POLLINATION OF THE ARROYO WILLOW,SALIX LASIOLEPIS: ROLE OF INSECTS AND WIND

We studied the relative roles of insects and wind as pollen vectors for the arroyo willow, Salix lasiolepis Benth. through insect exclosure studies at two elevational sites. Insect pollination resulted in production of 99.9% of seeds at a low elevation site (2,130 m) and at a high elevation site (2,450 m) with wind playing an exceedingly small role. Few pollen grains reached individual pistillate catkins despite the presence of abundant airborne willow pollen near male plants. In flowers that showed evidence of receipt of wind transferred pollen, due to their swollen appearance, we found only 1.74 fertilized ovules per flower and that 90.9% of these fertilized ovules were aborted by the maternal plant. In contrast, flowers open to insect visitation possessed 7.00 fertilized ovules per flower, 83.3% of which became filled seeds. We tested for the existence of apomictic seed production through use of closed bags that excluded both insect-and wind-transferred pollen but found no evidence of seed produced in this treatment. Additional control studies documented that wind treatment bags did not impede pollen flow while neither wind or closed bags influenced seed set in our treatments.     

PATTERNS OF GENETIC VARIATION IN SALIX SECTION LONGIFOLIAE (SALICACEAE)

Enzyme electrophoresis was used to examine genetic relationships within a taxonomically complex group of diploid willows (Salix section Longifoliae). Forty-eight populations representing seven morphogeographic taxa were analyzed for 15 putative enzyme loci. Levels of genetic variation (A, P, H_e, and H_T) differed greatly among populations, but were generally lower than those reported for other species with similar life history traits. In populations and species from southern latitudes, where populations tend to be smaller and more isolated, genetic variation decreased and population differentiation increased. Genetic identity values suggest that the section is divisible into four major elements: 1) Salix interior, 2) S. taxifolia, 3) the S. exigua group, and 4) S. melanopsis. The first three occupy eastern, southern, and western portions of the continent, respectively. Salix melanopsis is sympatric with the S. exigua group in western North America, but is ecologically distinct. The S. exigua group is differentiated into four major geographic entities: S. hindsiana from west of the Sierras in California and southwest Oregon, S. sessilifolia from west of the Cascades in central Oregon to British Columbia, and a northern and southern race of S. exigua in the Intermountain West. The relationships suggested by allozyme data do not agree with traditional taxonomic concepts. Evidence of hybridization was obtained at five points where taxa come into contact, and Salix fluviatilis may be of hybrid origin. Taxonomic confusion appears to be largely the result of frequent hybridization events and overemphasis of a few highly variable morphological traits. The section appears to be a syngameon undergoing geographic speciation, with one taxon gaining reproductive isolation by a shift in habitat preference.     

EFFECTS OF SOIL WATERLOGGING ON THE ENERGY STATUS AND DISTRIBUTION OF SALIX NIGRA AND S. EXIGUA (SALICACEAE) IN THE ATCHAFALAYA RIVER BASIN OF LOUISIANA

The influence of soil waterlogging on the distribution and energy status of Salix nigra and S. exigua were studied in the field and laboratory. Differential flooding in natural stands of Salix spp. and under controlled conditions in the greenhouse demonstrated that S. nigra was more tolerant of waterlogged soil conditions than was S. exigua. Salix nigra exhibited a significantly higher leaf energy status and greater ability to oxidize waterlogged soil when flooded than did S. exigua. However, S. exigua appeared to be more tolerant of moisture stress than S. nigra as indicated by the significantly higher leaf resistances to water vapor exchange and higher leaf water potentials of S. exigua. These data support the theory that the reason S. nigra inhabits lower elevation sites than S. exigua in the Atchafalaya River Basin may be the differential tolerance of these two species to soil waterlogging on one extreme and moisture deficiencies on the other.     

THE ELASTIC MODULI AND MECHANICS OF POPULUS TREMULOIDES (SALICACEAE) PETIOLES IN BENDING AND TORSION

Changes attending leaf development in the mechanical behavior and elastic moduli of the petioles of Populus tremuloides Michx. were examined in terms of total leaf weight (Wt), lamina and petiole weight (Wl and Wp), petiolar length (L), and lamina surface area (A). A primary concern was the extent to which two elastic moduli (Young's modulus E and the shear modulus G) of petioles changed over time and were correlated with one another. E and G were measured by means of multiple resonance frequency spectra, and together with the dimensions of cross sections through petioles, these two elastic moduli were used to estimate the stiffness of petioles in simple bending and torsion. Petiolar bending was predicted by means of a model incorporating expressions for both the bending stiffness (El) and torsional rigidity (GJ), where I is the second moment of area and J is the torsional constant. The predictions from these models were compared to observed petiolar bendings due to Wl. Additionally, the frequency of the oscillatory motion of leaves (placed in a wind tunnel with a 1 m sec^-1 ambient wind speed, directed normal to the blade of the leaf) was determined. Results indicate that L, A, Wl, Wp, and Wt were positively correlated with the age of leaves (crudely estimated as a function of leaf plastochron index, LPI); these morphometric parameters were also correlated with the magnitudes of E and G. Also, G was positively and linearly correlated with E, and was, on the average, an order of magnitude less than E. EI and GJ were positively correlated with LPI. The relationships among E, G, EI, C and Wl, Wp, Wt are discussed in terms of leaf allometries.     

GROWTH AND DEMOGRAPHY OF POPULATIONS OF BIOMPHALARIA PFEIFFERI (GASTROPODA, PLANORBIDAE) IN THE LABORATORY

The growth and demography of the freshwater snail Biomphalariapfeifferi, an intermediate host of the parasitic worm Schistosomamansoni in Eastern Zaire, were investigated in the laboratoryunder two conditions of crowding. Both individual and populationgrowth were depressed by crowding. Crowding reduced growth rateand all the variables and parameters related to reproduction(fecundity, instantaneous birth rate, net reproduction rate),but did not affect maximum size, survivorship, instantaneousdeath rate and life expectancy. The mechanism involved in theseeffects is likely to be exploitation competition for especiallyprofitable food items such as diatoms. (Received 10 November 1986;     

THE ROLES OF HETEROCHRONY AND HETEROBLASTY IN THE DIVERSIFICATION OF LEAF SHAPES IN BEGONIA DREGEI (BEGONIACEAE)

The relationship between shape variation in the transitional series of leaves and in adult leaves was examined in seedlings of seven morphs of Begonia dregei using several quantitative methods of shape analysis. There is variation in the shape of adult leaves among individuals as well as in juvenile leaves within individuals in B. dregei. As an individual grows, there is a gradual transition in leaf shape from the symmetrical, oval, smooth-margined leaves through a series of more than ten transitional leaves to a stable adult leaf shape. There appear to be two basic patterns to the acquisition of adult traits. Traits that differ among morphs are acquired gradually throughout the entire transitional series while those that are similar among morphs are acquired by about leaf 5 and remain stable through the later juvenile leaves. There is no identity of leaf shape between the earlier leaves of some morphs and the later leaves of others. Evolutionary diversification in adult leaf morphology in this species is not related to simple changes in ontogeny of the whole plant.     

STUDIES ON THE LEAF OF POPULUS DELTOIDES (SALICACEAE): MORPHOLOGY AND ANATOMY

Mature field- and growth-chamber-grown leaves of Populus deltoides Bartr. ex Marsh. were examined with light and scanning electron microscopes to determine their vasculature and the spatial relationships of the various orders of vascular bundles to the mesophyll. Three leaf traces, one median and two lateral, enter the petiole at the node. Progressing acropetally in the petiole these bundles are rearranged and gradually form as many as 13 tiers of vascular tissue in the petiole at the base of the lamina. (Most leaves contained seven vertically stacked tiers.) During their course through the midrib the tiers “unstack” and portions diverge outward and continue as secondary veins toward the margin on either side of the lamina. As the midvein approaches the leaf tip it is represented by a single vascular bundle which is a continuation of the original median bundle. Tertiary veins arise from the secondary veins or the midvein, and minor veins commonly arise from all orders of veins. All major veins–primaries, secondaries, intersecondaries, and tertiaries–are associated with rib tissue, while minor veins are completely surrounded by a parenchymatous bundle sheath. The bundle sheaths of tertiary, quaternary, and portions of quinternary veins are associated with bundle-sheath extensions. Minor veins are closely associated spatially with both ad- and abaxial palisade parenchyma of the isolateral leaf and also with one or two layers of paraveinal mesophyll that extend horizontally between the veins. The leaves of growth-chamber-grown plants had thinner blades, a higher proportion of air space, and greater interveinal distances than those of field-grown plants.     

Secreted Versus Membrane-anchored Collagenases: RELATIVE ROLES IN FIBROBLAST-DEPENDENT COLLAGENOLYSIS AND INVASION*

Fibroblasts degrade type I collagen, the major extracellular protein found in mammals, during events ranging from bulk tissue resorption to invasion through the three-dimensional extracellular matrix. Current evidence suggests that type I collagenolysis is mediated by secreted as well as membrane-anchored members of the matrix metalloproteinase (MMP) gene family. However, the roles played by these multiple and possibly redundant, degradative systems during fibroblast-mediated matrix remodeling is undefined. Herein, we use fibroblasts isolated from Mmp13^−/−, Mmp8^−/−, Mmp2^−/−, Mmp9^−/−, Mmp14^−/− and Mmp16^−/− mice to define the functional roles for secreted and membrane-anchored collagenases during collagen-resorptive versus collagen-invasive events. In the presence of a functional plasminogen activator-plasminogen axis, secreted collagenases arm cells with a redundant collagenolytic potential that allows fibroblasts harboring single deficiencies for either MMP-13, MMP-8, MMP-2, or MMP-9 to continue to degrade collagen comparably to wild-type fibroblasts. Likewise, Mmp14^−/− or Mmp16^−/− fibroblasts retain near-normal collagenolytic activity in the presence of plasminogen via the mobilization of secreted collagenases, but only Mmp14 (MT1-MMP) plays a required role in the collagenolytic processes that support fibroblast invasive activity. Furthermore, by artificially tethering a secreted collagenase to the surface of Mmp14^−/− fibroblasts, we demonstrate that localized pericellular collagenolytic activity differentiates the collagen-invasive phenotype from bulk collagen degradation. Hence, whereas secreted collagenases arm fibroblasts with potent matrix-resorptive activity, only MT1-MMP confers the focal collagenolytic activity necessary for supporting the tissue-invasive phenotype.In the postnatal state, fibroblasts are normally embedded in a self-generated three-dimensional connective tissue matrix composed largely of type I collagen, the major extracellular protein found in mammals (1–3). Type I collagen not only acts as a structural scaffolding for the associated mesenchymal cell populations but also regulates gene expression and cell function through its interactions with collagen binding integrins and discoidin receptors (2, 4). Consistent with the central role that type I collagen plays in defining the structure and function of the extracellular matrix, the triple-helical molecule is resistant to almost all forms of proteolytic attack and can display a decades-long half-life in vivo (4–6). Nonetheless, fibroblasts actively remodel type I collagen during wound healing, inflammation, or neoplastic states (2, 7–13).To date type I collagenolytic activity is largely confined to a small subset of fewer than 10 proteases belonging to either the cysteine proteinase or matrix metalloproteinase (MMP)² gene families (4, 14–18). As all collagenases are synthesized as inactive zymogens, complex proteolytic cascades involving serine, cysteine, metallo, and aspartyl proteinases have also been linked to collagen turnover by virtue of their ability to mediate the processing of the pro-collagenases to their active forms (13, 15, 19). After activation, each collagenase can then cleave native collagen within its triple-helical domain, thus precipitating the unwinding or “melting” of the resulting collagen fragments at physiologic temperatures (4, 15). In turn, the denatured products (termed gelatin) are susceptible to further proteolysis by a broader class of “gelatinases” (4, 15). Collagen fragments are then either internalized after binding to specific receptors on the cell surface or degraded to smaller peptides with potent biological activity (20–24).Previous studies by our group as well as others have identified MMPs as the primary effectors of fibroblast-mediated collagenolysis (20, 25, 26). Interestingly, adult mouse fibroblasts express at least six MMPs that can potentially degrade type I collagen, raising the possibility of multiple compensatory networks that are designed to preserve collagenolytic activity (25). Four of these collagenases belong to the family of secreted MMPs, i.e. MMP-13, MMP-8, MMP-2, and MMP-9, whereas the other two enzymes are members of the membrane-type MMP subgroup, i.e. MMP-14 (MT1-MMP) and MMP-16 (MT3-MMP) (13, 27–29). From a functional perspective, the specific roles that can be assigned to secreted versus membrane-anchored collagenases remain undefined. As such, fibroblasts were isolated from either wild-type mice or mice harboring loss-of-function deletions in each of the major secreted and membrane-anchored collagenolytic genes, and the ability of the cells to degrade type I collagen was assessed. Herein, we demonstrate that fibroblasts mobilize either secreted or membrane-anchored MMPs to effectively degrade type I collagen in qualitatively and quantitatively distinct fashions. However, under conditions where fibroblasts use either secreted and membrane-anchored MMPs to exert quantitatively equivalent collagenolytic activity, only MT1-MMP plays a required role in supporting a collagen-invasive phenotype. These data establish a new paradigm wherein secreted collagenases are functionally limited to bulk collagenolytic processes, whereas MT1-MMP uniquely arms the fibroblast with a focalized degradative activity that mediates subjacent collagenolysis as well as invasion.     

THE RELATIVE COSTS OF SELF- AND CROSS-FERTILIZED SEEDS IN IMPATIENS CAPENSIS (BALSAMINACEAE)

By using a generally applicable technique that involves monitoring the development and survivorship of flowers and seed capsules, I estimated the material and energetic costs of producing self- and cross-fertilized seeds in Impatiens capensis. All flowers and fruits on six plants were censused intensively for the two-month period of reproduction. Cleistogamous (selfing) flowers ripened seed in about 24 days, compared to about 36 days for the chasmogamous (outcrossing) flowers. In terms of dry weight, selfed seeds cost about two-thirds as much as outcrossed seeds: 12.4 versus 18.4 mg dry weight per seed. When adjusted to the currency of calories, and including an independent estimate of pollen and nectar production in outcrossing flowers, I estimate the costs to be about 65 and 135 calories per selfed or outcrossed seed. Sources of error include the accuracy of the estimates of flower and fruit weight, and possible differences among the developmental stages in respiratory costs. The cost discrepancy implies that outcrossed seeds should possess a countervailing fitness advantage large enough to offset their greater energetic cost.     

一氧化氮和前列腺素在内毒素引起降钙素基因相关肽释放中的作用

本实验在离体灌流大鼠肠系膜动脉床研究内毒素引起降钙素基因相关肽（ＣＧＲＰ）释放的机制。内毒素（５０μｇ／ｍｌ）使ＣＧＲＰ释放增加１６倍,一氧化氮合成酶（ＮＯＳ）底物Ｌ－精氨酸（Ｌ－Ａｒｇ）能促进内毒素引起的ＣＧＲＰ释放（４１％）。ＮＯＳ抑制剂Ｎ￣Ｇ－硝基－Ｌ－精氨酸（Ｌ－ＮＮＡ）及鸟苷酸环化酶抑制剂甲基蓝（ＭＢ）能使内毒素的上述作用分别降低３５％与３６％,Ｌ－精氨酸（ｔ－Ａｒｇ）能逆转Ｌ－ＮＮＡ的作用。提示内毒素的作用机制中部分是通过一氧化氮引起细胞内ｃＧＭＰ升高而介导的。用化学方法破坏血管内皮细胞,Ｌ－ＮＮＡ与Ｌ－Ａｒｇ的上述作用依然存在。提示内毒素主要是激活血管周围感觉神经末梢的神经源ＮＯＳ,而非内皮源ＮＯＳ。环氧化酶抑制剂消炎痛（Ｉｎｄｏ）与布洛芬（Ｉｂｕ）也能使内毒素引起ＣＧＲＰ释放的作用分别降低３４％与３９％,但与Ｌ－ＮＮＡ的作用不能迭加。提示内毒素可能通过激活神经源ＮＯＳ进而引起环氧化酶活化而起作用。     

STUDIES ON THE LEAF OF POPULUS DELTOIDES (SALICACEAE): QUANTITATIVE ASPECTS,AND SOLUTE CONCENTRATIONS OF THE SIEVE-TUBE MEMBERS

The vascular system of the leaf of Populus deltoides Bartr. ex Marsh, was examined quantitatively, and plasmolytic studies were carried out to determine the solute concentrations of sieve-tube members at various locations in the leaf. Both the total number and total crosssectional area of each cell type decreases with decreasing vein size. Although the proportion of phloem occupied by sieve tubes varies considerably from location to location, a linear relationship exists between cross-sectional area of the vascular bundles and both total and mean cross-sectional area of sieve tubes. Collectively, the cross-sectional area of all tertiary and minor veins feeding into a secondary exceeds the total cross-sectional area of sieve tubes at the base of that secondary. Moreover, the total volume of sieve tubes in the “catchment area” of a secondary vein is much greater than the total sieve tube volume of the secondary itself. Both tracheary elements and sieve-tube members undergo a reduction in both total and mean crosssectional area in the constricted zone at the base of the leaf. The plasmolytic studies revealed the presence of positive concentration gradients in sieve tubes of the lamina from the minor veins and tips of the secondaries to the bases of the secondaries and their associated subjacent midvein bundles and from the upper to lower portions of the median bundle of the midvein.