Genetic Diversity,Genotype Discrimination,and Population Structure of Mexican <Emphasis Type="Italic">Opuntia</Emphasis> sp., Determined by SSR Markers

The Opuntia (prickly pear) genus, an important horticultural crop in Mexico, is essentially a fruit crop with two variants: sweet (“tunas”) or acid (“xoconostles”) fruits; it is also a source of vegetables “nopalitos” or fodder for livestock, among other uses. Its taxonomical classification has been reported as complex, although few studies on the genetic structure of Mexican Opuntia are available, and genetic differences between the two types of fruits are unknown. Opuntia genotype identification and classification are still mainly based on morphological characters. In this study, the genetic diversity of Mexican Opuntia germplasm with agronomic and economic importance was revealed, using 88 accessions and 13 SSR markers, in an attempt to explore the genetic relationships among them. A total of 159 alleles were detected ranging from 7 to 23 per locus with an average of 12.2. The SSR markers generated unique fingerprints for each Opuntia accession confirming their usefulness for genetic analysis. The accessions’ grouping was defined by several complementary clustering methods, and the moderate incongruences between the different methods did not influence the overall clustering. DAPC and STRUCTURE analyses grouped the accessions into five groups, thus confirming the incorrect delimitation of species in this genus. The following species had no clear boundaries: Opuntia ficus-indica, Opuntia albicarpa, Opuntia megacantha, Opuntia streptacantha, Opuntia lasiacantha, and Opuntia hyptiacantha. However, Opuntia robusta was separated from the rest of the species. Opuntia joconostle and Opuntia matudae, which produce acid fruits, tended to differ from the others. Median-joining simulation classified all genotypes into a complex network, and both linear and reticular ties between Mexican Opuntia genotypes were revealed. The genetic distance revealed in the present study shows the importance of Mexican accessions for conservation and use in breeding programs.     

Analysis of the community structure of yeasts associated with the decaying stems of cactus. II.Opuntia species

A survey was made of yeast species associated with the decaying pads of 3 prickly pear cacti (Opuntia phaeacantha, O. ficus-indica, andO. lindheimeri) in Arizona and Texas. Yeast communities from 12 localities were compared among localities, amongOpuntia species, and with previous data on yeast communities associated with columnar cacti. The results indicate thatOpuntia necroses contain relatively more yeast species with broader physiological abilities in their communities than columnar necroses. It is argued that differences in chemistry of the opuntias and columnar forms in concert with the insect vectors specific for these cacti account for the differences in yeast community structure. It is further hypothesized that the differences in yeast community structure have been important in the evolution and maintenance of species diversity forDrosophila species which live in the decaying stems or cladodes of various cacti. Most of the yeast community evolution in the cacti is postulated to have proceeded by evolution in situ and not by additions and replacements from outside of the system.     

Traditional knowledge and genetic diversity ofopuntia pilifera (cactaceae) in the tehuacán-cuicatlán valley, Mexico

Traditional Knowledge and Genetic Diversity of Opuntia pilifera (Cactaceae) in the Tehuacán-Cuicatlán Valley, Mexico. Economic Botany 59(4)366-376, 2005. Most studies of the genusOpuntia have focused on economically important species, and therefore more knowledge concerning the genetic diversity among wild and locally managedOpuntia species is needed for an expanded use of cacti in the future. The present study is part of ongoing ethnobotanical work in the Tehuacán-Cuicatlán Valley of Mexico and focuses on six traditionally classified forms ofOpuntia pilifera used as food by the indigenous Popoloca people in San Juan Atzingo. Traditional knowledge of how to distinguish these forms based on fruit flavor, color, size, and number of spines on the fruits and cladodes is preserved in the local community. Genetic fingerprinting with 129 AFLPs did not correlate with this traditional morphological classification of 67 cacti. Yet, these AFLPs distinguished the analyzed 67Opuntia pilifera cacti easily from the out-group comprising 17 wildOpuntia velutina.     

Which Morphological Characteristics Are Most Influenced by the Host Matrix in Downy Mildews? A Case Study in Pseudoperonospora cubensis

Before the advent of molecular phylogenetics, species concepts in the downy mildews, an economically important group of obligate biotrophic oomycete pathogens, have mostly been based upon host range and morphology. While molecular phylogenetic studies have confirmed a narrow host range for many downy mildew species, others, like Pseudoperonospora cubensis affect even different genera. Although often morphological differences were found for new, phylogenetically distinct species, uncertainty prevails regarding their host ranges, especially regarding related plants that have been reported as downy mildew hosts, but were not included in the phylogenetic studies. In these cases, the basis for deciding if the divergence in some morphological characters can be deemed sufficient for designation as separate species is uncertain, as observed morphological divergence could be due to different host matrices colonised. The broad host range of P. cubensis (ca. 60 host species) renders this pathogen an ideal model organism for the investigation of morphological variations in relation to the host matrix and to evaluate which characteristics are best indicators for conspecificity or distinctiveness. On the basis of twelve morphological characterisitcs and a set of twelve cucurbits from five different Cucurbitaceae tribes, including the two species, Cyclanthera pedata and Thladiantha dubia, hitherto not reported as hosts of P. cubensis, a significant influence of the host matrix on pathogen morphology was found. Given the high intraspecific variation of some characteristics, also their plasticity has to be taken into account. The implications for morphological species determination and the confidence limits of morphological characteristics are discussed. For species delimitations in Pseudoperonospora it is shown that the ratio of the height of the first ramification to the sporangiophore length, ratio of the longer to the shorter ultimate branchlet, and especially the length and width of sporangia, as well as, with some reservations, their ratio, are the most suitable characteristics for species delimitation.     

Pre‐zygotic reproductive isolation between two synchronopatric Opuntia (Cactaceae) species in the Brazilian Chaco

• Opuntia (Cactaceae) is known for high rates of hybridization and ploidisation, resulting in the formation of new species. The occurrence of two sympatric and closely related species of Opuntia, O. elata and O. retrorsa, in Brazilian Chaco enabled us to test the hypothesis that pre‐zygotic reproductive isolation mechanisms operate in both species.
• We monitored the flowering period, as well as floral biology, and compared the morphological variation of floral structures through measurements, performed intra‐ and interspecific cross‐pollination tests, and recorded the guild of floral visitors and pollinators.
• Flowering was seasonal and highly synchronous. Floral biology exhibits similar strategies, and although floral morphology differs significantly in many of the compared structures, such morphological variation does not result in the selection of exclusive pollinators. Floral visitors and pollinators are oligolectic bees shared by both species. Opuntia elata and O. retrorsa are self‐compatible. While interspecific cross‐pollination (bidirectional) resulted in germination, the pollen tube did not penetrate the stigma.
• Opuntia elata and O. retrorsa are closely related; however, they are isolated and do not hybridise in Brazilian Chaco. We found that both have weak pre‐pollination barriers, but that they are strongly isolated by pollen–pistil incompatibility, i.e. post‐pollination barrier.

     

Phylogenetic relationships of five Oriental Apanteles species-groups (Hymenoptera: Braconidae: Microgastrinae) by concatenating four molecular markers

Genus Apanteles is a cosmopolitan and largest genus in Microgastrinae that can be set as chief example of the subfamily to be studied for. However, highly similar morphological characters intraspecifically lead to difficulty in identifying this genus. In this study, 25 individuals of Apanteles were identified morphologically and molecularly by using error rates approach of barcoding analysis. The phylogenetic of the Apanteles that consisted of five species-groups were studied together with their molecular time divergences by using four molecular markers i.e., COI, 16S, 28S and Cyt B. The species groups are mycetophilus, ultor, merula, merope and ater. The phylogenetic analyses show that each species groups were monophyletic with formation of sister clad for merula and merope-group. The phylogenetic analyses of Apanteles species-group were well resolved and supported by their morphological and taxonomical status. This study is a first record on the phylogenetic of five Oriental Apanteles species-group.     

Orbilia tianmushanensis sp. nov., a new member of the O. luteorubella group with an unusual asexual morph

A new species of Orbilia related to O. luteorubella is described mainly based on morphological characters of its asexual morph and molecular data. The sexual morph does not significantly differ from O. luteorubella, whereas the asexual morph obtained from its ascospore isolate resembles members of the non-predacious genus Dactylella, because it has fusiform phragmoconidia borne singly at the apex of conidiophores. Phylogenetic analysis showed that this strain clustered with a clade that included available strains of the O. luteorubella aggregate and was distant from all analysed Dactylella species. Within this clade, the new strain fell between species with filiform conidia and those of a Pseudotripoconidium anamorph. By combining morphological and phylogenetic analyses, we conclude that our isolate belongs to a new taxon. Pleomorphism of the new taxon is described and discussed.     

What helps Opuntia stricta invade Kruger National Park,South Africa: Baboons or elephants?

Question : Is Opuntia stricta more frequent, and its patches larger, under trees suitable for baboon roosting? If so, does it mean that baboons are major dispersal agents and that plants established under these trees are important foci of Opuntia stricta spread? Location: Skukuza, Kruger National Park, South Africa. Method: We surveyed an area invaded by Opuntia stricta in the Skukuza region of KNP. The survey included plots under potential baboon roosting trees,plots under trees unlikely to support baboons,and paired randomly located open sites. Results: The null hypothesis ‐tree‐Opuntia spatial independence – can be rejected for Acacia nilotica, but not for Spirostachys africana. Opuntia plants are positively associated with Acacia trees suitable for baboon roosting. However, there is no significant difference between frequency of Opuntia under Acacia trees suitable and unsuitable for baboon roosting.It appears that all Acacia trees can serve as nurse trees for Opuntia. Compared to plots under Acacia trees, frequencies of old and robust Opuntia plants are significantly higher in open areas and under dead trees. Conclusions: While baboons may be responsible for long distance Opuntia dispersal (over km),their role is not detectable at a local scale.On the other hand, elephants seem to contribute substantially to the local vegetative propagation of this species. Opuntia establishment and growth are more influenced by micro‐habitat than previously thought.     

Integration of invasive <Emphasis Type="Italic">Opuntia</Emphasis> spp. by native and alien seed dispersers in the Mediterranean area and the Canary Islands

The success of many alien plant species depends on mutualistic relationships with other species. We describe the assemblage of seed dispersers on three species of alien Opuntia invading Mediterranean and Macaronesian habitats, and examine the quality of such plant-animal interactions. We identified vertebrates consuming O. maxima, O. dillenii and O. stricta fruits by direct observation and collecting droppings and pellets. Phenology of the alien species, as well as that of coexisting native species, was monitored for an entire year. Germination tests of ingested and non-ingested seeds were performed both in the greenhouse and in the field. Seed coat thickness and viability were also measured for all treatments. A great variety of taxa, including reptiles, birds and mammals actively participate in the seed dispersal of Opuntia. Phenology of Opuntia fruits in Menorca and Tenerife overlaps with only a few native fleshy-fruited plants present in the study areas, which suggests an advantage for the invader. Most seeds germinated during the second year of the experiment, independently of the effect produced by the dispersers’ guts. We found great variation in the germination percentage of Opuntia after gut passage and in the effects of ingestion on seed coat thickness. Seed viability was somewhat reduced after gut passage compared to manually depulped seeds. Our results show how different Opuntia species are integrated into native communities by means of mutualistic interactions, with both native and alien dispersers. Although with heterogeneous effects, either type of disperser potentially contributes to the spread of these alien cacti in the recipient areas.     

Aspergillus yunnanensis,a new and rare species in the Aspergillus section Jani

Aspergillus is a monophyletic genus comprising the subgenera Aspergillus, Circumdati, Cremei, Fumigati, Nidulantes and Polypaecilum. The subgenus Circumdati contains many economically important species and mycotoxin producers. Section Jani was recently introduced with morphological and molecular support. In the present study, two strains isolated from farmland soil were assigned in section Jani based on multi-locus phylogenetic analyses but showed low similarity with existing species. Further morphological observation found they had wider vesicles and conidia connections which were different from the known species. Based on phylogenetic and morphological data, Aspergillus yunnanensis was introduced as the third species in section Jani. Members in section Jani are rarely distributed, this is the first report of this section in China.     

Species delimitation in the European species of Clavulina (Cantharellales, Basidiomycota) inferred from phylogenetic analyses of ITS region and morphological data

The identification of the conventionally accepted species of Clavulina (Cantharellales, Basidiomycota) in Europe (Clavulina amethystina, Clavulina cinerea, Clavulina cristata, and Clavulina rugosa) is often difficult and many specimens are not straightforwardly assignable to any of those four species, which is why some authors have questioned their identity. In order to assess the status of those species, a morphological examination was combined with the molecular analysis of the ITS region. The same six major clades were obtained in the Bayesian and parsimony phylogenetic analyses, and all six clades were well-supported at least by one of the analyses. Morphological characters, such as the overall branching pattern, the presence and intensity of grey colour, the cristation of the apices, and basidiospore size and shape were to various extents correlated with the phylogenetic signal obtained from the ITS region. The congruence between the molecular analyses and morphology, rather than geographical origin, suggests the existence of several species that can be delimited using a combined phylogenetic and morphological species recognition. The analyses revealed that C. cristata and C. rugosa are well-delimited species. In contrast, more than one taxa could be subsumed under the names C. amethystina and C. cinerea, the taxonomical complexity of which is discussed. The ITS region is proved to be adequate to separate phylogenetic species of Clavulina.     

Characterization of 16SrII-D subgroup associated phytoplasmas in new host plants in Egypt

Samples of three plant species displaying phytoplasma symptoms were collected from Kafrelsheikh and Al-Gharbia governorates during 2014. Witches’ broom and virescence symptoms were observed in periwinkle (Catharanthus roseus). Onion (Allium cepa) plants showed yellowing, streaks and twisting and Opuntia abjecta with proliferation and cylindrical of cladodes. Total DNA was extracted from symptomatic and asymptomatic plants, and phytoplasma were detected in all 12 symptomatic plants collected through direct and nested PCR assays with primers P1/P7 and R16F2n/R16R2. The results of phylogenetic analysis revealed that the phytoplasma isolates belong to 16SrII group. With a nucleotide identity greater than 98.7% with three members of 16SrII group, Papaya yellow crinkle, Y10097; “Ca. P. aurantifolia”, U15442; and peanut witches’ broom, Al33765, the strains identified in this study are “Ca. P. aurantifolia”-related strains. Virtual RFLP analysis of the 16S rRNA gene sequences with 17 restriction enzymes confirmed that the phytoplasma isolates belong to the “Candidatus Phytoplasma australasia” 16SrII-D subgroup. To the best of our knowledge, periwinkle, onion and Opuntia abjecta are considered new hosts for 16SrII group in Egypt.     

Antigenic patterns of seed proteins in Opuntioideae (Cactaceae)

The antigenic patterns recognised by Western blotting in seed proteins of species of Opuntioideae (Cactaceae) were analysed in an attempt to evaluate their usefulness in systematics. Total protein profiles were also analysed by SDS–PAGE. The resulting similarity and distance matrices were further used to carry out Cluster Analysis (UPGMA) and Principal Coordinates Analysis. Populations of Opuntia cardiosperma were found to exhibit a prominent morphological uniformity, a unique electrophoretic pattern and a uniform antigenic pattern. The latter was obtained using anti-O. cardiosperma as antiserum. Results from the qualitative and quantitative interspecific analyses of antigenic profiles helped to characterise all the species studied. Tephrocactus articulatus and Cylindropuntia imbricata evidenced lower affinity with O. cardiosperma than the species of Opuntia s.s. Our results demonstrate that in Cactaceae, Western blotting analysis broadens the usefulness range of immunological techniques at the specific level and complements the information collected from electrophoretic profiles.     

Microsatellite markers help to assess genetic diversity among Opuntia ficus indica cultivated genotypes and their relation with related species

Opuntia spp. belong to the Cactaceae family and are native to Central America. The most economically important species is O. ficus indica, cultivated both for fruits and cladodes. The genus includes other important edible species (from diploid to octoploid) that occur worldwide as either wild or cultivated species in many arid or semiarid areas (e.g., the Mediterranean region). Several accessions are cultivated in different growing regions, but little is known about their ancestries and levels of genetic diversity. The aim of this study was to investigate the level of intraspecific genetic diversity among O. ficus indica cultivated varieties and some related species. Specifically, six highly polymorphic simple sequence repeats (SSR) and two expressed sequence tag (EST)-SSR loci were investigated in 62 wild and cultivated genotypes belonging to 16 Opuntia species. The clusters identified by the distance and model-based analyses clearly separated the wild opuntias from the cultivated ones. However, the O. ficus indica accessions did not cluster separately from other arborescent cactus pear species, such as O. amyclaea, O. megacantha, O. streptacantha, O. fusicaulis, and O. albicarpa, indicating that their current taxonomical classifications do not fit with their genetic variability. In general, the genotypes cultivated in Mexico showed high levels of diversity, whereas most of the spineless accessions collected in other countries had a very narrow genetic base. This study increases our knowledge of the variability among some of the most diffused Opuntia cultivated accessions. This study also points to the inconsistencies of previous taxonomical genotype assignments that were based solely on morphological characteristics.     

CHROMOSOME NUMBERS IN CHIHUAHUAN DESERT CACTACEAE. TRANS-PECOS TEXAS

Chromosome numbers are presented for 100 collections of Cactaceae from the Trans-Pecos region of Texas. A total of 65 taxa representing 52 species and 12 genera were counted, including first reports for 45 taxa and new ploidy levels for four taxa. Notable among those listed are counts for Opuntia schottii var. grahamii (n = 11, ca. 22), O. stanlyi (n = 22), O. arenaria (n = 11), O. phaeacantha var. spinosibacca (n = 22), O. lindheimeri var. lindheimeri (n = 11), O. strigil (n = 11), Echinocereus enneacanthus var. cf. dubius (n = ca. 22), E. pectinatus var. neomexicanus (n = 22), and Thelocactus bicolor var. bicolor (n = 22). Endomitosis was found to be present in O. phaeacantha var. spinosibacca, Mammillaria pottsii, and Neolloydia intertexta var. dasyacantha. Meiotic irregularities were noted in some species belonging to the genera Opuntia and Echinocereus. Phytogeographic considerations are inferred from the chromosomal data for O. polyacantha, O. lindheimeri, and O. ficusindica. A base number of x = 11 in Cactaceae is supported.     

THE COMPARATIVE MORPHOLOGY OF THREE CYLINDROPUNTIAS

The three species discussed in this article have at one time been considered congeneric in Grusonia. Present evidence indicates the three are specifically distinct and should be congeneric, but as cylindropuntias in Opuntia. There seems to be no valid reason for retaining the genus Grusonia. Except for branching and decomposition, Opuntia bradtiana and O. santamaria are more similar anatomically and morphologically than O. kunzei is to either of the two.     

Morphological and phylogenetic studies of Cytospora (Valsaceae,Diaporthales) isolates from Chinese scholar tree,with description of a new species

Cytospora species are the most serious and widespread pathogens associated with canker disease on multiple plants. In this study, three species, i.e., Cytospora sophoricola, C. chrysosperma, and C. sophorae, which were isolated from Sophora in China, are described and illustrated based on their morphological characteristics and phylogenetic analyses. Cytospora sophoricola was distinguished clearly by its larger disc, multiple ostioles, cystic and multiple locules, and specific cultural characteristics, i.e., protruding fruiting bodies. Maximum parsimony and maximum likelihood analysis showed that it did not cluster with any known species of Cytospora, so it is described as a new species. Cytospora sophorae is a previously reported species from Sophora, which is redescribed based on new isolates and additional observations. Another species was identified as C. chrysosperma, which is reported for the first time on Sophora, so Papilionaceae is shown to be a new host family for C. chrysosperma. The morphological affinities of these species with related taxa are discussed, while the phylogenetic relationships of these species with other fungus in the genus Cytospora were elucidated based on their internal transcribed spacer (ITS) rRNA region sequences.     

Conspecificity of <Emphasis Type="Italic">Endothia eugeniae</Emphasis> and <Emphasis Type="Italic">Cryphonectria cubensis</Emphasis>: a re-evaluation based on morphology and DNA sequence data

Cryphonectria cubensis and Endothia eugeniae are fungal pathogens of Eucalyptus and clove that were reduced to synonymy on the basis of results of cross-inoculation studies, isozyme analysis, cultural studies, and morphology. A previous phylogenetic study on Cryphonectria, based on sequence variation in the ITS region of the ribosomal RNA operon, also supported the conspecificity of C. cubensis and E. eugeniae, but was based on only one E. eugeniae isolate. New collections from clove in Brazil and Indonesia have become available, providing the opportunity to reconsider the conspecificity of C. cubensis and E. eugeniae. The occurrence of C. cubensis on clove was confirmed based on morphological comparisons and phylogenetic analyses of ribosomal DNA and β-tubulin gene sequence data. In addition to C. cubensis, other fungi morphologically similar to Cryphonectria species on the basis of their orange stromata were present on some clove specimens, but no isolates were available for these fungi. Furthermore, some isolates, for which no herbarium material exists, grouped separately from the C. cubensis clade and closer to the Cryphonectria clade. The presence of more than one closely related fungus on clove raises questions relating to the legitimacy of the synonymy of E. eugeniae and C. cubensis. Based on the presence of C. cubensis on the type specimen of E. eugeniae, we recognize the synonymy of the two fungi but provide evidence that other fungi, more closely related to Cryphonectria spp. than to C. cubensis, are present on clove.     

Phylogenetic position of Octosporea muscaedomesticae (Microsporidia) and its relationship to Octosporea bayeri based on small subunit rDNA analysis

Comparative phylogenetic analysis of the small subunit rDNA sequence of Octosporea muscaedomesticae (Flu, 1911) (type species) (Microsporidia) isolated from the blowfly Phormia regina (Diptera:Calliphoridae) is presented. Neighbor Joining bootstrap, Maximum Parsimony and Maximum Likelihood analyses with 38 microsporidian taxa representing five major clades of Microsporidia placed O. muscaedomesticae on a separate branch within a clade containing parasites of freshwater hosts. O. muscaedomesticae differed from Octosporea bayeri, a parasite of the microcrustacean, Daphnia magna (Cladocera:Daphniidae) by 29% demonstrating that the latter microsporidium is not closely related to the type species at the generic level, and should not be placed within the genus Octosporea, a conclusion that is further supported by morphological and developmental differences. Considering the number of disparately related hosts from which Octosporea species have been previously described based mostly on developmental and morphological characters it is likely that many will not fit the current definition of the genus, and it is possible that molecular analysis of these species will show that this genus as defined represents a polyphyletic grouping of unrelated taxa.