Evidence that Blastocladiella emersonii zoospore maintenance factor is a sulfhydryl group-containing cyclic ribotide

Blastocladiella emersonii zoospore maintenance factor (ZMF), released into the medium during zoospore production, mediates a reversible developmental block to zoospore encystment (Gottschalk, W. K., and Sonneborn, D. R. (1981) Exp. Mycol. 5, 1-14 and (1982) Dev. Biol. 93, 165-180). Crude ZMF and purified ZMF display indistinguishable sensitivities/insensitivities to inactivations by several different chemical or enzymatic treatments. Such data have provided additional support for the conclusion (Gottschalk, W. K., and Sonneborn, D. R. (1985) J. Biol. Chem. 260, 6588-6591) that ZMF biological activity resides in a single molecular species. The inactivation analyses have provided substantial evidence that ZMF is a newly discovered SH-containing cyclic ribotide. At least one SH-containing side group and at least one free amino group linked to an imidazole, as well as a ribosyl moiety containing a cyclic 3',5'-phosphate, a 2'-free hydroxyl, and a 1'-linkage to the imidazole, appear to be essential structural requirements for ZMF-mediated encystment blockage. The proposed structure of biologically functional ZMF is similar to that of a key intermediate in the de novo pathway of purine nucleotide biosynthesis (5'-phosphoribosyl-5-aminoimidazole-4-N-succino-carboxamide), except that ZMF, and not 5'-phosphoribosyl-5-aminoimidazole-4-N-succinocarboxamide, contains a cyclic phosphate and at least one reduced SH group.     

Phosphorylation-dependent regulation of amidotransferase during the development of Blastocladiella emersonii

The enzyme amidotransferase [2-amino-2-deoxy-D-glucose-6-phosphate ketol isomerase (amino-transferring); EC 2.6.1.16] catalyzes the first step in the hexosamine biosynthetic pathway. In Blastocladiella emersonii the sensitivity of the enzyme to the inhibitor uridine-5'-diphospho-N-acetylglucosamine (UDP-GlcNAc) is developmentally regulated. The inhibitable form of amidotransferase activity present in the zoospore is converted to a noninhibitable form during germination. The latter form is present throughout the growth phase and sensitivity to UDP-GlcNAc gradually returns to the zoospore level during sporulation [C.P. Selitrennikoff, N.E. Dalley, and D.R. Sonneborn (1980) Proc. Natl. Acad. Sci. USA 77, 5998-6002]. The following evidence suggests that a phosphorylation/dephosphorylation mechanism underlies this interconversion: (i) Both the vegetative and zoospore enzymes have the same molecular weight of 140,000, but the vegetative enzyme elutes significantly earlier on a DEAE-cellulose column than does the zoospore enzyme. (ii) The increased sensitivity to UDP-GlcNAc occurring in vivo and in vitro correlates with increased phosphorylation of a polypeptide of apparent Mr 76,000. This component copurifies with amidotransferase activity through ion-exchange chromatography and sucrose density gradient centrifugation. (iii) Desensitization and concurrent dephosphorylation of sensitive amidotransferase can be observed in vitro after treatment with a partially purified magnesium-dependent phosphoprotein phosphatase from zoospores.     

COMPARATIVE CYTOLOGY AND TAXONOMY OF THE ULVAPHYCEAE. I. THE ZOOSPORE OF ULOTHRIX ZONATA (CHLOROPHYTA)1

This fine structural study of the quadriflagellate zoospore of Ulothrix zonata (Weber & Mohr) Kützing, with special attention to the flagellar root system, demonstrates that it is very similar to the zoospore of Ulva lactuca L. in several aspects. Common features include the presence of a cruciate root system (4-2-4-2 type), a non-striated band that connects basal bodies, a so-called terminal cap, and system I and system II striated root components. Only slight differences exist, i.e. in the shape of the terminal cap, and in the number and position of the system II root components. It is concluded that the taxonomic affinities of U. zonata lie with the Ulvaphyceae sensu Stewart and Mattox rather than with the Chlorophyceae. Additional support for this conclusion is the discovery of tiny, flat body scales on the zoospore of U. zonata. A summary of the distinctive characteristics of the Chlorophyceae, Charophyceae and Ulvaphyceae reflecting the current state of knowledge is given.     

3' cleavage and polyadenylation of mRNA precursors in vitro requires a poly(A) polymerase, a cleavage factor, and a snRNP

We have separated and purified three factors from HeLa cell nuclear extracts that together can accurately cleave and polyadenylate pre-mRNAs containing the adenovirus L3 polyadenylation site. One of the factors is a poly(A) polymerase with a molecular weight of approximately 50-60 kd. The second activity is a cleavage factor with a native molecular weight in the range of 70-120 kd. The third component is a factor (cleavage and polyadenylation factor, CPF) that is needed for the cleavage reaction and, in addition, confers specificity to the poly(A) polymerase activity; the native molecular weight of CPF is approximately 200 kd. Poly(A) polymerase together with CPF is sufficient to specifically polyadenylate pre-mRNA substrates that have been precleaved at the poly(A) addition site. In contrast, all three components are required for accurate cleavage and polyadenylation of pre-mRNA substrates. Further purification of CPF by buoyant density centrifugation, ion exchange, and affinity column chromatography or by gel filtration demonstrates that CPF activity resides in a ribonucleoprotein and copurifies with U11 snRNP.     

Blastocladiella emersonii zoospore maintenance factor: A quantitative bioassay used to characterize extracellular appearance and maintenance of factor and interactions between factor,zoospores, and salt

A quantitative bioassay forBlastocladiella emersonii zoospore maintenance factor (ZMF) is documented analyzed. The basis of the assay is an antagonistic interaction between NaCl and ZMF; the former elicits rapid encystment in zoospore populations whereas the latter counteracts this effect. Dilution techniques are used to quantitate the effects of each variable in terms of percentage rapid encystment. Each conditional bioassay variable examined (NaCl concentration, assay pH, cell density, zoospore age, time of assay) appears to affect the sensitivity of zoospore populations to ZMF rather than the level of ZMF activity per se. In particular, zoospore populations “age,” in terms of lowered sensitivity to ZMF, in the following ways: (a) “zoospore aging” without NaCl added and either with or without ZMF added; and (b) “delayed encystment” with NaCl and different fixed levels of ZMF added. ZMF activity in the medium (buffered CaCl₂) accumulates gradually during sporulation; under bioassay conditions and even at subsaturating levels of ZMF activity, activity recoverable from the medium remains stable over long time intervals (4.5–24 h) whether populations remain zoospores or germinate. Populations incubated without added ZMF, whether they remain zoospores or germinate, do not detectably release ZMF activity into the medium. We discuss the following proposals concerning the functions of ZMF: (a) ZMF acts as a negative regulator of themechanics of zoospore encystment; and (b) ZMF acts as a self-generated, natural dispersal signal for the organism.     

Protein synthesis in Artemia salina

It is thought that eucaryotic elongation factor eEF-Ts catalyzes the replacement of GDP for GTP on eucaryotic elongation factor eEF-Tu. We have found that eEF-Ts displays a strong nucleoside diphosphate phosphotransferase activity. This transferase activity resides in a dimer molecule of a subunit molecular weight close to 30,000. The transfosforylating activity of eEF-Ts results in a stimulatory effect of ATP, GTP, UTP and CTP on protein synthesis provided that GDP is present. The specificity for guanine nucleotides in protein synthesis resides only in eEF-Tu.     

The last two pathway-specific enzyme activities of hexosamine biosynthesis are present in Blastocladiella emersonii zoospores prior to germination

Forward direction assays have been developed for the last two pathway-specific enzymes of hexosamine biosynthesis using crude extracts from Blastocladiella emersonii zoospores. The specific enzyme activities measured are substantially higher than those reported with enzyme preparations from other organisms. During the development of one of the assays, another enzyme activity was observed which converts one of the intermediates of the pathway, N-acetylglucosamine-6-phosphate, to N-acetylglucosamine. The finding of these three enzyme activities in zoospore extracts completes the demonstration that all the enzyme activities necessary to synthesize some 2% by weight as chitin early during zoospore germination (de novo cell wall formation) pre-exist in the zoospore. This demonstration is consistent with the conclusion that the hexosamine pathway is regulated at the post-translational level during zoospore germination.     

Ovine myometrial cytosol inhibits phospholipase A activity, in vitro

Myometrium obtained from pregnant ewes (30-80 days gestation) contains a factor which inhibits phospholipase A2 (PLA2) activity. The activity of this moiety was assessed using an in vitro porcine pancreatic PLA2 assay system. Inhibitory activity was associated with a 35-45000 dalton molecular weight fraction, heat-labile, sensitive to protease degradation and did not partition into organic solvents. These data are indicative that PLA2-inhibitory activity resides in a protein moiety. Dixon-plot analysis of myometrial-inhibitory activity was indicative that the inhibition of PLA2 activity was of a non-competitive nature (Ki = 4.1 +/- 0.7 micrograms/ml, ca 118 nmol/l). Myometrial phospholipase-inhibitory protein(s) may be involved in the suppression of eicosanoid biosynthesis by the uterine tissues throughout gestation thus inhibiting uterine contractile activity.     

BMP-binding modules in chordin: a model for signalling regulation in the extracellular space

A number of genetic and molecular studies have implicated Chordin in the regulation of dorsoventral patterning during gastrulation. Chordin, a BMP antagonist of 120 kDa, contains four small (about 70 amino acids each) cysteine-rich domains (CRs) of unknown function. In this study, we show that the Chordin CRs define a novel protein module for the binding and regulation of BMPs. The biological activity of Chordin resides in the CRs, especially in CR1 and CR3, which have dorsalizing activity in Xenopus embryo assays and bind BMP4 with dissociation constants in the nanomolar range. The activity of individual CRs, however, is 5- to 10-fold lower than that of full-length Chordin. These results shed light on the molecular mechanism by which Chordin/BMP complexes are regulated by the metalloprotease Xolloid, which cleaves in the vicinity of CR1 and CR3 and would release CR/BMP complexes with lower anti-BMP activity than intact Chordin. CR domains are found in other extracellular proteins such as procollagens. Full-length Xenopus procollagen IIA mRNA has dorsalizing activity in embryo microinjection assays and the CR domain is required for this activity. Similarly, a C. elegans cDNA containing five CR domains induces secondary axes in injected Xenopus embryos. These results suggest that CR modules may function in a number of extracellular proteins to regulate growth factor signalling.     

Purification of melanoma growth stimulatory activity

The Hs0294 human malignant melanoma cell line produces a monolayer mitogen that stimulates the serum free growth of low-density cultures of Hs0294 cells. This report describes the purification of that mitogen, termed MGSA for melanoma growth stimulatory activity, from serum-free conditioned medium from the Hs0294 cultures. MGSA has been purified from acetic acid extracts of lyophilized conditioned medium by gel filtration, reverse-phase high-pressure liquid chromatography (RP-HPLC), and preparative electrophoresis, resulting in a greater than 400,000-fold purification. MGSA bioactivity resides in acid- and heat-stable polypeptides of high and low molecular weight (24-28 kd and less than 14-16 kd). However, the majority of the activity is reproducibly associated with the approximately 16-kd moiety eluting from RP-HPLC at approximately 35% acetonitrile. Reduction with dithiothreitol or B-mercaptoethanol results in a loss of biological activity but does not convert the 24-28-kd moieties to the less than 14-16-kd forms of MGSA. 125I-MGSA that has been purified by preparative electrophoresis (16 kd) specifically binds to Hs0294 melanoma cells and retains 100% of the growth-stimulatory activity. The 16-kd MGSA stimulates the proliferation of Hs0294 cells at concentrations of 0.3-30 pM. The electrophoretic mobility of MGSA is also unaltered by the preparative electrophoresis procedure, further demonstrating that this procedure does not alter the biochemical integrity of the growth factor. Purified MGSA does not enable anchorage-independent growth of normal rat kidney (NRK) cells and is therefore different from the previously described transforming growth factors. The amino acid composition of MGSA differs from that of other previously described growth factors. These data demonstrate that MGSA represents a separate class of growth factors with biological and biochemical properties different from other growth factors.     

Purification of a factor inducing differentiation of mouse myeloid leukemic M1 cells from conditioned medium of mouse fibroblast L929 cells

A procedure is described for purification of a factor (D-factor)-inducing differentiation of mouse myeloid leukemic cells (M1) into macrophages from serum-free mouse L929 cell-conditioned medium. The procedure included ammonium sulfate precipitation, DEAE-cellulose, Sephadex G-200 and phenyl-Sepharose column chromatographies, reverse-phase high-performance liquid chromatography on a C18 hydrophobic support, and high-performance liquid chromatography on a gel-filtration column. The purified factor gave a single band of protein with a molecular weight of 62,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis which coincided with biological activity. Its half-maximal concentration for inducing differentiation of M1 cells into macrophages was 1.7 X 10(-11) M. Even at 2.6 X 10(-9) M, it did not induce colony formation of normal bone marrow cells, suggesting that it was distinct from the growth factor for normal precursors of macrophages and/or granulocytes.     

Evidence for differential regulation of two classes of poly(A) RNA inBlastocladiella emersonii zoospores

The poly(A) RNA in zoospores ofBlastocladiella emersonii contains RNA synthesized during the growth phase (GP poly(A) RNA) and late sporulation (LS poly(A) RNA). LS poly(A) RNA synthesized during the final 30 minutes of sporulation is bound exclusively to polyribosomes which comprise approximately 50% of the total zoospore ribosome population. In contrast, GP poly(A) RNA is bound to zoospore monoribosomes. During the final 30 minutes of sporulation, GP poly(A) RNA which is bound to polyribosomes makes a transition to monoribosomes. Zoospore monoribosomes and RNA extracted from zoospore monoribosomes are inactivein vitro while both zoospore polyribosomes and RNA extracted from zoospore polyribosomes stimulate protein synthesis in the wheat germin vitro system. Sedimentation of poly(A) RNA from zoospore monoribosomes on dimethyl sulfoxide gradients revealed that the GP poly(A) RNA was of sufficiently high molecular weight to code for average-sized proteins. These denaturing gradients failed to activate the zoospore monoribosome RNA. The results suggest that the inability to translate zoospore monoribosomesin vitro is due to some property or modification of the zoospore monoribosome poly(A) RNA. Zoospore monoribosomes bound to poly(A) RNA contain an average of two tRNA molecules while zoospore polyribosomes have an average of less than one tRNA bound. This suggests the two classes of ribosomes are blocked at different steps in the elongation process.     

Zoospore ultrastructure and phylogenetic position of Phlyctochytrium aureliae Ajello is revealed (Chytridiaceae, Chytridiales, Chytridiomycota)

From forest soils in Scotland Phlyctochytrium aureliae was observed and brought into pure culture. Previously included in a molecular phylogenetic study of Chytridiales as Phlyctochytrium sp. KP 061, the organism groups with Phlyctochytrium planicorne, P. bullatum, Chytridium olla and C. lagenaria in the family Chytridiaceae. Thallus morphology and development as well as zoospore ultrastructure are detailed herein. The sporangium is epibiotic, spherical or subspherical, apophysate or non-apophysate, and ornamented with dentate enations. The overall zoospore ultrastructural features are consistent with the Group II type zoospore that characterizes family Chytridiaceae in the Chytridiales, although the zoospore also has two character states unique to this taxon: the MLC cisterna fenestrations are one-third to one-half the diameter of fenestrations in other Chytridiaceae zoospores and an accumulation of electron-dense material (a kinetosome-associated structure, or KAS) proximal to the kinetosome and non-flagellated centriole is extensive and unique. This study verifies that zoospore ultrastructure of P. aureliae zoospores places this species in the Chytridiales and Chytridiaceae, as indicated in a previous molecular phylogenetic study.     

A catalytic domain exosite (Cys527-Cys542) in factor XIa mediates binding to a site on activated platelets

The zymogen, factor XI, and the enzyme, factor XIa, interact specifically with functional receptors on the surface of activated platelets. These studies were initiated to identify the molecular subdomain within factor XIa that binds to activated platelets. Both factor XIa (Ki approximately 1.4 nM) and a chimeric factor XIa containing the Apple 3 domain of prekallikrein (Ki approximately 2.7 nM) competed with [125I]factor XIa for binding sites on activated platelets, suggesting that the factor XIa binding site for platelets is not located in the Apple 3 domain which mediates factor XI binding to platelets. The recombinant catalytic domain (Ile370-Val607) inhibited the binding of [125I]factor XIa to the platelets (Ki approximately 3.5 nM), whereas the recombinant factor XI heavy chain did not, demonstrating that the platelet binding site is located in the light chain of factor XIa. A conformationally constrained cyclic peptide (Cys527-Cys542) containing a high-affinity (KD approximately 86 nM) heparin-binding site within the catalytic domain of factor XIa also displaced [125I]factor XIa from the surface of activated platelets (Ki approximately 5.8 nM), whereas a scrambled peptide of identical composition was without effect, suggesting that the binding site in factor XIa that interacts with the platelet surface resides in the catalytic domain near the heparin binding site of factor XIa. These data support the conclusion that a conformational transition accompanies conversion of factor XI to factor XIa that conceals the Apple 3 domain factor XI (zymogen) platelet binding site and exposes the factor XIa (enzyme) platelet binding site within the catalytic domain possibly comprising residues Cys527-Cys542.     

Studies on the vegetative life cycle of Chlamydomonas reinhardi Dangeard in synchronous culture III. Some notes on the process of zoospore liberation

When Chlamydomonas reinhardi cells liberate zoospores, theyexcrete into the medium a factor(s) which induces zoospore liberationof other cells that are not yet ready to liberate zoosporesby themselves. The "factor" is contained within cells at laterstages of the cell cycle, but its action is suppressed untilthe regular time of zoospore liberation in the cell cycle. (Received November 18, 1974; )     

Characterization and Partial Purification of a Novel Neuronotrophic Factor from Bovine Seminal Vesicle

Extracts from bovine seminal vesicles have been shown to contain high concentrations of nerve growth factor (NGF)-like biological activity and of the NGF protein with properties corresponding to that of NGF from other sources. We now demonstrate that a second neuronotrophic protein, termed seminal vesicle-derived neuronotrophic factor (SVNF), is present in seminal vesicle extracts (SVEs), which could not be distinguished from NGF on the basis of biological activity. SVNF has neuronotrophic activity on NGF target cells like embryonic chicken-sensory and sympathetic neurons, sympathetic neurons, and chromaffin cells from neonatal rats, but it is inactive on embryonic chicken ciliary or neonatal rat nodose ganglion neurons. It also stimulates fiber outgrowth from rat pheochromocytoma (PC 12) cells. In gel filtration chromatography on Biogel A 1.5 m, the activity is eluted with an apparent molecular weight of 40 kilodaltons, and by preparative isoelectric focusing, the isoelectric point was determined to be in the neutral range (6.8-7.8). The biological activity of SVNF, in contrast to that of NGF, is partially retained after preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis and can be electrophoretically eluted with an apparent molecular weight of 16-20 kilodaltons. Electrophoretically purified SVNF is not inhibited by antisera to mouse NGF, but its activity is increased greater than 10-fold in the presence of very low concentrations of NGF. For partially purified SVNF, a specific activity of 2.9-5.8 X 10(5) biological units/mg of protein was determined in the presence of subthreshold NGF concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Epidermal growth factor-like transforming growth factor. I. Isolation, chemical characterization, and potentiation by other transforming factors from feline sarcoma virus-transformed rat cells

An acid-stable transforming growth factor (TGF) that interacts with epidermal growth factor (EGF) receptors and is structurally related to EGF was isolated from serum-free culture fluids of Snyder-Theilen feline sarcoma virus-transformed rat embryo (FeSV-Fre) cells. Purification of this EGF-like TGF (eTGF) was achieved by molecular filtration chromatography and successive reverse-phase high pressure liquid chromatography steps on octadecyl support eluted with acetonitrile and 1-propanol gradients, respectively. Rat eTGF consists of a 7.4-kD single polypeptide chain that co-migrates with biological activity in dodecyl sulfate-polyacrylamide electrophoresis gels. Like preparations of a related TGF from human melanoma cells (Marquardt, H., and Todaro, G.J. (1982) J. Biol. Chem. 257, 5220-5225), but unlike EGF from rat, human, or mouse, rat eTGF has phenylalanine and lacks methionine. However, the sequence of the first 30 amino acid residues in rat eTGF is H2N-Val-Val-Ser-His-Phe-Asn-Lys-Cys-Pro-Asp-Ser-His-Thr-Gln-Tyr-Cys-Phe-His-Gly - Thr-(x)-Arg-Phe-Leu-Val-Gln-Glu-Glu-(Lys)-(Lys)-, which is significantly (20% and 28%) homologous to the NH2-terminal region of mouse EGF and human EGF, respectively. In addition to eTGF, molecular filtration chromatography of acid-soluble extracts from medium conditioned by FeSV-Fre cells resolved a 14-kD transforming factor(s) apparently devoid of intrinsic mitogenic activity but able to elicit a strong anchorage-independent growth response in the presence of eTGF or EGF. These results show that: 1) a 7.4-kDa TGF structurally and functionally related to EGF has been isolated from FeSV-Fre cells and 2) the full anchorage-independent growth-promoting activity of medium conditioned by FeSV-Fre cells is due to the coordinate action of at least two types of factors, the 7.4-kDa eTGF and a second 14-kDa transforming factor(s).     

Cholecystokinin octapeptide in rat central nervous system: Immunocytochemical studies using a monoclonal antibody that does not react with CGRP

The biological activity of the important neuropeptide cholecystokinin octapeptide (CCK8) resides at the C-terminus. Antibodies with C-terminal specificity have been reported to cross-react with a different neuropeptide, calcitonin gene related peptide (CGRP) and this has frustrated the interpretation of immunohistochemical studies. We describe here the properties of a monoclonal antibody to the CCK-related peptide, caerulein, that reacts with the C-terminal region of CCK8, but does not react with CGRP in radioimmunoassay or immunohistochemistry. The distribution of CCK-like activity revealed by immunohistochemistry using this antibody broadly resembles that described previously with a single major exception: in the dorsal horn of the spinal cord. The results support the suggestion that apparent CCK activity in the terminals of rat primary sensory neurones is due to cross-reactivity with CGRP.     

THE SECRET LIFE OF KELPS: PLANKTONIC PROCESSES AND POPULATION DYNAMICS

Understanding spatio‐temporal variability in recruitment is vital to studies of kelp population dynamics. Research on settlement and post‐settlement processes has suggested that arrival of kelp zoospores to suitable substrate is important in limiting kelp recruitment, yet the role of planktonic processes in kelp population dynamics has not been studied due to difficulties in sampling and identifying zoospores. I developed a method to estimate kelp zoospore abundance from in situ plankton samples and used it to study various processes regulating the availability of giant kelp (Macrocystis pyrifera) zoospores for settlement. My studies focused on (1) identifying temporal scales over which zoospore abundance is most variable, (2) describing physical and biological processes that regulate this variability, and (3) determining the relationship between zoospore abundance and settlement. I found that short‐term variability in zoospore abundance (<24 hrs) was not due to changes in supply but rather dispersion, caused by oscillating hydrodynamic forces (e.g. waves). Long‐term variability in zoospore abundance, however, was best explained by the size and density of reproductive adult plants, with zoospore abundance being most variable at the scale of days to months. Changes in adult reproductive condition caused rapid changes in zoospore abundance suggesting that the supply of kelp zoospores is sensitive to environmental regulation of adult physiology. Thus, unlike with marine animals, these results indicate that variability in kelp propagule supply, over scales most likely to affect subsequent settlement and recruitment, is more tightly coupled to demographic and reproductive mechanisms than to physical transport processes.     

LET THERE BE LIGHT (BUT NOT TOO MUCH): MOLECULAR EVOLUTION OF LIGHT-HARVESTING COMPLEXES

Understanding spatio-temporal variability in recruitment is vital to studies of kelp population dynamics. Research on settlement and post-settlement processes has suggested that arrival of kelp zoospores to suitable substrate is important in limiting kelp recruitment, yet the role of planktonic processes in kelp population dynamics has not been studied due to difficulties in sampling and identifying zoospores. I developed a method to estimate kelp zoospore abundance from in situ plankton samples and used it to study various processes regulating the availability of giant kelp ( Macrocystis pyrifera ) zoospores for settlement. My studies focused on (1) identifying temporal scales over which zoospore abundance is most variable, (2) describing physical and biological processes that regulate this variability, and (3) determining the relationship between zoospore abundance and settlement. I found that short-term variability in zoospore abundance (<24 hrs) was not due to changes in supply but rather dispersion, caused by oscillating hydrodynamic forces (e.g. waves). Long-term variability in zoospore abundance, however, was best explained by the size and density of reproductive adult plants, with zoospore abundance being most variable at the scale of days to months. Changes in adult reproductive condition caused rapid changes in zoospore abundance suggesting that the supply of kelp zoospores is sensitive to environmental regulation of adult physiology. Thus, unlike with marine animals, these results indicate that variability in kelp propagule supply, over scales most likely to affect subsequent settlement and recruitment, is more tightly coupled to demographic and reproductive mechanisms than to physical transport processes.