Agave as a biofuel feedstock in Australia

The opportunity for commercial production of Agave in Australia stems from the substantial carbohydrate and fibre content of Agave, the nature of carbohydrates stored, the pre‐existence of an Agave ethanol‐producing industry in Mexico, demand for biofuel feedstocks, impressive water‐use efficiencies of plants with the CAM pathway, and legislation mandating the ethanol content of fuel in Australia, where an estimated 748 ML will be required for blending with petrol by 2010–2011, compared with about 440 ML in 2009. Agave has potential as a crop for areas of seasonally limited rainfall in Australia, a judgement based upon desktop analyses and agronomic experience of growing Agave in Australia before 1915. Development of a viable Australian Agave farming system requires production be located in suitable regions, efficient propagation methods, mechanized production, and viable business plans at grower, processor and marketing levels. Growers and processors agree that Agave will not be grown commercially until plants are grown, maintained and harvested in Australia, product is produced and tested, and yield and risks are evaluated. To this end, Agave were imported into Australia from Mexico and a tissue culture propagation technique developed. A trial crop of Agave tequilana was planted in northern Queensland and CO₂ exchange, nitrogen and water dynamics, carbohydrate content, and system inputs and outputs are being monitored. The experience will be used to fine‐tune the farming system, assess production costs and develop robust life‐cycle assessments. Processing of plants from trials will test harvesting and transport infrastructure and will provide material to processors for testing. Samples will be provided to potential producers of value‐added products. An Australian Agave industry should provide opportunities for stimulating agronomic, scientific and commercial exchange between Australia and Mexico. Successful integration of Agave into Australian agriculture will require a biofuels‐focussed breeding programme in collaboration with Mexican researchers.     

The global potential for Agave as a biofuel feedstock

Large areas of the tropics and subtropics are too arid or degraded to support food crops, but Agave species may be suitable for biofuel production in these regions. We review the potential of Agave species as biofuel feedstocks in the context of ecophysiology, agronomy, and land availability for this genus globally. Reported dry biomass yields of Agave spp., when annualized, range from <1 to 34 Mg ha^?1 yr^?1 without irrigation, depending on species and location. Some of the most productive species have not yet been evaluated at a commercial scale. Approximately 0.6 Mha of land previously used to grow Agave for coarse fibers have fallen out of production, largely as a result of competition with synthetic fibers. Theoretically, this crop area alone could provide 6.1 billion L of ethanol if Agave were re‐established as a bioenergy feedstock without causing indirect land use change. Almost one‐fifth of the global land surface is semiarid, suggesting there may be large opportunities for expansion of Agave crops for feedstock, but more field trials are needed to determine tolerance boundaries for different Agave species.     

Historical,temporal, and geographic dynamism of the interaction between Agave and Leptonycteris nectar-feeding bats

Premise

The interaction between ecological and evolutionary processes has been recognized as an important factor shaping the evolutionary history of species. Some authors have proposed different ecological and evolutionary hypotheses concerning the relationships between plants and their pollinators; a special case is the interaction and suspected coevolution among Agave spp. and their main pollinators, the Leptonycteris bats. Agave spp. have, in general, a pollination syndrome compatible with chiropterophily including floral shape and size, nocturnal nectar production, and nectar quality and sugar concentration. Our goal was to analyze the interaction Agave–Leptonycteris and its dynamics during three different climate scenarios.

Methods

We modeled the Agave–Leptonycteris interaction in its spatial and temporal components during the Pleistocene using Ecological Niche Models (ENMs) and three climate scenarios: Current, Last Glacial Maximum (LGM), and Last InterGlacial (LIG). Furthermore, we analyzed the geographic correlation between 96 Agave spp. and two of the Mexican Tequila bats, genus Leptonycteris.

Results

We found that Leptonycteris spp. interact with different Agave spp. over their migratory routes. We propose an interaction refuge in Metztitlán and Tehuacán-Cuicatlán areas, where Agave- Leptonycteris interaction has probably remained active. During the nonmigratory season, both bat species consume nectar of almost the same Agave spp., suggesting the possibility of a diffuse coevolution among Agave and Leptonycteris bats.

Conclusions

We propose that in the areas related to migratory bat movements, each bat species interacts with different Agave spp., whereas in the areas occupied by nonmigrant individuals, both bat species consume nectar of almost the same Agave taxa.

     

Fuzzy GIS‐based multi‐criteria evaluation for US Agave production as a bioenergy feedstock

In the United States, renewable energy mandates calling for increased production of cellulosic biofuels will require a diversity of bioenergy feedstocks to meet growing demands. Within the suite of potential energy crops, plants within the genus Agave promise to be a productive feedstock in hot and arid regions. The potential distributions of Agave tequilana and Agave deserti in the United States were evaluated based on plant growth parameters identified in an extensive literature review. A geospatial suitability model rooted in fuzzy logic was developed that utilized a suite of biophysical criteria to optimize ideal geographic locations for this new crop, and several suitability scenarios were tested for each species. The results of this spatially explicit suitability model suggest that there is potential for Agave to be grown as an energy feedstock in the southwestern region of the United States – particularly in Arizona, California, and Texas – and a significant portion of these areas are proximate to existing transportation infrastructure. Both Agave species showed the highest state‐level renewable energy benefit in Arizona, where agave plants have the potential to contribute 4.8–9.6% of the states' ethanol consumption, and 2.5–4.9% of its electricity consumption, for A. deserti and A. tequilana, respectively. This analysis supports the feasibility of Agave as a complementary bioenergy feedstock that can be grown in areas too harsh for conventional energy feedstocks.     

Species of Agave with antimicrobial activity against selected pathogenic bacteria and fungi

The in vitro antimicrobial activity against pathogenic bacteria, yeast, and molds were examined in extracts of the Agave species A. lecheguilla, A. picta, A. scabra and A. lophanta using an agar diffusion technique. The extracts of A. picta produced zones of inhibition of 9–13 mm for E. coli, L. monocytogenes, S. aureus, and V. cholerae, while B. cereus and Y. enterocolitica were not inhibited. The other Agave species did not show any detectable inhibitory activity against the bacteria tested; however, all four Agave sp. were inhibitory against all yeast and molds analyzed as evident by 9–20 mm zones of inhibition. The minimum microbicidal concentration (MMC) of the active extract ranged from 1.8 to 7.0 mg/ml for the sensitive bacteria, and 2.0–3.0 mg/ml for yeast. In the case of molds, the minimum inhibitory concentration (MIC) of the active extracts ranged from 3.0 to 6.0 mg/ml. Together, these data suggest that the Agave sp. analyzed are potential antimicrobial candidates with a broad range of activity.     

Agave for tequila and biofuels: an economic assessment and potential opportunities

This paper explores the economic viability of producing biofuels from Agave in Mexico and the potential for it to complement the production of tequila or mescal. We focus on Agave varieties currently being used by the tequila industry to produce two beverages, tequila and mescal, and explore the potential for biofuel production from these plants. Without competing directly with beverage production, we discuss the economic costs and benefits of converting Agave by‐products to liquid fuel as an additional value‐added product and expanding cultivation of Agave on available land. We find that the feedstock cost for biofuel from the Agave piña alone could be more than US$3 L^?1 on average. This is considerably higher than the feedstock costs of corn ethanol and sugarcane ethanol. However, there may be potential to reduce these costs with higher conversion efficiencies or by using sugar present in other parts of the plant. The costs of cellulosic biofuels using the biomass from the entire plant could be lower depending on the conversion efficiency of biomass to fuel and the additional costs of harvesting, collecting and transporting that biomass.     

Productivity and water use efficiency of Agave americana in the first field trial as bioenergy feedstock on arid lands

Agave species are high‐yielding crassulacean acid metabolism (CAM) plants, some of which are grown commercially and recognized as potential bioenergy species for dry regions of the world. This study is the first field trial of Agave species for bioenergy in the United States, and was established to compare the production of Agave americana with the production of Agave tequilana and Agave fourcroydes, which are produced commercially in Mexico for tequila and fiber. The field trial included four experimental irrigation levels to test the response of biomass production to water inputs. After 3 years, annual production of healthy A. americana plants reached 9.3 Mg dry mass ha^?1 yr^?1 (including pup mass) with 530 mm of annual water inputs, including both rainfall and irrigation. Yields in the most arid conditions tested (300 mm yr^?1 water input) were 2.0–4.0 Mg dry mass ha^?1 yr^?1. Agave tequilana and Agave fourcroydes were severely damaged by cold in the first winter, and produced maximum yields of only 0.04 Mg ha^?1 yr^?1 and 0.26 Mg ha^?1 yr^?1, respectively. The agave snout weevil (Scyphophorus acupunctatus) emerged as an important challenge for A. americana cropping, killing a greater number of plants in the higher irrigation treatments. Physiological differences in A. americana plants across irrigation treatments were most evident in the warmest season, with gas exchange beginning up to 3 h earlier and water use efficiency declining in treatments with the greatest water input (780 mm yr^?1 water input). Yields were lower than previous projections for Agave species, but results from this study suggest that A. americana has potential as a bioenergy crop and would have substantially reduced irrigation requirements relative to conventional crops in the southwestern USA. Challenges for pest management and harvesting must still be addressed before an efficient production system that uses Agave can be realized.     

Prospecting for Energy-Rich Renewable Raw Materials: Agave Leaf Case Study

Plant biomass from different species is heterogeneous, and this diversity in composition can be mined to identify materials of value to fuel and chemical industries. Agave produces high yields of energy-rich biomass, and the sugar-rich stem tissue has traditionally been used to make alcoholic beverages. Here, the compositions of Agave americana and Agave tequilana leaves are determined, particularly in the context of bioethanol production. Agave leaf cell wall polysaccharide content was characterized by linkage analysis, non-cellulosic polysaccharides such as pectins were observed by immuno-microscopy, and leaf juice composition was determined by liquid chromatography. Agave leaves are fruit-like—rich in moisture, soluble sugars and pectin. The dry leaf fiber was composed of crystalline cellulose (47–50% w/w) and non-cellulosic polysaccharides (16–22% w/w), and whole leaves were low in lignin (9–13% w/w). Of the dry mass of whole Agave leaves, 85–95% consisted of soluble sugars, cellulose, non-cellulosic polysaccharides, lignin, acetate, protein and minerals. Juice pressed from the Agave leaves accounted for 69% of the fresh weight and was rich in glucose and fructose. Hydrolysis of the fructan oligosaccharides doubled the amount of fermentable fructose in A. tequilana leaf juice samples and the concentration of fermentable hexose sugars was 41–48 g/L. In agricultural production systems such as the tequila making, Agave leaves are discarded as waste. Theoretically, up to 4000 L/ha/yr of bioethanol could be produced from juice extracted from waste Agave leaves. Using standard Saccharomyces cerevisiae strains to ferment Agave juice, we observed ethanol yields that were 66% of the theoretical yields. These data indicate that Agave could rival currently used bioethanol feedstocks, particularly if the fermentation organisms and conditions were adapted to suit Agave leaf composition.     

Varietal difference in antifungal activity of some species of Agave

Crude leaf extracts of different Agave species were eluted out with the help of 80% methanol for their antifungal activity. The activity of A. americana, A. ferox, A. montana, A. marginata, and A. scabra were evaluated against brown rot fungi Postia placenta, the causal agent of wood decay of some economically important plants using a media poising method. The percent inhibition of hyphal growth was measured after the seventh day of incubation. A. montana shows the highest percent inhibition, 69.31%, of hyphal growth. These data suggested that the different Agave species analyzed have potential as antifungal agents with a broad range of activity. In the future they may be used as biocontrol agents to cure plant diseases without harming them.     

Culture-Independent Analysis of Lactic Acid Bacteria Diversity Associated with Mezcal Fermentation

Mezcal is an alcoholic beverage obtained from the distillation of fermented juices of cooked Agave spp. plant stalks (agave must), and each region in Mexico with denomination of origin uses defined Agave species to prepare mezcal with unique organoleptic characteristics. During fermentation to produce mezcal in the state of Tamaulipas, not only alcohol-producing yeasts are involved, but also a lactic acid bacterial community that has not been characterized yet. In order to address this lack of knowledge on this traditional Mexican beverage, we performed a DGGE-16S rRNA analysis of the lactic acid bacterial diversity and metabolite accumulation during the fermentation of a typical agave must that is rustically produced in San Carlos County (Tamaulipas, Mexico). The analysis of metabolite production indicated a short but important malolactic fermentation stage not previously described for mezcal. The denaturing gradient gel electrophoresis (DGGE) analysis of the 16S rRNA genes showed a distinctive lactic acid bacterial community composed mainly of Pediococcus parvulus, Lactobacillus brevis, Lactobacillus composti, Lactobacillus parabuchneri, and Lactobacillus plantarum. Some atypical genera such as Weissella and Bacillus were also found in the residual must. Our results suggest that the lactic acid bacteria could strongly be implicated in the organoleptic attributes of this traditional Mexican distilled beverage.     

In situ diversity and maintanenace of traditional Agave landraces used in spirits production in West-Central Mexico

Extensive monovarietal cultivation of Age tequilana Weber var. azul is threatening the diversity of the germplasm used in traditional Agave spirits production in west-central Mexico. To promote the preservation, use, and management of this germplasm, an ethnobotanical and morphological study was done in the center and south of the state of Jalisco, Mexico. The richness, distribution, and morphological variation of wild and cultivated Agave populations were characterized, and producers’ roles in germplasm maintenance and diversification were analyzed. Results indicated that: 1) A. angustifolia and A. rhodacantha are the primary gene pools used for selection; 2) Traditional landraces are differentiated morphological entities; and 3) In situ maintenance and increase of Agave germplasm diversity are the result of constant selection of wild germplasm, producer management of populations in the wild-domesticated gradient, and preservation of old landraces. Preservation of Agave germplasm diversity in west-central Mexico requires increased cultivation and valuation of traditional landraces.     

Investigations on chemical control of zebra disease in Agave hybrid no. 11648

In laboratory, greenhouse and field experiments, mancozeb and copper compounds controlled the leaf-spot phase of zebra disease caused by Phytophthora arecae, one of the pathogens responsible for this serious disease in Tanzania's high-yielding Agave hybrid no. 11648. As the copper compounds were phytotoxic, Dithane M-45 (Mancozeb) is recommended as a protectant during the rainy season.     

Seed germination of Agave species as influenced by substrate water potential

Background

Plants of Agave spp. perform Crassulacean acid metabolism (CAM) and are highly drought-tolerant, but little is known concerning seed germination under low water availability. The aim of this study was to assess the effect of substrate water potential (Ψ_W) on seed germination and contrast hydrotime parameters of seven valuable and commercially-important Agave species from different geographical distributions and climatic regions of Mexico. Our hypothesis was that seed germination of Agave species is not affected by low water availability independently of seed biomass and the climate of their distribution area.

Results

Seed germination (at 25°C and in the dark) between 85 and 100% for all species occurred within 80–180 h at -0.03 MPa and 250–430 h at -1.0 MPa. Seed germination at -1.5 MPa declined to less than 50% (p < 0.05) for A. asperrima and A. cupreata but did not change significantly for A. americana var. marginata, A. lechuguilla and A. striata, although they showed the lowest mean base water potential (-2.01 to -2.64 MPa). Seed germination of 40% Agave species, from arid and semi-arid climates in this study, was not affected by the lower Ψ_W.

Conclusion

Germination of seeds of Agave species is moderately affected by low water availability, is partially dependent of their ecological distribution, and is independent of seed mass.     

From Isozymes to Genomics: Population Genetics and Conservation of Agave in México

Mexico is a megadiverse country, but less than 54 % of its original vegetation still remains. In particular Mexican deserts and arid and semiarid ecosystems harbor a large number of endemic taxa, and the genus Agave is an outstanding example. Agave is one of the largest genera of the Mexican flora, including a total estimated number of 200 species, 74 % of them endemic to the country. Agave is also one of the Mexican plant genera with more population genetic studies. We describe here studies in 22 Agave species using different genetic markers. For the genus we found on average a high level of genetic variation, H _s ?=?0.19, and a low genetic differentiation, F _st ?=?0.15. We identify some species that should be subject to special conservation genetic efforts, in particular the endangered A. victoriae-reginae and both wild populations and landraces of A. angustifolia, including the cultivated A. tequilana.     

Potential of Plants from the Genus Agave as Bioenergy Crops

Agave is a succulent genus within the monocot family Agavaceae. The plants have a large rosette of thick fleshy leaves, each ending generally in a sharp point, and are native to arid and semi-arid regions from the southern USA to northern South America. The most important commercial species is Agave tequilana grown for production of tequila. Several cultivated species of Agave such as Agave sislana and Agave salmiana can perform well in areas where rainfall is insufficient for the cultivation of many C₃ and C₄ crops. The key feature of the crassulacean acid metabolism photosynthetic pathway used by agaves is the stomata opening and CO₂ uptake during the night, thus allowing less water to be lost by transpiration. Alcoholic beverages, sweeteners, fibers, and some specialty chemicals are currently the main products coming from agave plants. The recovered information related to productivity, biofuel processability, by-products, etc. suggests that some Agave species have a real potential to compete economically with other bioenergy crops. But more than compete, it could complement the list of bioenergy crops due to its capacity to grow with very little rainfall and/or inputs and still reach good amount of biomass, so unused semi-arid land could be productive. Although Agave has great potential to be developed as a bioenergy crop, more laboratory and field research are needed.     

Wild and agronomically important Agave species (Asparagaceae) show proportional increases in chromosome number,genome size,and genetic markers with increasing ploidy

Agave (Asparagaceae) includes cultivated and wild varieties of henequen used for hard fibre production. As part of a breeding programme to improve Agave production, species with different ploidy levels were genetically characterized: two diploids [A. tequiliana Weber and the hybrid H11648 ((A. amaniensis Trel. & Nowell × A. angustifolia Haw.) × A. amaniensis)], a triploid (A fourcroydes Lem. var. kitam ki), a tetraploid (A. angustifolia var. letona), three pentaploids (A. fourcroydes var. sac ki, A. fourcroydes var. yaax ki, and A. sisalana Perrine), and two hexaploids (A. angustifolia var. chelem ki from two locations). Chromosome spreading was used to determine the chromosome number, flow cytometry was employed to measure the genome size, and fluorescent in situ hybridization was performed using 45S and 5S ribosomal DNA (rDNA) and the telomeric sequences (TTAGGG)_n and (TTTAGGG)_n as genetic markers. There were proportional increases with ploidy level of the following: (1) chromosome number (from diploid 2n = 2x = 60 to hexaploid 2n = 6x = 180), including the number of large and small chromosomes in the bimodal karyotype of Agave; (2) genome size, with a mean monoploid genome size (1Cx) of 7.5 pg (range, 7.36–7.61 pg); and (3) the number and distribution of 45S and 5S rDNA loci, with one locus of each per basic, monoploid genome. Thus there was complete additivity in genome structure with increasing ploidy, as reported in some angiosperm polyploids. However, as other analyses of polyploids have revealed a decrease in 1Cx values with increased ploidy, possible explanations for the observed genomic stability were considered. With the (TTAGGG)_n probe, the signal was localized at the telomeres, consistent with published data showing that many species in the order Asparagales have this type of telomere sequence. It is speculated that sporadic telomeric signals using the (TTTAGGG)_n probe are probably derived from either errors in telomerase activity or relic ancestral‐type telomeric sequences. © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society, 2008, 158 , 215–222.     

Evolutionary ecology of Agave: distribution patterns,phylogeny, and coevolution (an homage to Howard S. Gentry)

With more than 200 species, the genus Agave is one of the most interesting and complex groups of plants in the world, considering for instance its great diversity and adaptations. The adaptations include the production of a single, massive inflorescence (the largest among plants) where after growing for many years, sometimes more than 30, the rosette dies shortly afterward, and the remarkable coevolution with their main pollinators, nectarivorous bats, in particular of the genus Leptonycteris. The physiological adaptations of Agave species include a photosynthetic metabolism that allows efficient use of water and a large degree of succulence, helping to store water and resources for their massive flowering event. Ecologically, the agaves are keystone species on which numerous animal species depend for their subsistence due to the large amounts of pollen and nectar they produce, that support many pollinators, including bats, perching birds, hummingbirds, moths, and bees. Moreover, in many regions of Mexico and in the southwestern United States, agaves are dominant species. We describe the contributions of H. S. Gentry to the understanding of agaves and review recent advances on the study of the ecology and evolution of the genus. We analyze the present and inferred past distribution patterns of different species in the genus, describing differences in their climatic niche and adaptations to dry conditions. We interpret these patterns using molecular clock data and phylogenetic analyses and information of their coevolving pollinators and from phylogeographic, morphological, and ecological studies and discuss the prospects for their future conservation and management.     

Two new Agave species (Agavaceae) from central Arizona and their putative pre-Columbian domesticated origins

Recent fieldwork in central Arizona resulted in the discovery of two agaves that display low seed set, reproduce mainly by vegetative means, have restricted distributions, and cannot be readily assigned to any existing species. These agaves are closely associated with archaeological structures and features and can be found growing with other previously described pre-Columbian Agave domesticates. Herein we describe Agave verdensis and A. yavapaiensis, two species that can be placed within Gentry’s informal Ditepalae Agave group, and propose that they are clonal relict domesticates. The two species have affinities with A. chrysantha, A. shrevei, and A. delamateri, the latter also a central Arizona pre-Columbian domesticate. We provide a key to distinguish these species from other agaves in central Arizona and adjacent northern Mexico with which they may be confused. The discovery of these two new species brings the total number of putative Arizona Agave domesticates to five species.     

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.     

Pollen viability and pollen vigor

Summary Investigations were carried out to correlate pollen viability, assessed on the basis of a fluorochromatic reaction (FCR) test, with pollen vigor, assessed on the basis of the time taken for in vitro germination in pollen grains subjected to high humidity (>95% RH) and temperature (38 °C) or storage stress of Nicotiana tabacum, Agave sp., Tradescantia virginiana, and Iris sp. Both high RH and temperature, as well as storage stresses, affected pollen vigor before affecting pollen viability. The results are discussed in the light of available data on the viability and vigor of stressed pollen and of aged seeds. The need for consideration of pollen vigor, particularly in stored pollen, the inadequacy of the methods presently used, and some of the methods suitable to assess pollen vigor are elaborated.