the combined result of flowering time differentiation and cross‐incompatibility. In contrast to the parental species, hybrids were fertilized evenly by the two parental species and themselves. The breakdown of prezygotic barriers (intermediate flowering phenology) and cross‐incompatibility may account for the unselective mating. It is suggested that introgression is ongoing on Mount Apoi through backcrossing between hybrids and parental species, despite strong isolation barriers between the parental species.