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Spot test and spore count assay data for combinations of dictyostelid hosts infected by Paraburkholderia spp.
<p class="MsoNormal">The social amoeba <em>Dictyostelium discoideum </em>engages in a complex relationship with bacterial endosymbionts in the genus <em>Paraburkholderia, </em>which can benefit their host by imbuing it with the ability to carry prey bacteria throughout its life cycle.<span> </span>The relationship between <em>D. discoideum </em>and <em>Paraburkholderia </em>has been shown to take place across many strains and a large geographical area, but little is known about <em>Paraburkholderia's </em>potential interaction with other dictyostelid species.<span> </span>We explore the ability of three <em>Paraburkholderia </em>species to stably infect and induce bacterial carriage in other dictyostelid hosts.<span> </span>We found that all three <em>Paraburkholderia </em>species successfully infected and induced carriage in seven species of <em>Dictyostelium </em>hosts.<span> </span>While the overall behavior was qualitatively similar to that previously observed in infections of <em>D. discoideum, </em>differences in the outcomes of different host/symbiont combinations suggest a degree of specialization between partners.<span> </span><em>Paraburkholderia </em>was unable to maintain a stable association with the more distantly related host <em>Polysphondylium violaceum</em>.<span> </span>Our results suggest that the mechanisms and evolutionary history of <em>Paraburkholderia's </em>symbiotic relationships may be general within <em>Dictyostelium </em>hosts, but not so general that it can associate with hosts of other genera.<span> </span>Our work further develops a emerging model system for the study of symbiosis in microbes.</p>
Spot test and spore count assay data for combinations of dictyostelid hosts infected by Paraburkholderia spp.
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Diversity of cellular slime molds (dictyostelids) in the Fanjing Mountain Nature Reserve and geographical distribution comparisons with other representative nature reserves in different climate zones of China
<p>Protected areas are widely considered an essential strategy for biodiversity conservation. Dictyostelids are unique soil protists known to have important ecological functions, but the relationship between dictyostelid diversity within protected areas remains poorly understood, especially on a large scale. Herein, we report data on the distribution of dictyostelids, identified with ITS + SSU rRNA molecular and morphology-based taxonomy, from soil samples collected in the Fanjing Mountain protected area of Guizhou Province, Southwest China. We compared the dictyostelids biodiversity data with similar data from previously sampled sites in other protected areas in China. We identified four species of dictyostelids belonging to three genera (<em>Dictyostelium</em>, <em>Heterostelium</em>, and <em>Polysphondylium</em>)and herein provide information on the taxonomy of these species information. Two species (<em>Heterostelium pallidum</em> and <em>Dictyostelium purpureum</em>) are common and widely distributed throughout the world, but one species (<em>Polysphondylium fuscans</em>) was new to China. Our data indicate that the distribution patterns of dictyostelids are most influenced by vegetation and temperature. Overall, the similarity index between Baiyun Mountain in Henan Province and Fanjing Mountain in Guizhou Province, located at approximately the same longitude, is the highest, and the similarity coefficients of family, genus and species are 100%, 100% and 12.5%, respectively. From a species perspective, species in the same climate zone are not closely related, but obvious geographical distributions are evident in different climate zones. This preliminary study provided evidence of the ecological adaptation of dictyostelids to different biological niches.</p>
FIGURE. 1 Dictyostelium recurvibasicum and D. longosporum. D in Dictyostelids from Jilin Province, China II
FIGURE. 1 Dictyostelium recurvibasicum and D. longosporum. D. recurvibasicum: A, B, sorocarps; C, cell aggregation; D, spores; E, sorophore tip; F, G, H, sorophore base. D. longosporum: I, sorocarps; J, cell aggregation; K, sorophore tip; L, sorophore base; M, spores; N, macrocysts. Bars: A = 5 mm; B = 3 mm; C, J = 2 mm; D–H, K–N = 10 μm; I = 10 mm.
FIGURE. 2 in Dictyostelids from Jilin Province, China II
FIGURE. 2 Polysphondylium tenuissimum (A–F) and Dictyostelium brefeldianum (G–K): A, G, sorocarps; B, H, sorophore base; C, sorophore branch; D, I, sorophore tip; E, J, spores; F, macrocysts; K, myxamoebae. Bars: A = 2 mm; B, C, D, E, F, H, I, J, K = 10 μm; G = 3 mm.
FIGURE 11 in New dictyostelid cellular slime molds from South Africa
FIGURE 11. Features of Raperostelium crispum (Eden 2). A. Asymmetrical small aggregation with typical curled, unfinished streams, curled streams anastomose soon after leaving clear areas, sometimes streams fork at acute angles and give the appearance of drastic change of direction, stream diameters vary from thin to very ample at any point of their length, leaving minute masses of streamed pseudoplasmodia progressively unconnected, the feeding front is conspicuous and continuously groups of myxamoebae migrate to larger streams or pseudoplasmodia (above), larger streamed pseudoplasmodia fragment at the periphery, leaving behind a number of pseudoplasmodial masses, which become irregular mounds, the entrance of myxamoebae is constant in the smaller mounds (below), B. Early (left) and late (right) sorogens in clusters, two of them rise up with a coremiform habit (right), the curled streamed basal pseudoplasmodia persist when mounds are not formed, C. Three models of basal section of late sorogens: with striated sheath (left) and with an ample slime mass (center, right), all with halos of myxamoebae, D. Young solitary unbranched sorocarp, the sorophore consists of one tier of cells, untapered, a tiny blob tightly joined to the base is a very early sorogen, almost round, two or three clouds of myxamoebae are the relicts of streams, E. Tightly clustered sorocarps with the typical curled streams at their bases, these formations are several times curved and sometimes parallel with each other, sorophores are curved, broken, sori globose, of different diameters, that may slide down; bases are also covered by lobed slime, masses of rising pseudoplasmodia settle at different points of the sorophore and form small to large branches at acute or right angles, F. Four models of bases: multicellular clavate (above, left), three bases of one tier of cells in a cluster with their curved termina in contact and resting on a dense darker slime mass (above, right), a multicellular base with protruding cells, with strong crampon of sheath (left, below), a base with a bifid terminus on a cushion of darker, dense slime and some undifferentiated cells (below, right), G. Very thin terminal segments with amorphous flexuous capitate cells (left) and piliform cells (right), in most cases with sticky slime, H. Myxamoebae, I. Small short elliptical oblong spores with consolidated granules at the poles and subpolar, heterogeneous content with small granules and vacuoles within the spore body, J. Spore case in two longitudinal halves. Bars: A–E = 150 μm, F–G = 15 μm, H = 10 μm, I–J = 5 μm.
FIGURE 7 in New dictyostelid cellular slime molds from South Africa
FIGURE 7. Features of Hagiwaraea irregularibrachiatum (Krug 6–5A). A. Streamed aggregation of thin streams and dense center, with short secondary unfinished streams (forked), sometimes anastomosed at the end, small circles are microcysts, B. Early (above) and later aggregations with elongated coremiform sorogens in a cluster (below), C. Mature cluster of whorled sorocarps (including a late sorogen), some appear coremiform (left), others are decumbent and become prostrate (right), abundant microcysts within and surrounding the cluster, D. Solitary branched sorocarp, each branch may hold 1 to 4 whorls, or two groups of few whorls, E. Four models of bases: clavate, of several rounded enlarged cells of a solitary sorocarp (above left), coremiform bases consisting of one tier of cells, these are very uniform and rest on a dark cushion of dense slime (above, right), curved base with a stolon also angled (below left), curved base with protruding cells and a dense matrix of slime; in all cases, sorophores above the basal termina consist of one tier of cells, F. Model of a whorl, with its node and two curved branches, one is broken, a collapsed cell sustained on the other side by a cellular mass and slime (left), G. Microcysts, H. Capsulated small and broad spores with polar to subpolar unconsolidated granules, sometimes granules are widely scattered within the spore body, also with tiny vacuoles. Bars: A–B = 100 μm, C–D = 200 μm, E–F = 20 μm, G = 4 μm, H = 6 μm.
FIGURE 6 in New dictyostelid cellular slime molds from South Africa
FIGURE 6. Features of Hagiwaraea tenebrica important for identification: A. Irregular off-centered streamed aggregations, the streams are broad, with lobed margins and ends, also with anastomosed termina, which then disrupt and form tiny masses of pseudoplasmodia that tend to surround the center, streams also open to incoming myxamoebae, B. Clustered (left) to solitary mostly unbranched sorocarps (right), sorocarps are gently curved, sori globose, one sorus has slid down a little, sorophores appear to be coremiform in part, branches few and very small, C. Main feature of the species is a crampon base immersed into its dense slime and pleated sheath, with visible dark large granules, the number of crampon "roots" is usually 4 or 5, characteristically incased in a slime sheath, D. Small elliptical-oblong spores with polar to subpolar consolidated granules, some large granules are also apparent in the spore body. Bars: A = 150 μm, B = 500 μm, C = 20 μm, D = 6 μm.
FIGURE 2 in New dictyostelid cellular slime molds from South Africa
FIGURE 2. Features of Cavenderia fulva important for identification. A. Streamed aggregation with few and strong, thin forked streams of open ends, the "violaceum" type of aggregation is evident and at the center rise up two dichotomous sorogens that diverge, B. Clustered mature branched sorocarps with most sorophores coremiform in habit (right), solitary unbranched sorcarps on the left and right, C. Late dichotomous sorogens, the irregular, sometimes tortoise sorophores are visible, branches achieve a length to the top of the structure, D. Elliptical spores with prominent refractive consolidated granules at their poles. Bars: A = 40 μm, B = 30 μm, C = 140 μm, D = 5 μm.
FIGURE 10 in New dictyostelid cellular slime molds from South Africa
FIGURE 10. Features of Raperostelium cymosum important for identification. A. Minute aggregations at different stages of progress: early-mature aggregation with short anastomosed streams that cover almost the whole area surrounding the centers (above, left), cupuliform, button-like pseudoplasmodium with a tiny very early sorogen (above, right), three closer late aggregations, almost mound-like, with one to three streams (below, right); streams are very short and ample, expanded, lobed, flattened progressively, inconspicuous and open at the end to incoming myxamoebae, B. Three tight clusters of sorogens, early curved, irregular sorogens that rise up in close contact each other, bend over and separate at tips, in this case, almost at the same time, C. Early and late solitary sorogens (above, right and left) and mature fruiting bodies tightly clustered; a majority of sorocarps have well separated sorophores and sori, one is coremiform (center, left), two clusters show the on-going production of early sorogens at base (below, left), sorophores are of different heights, delicate, irregular, curved, somewhat broken, sometimes branched, few in number; sori are globose, one sorocarp has a collapsed its sorus, D. Large elliptical, mostly reniform spores with conspicuous consolidated polar to subpolar granules. Bars: A = 50 μm, B = 50 μm, C = 350 μm, D = 6 μm.
FIGURE 1 in New dictyostelid cellular slime molds from South Africa
FIGURE 1. Features of Cavenderia fulva (Krug 6–5A). A. Radiate irregular streamed "violaceum" type of aggregation, streams are forked, B. Four habits of early to late sorogens: coremiform in an old culture (left), clustered dichotomous (center), solitary curved elongated (right), a migrating sorogen with stalk formation that leaves behind a net or framework of sheath fibers as it advances (below), C. Tight cluster of late branched sorogens, a solitary creeping sorogen on the left, D. Solitary unbranched sorocarp, the sorus has regrown, the base is covered by abundant mucilage and a typical reticulate or net-like sheath, E. Tight cluster of unbranched to branched sorocarps, large branches tend to achieve the level of the main axis, the bases are tightly united, with clear and dark areas, and fibers in net are attached to marginal undifferentiated cells, on the right a decumbent collapsed sorocarp with immediate regrowth; sorophores are irregular, curved, sometimes broken (in all cases), F. Four bases: multicellular, with protruding cells, the sheath is cone-like, within the mucilage there are undifferentiated cells (above, left); base with a large curved clavate terminal cell, the sheath is represented as a net of fibers (above, right); one to two celled base resting on and in touch with a large globoid cellular mass, the mucilage is dense and dark below the base (below, left); one-celled round to clavate base with its enlarged terminal cell (below, right), G. Two tips: simple, with a broad subtip (left) and a capitate tip with flexuous cells (right), both with dense sticky mucilage, H. Elliptical slightly broad spores with conspicuous consolidated polar granules, sometimes with halos. Bars: A–C = 100 μm, D–E = 200 μm, F–G = 25 μm, H = 5 μm.
FIGURE 13 in New dictyostelid cellular slime molds from South Africa
FIGURE 13. An 18S ribosomal RNA phylogeny including all new species of dictyostelids. The figure shows portions of a single tree expanded separately for the (A) Cavenderiaceae, (B) the Acytosteliaceae and (C) the Raperosteliaceae. The tree was derived by maximum likelihood analysis of ~1500 aligned positions of the SSU rDNA molecule using the program RAxML 8.2.10 with a GTR+gamma substitution model. The sequence alignment was staggered for two hypervariable regions so that only relationships within families were analyzed for these regions. New species described here are indicated in red and environmental (culture-independent) sequences in purple (Baldauf et al. 2018). Bootstrap values of 60-95% are shown on or adjacent to the relevant branches, and values of 95-100% are indicated by filled circles. The scale bar at the bottom indicates substitutions per site.
FIGURE 12 in New dictyostelid cellular slime molds from South Africa
FIGURE 12. Features of Raperostelium crispum important for identification. A. Irregular asymmetrical small aggregation with typical curled, unfinished streams, streams soon anastomose, leaving very small empty areas that sometimes give the appearance of acute angles; stream diameters vary from very thin to relatively wide at any point of their length, leaving minute masses of streamed pseudoplasmodia progressively isolated (above), larger streamed pseudoplasmodia fragment and form irregular close, sometimes connected, mounds, the peripheral ones are open to incoming myxamoebae (below), B. Clustered and solitary sorocarps with the remaining expanded formation of curled streams at their bases, often streams are annular, sorophores are curved, broken, sori globose, small to large, C. Two late pseudoplasmodia with a very early sorogen (right) and with a bifid early-late sorogen (left), small pseudoplasmodia that surround the main mass soon condense into mounds, D. Small short elliptical or oblong spores with consolidated granules at the poles. Bars: A = 40 μm, B = 300 μm, C = 150 μm, D = 6 μm.
FIGURE 4 in New dictyostelid cellular slime molds from South Africa
FIGURE 4. Features of Cavenderia minima. A. Irregular shortly streamed aggregation, streams are ample, with lobular margins and ends, also open to incoming myxamoebae, B. Solitary unbranched, sometimes curved small sorocarps, at the top, a twisted late sorogens; sorocarps are mostly scarce and very small, hence the name C. minima, C. Short early solitary sorogens and the minute mound-like stage of the aggregations, a sorocarp has collapsed (bottom), D. Small elliptical irregular spores with polar to subpolar consolidated granules, generally the cluster of granules appears larger at one of the poles. Bars: A = 30 μm, B = 200 μm, C = 100 μm, D = 6 μm.
FIGURE 3 in New dictyostelid cellular slime molds from South Africa
FIGURE 3. Features of Cavenderia minima (Eden 1). A. Irregular small aggregation with few ample streams, the streams have lobular margins and ends, open to incoming myxamoebae; generally, one smaller stream appears surmounted over the body of the others, the center is elevated and granulose (above), three minute and irregular mound-like aggregations, with a flattened edge of incoming myxamoebae (below), B. A sequence of development of small solitary early sorogens from left to right, a very early sorogen rises up from the center of a cupuliform pseudoplasmodium, one to three old flattened streams appear as clouds of myxamoebae (left) and may remain until further development, a sorus-like, rounded terminus, sorogen may be formed (right), C. A tight cluster of sorocarps with a late small sorogen and a mature sorocarp with rising up tiny masses of pseudoplasmodia and slime (right), on the left a solitary decumbent marginal sorocarp has collapsed, with a stoloniferous habit; at the center a curved sorophore with no sorus; the relicts of three ample streams remain as clouds of myxamoebae and mucilage at bases, D. Two solitary unbranched sorocarps, straight prone with a globoid sorus (left) and a sorocarp with two curves of the sorophore, with a globose small sorus, bases inconspicuous but surrounded by granules, slime, and relicts of the pseudoplasmodia, E. Three bases with one tier of cells: clavate, with smaller terminal cell, the sheath is cone-like (left), curved, with a curved enlarged terminal cell (center), a base with its last four cells enlarged, showing the mucilage and a small mass of more dense and darker mucilage (right), F. Different simple one-celled tips: with an amorphous cell (left), three with an enlarged granulated cell and/or abundant slime (center), with a piliform elongated terminus (right); the last sorophore segment is always flexuous, most of tips are curved at the end, G. Myxamoebae with an enlarged vacuole, H. Elliptical irregular spores with clusters of consolidated and unconsolidated (spaced) granules mostly at poles, one cluster of granules is larger than the opposite one, halos are common around the clusters of granules, I. Spore break-line and the open spore capsule (below) in two halves. Bars: A–B = 50 μm, C–D = 100 μm, E = 15 μm, F = 3 μm, G = 10 μm, H–I = 6 μm.
FIGURE 5 in New dictyostelid cellular slime molds from South Africa
FIGURE 5. Features of Hagiwaraea tenebrica (ONG2). A. Irregular off-centered streamed aggregations, streams are broad, with lobular margins and ends, streams termina separate and can conform flattened tiny masses of pseudoplasmodia that surround the aggregation, streams also are open to incoming myxamoebae, the center is elevated, somewhat cupuliform, B. Condensed pseudoplasmodia from which arise solitary centered very early sorogens, at the basal part there are rudimentary streams (above, left), clustered early sorogens tightly joined, clustered early-late sorogens which separate (center, left), solitary late sorogen with the lower sorophore prostrate (center, right), solitary typically curved late sorogen; surrounding the base is a disrupted circle of mucilage and cells (below), C. Cluster of mature unbranched sorocarps, curved and straight, one of them decumbent with its sorus collapsed and with regrowth; sori globose, pigmented different from the sorophore and the crampon base, more obscure, D. Solitary erect sorocarp with its sorus and cramponated base and surrounded by tiny masses of pseudoplasmodia and slime, E. Crampon base immersed within its dense slime matrix full of large and small pigmented granules; the sheath is strong and attaches the structure to the substrate; the crampon base is of a darker tone of fuscus color, each crampon "root" shown is borne at a different level of the sorophore base with respect to the others and generally consists of one tier of cells; the crampon terminal "root" cell commonly clavate (left), the bases are encased in a slime sheath which is cone-like (above, left) and sometimes the base is positioned in a cavity on the substrate (center, right), a crampon base early in formation with protruding terminal cells (below, left), F. Two tips: a capitate tip with amorphous cells and flexuous subtip (above, left), a very thin one tier of cell terminus with its last section covered by a dense hyaline persistent slime that attaches undifferentiated unconnected small cells and many large and small pigmented granules, G. Microcysts, H. Spore capsule opens irregularly at the equator (left), dense myxamoebae with many dark granules (right), I. Elliptical irregular spores with polar to subpolar consolidated granules, some at the center of the spore body, of different sizes with vacuoles and heterogeneous content. Bars: A–B = 200 μm, C–D = 300 μm, E = 25 μm, F = 7 μm, G = 2 μm, H = 10 μm, I = 5 μm.
FIGURE 9 in New dictyostelid cellular slime molds from South Africa
FIGURE 9. Features of Raperostelium cymosum (Krug 15A). A. Very small aggregations at different stage of progress, early-mature aggregation with very short flattened progressively expanded streams that may partitioned, streams are ample, with irregular ill-defined margins, sometime lobed, thin in height, unfinished and open to incoming myxamoebae (left), cupuliform pseudoplasmodium with a remnant of an expanded flattened stream (right), B. Three early tight clusters of sorogens, a common pair of sorogens (early and late sorogens) rise up at different times after resumption of the pseudoplasmodium formation (left), an early curved sorogen close to a branched late sorogen (center), a cluster with sorogens of different stages of development, one of them bent over drastically, along with a mature unbranched sorocarp, short lobed streamed pseudoplasmodia are common at base (right), C. Cluster of open branched and unbranched sorocarps with an early late sorogen, some minute pseudoplasmodial masses remain close to the cluster (right), D. Solitary unbranched sorocarp with lobules of pseudoplamodium and mucilage at the base, E. Tight cluster of unbranched, branched and rebranched sorocarps, in almost all cases sorophores are of different heights but decreasing from the oldest to the younger joined sorocarps; they are delicate, irregular, broken, with an expanded reticulate slimy basal part, at the point of breakage there are marginal tiny masses of slime and undifferentiated cells; also, there are minute ascending masses of pseudoplasmodia on these sorophores of one tier of cells, with abundant slime, one (or more) tightly joined early sorogens as is typical for this species, a secondary sorogen may ascend as a branch or not, sori are of different diameters, one branch has collapsed, giving rise to a new sorogen (below, right), most bases are surrounded by a halo, F. Three representations of bases: clavate, with several cells above a larger terminal cell (above, left), three one-celled bases with round to clavate terminal cells in touch with each other and resting on a darker dense discrete granular structure (center, left), a clavate base with protruding cells as crampons, within a dense and abundant slime, G. Group of very variable tips: with an amorphous flexuous terminal cell, with elongated or short piliform terminus, with an elongated curved cell, in most cases tips are simple, single-celled, curved, thin and with a dense matrix of sticky granulated slime (attached spores), H. Microcysts, I. Large elliptical, mostly reniform spores with conspicuous consolidated polar to subpolar granules, spores are also vacuolated and with heterogeneous content. Bars: A–C = 50 μm, D–E = 100 μm, F–G = 15 μm, H = 3 μm, I = 6 μm.
FIGURE 8 in New dictyostelid cellular slime molds from South Africa
FIGURE 8. Features of Hagiwaraea irregularibrachiatum important for identification. A. Regular radiate streamed aggregation of the "violaceum" type with thin regular sections of the streams and branched or anastomosed ends, abundant microcysts on the surface, B. Two late sorogens, with a small globular mass of psudoplasmodia below the terminus that will form the first whorl, C. Irregular broken sorophores and noded 3–4 whorled sorocarps, each whorled branch curved, D. Elliptical short spores with small unconsolidated polar to subpolar granules. Bars: A = 120 μm, B = 60 μm, C = 45 μm, D = 6 μm.
FIGURE 14 in Distribution and ecology of dictyostelids in Madagascar
FIGURE 14. Total number of clones and number of clones per sample for collecting sites in Madagascar.
FIGURES 1–12 in Distribution and ecology of dictyostelids in Madagascar
FIGURES 1–12. Madagascar vegetation at some of the collecting sites. 1. Ranomafana National Park, subhumid tropical forest. 2. Ranomafana, subhumid tropical forest. 3. Mandraka, primary montane tropical wet forest. 4. Grassland. 5. L'Isalo National Park with Pandanus sp. present. 6. Road to Betroka, grassland with Eucalyptus sp. present. 7. L'Isalo National Park, sclerophyllous forest. 8.Andringitra National Park, fallow grassland with shrubs. 9. Ambositra, sclerophyllous Uapaca bojerii community. 10. Succulent scrub with Pachypodium sp. 11. Betroka-Beraketa xerophytic succulent scrub. 12. Near Berenty National Reserve, dry forest with Alluaudia sp
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