Stolon Tips Research Articles

Cytokinins: Determinants of Sink Storage Ability

A new study demonstrates that storage organ formation can be induced in the axillary meristem of non-tuberizing plants by ectopic expression of the cytokinin biosynthetic gene LONELY GUY (LOG1).

Down regulation of StGA3ox genes in potato results in altered GA content and affect plant and tuber growth characteristics

GA biosynthesis and catabolism has been shown to play an important role in regulating tuberization in potato. Active GAs are inactivated in the stolon tips shortly after induction to tuberization. Overexpression of a GA inactivation gene results in an earlier tuberization phenotype, while reducing expression of the same gene results in delayed tuberization. In addition, overexpression of genes involved in GA biosynthesis results in delayed tuberization, while decreased expression of those genes results in earlied tuberization. The final step in GA biosynthesis is catalysed by StGA3ox1 and StGA3ox2 activity, that convert inactive forms of GA into active GA1 and GA4. In this study we cloned StGA3ox2 gene in an RNAi construct and used this construct to transform potato plants. The StGA3ox2 silenced plants were smaller and had shorter internodes. In addition, we assayed the concentrations of various GAs in the transgenic plants and showed an altered GA content. No difference was observed on the time point of tuber initiation. However, the transgenic clones had increased number of tubers with the same yield, resulting in smaller average tuber weight. In addition, we cloned the promoter of StGA3ox2 to direct expression of the GUS reporter gene to visualize the sites of GA biosynthesis in the potato plant. Finally, we discuss how changes of several GA levels can have an impact on shoot, stolon and tuber development, as well as the possible mechanisms that mediate feed-forward and feed-back regulation loops in the GA biosynthetic pathway in potato.

The Impact of the Long-Distance Transport of aBEL1-Like Messenger RNA on Development

BEL1- and KNOTTED1-type proteins are transcription factors from the three-amino-loop-extension superclass that interact in a tandem complex to regulate the expression of target genes. In potato (Solanum tuberosum), StBEL5 and its Knox protein partner regulate tuberization by targeting genes that control growth. RNA movement assays demonstrated that StBEL5 transcripts move through the phloem to stolon tips, the site of tuber induction. StBEL5 messenger RNA originates in the leaf, and its movement to stolons is induced by a short-day photoperiod. Here, we report the movement of StBEL5 RNA to roots correlated with increased growth, changes in morphology, and accumulation of GA2-oxidase1, YUCCA1a, and ISOPENTENYL TRANSFERASE transcripts. Transcription of StBEL5 in leaves is induced by light but insensitive to photoperiod, whereas in stolon tips growing in the dark, promoter activity is enhanced by short days. The heterodimer of StBEL5 and POTH1, a KNOTTED1-type transcription factor, binds to a tandem TTGAC-TTGAC motif that is essential for regulating transcription. The discovery of an inverted tandem motif in the StBEL5 promoter with TTGAC motifs on opposite strands may explain the induction of StBEL5 promoter activity in stolon tips under short days. Using transgenic potato lines, deletion of one of the TTGAC motifs from the StBEL5 promoter results in the reduction of GUS activity in new tubers and roots. Gel-shift assays demonstrate BEL5/POTH1 binding specificity to the motifs present in the StBEL5 promoter and a double tandem motif present in the StGA2-oxidase1 promoter. These results suggest that, in addition to tuberization, the movement of StBEL5 messenger RNA regulates other aspects of vegetative development.

A crosstalk of auxin and GA during tuber development

Several hormones have been studied for their effect on tuber initiation and development. Until recently, the hormone with the most prominent role in tuber initiation was attributed to GA. Genes involved in GA degradation do exhibit an upregulated profile during early stages of tuber development, leading to a rapid decrease of active GA content, thereby facilitating stolon-tip swelling. While GA is known to be involved in shoot and stolon elongation, the development of the new tuberorgan requires changes in meristem identity and the reorientation ofthe plane of cell division. In other developmental processes, such as embryo patterning, flower development and lateral root initiation auxin plays a key role. Recent evidence on the involvement of auxin in tuber formation was providedby the measurement of auxin content in swelling stolons. Auxin content in the stolon tips increased several fold prior to tuber swelling. In vitro tuberisation experiments with auxin applications support the role of auxin during tuber initiation. Taken together, it is becoming clear that the initiation and induction of tubers in potato is a developmental process that appears to be regulated by a crosstalk between GA and auxin.

Dynamic proteomic profile of potato tuber during its in vitro development

Potato tuberization is a complicated biochemical process, which is dependent on external environmental factors. Tuber development in potato consists of a series of biochemical and morphological processes at the stolon tip. Signal transduction proteins are involved in the source-sink transition during potato tuberization. In the present study, we examined protein profiles under in vitro tuber-inducing conditions using a shotgun proteomic approach involving denaturing gel electrophoresis and liquid chromatography–mass spectrometry. A total of 251 proteins were identified and classified into 9 groups according to distinctive expression patterns during the tuberization stage. Stolon stage-specific proteins were primarily involved in the photosynthetic machinery. Proteins specific to the initial tuber stage included patatin. Proteins specific to the developing tuber stage included 6-fructokinase, phytoalexin-deficient 4-1, metallothionein II-like protein, and malate dehydrogenase. Novel stage-specific proteins identified during in vitro tuberization were ferredoxin–NADP reductase, 34kDa porin, aquaporin, calmodulin, ripening-regulated protein, and starch synthase. Superoxide dismutase, dehydroascorbate reductase, and catalase I were most abundantly expressed in the stolon; however, the enzyme activities of these proteins were most activated at the initial tuber. The present shotgun proteomic study provides insights into the proteins that show altered expression during in vitro potato tuberization.

The effects of auxin and strigolactones on tuber initiation and stolon architecture in potato

Various transcriptional networks and plant hormones have been implicated in controlling different aspects of potato tuber formation. Due to its broad impact on many plant developmental processes, a role for auxin in tuber initiation has been suggested but never fully resolved. Here, auxin concentrations were measured throughout the plant prior to and during the process of tuber formation. Auxin levels increase dramatically in the stolon prior to tuberization and remain relatively high during subsequent tuber growth, suggesting a promoting role for auxin in tuber formation. Furthermore, in vitro tuberization experiments showed higher levels of tuber formation from axillary buds of explants where the auxin source (stolon tip) had been removed. This phenotype could be rescued by application of auxin on the ablated stolon tips. In addition, a synthetic strigolactone analogue applied on the basal part of the stolon resulted in fewer tubers. The experiments indicate that a system for the production and directional transport of auxin exists in stolons and acts synergistically with strigolactones to control the outgrowth of the axillary stolon buds, similar to the control of above-ground shoot branching.

Molecular characterisation of the Cu/Zn superoxide dismutase gene (ZjSOD1) induced by salt stress in Zoysia japonica

SummaryNaCl was used to evaluate the molecular response to salt stress in Zoysia japonica. A 452-bp cDNA clone, which showed high similarity to other plant Cu/Zn superoxide dismutase (SOD) gene sequences, was selected and designated Zoysia japonica Cu/Zn superoxide dismutase 1 (ZjSOD1). Southern blot analysis showed the presence of at least two copies of the ZjSOD1 gene in Z. japonica. ZjSOD1 transcripts increased with exposure to a high salt concentration (300 mM NaCl). ZjSOD1 transcript levels increased in non-immersed tissues such as stolons and their tips, as well as in stolon tissue that had been immersed in 300 mM NaCl for 36 h. The highest level of ZjSOD1 gene expression was observed in stolon tips. However, ZjSOD1 transcript levels were higher in immersed stolons than in non-immersed stolons, or their tips, during the salt stress treatment. Molecular characterisation of the ZjSOD1 gene may provide clues about the response mechanism to salt stress in the stolons of Z. japonica and other turfgrasses.

A Model System of Development Regulated by the Long‐distance Transport of mRNA

BEL1-like transcription factors are ubiquitous in plants and interact with KNOTTED1-types to regulate numerous developmental processes. In potato, the RNA of several BEL1-like transcription factors has been identified in phloem cells. One of these, StBEL5, and its Knox protein partner regulate tuber formation by targeting genes that control growth. RNA detection methods and grafting experiments demonstrated that StBEL5 transcripts move across a graft union to localize in stolon tips, the site of tuber induction. This movement of RNA originates in source leaf veins and petioles and is induced by a short-day photoperiod, regulated by the untranslated regions, and correlated with enhanced tuber production. Addition of the StBEL5 untranslated regions to another BEL1-like mRNA resulted in its preferential transport to stolon tips leading to increased tuber production. Upon fusion of the untranslated regions of StBEL5 to a beta-glucuronidase marker, translation in tobacco protoplasts was repressed by those constructs containing the 3' untranslated sequence. The untranslated regions of the StBEL5 mRNA are involved in mediating its long-distance transport and in controlling translation. The 3' untranslated sequence contains an abundance of conserved motifs that may serve as binding motifs for RNA-binding proteins. Because of their presence in the phloem sieve tube system, their unique untranslated region sequences and their diverse RNA accumulation patterns, the family of BEL1-like RNAs from potato represents a valuable model for studying the long-distance transport of full-length mRNAs and their role in development.

Untranslated Regions of a Mobile Transcript Mediate RNA Metabolism

BEL1-like transcription factors are ubiquitous in plants and interact with KNOTTED1 types to regulate numerous developmental processes. In potato (Solanum tuberosum subsp. andigena), the BEL1-like transcription factor StBEL5 and its Knox protein partner regulate tuber formation by targeting genes that control growth. RNA detection methods and heterografting experiments demonstrated that StBEL5 transcripts are present in phloem cells and move across a graft union to localize in stolon tips, the site of tuber induction. This movement of RNA originates in leaf veins and petioles and is induced by a short-day photoperiod, regulated by the untranslated regions, and correlated with enhanced tuber production. Assays for RNA mobility suggest that both 5' and 3' untranslated regions contribute to the preferential accumulation of the StBEL5 RNA but that the 3' untranslated region may contribute more to transport from the leaf to the stem and into the stolons. Addition of the StBEL5 untranslated regions to another BEL1-like mRNA resulted in its preferential transport to stolon tips and enhanced tuber production. Transcript stability assays showed that the untranslated regions and a long-day photoperiod enhanced StBEL5 RNA stability in shoot tips. Upon fusion of the untranslated regions of StBEL5 to a beta-glucuronidase marker, translation in tobacco (Nicotiana tabacum) protoplasts was repressed by those constructs containing the 3' untranslated sequence. These results demonstrate that the untranslated regions of the mRNA of StBEL5 are involved in mediating its long-distance transport, in maintaining transcript stability, and in controlling translation.

Accumulation of BEL1-like transcripts in solanaceous species

Although numerous RNAs have been detected in the phloem, only a few have been confirmed to move long distances. In potato, full-length mRNA of the BEL1-like transcription factor, StBEL5, moves from leaf veins through the phloem to stolon tips to activate tuber formation. BEL1-like transcription factors are ubiquitous in plants and interact with KNOTTED1-types to regulate numerous developmental processes. To explore the range of KNOTTED1- and BEL1-like mRNAs present in phloem, an analysis of the transcript profile of phloem sap was undertaken. Using a modified technique for the collection of phloem-enriched exudate from excised stems, numerous RNAs encoding these transcription factors were detected in the phloem sap from several solanceous species. All seven known BEL1-like RNAs of potato were detected in the phloem-enriched exudates of stem, whereas several stolon-abundant RNAs were not. After refining the technique to minimize the contamination from RNA arising from wounded cells, KNOTTED1-like RNAs were detected in phloem-enriched sap of potato and BEL5 RNA was detected in the sap collected from two closely related nontuber-bearing potato species and tomato. BEL5 RNA was also detected in RNA extracted from leaf veins of tobacco. The detection of these full-length mRNAs from the KNOTTED1- and BEL1-like families in phloem sap indicates that their potential role as long-distance signals seems to be much more extensive than previously known.

The Canon of Potato Science: 37. Stolonization, Tuber Induction and Tuberization

What is it? Stolon growth, tuber induction and tuberization are key developmental events in the potato plants life as they lead to the formation of the harvestable products of the potato crop. Tubers initiate at the sub-apical region of stolons, which are lateral shoots usually emerging from the basal, below-ground internodes of plant main stems and displaying a diageotrophic growth habit, elongated internodes, spirally arranged scale leaves and a hooked tip with an apical meristem. The complex series of processes involved in the developmental switch from stolon elongation to tuber growth are amongst the most intensively researched topics in potato science. Stolons are generally initiated first at the most basal stem nodes, and then progressively at higher nodes. Stolons may be initiated from above-ground stem nodes, and this capacity is evident in the formation of aerial tubers as well as the use of stem cuttings comprising one or more nodes as a model system to study tuberization. Stolonization at above-ground nodes rarely occurs in field grown potato plants, with leafy shoot development more prevalent where growth from axillary buds occurs. Adequate moisture and darkness promote stolonization at basal nodes, but only in the presence of a dominant shoot apex. The developmental switch associated with tuber formation commences with an induction phase where the stolon tip acquires the competence to form a tuber. Induction involves shoot derived signals which have been shown to be graft transmissible. The first visible sign of tuberization is swelling of the sub-apical region of the stolon tip associated with cell expansion preceding or coinciding with radial cell division. Reorientation of the cytoskeleton from the longitudinal to the radial axis of the stolon occurs during induction and permits transverse cell expansion. Increased tissue turgor occurs at the onset of radial expansion of the stolon and may be part of the tuberization stimulus. Apical meristem mitotic activity ceases at tuberization, while changes in carbohydrate metabolism result in

Somatic Embryogenesis and Plant Development in Centella asiatica L., a Highly Prized Medicinal Plant of the Tropics

Past research experience with Centella asiatica micropropagation suggests a very high rate of contamination during the culture establishment stage. We demonstrate protocols for successful sterilization of Centella explants prepared from field-grown plants with an abundance of fungal and bacterial contamination. Sequential steps during sterilization and explant preparation process included a dip for 30 s in 70% ethyl alcohol, weak bleach treatment for 12 min, and a 60-min soak in plant preservative mixture before establishing cultures. We also report a reproducible system for somatic embryogenesis in Centella using leaf and stolon tip explants collected from naturally growing populations. Somatic embryos were induced within 3 to 4 weeks of culture in the dark on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). Initial embryogenic mass appeared as nodular callus, which eventually developed into actual somatic embryos exhibiting globular, heart-shaped, and cotyledonary stages. Leaves produced embryogenic calli at 2.26 and 4.52 μm 2,4-D, whereas stolon tips were responsive only in the 9.04 μm 2,4-D treatment. Withdrawal of 2,4-D/growth regulators from the induction medium resulted in the maturation and further development of the embryos into plantlets. Regular subculturing of the embryogenic calli into MS medium sustained their regenarability for more than 1 year. Somatic embryos were individually encapsulated in sodium alginate and calcium chloride-based encapsulation matrix to produce artificial or synthetic seeds (synseeds). Synseeds with 2% sodium alginate were found best for the survival and germination recorded after their storage at 5 to 8 °C for 30 and 60 days. We report protocols for C. asiatica to reduce explant contamination before establishment of cultures on somatic embryo induction medium and efficient somatic embryogenesis to facilitate conservation and mass production of elite germplasm. This may further assist rapid dissemination of superior clones needed for research and commercial production.

Tissue integrity and RNA quality of laser microdissected phloem of potato

The phloem is an important conduit for the transport of signaling molecules including RNA. Phloem sap has served as a source of RNA to profile uncontaminated phloem transcriptomes but its collection is difficult in many species. Laser capture microdissection techniques offer a valuable alternative for isolating RNA from specific vascular cells. In potato (Solanum tuberosum L.), there are seven BEL1-like transcription factors expressed throughout the plant with diverse functions. The RNA of one of these, StBEL5, moves through the phloem from the leaf to stolon tips to regulate tuber formation. In this study, the presence of several BEL RNAs and one Knotted1-like RNA was determined in phloem cells collected by laser microdissection coupled to laser pressure catapulting (LMPC). Three fixatives were compared for their effect on cell morphology and RNA quality in transverse sections of stems of potato. For optimum tissue integrity and quality of RNA from potato stem sections, the best results were achieved using ethanol acetate as the fixative. In addition, the RT-PCR results demonstrated the presence of six out of seven of the StBEL RNAs and a potato Knox RNA in phloem cells.

Dynamics of a mobile RNA of potato involved in a long-distance signaling pathway.

BEL1-like transcription factors interact with Knotted1 types to regulate numerous developmental processes. In potato (Solanum tuberosum), the BEL1 transcription factor St BEL5 and its protein partner POTH1 regulate tuber formation by mediating hormone levels in the stolon tip. The accumulation of St BEL5 RNA increases in response to short-day photoperiods, inductive for tuber formation. RNA detection methods and heterografting experiments demonstrate that BEL5 transcripts are present in phloem cells and move across a graft union to localize in stolon tips, the site of tuber induction. This movement of RNA to stolon tips is correlated with enhanced tuber production. Overexpression of BEL5 transcripts that include the untranslated sequences of the BEL5 transcript endows transgenic lines with the capacity to overcome the inhibitory effects of long days on tuber formation. Addition of the untranslated regions leads to preferential accumulation of the BEL5 RNA in stolon tips under short-day conditions. Using a leaf-specific promoter, the movement of BEL5 RNA to stolon tips was facilitated by a short-day photoperiod, and this movement was correlated with enhanced tuber production. These results implicate the transcripts of St BEL5 in a long-distance signaling pathway that are delivered to the target organ via the phloem stream.

Mechanics of growth pulsations as the basis of growth and morphogenesis in colonial hydroids

Growth and shaping in colonial hydroids (Hydrozoa, Cnidaria) are realized due to the functioning of special colony elements, growing tips located at the terminuses of branched colony body. Unlike in plants, the growing tips of colonial hydroids are sites of active cell movements related to morphogenesis and lacking proliferation. The activity of hydroid growing tips is expressed as growth pulsations: cyclic repetitions of their apex extensions and retractions. The parameters of growth pulsations are species specific and related to the shape of a forming element. Here, the succession of cell movements and changes in mutual arrangement within the growing tip are described in detail at all pulsation phases. The role of the inner cell layer in the tip activity was demonstrated for the first time. Relationships between the growing tip parameters, length and diameter, and pulsations are discussed. A scheme is proposed for cyclic processes in both epithelial layers. An explanation is provided for the two-step mode of growth pulsations with relative independence of the main phases. It was proposed that successive activities of the tip ecto- and endoderm serve as driving forces provided there is a hard outer skeleton. This scheme makes it possible to explain some patterns of growth and morphogenesis in colonial hydroids, such as gradually increasing growth rate of a new tip and its maximum growth rate, differences in the parameters of growth pulsations between shoot and stolon tips, shoot base inclination towards the stolon tip, etc., and provides a basis for further improvement of the model of morphogenesis in hydroids.

Physiological and genetic aspects of a diploid potato population in the Netherlands and Northern Finland

Tuberization of potatoes exposed to different photoperiod regimes has been earlier investigated in several studies. However, there is still a limited understanding of the entire tuberization process and the factors influencing this process. One of the constraints of the previous studies has been the use of only one or a few genotypes. Furthermore, the experimental designs have not utilised the natural growing conditions with continuous changes in daylength during the growing season. The general aim of the project was to study the developmental dynamics of the broad-based potato (Solanum tuberosum L.) (CxE) population development at different climatical regimes under the very long-day, long-day and short conditions in Finland, the Netherlands and Ecuador/Venezuela, respectively. In this paper we are presenting some of the results achieved in the studies in Finland during the growing season 2004. In addition to population level trait characteristics we also describe here some of the identified QTLs (quantitative trait loci) for stolon related and tuber formation traits. In some cases we also compare the expression of some of the traits both in Finland and Netherlands.The main difference between the CxE population grown in Finland and the Netherlands was that the onset of flowering took place one week later in Finland. The relationship between tuber initiation and onset of flowering differed depending on the day length. In Finland approximately 70 % of the genotypes had swollen stolon tips before the onset of flowering, while in the Netherlands only 30 % of the genotypes had reached this condition. We also found numerous different trait linked QTLs, for example, a QTL associated with tuber formation was identified on chromosome E5, and QTLs associated with stolon characteristics on chromosomes E1, E4, E10 and E12. As a conclusion, the present preliminary results provide a good basis for determining the influence of different environmental conditions on potato development. In addition, the QTLs obtained in this study give a better understanding of the genetics of complex characters, and can be used in improving the potato crop in breeding programs.

Expression of aGsx parahox gene,Cnox-2, in colony ontogeny inHydractinia (Cnidaria: Hydrozoa)

The ontogeny of colonial animals is markedly distinct from that of solitary animals, yet no regulatory genes have thus far been implicated in colonial development. In cnidarians, colony ontogeny is characterized by the production of a nexus of vascular stolons, from which the feeding and reproductive structures, called polyps, are budded. Here we describe and characterize the Gsx parahox gene, Cnox-2, in the colonial cnidarian Hydractinia symbiolongicarpus of the class Hydrozoa. Cnox-2 is expressed in prominent components of the colony-wide patterning system; in the epithelia of distal stolon tips and polyp bud rudiments. Both are regions of active morphogenetic activity, characterized by cytologically and behaviorally distinct epithelia. Experimental induction and elimination of stolonal tips result in up- and down-regulation, respectively, of Cnox-2 expression. In the developing polyp, Cnox-2 expression remains uniformly high throughout the period of axial differentiation. The differential oral-aboral Cnox-2 expression in the epithelia of the mature polyp, previously described for this and another hydrozoan, arises after oral structures have completed development. Differential Cnox-2 expression is, thus, associated with key aspects of patterning of both the colony and the polyp, a finding that is particularly striking given that polyp and colony form are dissociable in the evolution of Hydrozoa.

The effect of root and shoot temperature of 8°C or 24°C on the uptake and distribution of nitrogen in white clover (Trifolium repens L.)

Limited information is available on the factors influencing the uptake and distribution of nitrogen (N) at low temperatures. This experiment quantified the distribution of N in white clover at root and shoot temperatures of 8°C or 24°C. Stolon tip cuttings of white clover (Trifolium repens L.) were grown in silica sand. After 62 days, plants were transferred to an 8°C or 24°C controlled environment room and, to quantify the distribution of N, a pulse of 15KNO3 was applied. Thereafter, plants were supplied with a complete nutrient solution containing NH4NO3 at a concentration calculated to provide plants with 20% of their N requirement. Plants were harvested at 0, 1, 4, 8, 24, 168, or 336 h. Leaf area and dry weights plus 14N/15N distribution in all fractions and total N concentration were measured. At both temperatures, the dry weights in all fractions increased significantly (P < 0.05) with time. After 336 h the amount of labelled 15N contained in the laminae and petioles was lower at 8°C than at 24°C. The higher 15N recovery in the laminae and petioles, and the higher lamina N%, indicated more N had been transported from the roots to the laminae at 24°C. This investigation suggests that temperature does affect the movement of N around the plant, with a consequent effect on N pool sizes and, hence, growth.

Biology and life history of Cheilosia urbana (Meigen) and Cheilosia psilophthalma (Becker), two sympatric hoverflies approved for the biological control of hawkweeds (Hieracium spp.) in New Zealand

Abstract The biology of the sympatric hoverflies Cheilosia urbana (Meigen) and Cheilosia psilophthalma (Becker) was studied at Delemont, Switzerland. Both species are associated with mouse-ear hawkweed, Hieracium pilosella L., in Central Europe and were chosen as potential biological control agents of alien invasive hawkweeds in New Zealand. Cheilosia urbana and C. psilophthalma are univoltine species which overwinter in the pupal stage. Adults start emerging in early April and have a protandric emergence pattern. Gravid C. urbana and C. psilophthalma females were collected on Hieracium between 10 April and 16 May. Females of both species oviposit onto leaf axils of H. pilosella rosettes during April and May, but their larvae occupy different feeding niches. Freshly hatched C. urbana larvae move into the soil and feed externally on the roots, creating small holes. Cheilosia psilophthalma larvae remain on the aerial plant parts to feed in leaf axils, rosette centers, stolon tips, and leaves. Fecundity was estimated by keeping field-collected females in small vials provided with food. Field-collected C. urbana females laid on average 74 eggs with a maximum of 184 eggs per female, C. psilophthalma females laid 54.3 eggs with a maximum of 158 eggs. At 20 °C, C. urbana eggs hatch within five days, whereas C. psilophthalma eggs need four days. Both species have three larval instars. Mature C. urbana larvae pupate in the soil, very close to the surface, whereas C. psilophthalma larvae pupate on the soil surface. Two Phygadeuon spp. (Hymenoptera, Ichneumonidae) were reared from puparia of C. urbana and C. psilophthalma , and a braconid from mummified C. psilophthalma larvae.

First North American record of the ascidian Perophora japonica

Colonies of the phlebobranch ascidian Perophora japonica were found in August 2003 in Humboldt Bay, northern California, growing on a submerged settling panel. This species has a unique terminal budding in addition to the usual subterminal stolonial budding: bright yellow stellate plaques form at the tips of many stolons. The plaques break off, float away and reattach, subsequently forming new stolons and new colonies. The presence of these plaques, and the larger size of the zooids, distinguish it from the native Perophora annectens. This is the first North American record for this species.