Isozymatic variation and phylogenetic relationships between henequen (Agave fourcroydes) and its wild ancestor A. angustifolia (Agavaceae)

Isozymatic variation and phylogenetic relationships among extant henequén (Agave fourcroydes) germplasm and wild populations of its ancestor A. angustifolia in the Yucatan Peninsula in Mexico were analyzed. Analysis of three isozyme systems using starch gel electrophoresis indicated that while A. angustifolia populations have relatively high levels of variation, within each henequén cultivar all individuals were identical. This result corresponds to previous ethnobotanical and morphological analyses, which indicated severe loss of genetic variation of this domesticated plant as a consequence of the promotion by means of asexual propagation of only one cultivar since the middle of the last century. The three extant cultivars of henequén were distinct from each other. Two of them, Sac Ki (SK) and Yaax Ki (YK), could be matched within the progenitor, but Kitam Ki (KK) has a MDH electrophenotype not found in any of the plants growing inside the Yucatan Peninsula, but found in some A. angustifolia plants growing in the Mexican states of Oaxaca and Veracruz. A parsimony analysis of the morphological data indicated two lineages: that of SK and YK, cultivated cordage plants selected for stronger and longer fibers, whose sister group is the Tropical subdeciduous forest ecotype (SF), and that of all the other wild populations, which also included KK, the cultivated textile plants selected for finer fibers and nearly extinct in Yucatan. These results support the hypothesis of the yucatecan origin of SK and YK from the SF ecotype, as well as the hypothesis of a recent introduction of KK to the Yucatan Peninsula in a domestication trend that probably included also Chelem White (its cultivation being abandoned later).     

Morphological variation of henequen (Agave fourcroydes, Agavaceae) germplasm and its wild ancestor (A. angustifolia) under uniform growth conditions: diversity and domestication

Extant variants of henequen (Agave fourcroydes Lem.) and wild populations of its putative ancestor A. angustifolia Haw. were grown in the Mexican state of Yucatan for 10 yr under homogeneous conditions. A statistical and numerical analysis of their patterns of morphological variation was performed as part of broader research to provide evidence of its genetic diversity, evolutionary relationships and changes under human selection. A comparison with results of a similar analysis under natural growing conditions was also made. The study indicated the following. (1) Under natural growth conditions, the three putative wild ecotypes are morphologically distinct, but under uniform conditions only populations growing in Tropical subdeciduous forest may be distinguished from the other two, thus indicating the probable existence of only two ecotypes: one growing in Coastal dunes and Tropical subdeciduous forest, and the other growing in Tropical deciduous forest. (2) This last ecotype is the most similar to cultivated variants. Within its populations, the most similar to the cultivated is that known as Chelem White, gathered by artisans for its textile use. (3) The cordage-cultivated Sac Ki and Yaax Ki differ from wild populations in four syndromes of domestication: gigantism, greater fibrosity, less thorniness, and less reproductive capacity. The lower cv of their characteristics compared with those of wild populations suggest less genetic diversity. (4) Kitam Ki is probably a textile-cultivated variant of recent introduction and/or a variant in which the artificial selection process has had different direction and intensity. (5) Improved growth conditions in the botanic garden resulted in a decreased cv, an increase in size and fiber content, and a reduction of thorniness for both wild and cultivated variants. Given that wild populations with desirable characteristics exist and that these characteristics are highly plastic and respond positively to cultivation, then selection and cultivation of populations such as those from Tropical deciduous forest may well have been the path taken by the ancient Maya during henequen domestication.     

In vitro induction of a trisomic of Agave tequilana Weber var. Azul (Agavaceae) by para-fluorophenylalanine treatment

The effect of para-fluorophenylalanine (PFP) on the production of trisomic plants of Agave tequilana Weber var. Azul produced through somatic embryogenesis was investigated. Normal diploid plants with 2n = 2x = 60 were obtained in the control treatment and with 4 mg L⁻¹ PFP exposure, while use of 8 and 12 mg L⁻¹ PFP led to production of trisomics with 2n = 2x = 61. Normal diploid plants showed a bimodal karyotype with five pairs of large chromosomes and 25 pairs of small chromosomes. Trisomic plants also had a bimodal karyotype with a group of three chromosomes in position five of the chromosome set. More than 13 homologous chromosome pairs exhibited structural changes. Differences in chromosome arm ratio (long arm/short arm) were also found in eight chromosome pairs; all these aberrations in the chromosome complement of trisomic plants were probably caused by inversions, deletions, and/or duplications produced by high concentrations of PFP. The gross chromosome structural changes and the presence of a single extra chromosome could have been induced by the effect of PFP on the mitotic spindle by inducing nondisjunction of sister chromatids, resulting in hyperploids (2n + x) and hypoploids (2n − x). Flow cytometric analysis of nuclear DNA content was performed using nuclei isolated from young leaves of normal and trisomic plants. The 2C DNA content of 8.635 pg (1Cx = 4,223 Mbp of trisomic plants was different (p < 0.001) than that of normal plants (2C DNA = 8.389 pg (1Cx = 4,102 Mbp). The difference in genome size was correlated with the large structural changes in the trisomic plant genomes.     

Reproductive biology of henequen (Agave fourcroydes) and its wild ancestor Agave Angustifolia (Agavaceae). i. Gametophyte development

The pathways of micro- and megagametophyte development in Agave fourcroydes (henequén) and A. angustifolia were studied. We used histology and light microscopy to observe anther ontogeny and ovary differentiation in relation to flower bud size. Both species have the same sexual reproductive strategies and gametophyte development that may be divided into three phases: (1) premeiotic, which includes the establishment of the megaspore mother cell and the pollen mother cell; (2) meiotic, the formation of mature microspores and functional megaspores; (3) postmeiotic, which encompasses the development of mature pollen grains and the formation of the embryo sac. A successive type microsporogenesis was found in both species with formation of T-shaped tetrads and binuclear pollen grains. In vitro germination tests revealed very low pollen fertility. The female gametophyte is formed from two micropylar megaspore cells after the first meiotic division (bisporic type). Male and female gametogenesis occur asynchronously with microsporogenesis finishing before macrosporogenesis. The results so far show that the formation of male and female gametophytes in henequén is affected at different stages and that these alterations might be responsible for the low fertility shown by this species.     

Genetic consequences of pre-Columbian cultivation for Agave murpheyi and A. delamateri (Agavaceae)

Pre-Columbian farmers cultivated several species of agave in central Arizona from ca. A.D. 600-1350. Because of the longevity and primarily asexual reproduction of these species, relict agave clones remain in the landscape and provide insights into pre-Columbian agricultural practices. We analyzed variation in allozyme allele frequencies to infer genetic effects of prehistoric cultivation on Agave murpheyi and A. delamateri, specifically to estimate genetic diversity and structure, to determine whether cultivated populations descended from a single clone, and to examine regional-scale genetic variation. Agave murpheyi maintained more genetic diversity at the species and population levels than A. delamateri, and A. murpheyi populations typically included more multilocus genotypes. Relict plants from prehistoric fields reflect a more complex history than descent from a single clone; A. murpheyi populations may have included more diversity initially because bulbils (produced routinely in A. murpheyi but not A. delamateri) and possibly seed would have facilitated transport of genetically diverse planting stock. Genetic variation in both cultigens was lower than in most contemporary commercial crops but similar to that observed in modern traditional agricultural systems.     

Technological and Structural Properties of Oat Cookies Incorporated with Fructans (Agave tequilana Weber)

Food Biophysics - The beneficial effect of agave fructans on health has been demonstrated gaining popularity as a new prebiotic and functional food ingredient, however, their role as an ingredient...     

A taxonomic revision of the genus Agave (Agavaceae) in the Lesser Antilles, with an ethnobotanical hypothesis

The genusAgave (Agavaceae) is revised taxonomically for the Lesser Antilles. All 12 previously recognized species are placed into synonymy underAgave karatto Miller. A neotype is selected for the nameAgave karatto, and lectotypes are designated forA. barbadensis Trel.,A. medioxima Trel.,A. obducta Trel.,A. vangrolae Trel., andA. ventum-versa Trel. Specimens are cited for most of the Lesser Antillean islands. That the Caribbean agaves are similar to the MexicanAgave nayaritensis Gentry is supported. The hypothesis that the pattern of partial sterility, clonality, and minimal morphological differentiation of the Lesser Antillean agaves is related to prehistoric human activity is developed based on similar patterns among mainland agaves and historical information. Uses for the archaeological “Barbados shell tool” may have included decorticating agave leaves for fiber extraction.     

The comparative leaf anatomy of Agave, Beschorneria, Doryanthes and Furcraea species (Agavaceae: Agaveae)

The leaf anatomy is compared of 35 ppecies and one variety of Agave , three species of Furcraea , one species of Beschorneria and one species of Doryanrhes , four genera assigned to the tribe Agaveae by Hutchinson in his classification of the family Agavaceae. The genera can be readily differentiated by leaf anatomy. There are close similarities between Agave, Beschorneria and Furcraea , but Doryanthes differs widely from these genera. Sufficient anatomical differences exist to differentiate the Agave species examined.     

Spatial and temporal dynamics of blue agave (Agave tequilana Weber var. azul) wilt in Jalisco Mexico

Tequila is a Mexican alcoholic beverage made from the fermentation and distillation of the blue agave (Agave tequilana Weber var. azul) stem. This crop is affected by a wilt associated mainly with Fusarium oxysporum. This disease can produce considerable yield losses. Little is known about the spatial and temporal behaviour of blue agave wilt. In this work, the spatial and temporal dynamics of the disease in commercial blue agave plantations in the state of Jalisco, Mexico, were analysed. Four fields of approximately 1 ha were selected in the municipalities of Arandas and Magdalena, in which disease assessments were carried out over a year of evaluation. Each plant was categorized based on a scale with four severity classes (healthy plant, severity class 1, severity class 2 and severity class 3). Maps of disease distribution were made. The spatial pattern was analysed by means of four indicators of spatial variation for binomial data; the spatial and temporal variation was analysed by means of transition matrices. An aggregate spatial pattern was observed in all fields. The transitions in severity classes were not completely unidirectional; some plants showed symptom remission between the date of first and second disease assessment, while others remained at their original severity. Severity class 1 occurred most frequently in Arandas fields (from 12.9% to 40.3%). There was a notable increase in wilt severity to class 2 in the Magdalena fields (from 4% to 50.6%). The rates of disease development towards severity class 3 are low and do not suppose a significant loss for the crop; nevertheless, the rates of disease development towards the wilt severity class 2 do put in risk the health of the crop and the availability of the raw material for the making of tequila.     

Pollination ecology of Agave macroacantha (Agavaceae) in a Mexican tropical desert. I. Floral biology and pollination mechanisms

In a study of sexual reproduction in long-lived semelparous plants, we observed Agave macroacantha in the tropical desert of Tehuacán-Cuicatlán, Mexico, describing duration of flowering, flower phenology, and nectar production patterns. We also performed two manipulative experiments evaluating (a) the seed production efficiency of different crossing systems (selfing, cross-pollination, apomixis, and control), and (b) the effect of different pollinators (diurnal exposure to pollinators, nocturnal exposure, exclusion, and control) on the seeds produced. Flowering occurred from early May to late July and had a mean duration of 29 days in the individual rosettes. The flowers were protandrous; anthesis occurred in the afternoon of the third day after floral opening, and the pistils matured in the afternoon of the fifth day. The stigmas remained receptive from dusk to the following morning. Pollination was mostly allogamous. Nectar was produced principally during the night, from the first stages of floral aperture until the stigmas wilted and flowering ceased. The flowers were visited during the day by hymenoptera, butterflies, and hummingbirds and during the night by bats and moths. Only the nocturnal visitors, however, were successful pollinators. Agave macroacantha is extremely dependent on nocturnal pollinators for its reproductive success.     

Temperature influences on root growth for Encelia farinosa (Asteraceae), Pleuraphis rigida (Poaceae), and Agave deserti (Agavaceae) under current and doubled CO2 concentrations

To help evaluate root distribution patterns, elongation rates of individual roots were measured as a function of soil temperature for Encelia farinosa (a C₃ species), Pleuraphis rigida (C₄), and Agave deserti (CAM), sympatric codominants in the northwestern Sonoran Desert. Measurements were made at current and doubled CO₂ concentrations under winter and summer conditions of air temperature (day/night temperatures of 17 C/10 C and 33 C/22 C, respectively). The three species had different optimal temperatures for root elongation (T_opt) under winter conditions (25 C for E. farinosa, 35 C for P. rigida, and 30 C for A. deserti); T_opt increased by 2-3 C under summer conditions for all three species. The limiting temperatures for elongation also acclimated from winter to summer conditions. The rate of root elongation at T_opt was higher under summer than winter conditions for E. farinosa (9 vs. 6 mm d⁻¹) and P. rigida (20 vs. 14 mm d⁻¹), reflecting conditions for maximum photosynthesis; no difference occurred for A. deserti (9 vs. 10 mm d⁻¹). Decreased elongation rates at extreme temperatures were associated with less cell division and reduced cell extension. The doubled CO₂ concentration increased average daily root elongation rates for A. deserti under both winter (7%) and summer (12%) conditions, reflecting increased cell extension, but had no effect for the other two species. Simulations of root elongation as a function of soil temperatures showed that maximum elongation would occur at different depths (16-20 cm for E. farinosa, 4-8 cm for P. rigida, and 0-4 cm for A. deserti) and during different seasons (winter to spring for E. farinosa, spring to summer for P. rigida, and all year for A. deserti), contributing to their niche separation. Shading of the soil surface moderated daily variations in soil temperature, reducing seasonal root elongation for winter and spring and increasing elongation for summer. Shading also altered root distribution patterns, e.g., optimal rooting depth for A. deserti and especially P. rigida increased for a hot summer day.     

Population structure and genetic diversity distribution in wild and cultivated populations of the traditional Chinese medicinal plant Magnolia officinalis subsp. biloba (Magnoliaceae)

Magnolia officinalis subsp. biloba, a traditional Chinese medicinal plant, experienced severe declines in the number of populations and the number of individuals in the late 20th century due to the widespread harvest of the subspecies. A large-scale cultivation program was initiated and cultivated populations rapidly recovered the loss in individual plant numbers, but wild populations remained small as a consequence of cutting. In this study, the levels of genetic variation and genetic structure of seven wild populations and five domestic populations of M. officinalis subsp. biloba were estimated employing an AFLP methodology. The plant exhibited a relatively high level of intra-population genetic diversity (h = 0.208 and H _j = 0.268). The cultivated populations maintained approximately 95% of the variation exhibited in wild populations, indicating a slight genetic bottleneck in the cultivated populations. The analysis of genetic differentiation revealed that most of the AFLP diversity resided within populations both for the wild group (78.22%) and the cultivated group (85.92%). Genetic differentiation among populations in the wild group was significant (F _ST = 0.1092, P < 0.005), suggesting wild population level genetic structure. Principal coordinates analysis (PCO) did not discern among wild and cultivated populations, indicating that alleles from the wild population were maintained in the cultivated gene pool. Results from the present study provide important baseline data for effectively conserving the genetic resources of this medicinal subspecies.     

Diversity of S-alleles and mate availability in 3 populations of self-incompatible wild pear (Pyrus pyraster)

Small populations of self-incompatible plants may be expected to be threatened by the limitation of compatible mating partners (i.e., S-Allee effect). However, few empirical studies have explicitly tested the hypothesis of mate limitation in small populations of self-incompatible plants. To do so, we studied wild pear (Pyrus pyraster), which possesses a gametophytic self-incompatibility system. We determined the S-genotypes in complete samplings of all adult trees from 3 populations using a PCR-RFLP approach. We identified a total of 26 different S-alleles, homologous to S-alleles of other woody Rosaceae. The functionality of S-alleles and their Mendelian inheritance were verified in artificial pollination experiments and investigations of pollen tube growth. The smallest population (N = 8) harbored 9 different S-alleles and showed a mate availability of 92.9%, whereas the 2 larger populations harbored 18 and 25 S-alleles and exhibited mate availabilities of 98.4% and 99.2%, respectively. Therefore, we conclude that even small populations of gametophytic self-incompatible plants may exhibit high diversity at the S-locus and are not immediately threatened owing to reduced mate availability.     

Diversity and genetic structure in populations of Pseudotsuga menziesii (Pinaceae) at chloroplast microsatellite loci.

Genetic variation was compared between uniparentally-inherited (chloroplast simple sequence repeats, cpSSRs) vs. biparentally-inherited (isozyme and random amplified polymorphic DNA, RAPD) genetic markers in Douglas-fir (Pseudotsuga mensiezii) from British Columbia. Three-hundred twenty-three individuals from 11 populations were assayed. In Douglas-fir, the cpSSR primer sites were well-conserved relative to Pinus thunbergii (11 of 17 loci clearly amplified), but only 3 loci were appreciably polymorphic. At these cpSSR loci, we found an unexpectedly low level of polymorphism within populations, and no genetic differentiation among populations. By contrast, the nuclear markers showed variation typical of conifers, with significant among-population differentiation. This difference is likely the outcome of both historical factors and high pollen dispersal.     

Genetic structure of wild populations of the endangered Desert Pupfish complex (Cyprinodontidae: Cyprinodon)

Data from 10 microsatellite DNA loci were used to describe the genetic structure of the two extant species (Cyprinodon macularius and C. eremus) of the endangered Desert Pupfish complex of southwestern United States and northwestern Mexico. Variation at microsatellite loci was significantly correlated (Mantel test) with that of previous mtDNA results, both for the complex and for the relatively wide-ranging C. macularius alone. Both species showed unusually high levels of microsatellite diversity for non-marine fish (H _e = 0.84–0.93; A_R = 11.9–17.0). There was evidence (R _ST > F _ST) that the two extant populations of C. eremus have been isolated sufficiently long for mutation to contribute significantly to genetic divergence, whereas divergence among the nine assayed populations of C. macularius could be attributed to genetic drift alone. Correspondingly, 10% of the diversity in C. eremus was attributable to differences between the two populations, whereas, for C. macularius, only 2.7% was attributable to among-population variation. Within C. macularius, a small (0.8%), but statistically significant, portion was attributable to differences between populations in the Salton Sea area and those on the lower Colorado River delta. The two populations of C. eremus and five groups of populations of C. macularius are recommended as management units for conservation genetics management of the two species.     

Geographic patterns in the reproductive ecology of Agave lechuguilla (Agavaceae) in the Chihuahuan desert. I. Floral characteristics, visitors, and fecundity

Floral characteristics such as morphology and flower color have been interpreted as adaptive traits that evolved through selective pressures generated by pollinators. Differences among populations in the expression of floral characters could result from natural selection for their adaptive value to local conditions. We describe the patterns of variation of flower morphology, color, and fecundity of Agave lechuguilla in 11 populations along a latitudinal gradient encompassing the whole range of the species in the Chihuahuan desert. We found a latitudinal pattern in flower shape and color. Flowers tended to be shorter, more open, and colorful toward the northern part of the gradient. We also recorded flower visitation, discriminating between pollinators and floral robbers. The main pollinators seems to be nocturnal hawk moths (Hyles lineata) and diurnal large bees (Bombus pennsylvanicus and Xylocopa californica). In all populations large bees were the most abundant potential pollinators. However, the abundance of the potential pollinators varied along the gradient. We observed no bat visits along the gradient. The number of visits by all potential pollinators decreased significantly with latitude as did fruit set.     

Genetic structure of Araucaria angustifolia (Araucariaceae) populations in Brazil: implications for the in situ conservation of genetic resources

The distribution of the genetic variation within and among natural populations of A. ANGUSTIFOLIA growing in different regions in Brazil was assessed at microsatellite and AFLP markers. Both markers revealed high gene diversity ( H = 0.65; AR = 9.1 for microsatellites and H = 0.27; P = 77.8 % for AFLPs), moderate overall differentiation ( RST = 0.13 for microsatellites and FST = 0.10 for AFLPs), but high divergence of the northernmost, geographically isolated population. In a Bayesian analysis, microsatellite data suggested population structure at two levels: at K = 2 and at K = 3 in agreement to the geographical distribution of populations. This result was confirmed by the UPGMA dendrogram based on microsatellite data (bootstrap support > 95 %). Non-hierarchical AMOVA revealed high variation among populations from different A POSTERIORI defined geographical groups. The genetic distance between sample locations increased with geographical distance for microsatellites ( R = 0.62; P = 0.003) and AFLPs ( R = 0.32; P = 0.09). This pattern of population differentiation may be correlated with population history such as geographical isolation and postglacial colonization of highlands. Implications of the population genetic structure for the conservation of genetic resources are discussed.     

Clonal structure of wild populations and origins of horticultural stocks of Illicium parviflorum (Illiciaceae)

? Premise of the study: Habitat fragmentation is often assumed to result in limited genetic diversity across impacted plant communities. Central Florida has undergone extensive anthropogenic changes, while also harboring large numbers of endemic species. In this study, we assessed genetic structure and dependence on clonality in a central Florida endemic, Illicium parviflorum (Illiciaceae), as well as evaluated genetic diversity of this species in horticultural stocks. ? Methods: Six sites were sampled across the geographic range of I. parviflorum. A PCR-based assay using intersimple sequence repeats (ISSRs) was used to assess genetic structure. ? Key results: Results, based on 26 ISSR loci, suggest that clonal structure plays a role in all populations, with PD values ranging from 0.25 to 0.50. Only two populations exhibited unique genotypes, while the remaining four populations shared genotypes. Horticultural samples all shared one genotype, which can be traced back to a single natural population. ? Conclusions: Clonal reproduction is an important factor in the maintenance of natural populations of I. parviflorum, although the degree to which this is true varies by population. Horticultural samples likely represent a single or very few collection events, indicating the need for greater genetic diversity within horticultural stocks. Further analyses using microsatellites are planned to confirm these results.     

Geographic patterns in the reproductive ecology of Agave lechuguilla (Agavaceae) in the Chihuahuan desert. II. Genetic variation, differentiation, and inbreeding estimates

Plants with natural variation in their floral traits and reproductive ecology are ideal subjects for analyzing the effects of natural selection and other evolutionary forces on genetic structure of natural populations. Agave lechuguilla shows latitudinal changes in floral morphology, color, and nectar production along its distribution through north-central Mexico. Both the type and abundance of its pollinators also change with latitude. Using starch electrophoresis, we examined the levels and patterns of variation of 13 polymorphic allozyme loci in 11 populations of A. lechuguilla. The overall level of genetic variability was high (H(e) = 0.394), but the levels of genetic variation had no geographic pattern. However, the southern populations exhibited an excess of heterozygotes in relation to expectations for Hardy-Weinberg equilibrium, whereas the northern populations had an excess of homozygotes. Total differentiation among populations was low (θ = 0.083), although gene flow estimates (Nm) varied among groups of populations: southern populations had the lowest levels of genetic differentiation, suggesting high levels of gene flow; northern populations had greater levels of genetic differentiation (θ = 0.115), suggesting low gene flow among them. The patterns and inferences of the genetic structure of the population at the molecular level is consistent with variation in floral traits and pollinator visitation rates across the range of the species.