Bioprospecting for fungi with potential pathogenic activity on leaves of Agave tequilana Weber var. Azul

Thirty different fungal strains were isolated from A. tequilana leaves showing disease symptoms such as wilt and curled leaves, black, red and chlorotic spots. Ten genera were identified and confirmed by using the LSU D1/D2 rDNA and ITS1‐5.8S‐ITS2 regions, mainly of the Ascomycota phylum, where the Lasiodiploidia and Neoscytalidium genera were the more (46.6%) abundant. The other genera identified were Cladosporium, Cytospora, Epicoccum, Flavodon, Lasiodiplodia, Myrmaecium, Neoscytalidium, Penicillium, Peniophora, Purpureocillium, Trametes and Fusarium. Five strains of Lasiodiplodia and one of Fusarium were selected based on their representativeness and pathogenic potential on Agaves. Pathogenic potential was analysed by both, an infection assay, evidenced as necrosis, and by pectinolytic activity. Specifically, necrosis infection assay was conducted by puncture (wounded) infection and by direct mycelium contact. In general, Lasiodiplodia strains exhibited different pathogenic profiles according to their necrosis percentages, regardless of the infection method used. Fusarium strain analysed also showed a high necrosis infection (> 99%). Pectinolytic activity used as an indirect measurement of pathogenesis presented a high Fusarium extract activity (peaking at 23.9 U). Lasiodiplodia strains exhibited up 6 times more enzymatic activity (peaking at 143.5) than Fusarium strain analysed. In addition, Agave leaf extracts used totally or partially as carbon source during fungal induction culture may induce different pathogenic activities in these strains. In general, the two pathogenicity assays implemented evidenced differences in the pathogenicity profile of these analysed strains.     

Aberrant meiotic behavior in Agave tequilana Weber var. azul

Background  

Agave tequilana Weber var. azul, is the only one variety permitted by federal law in México to be used for tequila production which is the most popular contemporary alcoholic beverage made from agave and recognized worldwide. Despite the economic, genetic, and ornamental value of the plant, it has not been subjected to detailed cytogenetic research, which could lead to a better understanding of its reproduction for future genetic improvement.     

Stomatal physiology of a micropropagated CAM plant; Agave tequilana (Weber)

Experiments were designed to assess the capacity of an in vitro cultured CAM plant to control water loss and to examine the response of their stomata to various factors. Detached leaves of micropropagated Agave tequilana plants lost water at similar rates as did field-grown plantlets when dehydrated in air. This was consistent with the fact that stomata from micropropagated plants show similar morphology than field-grown plantlets. In addition, stomata from micropropagated plants responded to various factors in a manner similar to those from field-grown plantlets. It appears that in vitro culture does not affect the capacity of leaves to control water loss nor does it alters the nocturnal stomatal opening of this CAM plant.     

Active telomerase during leaf growth and increase of age in plants from Agave tequilana var. Azul

In plants, previous studies show that telomerase activity contributes to the maintenance of telomeric length for the proper development of organs and tissues. In this work, we investigated telomerase activity in A. tequilana during several years of cultivation. We found that during growth of the leaf there are two crucial phases: (1) the onset of cell elongation in 3 years and (2) differentiation of vascular bundles in 6 years. This coincides with the ages where the highest telomerase activity is seen. Therefore indicates that telomerase is associated with cellular activities such as; elongation, division, and cell differentiation. Likewise, we detected high activity during the period of vegetative growth, indicating that telomerase also contributes to telomeric maintenance on the leaf in A. tequilana.     

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.     

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...     

Thermal properties of agave fructans (Agave tequilana Weber var. Azul)

Thermal properties of agave (A. tequilana Weber var. Azul) at different water contents were investigated. HP-TLC results showed a complex mixture of mono-, di-, oligo, and polysaccharides in agave fructans samples. The thermal decomposition temperatures were observed below to 200 °C. Modulated-differential scanning calorimetry studies showed a glass transition and a relaxation enthalpy processes in agave fructans. Samples with the highest moieties of monosaccharides showed the lower glass transition temperatures (Tg). The moisture sorption isotherm of agave fructans was determined at 20 °C and fitted to the GAB model. Gordon-Taylor equation was used to fit the Tg experimental data as a function of water content. Agave fructans was found to be an amorphous material. At low water activity (a_w) values (<0.4), agave fructans remained in a powdered amorphous state; and at intermediate a_w (0.4-0.75) collapsed and caked; and at high a_w (>0.75) changed in a highly viscous liquid-like solution.     

Glutamate dehydrogenase activity in normal and vitrified plants of Agave tequilana Weber propagated in vitro

Agave tequilana stem explants were used to produce adventitious shoots under a set of different water potentials induced by different concentrations of gelrite in the medium. At high water potentials all shoots were vitrified; as the medium water potential became more negative the degree of vitrification decreased but the number of shoots per explant also diminished. The enzymes NADH and NAD-GDH (EC. 1.4.1.2) were measured along the water potential gradient. GDH activity was high in the non-vitrified tissues and decreased significantly in the vitrified ones.Abbreviations GDH glutamate dehydrogenase - MS Murashige and Skoog medium - MSO methionine sulfoximine - PVP polyvinylpolypyrrolidone - GS glutamine synthetase - GOGAT glutamine: oxoglutarate amino transferase     

In vitro propagation of Agave arizonica Gentry & Weber

Agave arizonica Gentry & Weber, an extremely rare and endangered species native to Arizona, was successfully propagated in vitro using modified Murashige and Skoog media. Adventitious shoots developed from callus which formed on bulbil explants grown in a medium supplemented with 1.4 μM 2,4-dichlorophenoxyacetic acid. These shoots proliferated by subculture in media supplemented with 44.4 μM 6-benzylaminopurine, and either 0.5 or 5.4 μM naphthaleneacetic acid. Rooting occurred on shoots transferred to a growth regulator free medium. Rooted plants transferred to potting soil could be established under greenhouse conditions following gradual acclimatization indoors.     

Interpopulation reproductive synchrony of Agave cocui (Agavaceae) in Venezuela

Agave cocui (Agavaceae) is a species with broad distribution in arid and semiarid areas of Venezuela and Colombia. Despite of its ecological importance as a source of food for wildlife, and its economic value for production of a spirit drink, studies on the reproductive ecology of the species are relatively rare. In this study, we conducted a one-year evaluation of the flowering and fruiting phenology of A. cocui in the eight representative localities of the species' distribution in Venezuela. Within each study site, we chose an area with a minimum of 50 reproductive individuals and followed their reproductive phenophases with the help of binoculars, using six qualitative cathegories (emerging reproductive stalk, flowers, inmature fruits, mature fruits, bulbils and dry stalk) every two months. Emergence of the reproductive stalk in most of the examined populations began in September (rainy season), although this event delayed two months in a few populations. We detected significant negative correlations between precipitation and the percentage of flowering occurrence in four of the eight populations. Floral resources are available for flower visitors during approximately five months of the year (January-May). In most populations production of flowers initiated in January (dry season), and for Western Venezuela and Andean regions, the flowering main peak occurred in January. Localities from the Central and Eastern Coast exhibited the flowering peak in March, showing a delay of approximately two months with respect to other populations. Beginning of fruit set varied among localities from January to May; however, peak production of mature fruits concentrated in May, and fruit occurrence varied broadly between 5.2 and 85%. Bulbil production was detected in all populations and varied greatly among them (maximum percentage per population: 26.19-92.10%). High flowering synchronicity (Phenophase Overlapping Index: 0.756 and 0.999) was observed among all populations monitored in Western Venezuela, including the Andean localities. This condition might facilitate the existence of a nectar corridor from the Western Coast and nearby islands, to the Andean arid patches, which could be potentially used by nectar-feeding bats and birds dependent on agave flowers during part of the year.     

In vitro propagation of Agave fourcroydes Lem. (Henequen)

In vitro plant regeneration of Agave fourcroydes Lem. (Agavaceae) is described. Results suggest that the NO₃ ^-:NH₄ ⁺ balance in the culture medium is a key factor controlling callus growth and organogenesis in rhizome cultures. Stem callus showed limited organogenic capacity, but high cytokinin concentrations induced adventitious shoot formation on stem explants. When these shoots were excised and subcultured, new callus formed at their base from which new shoots arose. The shoots from stem explants and rhizome callus formed extensive root systems in vitro and were transferred to pot culture with a 90% survival rate.     

Physiological activity in persistent bulbils of Agave vilmoriniana (Agavaceae)

Plants of Agave vilmoriniana are unique members of a reproductively diverse genus since they produce prolific numbers of both bulbils and viable seeds. Bulbils were observed to remain attached to the flowering stalk into a second summer, instead of abscising in the year they developed. Hence, a number of hypotheses were tested to establish the physiological basis for their longevity and evaluate their influence upon the plant's reproductive biology. Bulbils produced following anthesis were abundant (>2000 bulbils), large (0.44 g dry mass per individual), and their combined leaf surface area (one side) could exceed 3 m². Thus, the current-year inflorescence may act as a photosynthate source. Second-year, attached bulbils are healthy-looking, even after leaves of the parent plant have senesced, since maternal roots continue to absorb rain water. Rehydrated 2nd-yr bulbils resume Crassulacean acid metabolism, sometimes demonstrating net daytime tissue acidification. Water conservation by bulbils is very efficient, exhibiting very low cuticular conductances (0.016-0.0035 mm/s). Bulbils detach over all climatic seasons after having formed >3 and >18 root primordia in their 1st and 2nd year, respectively. About one-third of bulbils are capable of forming roots after a 1-yr storage at 35°C, when their water contents averaged 37%. Irrigated bulbils produce water-absorbing roots within 2 d, with de novo chlorophyll synthesis and nocturnal acidification commencing within 4 d. Two bulbils were observed to root and become established in shaded microhabitats during this 2-yr study of 29 plants.     

Heterologous expression of Fusarium oxysporum tomatinase in Saccharomyces cerevisiae increases its resistance to saponins and improves ethanol production during the fermentation of Agave tequilana Weber var. azul and Agave salmiana must

This paper describes the effect of the heterologous expression of tomatinase from Fusarium oxysporum f. sp lycopersici in Saccharomyces cerevisiae. The gene FoTom1 under the control of the S. cerevisiae phosphoglycerate kinase (PGK1) promoter was cloned into pYES2. S. cerevisiae strain Y45 was transformed with this vector and URA3 transformant strains were selected for resistance to α-tomatine. Two transformants were randomly selected for further study (designated Y45-1 and Y45-2). Control strain Y45 was inhibited at 50 μM α-tomatine, in contrast, transformants Y45-1 and Y45-2 did not show inhibition at 200 μM. Tomatinase activity was detected by HPLC monitoring tomatine disappearance and tomatidine appearance in the supernatants of culture medium. Maximum tomatinase activity was observed in the transformants after 6 h, remaining constant during the following 24 h. No tomatinase activity was detected in the parental strain. Moreover, the transformants were able to grow and produce ethanol in a mix of Agave tequilana Weber var. azul and Agave salmiana must, contrary to the Y45 strain which was unable to grow and ferment under these conditions.     

Utilization of by-products from the tequila industry. Part 2: Potential value of Agave tequilana Weber azul leaves

The leaves of the agave plant are left in the field after harvesting the heads for tequila production. Different types of agave leaves were isolated, classified, and their content in the total plant determined. The usable fractions were collected and their properties determined. Of the total wet weight of the agave plant, 54% corresponds to the agave head, 32% corresponds to materials which could be usable for sugar and fiber production which leaves 14% of the wet plant without apparent utility. The fractions with higher total reducing sugars (TRS) content were the fresh fraction of partially dry leaves stuck to the head and the leaf bases with a TRS content of 16.1% and 13.1%, respectively. The highest TRS concentration (16-28%) is in the agave head which is used for tequila production. The leaves are 90-120 cm long and 8-12 cm wide and contain fiber bundles that are 23-52 cm long and 0.6-13 mm wide. The ultimate fiber length is approximately 1.6 mm with an average width of 25 microns. There are several types of leaf fibers that can be utilized depending on what part of the plant they come from and what product is desired. Agave leaf fibers were pulped using a soda pulping process and the pulp was hand formed into test sheets. Test sheets made from pulped agave leaf fibers had a breaking length comparable to paper made from both pine and eucalyptus fibers, but the tear index and burst index were lower than the other two papers.     

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.     

Agave rzedowskiana, a new species in subgenus Littaea (Agavaceae) from western Mexico

Agave rzedowskiana is described and illustrated from the states of Sinaloa and Jalisco, Mexico. This species is a member of subgenusLittaea (Tagliabue) Baker “groupStriatae” of Baker. It is compared withA. petrophila García-Mend. & E. Martínez andA. dasylirioides Jacobi & Bouché.

---

Resumen  Se describe e ilustraAgave rzedowskiana de los estados de Sinaloa y Jalisco, México. La especie pertenece al subgéneroLittaea (Tagliabue) Baker “Grupo Striatae” de Baker. Se le compara conA. petrophila García-Mend. & E. Martínez yA. dasylirioides Jacobi & Bouché.

     

Effect of light quality and culture medium on somatic embryogenesis of <Emphasis Type="Italic">Agave tequilana</Emphasis> Weber var. Azul

Somatic embryogenesis in Agave tequilana Weber var. Azul was affected by the interaction between the light regimes applied during the induction phase and the expression phase. When embryogenic calli was exposed to white or red light during the expression phase, an average of two germinated embryos per explant was obtained regardless of the light treatment used for callus induction. Conversely, the highest number of germinated embryos, an average of 18 per explant, was obtained when applying either white or red light during the induction phase and then wide-spectrum light during the expression phase. Culture medium had also a great influence in this process, with embryo germination being reduced by up to 70%, depending on the light treatment, when using Schenk and Hildebrandt (SH) medium instead of Murashige and Skoog (MS) medium.