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Evidence for Pleistocene Population Divergence and Expansion of Anopheles albimanus in Southern Central America
The micro-geographic structure of Anopheles albimanus was studied in southern Central America using partial sequences of the mtDNA cytochrome oxidase subunit I gene (COI). Analysis of molecular variance supported significant genetic structure between populations from Costa Rica and western Panama versus those from centraleastern Panama (ΦCT = 0.33), whereas the within group divergence was shallow and statistically insignificant (ΦST = 0.08). Furthermore, a statistical parsimony network depicted three divergent groups of haplotypes that were not evenly distributed across the study area. Our findings are in partial agreement with previous studies, yet they do not support physical barriers to gene flow or contemporary isolation by distance in this region. Instead, three co-occurring groups of An. albimanus may be the result of multiple introductions, most likely caused by historical fragmentation and subsequent secondary contact. In addition, the molecular signature of population expansion of An. albimanus was detected in central-eastern Panama approximately 22,000 years ago (95% confidence interval [CI] 10,183–38,169). We hypothesize that the population structure of An. albimanus , as determined by our COI locus analysis, is the result of late Pleistocene climatic changes in northern South America.The micro-geographic structure of Anopheles albimanus was studied in southern Central America using partial sequences of the mtDNA cytochrome oxidase subunit I gene (COI). Analysis of molecular variance supported significant genetic structure between populations from Costa Rica and western Panama versus those from centraleastern Panama (ΦCT = 0.33), whereas the within group divergence was shallow and statistically insignificant (ΦST = 0.08). Furthermore, a statistical parsimony network depicted three divergent groups of haplotypes that were not evenly distributed across the study area. Our findings are in partial agreement with previous studies, yet they do not support physical barriers to gene flow or contemporary isolation by distance in this region. Instead, three co-occurring groups of An. albimanus may be the result of multiple introductions, most likely caused by historical fragmentation and subsequent secondary contact. In addition, the molecular signature of population expansion of An. albimanus was detected in central-eastern Panama approximately 22,000 years ago (95% confidence interval [CI] 10,183–38,169). We hypothesize that the population structure of An. albimanus , as determined by our COI locus analysis, is the result of late Pleistocene climatic changes in northern South America
Short Report: Anopheles darlingi (Diptera: Culicidae) in Panama
We report Anopheles darlingi in Darien Province in eastern Panama. Polymerase chain reaction–restriction fragment length polymorphism profiles of the single copy nuclear white gene and sequence comparisons confirmed the presence of 66 specimens of the northern lineage of An. darlingi . The parsimony network depicted 5 CO1 haplotypes in 40 specimens of An. darlingi , which connected through 7–8 mutational steps with sequences from Central and South America. Furthermore, the presence of haplotypes in Biroquera, Darien Province identical to those previously published from northern Colombia suggests that Panamanian samples originated in Colombia. Results of neutrality tests ( R 2 and Fu’s F S ) were not significant and the mismatch distribution was multimodal and did not fit the model of sudden population growth. These findings may indicate a long and stable presence of An. darlingi in eastern Panama.We report Anopheles darlingi in Darien Province in eastern Panama. Polymerase chain reaction–restriction fragment length polymorphism profiles of the single copy nuclear white gene and sequence comparisons confirmed the presence of 66 specimens of the northern lineage of An. darlingi . The parsimony network depicted 5 CO1 haplotypes in 40 specimens of An. darlingi , which connected through 7–8 mutational steps with sequences from Central and South America. Furthermore, the presence of haplotypes in Biroquera, Darien Province identical to those previously published from northern Colombia suggests that Panamanian samples originated in Colombia. Results of neutrality tests ( R 2 and Fu’s F S ) were not significant and the mismatch distribution was multimodal and did not fit the model of sudden population growth. These findings may indicate a long and stable presence of An. darlingi in eastern Panama
Species Composition and Distribution of Adult Anopheles (Diptera: Culicidae) in Panama
Anopheles (Diptera: Culicidae) species composition and distribution were studied using human landing catch data over a 35-yr period in Panama. Mosquitoes were collected from 77 sites during 228 Þeld trips carried out by members of the National Malaria Eradication Service. Fourteen Anopheles species were identiÞed. The highest average human biting rates were recorded from Anopheles (Nyssorhynchus) albimanus (Wiedemann) (9.8 bites/person/night) andAnopheles (Anopheles) punctimacula (Dyar and Knab) (6.2 bites/person/night). These two species were also the most common, present in 99.1 and 74.9%, respectively, of the sites. Anopheles (Nyssorhynchus) aquasalis (Curry) was encountered mostly in the indigenous Kuna Yala Comarca along the eastern Atlantic coast, where malaria case history and average human biting rate (9.3 bites/person/night) suggest a local role in malaria transmission. An. albimanus, An. punctimacula, and Anopheles (Anopheles) vestitipennis (Dyar and Knab) were more abundant during the rainy season (MayÐDecember), whereas An. aquasalis was more abundant in the dry season (JanuaryÐApril). Other vector species collected in this study were Anopheles (Kerteszia) neivai (Howard, Dyar, and Knab) and Anopheles (Anopheles) pseudopunctipenniss.l. (Theobald). High diversity of Anophelesspecies and six conÞrmed malaria vectors in endemic areas of Panama emphasize the need for more detailed studies to better understand malaria transmission dynamics.Anopheles (Diptera: Culicidae) species composition and distribution were studied using human landing catch data over a 35-yr period in Panama. Mosquitoes were collected from 77 sites during 228 Þeld trips carried out by members of the National Malaria Eradication Service. Fourteen Anopheles species were identiÞed. The highest average human biting rates were recorded from Anopheles (Nyssorhynchus) albimanus (Wiedemann) (9.8 bites/person/night) andAnopheles (Anopheles) punctimacula (Dyar and Knab) (6.2 bites/person/night). These two species were also the most common, present in 99.1 and 74.9%, respectively, of the sites. Anopheles (Nyssorhynchus) aquasalis (Curry) was encountered mostly in the indigenous Kuna Yala Comarca along the eastern Atlantic coast, where malaria case history and average human biting rate (9.3 bites/person/night) suggest a local role in malaria transmission. An. albimanus, An. punctimacula, and Anopheles (Anopheles) vestitipennis (Dyar and Knab) were more abundant during the rainy season (MayÐDecember), whereas An. aquasalis was more abundant in the dry season (JanuaryÐApril). Other vector species collected in this study were Anopheles (Kerteszia) neivai (Howard, Dyar, and Knab) and Anopheles (Anopheles) pseudopunctipenniss.l. (Theobald). High diversity of Anophelesspecies and six conÞrmed malaria vectors in endemic areas of Panama emphasize the need for more detailed studies to better understand malaria transmission dynamics
Leaf-inhabiting genera of the Gnomoniaceae, Diaporthales
The Gnomoniaceae are characterised by ascomata that are generally immersed, solitary, without a stroma, or aggregated with a rudimentary stroma, in herbaceous plant material especially in leaves, twigs or stems, but also in bark or wood. The ascomata are black, soft-textured, thin-walled, and pseudoparenchymatous with one or more central or eccentric necks. The asci usually have a distinct apical ring. The Gnomoniaceae includes species having ascospores that are small, mostly less than 25 μm long, although some are longer, and range in septation from non-septate to one-septate, rarely multi-septate. Molecular studies of the Gnomoniaceae suggest that the traditional classification of genera based on characteristics of the ascomata such as position of the neck and ascospores such as septation have resulted in genera that are not monophyletic. In this paper the concepts of the leaf-inhabiting genera in the Gnomoniaceae are reevaluated using multiple genes, specifically nrLSU, translation elongation factor 1-alpha (tef1-α), and RNA polymerase II second largest subunit (rpb2) for 64 isolates. ITS sequences were generated for 322 isolates. Six genera of leaf-inhabiting Gnomoniaceae are defined based on placement of their type species within the multigene phylogeny. The new monotypic genus Ambarignomonia is established for an unusual species, A. petiolorum. A key to 59 species of leaf-inhabiting Gnomoniaceae is presented and 22 species of Gnomoniaceae are described and illustrated.The Gnomoniaceae are characterised by ascomata that are generally immersed, solitary, without a stroma, or aggregated with a rudimentary stroma, in herbaceous plant material especially in leaves, twigs or stems, but also in bark or wood. The ascomata are black, soft-textured, thin-walled, and pseudoparenchymatous with one or more central or eccentric necks. The asci usually have a distinct apical ring. The Gnomoniaceae includes species having ascospores that are small, mostly less than 25 μm long, although some are longer, and range in septation from non-septate to one-septate, rarely multi-septate. Molecular studies of the Gnomoniaceae suggest that the traditional classification of genera based on characteristics of the ascomata such as position of the neck and ascospores such as septation have resulted in genera that are not monophyletic. In this paper the concepts of the leaf-inhabiting genera in the Gnomoniaceae are reevaluated using multiple genes, specifically nrLSU, translation elongation factor 1-alpha (tef1-α), and RNA polymerase II second largest subunit (rpb2) for 64 isolates. ITS sequences were generated for 322 isolates. Six genera of leaf-inhabiting Gnomoniaceae are defined based on placement of their type species within the multigene phylogeny. The new monotypic genus Ambarignomonia is established for an unusual species, A. petiolorum. A key to 59 species of leaf-inhabiting Gnomoniaceae is presented and 22 species of Gnomoniaceae are described and illustrated
ECOLOGICAL IMPLICATIONS OF ANTI-PATHOGEN EFFECTS OF TROPICAL FUNGAL ENDOPHYTES AND MYCORRHIZAE
We discuss studies of foliar endophytic fungi (FEE) and arbuscular mycorrhizal fungi (AMF) associated with Theobroma cacao in Panama. Direct, experimentally controlled comparisons of endophyte free (E—) and endophyte containing (E+) plant tissues in T. cacao show that foliar endophytes (FEE) that commonly occur in healthy host leaves enhance host defenses against foliar damage due to the pathogen (Phytophthora palmivora). Similarly, root inoculations with commonly occurring AMF also reduce foliar damage due to the same pathogen. These results suggest that endophytic fungi can play a potentially important mutualistic role by augmenting host defensive responses against pathogens. There are two broad classes of potential mechanisms by which endophytes could contribute to host protection: (1) inducing or increasing the expression of intrinsic host defense mechanisms and (2) providing additional sources of defense, extrinsic to those of the host (e.g., endophytebased chemical antibiosis). The degree to which either of these mechanisms predominates holds distinct consequences for the evolutionary ecology of host-endophyte-pathogen relationships. More generally, the growing recognition that plants are composed of a mosaic of plant and fungal tissues holds a series of implications for the study of plant defense, physiology, and genetics.We discuss studies of foliar endophytic fungi (FEE) and arbuscular mycorrhizal fungi (AMF) associated with Theobroma cacao in Panama. Direct, experimentally controlled comparisons of endophyte free (E—) and endophyte containing (E+) plant tissues in T. cacao show that foliar endophytes (FEE) that commonly occur in healthy host leaves enhance host defenses against foliar damage due to the pathogen (Phytophthora palmivora). Similarly, root inoculations with commonly occurring AMF also reduce foliar damage due to the same pathogen. These results suggest that endophytic fungi can play a potentially important mutualistic role by augmenting host defensive responses against pathogens. There are two broad classes of potential mechanisms by which endophytes could contribute to host protection: (1) inducing or increasing the expression of intrinsic host defense mechanisms and (2) providing additional sources of defense, extrinsic to those of the host (e.g., endophytebased chemical antibiosis). The degree to which either of these mechanisms predominates holds distinct consequences for the evolutionary ecology of host-endophyte-pathogen relationships. More generally, the growing recognition that plants are composed of a mosaic of plant and fungal tissues holds a series of implications for the study of plant defense, physiology, and genetics
I. eaf Litter and Leaf Age as Factors Affecting the Assemblage of Endophytes Associated with
Particular importance has been given to the potential effect of forest canopy on enhancing the rate of initial endophytic fungi colonization of seedlings. Here we compare the rate of endophyte accumulation when endophyte-free seedlings are placed under intact forest canopies with intact or removed leaf litter. The results show a much more rapid accumulation of endophytes in the presence of intact leaf litter. This not only suggests that dead leaves appear to be a primary source of inoculum of foliar endophytic fungi (FEF) for leaves in the understory, it further suggests that local FEF sources (ie, the local litter) can dominate the cornposition of colonizing spores, The results of this experiment suggest a mechanism explaining reports of relatively fine scale local differentiation of HEF communities within the same host plantParticular importance has been given to the potential effect of forest canopy on enhancing the rate of initial endophytic fungi colonization of seedlings. Here we compare the rate of endophyte accumulation when endophyte-free seedlings are placed under intact forest canopies with intact or removed leaf litter. The results show a much more rapid accumulation of endophytes in the presence of intact leaf litter. This not only suggests that dead leaves appear to be a primary source of inoculum of foliar endophytic fungi (FEF) for leaves in the understory, it further suggests that local FEF sources (ie, the local litter) can dominate the cornposition of colonizing spores, The results of this experiment suggest a mechanism explaining reports of relatively fine scale local differentiation of HEF communities within the same host plan
Fungal endophytes limit pathogen damage in a tropical tree
Every plant species examined to date harbors endophytic fungi within its asymptomatic aerial tissues, such that endophytes represent a ubiquitous, yet cryptic, component of terrestrial plant communities. Fungal endophytes associated with leaves of woody angiosperms are especially diverse; yet, fundamental aspects of their interactions with hosts are unknown. In contrast to the relatively species-poor endophytes that are vertically transmitted and act as defensive mutualists of some temperate grasses, the diverse, horizontally transmitted endophytes of woody angiosperms are thought to contribute little to host defense. Here, we document high diversity, spatial structure, and host affinity among foliar endophytes associated with a tropical tree (Theobroma cacao, Malvaceae) across lowland Panama. We then show that inoculation of endophyte-free leaves with endophytes isolated frequently from naturally infected, asymptomatic hosts significantly decreases both leaf necrosis and leaf mortality when T. cacao seedlings are challenged with a major pathogen (Phytophthora sp.). In contrast to reports of fungal inoculation inducing systemic defense, we found that protection was primarily localized to endophyte-infected tissues. Further, endophyte-mediated protection was greater in mature leaves, which bear less intrinsic defense against fungal pathogens than do young leaves. In vitro studies suggest that host affinity is mediated by leaf chemistry, and that protection may be mediated by direct interactions of endophytes with foliar pathogens. Together, these data demonstrate the capacity of diverse, horizontally transmitted endophytes of woody angiosperms to play an important but previously unappreciated role in host defense.Every plant species examined to date harbors endophytic fungi within its asymptomatic aerial tissues, such that endophytes represent a ubiquitous, yet cryptic, component of terrestrial plant communities. Fungal endophytes associated with leaves of woody angiosperms are especially diverse; yet, fundamental aspects of their interactions with hosts are unknown. In contrast to the relatively species-poor endophytes that are vertically transmitted and act as defensive mutualists of some temperate grasses, the diverse, horizontally transmitted endophytes of woody angiosperms are thought to contribute little to host defense. Here, we document high diversity, spatial structure, and host affinity among foliar endophytes associated with a tropical tree (Theobroma cacao, Malvaceae) across lowland Panama. We then show that inoculation of endophyte-free leaves with endophytes isolated frequently from naturally infected, asymptomatic hosts significantly decreases both leaf necrosis and leaf mortality when T. cacao seedlings are challenged with a major pathogen (Phytophthora sp.). In contrast to reports of fungal inoculation inducing systemic defense, we found that protection was primarily localized to endophyte-infected tissues. Further, endophyte-mediated protection was greater in mature leaves, which bear less intrinsic defense against fungal pathogens than do young leaves. In vitro studies suggest that host affinity is mediated by leaf chemistry, and that protection may be mediated by direct interactions of endophytes with foliar pathogens. Together, these data demonstrate the capacity of diverse, horizontally transmitted endophytes of woody angiosperms to play an important but previously unappreciated role in host defense