Pulque agave production systems and management categories. In El Cubo, the producers have small plots that have been used for producers for at least two generations. The agaves are cultivated in hileras (rows) interspersed with other crops or as living fences within producers' properties. The CCUB1 plot is a production system measuring 3.529 ha, where agaves are cultivated in rows within fields of seasonal crops, such as maize and beans. The CCUB2.1 plot is a backyard orchard with fruit trees and ornamental plants, occupying an area of 0.35 ha. In the CCUB2.2 and CCUB2.3 plots, agaves are cultivated in rows in within crops, such as maize and beans, and occupy a space of 1.309 and 0.781 ha, respectively, belonging to a single producer of Hñähñu origin. The agaves are propagated by transplanting 50- to 100 cm-tall suckers from the agaves from the plot into rows. Agaves purchased from other producers in the area are also eventually incorporated into the plots.
Agave suckers are classified for sale according to their vigor into first-rate plants, sold for about 50 Mexican pesos (2 USD) each, and second- and third-rate plants that sell for about 30 Mexican pesos (1.2 USD). Some families in the area sell their plants to pulque producers and barbacoyeros (people who use agave leaves to cook a regional dish: barbacoa de borrego) for 100 Mexican pesos (4 USD) per plant. In this locality we recorded eight traditional varieties of agave used for pulque production. There was one wild variety, known as 1) Corriente, Bronco or Verde de monte, and seven cultivated varieties: 2) Chino, 3) Xaminí, 4) Penca larga, 5) Mutá, 6) Hoc´uadá, 7) Poblano, and 8) Guanté (also known as Maguey blanco) (Table 2, Fig. 2). The producers affirm that their ancestors used wild agaves, like Corriente, but they currently only use cultivated agaves.
Table 2
Characteristics of traditional varieties of agaves identified by producers in the two study localities in Hidalgo state.
Traditional variety name (n = individuals measured) | Main Ethnobotany Features | Managed category | Location | Taxonomic identity | Voucher specimen |
1. Corriente, criollo, de cerro, verde, bronco (n = 44) | This is a wild species that is also managed in some localities. A medium to small plant with large and abundant lateral teeth. Produces abundant suckers. The roots are strong, so some prefer to use it for living fences, it is extremely easy to transplant. Currently it is generally not used for aguamiel because it produces a low volume per day, produces for fewer days, and sap is of lower quality (less sweet). It is also used for xanté (agave fiber), although this use is decreasing. It produces a high degree of guixe (contact dermatitis from sap). | W, T, TR | CCOY1, WCOY2, WSAUZ | Agave salmiana ssp. crassispina | CJFU010 CJFU011 CJFU015 CJFU016 |
2. Corriente-Cenizo (n = 2) | Similar in characteristics to the Corriente variety, but leaf color is glaucous. | W | WCOY2 | Agave salmiana ssp. crassispina | Photo record |
3. Corriente-Colorado (n = 1) | Similar in characteristics to the Corriente variety, but it is smaller, with reddish leaves. Produces very little aguamiel. | TR | CCOY1 | Agave salmiana ssp. crassispina | Photo record |
4. Corriente-Espina china (n = 1) | This variety is similar to the Corriente variety but has more pronounced leaf margins greater size and abundance of lateral teeth. | TR | CCOY1 | Agave salmiana var. ferox | Photo record |
5. Corriente-Penca ancha (n = 6) | Like Corriente, but was wider, more flexible, and more fibrous leaves. The leaves of this variety are used to prepare barbacoa. | W | WCOY2, WSAUZ | Agave salmiana ssp. crassispina | Photo record |
6. Corriente-Penca larga (n = 7) | Remarkably similar to Corriente but has a longer penca (leaf) and produces sweet aguamiel. | T, TR | CCOY1 | Agave salmiana | CJFU012 CJFU013 |
7. Manso de Zoqui o maguey fino (n = 21) | This variety is purchased in Zoquizoquipan, Metztitlán municipality. It has been grown for more than 40 years in the region. It is the largest variety and is highly valued because it produces a large amount of sweeter aguamiel. It can produce up to four liters a day for up to five months. | C | CCOY1 | Agave salmiana var. salmiana | CJFU014 |
8. Xaminí, Xa`mni grande y chico (n = 1) | This is a native variety from the Valle del Mezquital. It is of medium size is characterized by hook-shaped lateral teeth (to which the name Xaminí in Hñähñu refers; can also translated as “spike that scrapes”). More lateral teeth. Grows fast and is ready for harvest in 9 to 10 years. The sweetest and least viscous aguamiel called aguamiel clarito. | C | CCUB2.3 | Agave salmiana ssp. crassispina | CJFU007 |
9. Poblano (n = 13) | Arrived in the region 22 years ago through a state government program. The plants are long and have wide leaves (pencas). It is very susceptible to pests such as pinacatillo (maguey weevil). Produces a large amount of aguamiel. | C | CCUB1 | Agave salmiana var. salmiana | CJFU005 |
10. Guanté o maguey blanco (n = 3) | Cultivated species. Has some morphological similarities with Xaminí. It takes 15 years for the stalk to emerge, which is about two meters high. | C | CCUB2.1, CCUB2.2 | Likely Agave americana | CJFU008 |
11. Mutha, Mutá o penca ancha (n = 9) | Has a wide penca (leaf), it is larger than the Corrientes. Their pencas (leafs) are used for barbacoa. Takes about 12 to 13 years to be ready to harvest. | C | CCUB2.1, CCUB2.2, CCUB2.3 | Agave salmiana var. salmiana | CJFU006 |
12. Penca larga (n = 4) | It is a large variety, with long leaves and slightly rigid, takes about 14 years to be ready for harvest. Aguamiel is more viscous than other varieties. It used to take ayate (fiber). This is not good for barbacoa. Does produce aguamiel, but it is more viscous, so pulque from this variety is thicker and spoils quickly. | C | CCUB2.1, CCUB2.2, CCUB2.3 | Agave mapisaga | CJFU002 |
In this locality, each producer uses six to 20 plants simultaneously. The sap is collected with the elongated tool known as an acocote, essentially a large straw. One end is inserted into the cavity of the agave, and suction is generated by mouth at the other end, drawing the sap into the acocote which is then emptied into a bucket. Acocotes were traditionally made with a long, dried calabash fruit (Lagenaria siceraria), however due to the low availability of this material and its fragility, it has been replaced by plastic materials. Currently, acocotes are made by attaching a plastic hose (the end inserted into the plant) to a plastic bottle, which is drilled at the base (to allow suction). Pulque is produced with a combination of different sap varieties. The producers mix the fresh sap with old pulque, produced the day before. The fresh sap and pulque are used for self-consumption and sold locally at 10 Mexican pesos (0.4 USD) per liter, or to tourists for 25 Mexican Pesos (1 USD) per liter.
La Coyotera has been cultivating agaves for around 30 years. A total of 44 hectares are dedicated to agave production, where the plants are found at different densities and are managed in three ways: living fences, jilas, and nursery. For living fences, used to delimit spaces and paths, the wild Corriente variety is preferred, since it has more teeth, longer spines, is fast growing, and has deep roots. Jilas refers to rows of agaves that are interspersed with native vegetation and are planted perpendicular to the slope of the land. Juvenile agaves are cultivated and kept in nurseries to protect them from goat browsing and allow them to develop and are then transplanted into the jilas when they reach 100 cm in height.
The cultivated agaves come from various sources, including from the suckers of wild agaves from nearby areas and suckers bought from local producers and from other productive areas such as Zoquizoquiapan, Zotoltepec and Singuilucan in Hidalgo and Magdalena Contreras in Mexico City. In this locality, wild agaves are also used to produce pulque. Fourteen traditional varieties used for pulque production were listed: 1) Corriente, 2) Corriente-Cenizo, 3) Corriente-Colorado, 4) Corriente-Espina china 5) Corriente-Penca ancha, 6) Corriente-Penca larga, 7) Manso de Zoqui, 8) Manso del Altiplano, 9) Xaminí, 10) Púa larga, 11) Verde, 12) Penca larga 13) Blanco and 14) Sabililla (Table 2, Fig. 2). In this locality, 35 plants are used simultaneously, collecting, and preparing pulque similarly to the other locality and the price per liter is 10 Mexican pesos (0.4 USD).
In both localities there is a gradient of management practices that can be grouped into the following categories:
1) Wild (W). Agaves that inhabit the surrounding areas of native vegetation. These plants can eventually be collected and used for various purposes. According to the people interviewed, some 70 years ago there were no crops, and these agaves were used.
2) Tolerated (T). Agaves of wild origin, which in the conversion of an area with native vegetation to a productive system, have been maintained by the managers within the new system. These plants receive some management, such as removing dry leaves.
3) Transplanted (TR). Agaves removed from their original place, generally from nearby ecosystems, and placed within productive systems, forming part of the rows, and living fences.
4) Cultivated (C): Agaves cultivated within productive systems. This category generally contains the most used and valued traditional varieties. These are given greater care, including pruning and more intense propagation. These are the varieties that are bought or exchanged between localities.
Producers identify qualitative characteristics that make it possible to distinguish cerro and monte agaves (wild) from cultivated ones. These characteristics are the size of the plant and its leaves, the shape and length of the spines, and the number of lateral teeth. Another important characteristic for handlers is the color of the plant, which ranges from different shades of green to glaucous. Characteristics such as the time to reproductive maturity, the sap quantity, and qualities (such as sweetness or viscosity), and the duration of sap production are also considered (Table 2). Wild agaves can be distinguished from cultivated agaves because they are not aligned, have a smaller rosette with many lateral teeth, and are larger than cultivated ones. Of the traditional varieties described in Table 2, three were found in natural ecosystems and correspond to wild (W), two are tolerated within production systems (T), four varieties were transplanted from native vegetation to managed areas (TR) and six varieties are cultivated (C). The traditional variety Corriente is the most abundant and can be found in the most of management categories.
Morphological variability of pulque agaves. Of the 19 traditional varieties recorded, it was only possible to morphologically characterize 12 varieties because no measurable individuals were found in production of the other seven. Based on 23 morphometric variables, the first two PCA components together explained 47% of the variance (PC1: 29%, PC2: 18%; Fig. 3). The ordering of PC1 was related to the size of the plant, and the most important variables were leaf length (LL), General plant length (GPL), stem length (SL), the ratio of the length of lateral teeth to the length of the blade (LTEE / LL), and diameter (D). In PC2, the most important variables were associated with the plant’s thorniness: the ratio of the distance between teeth and the length of the leaf (DTEE / LL), the length of the lateral teeth (LTEE), number of teeth per 10 cm (TEE10), the length of the terminal spine (TTL), and the distance between teeth (DTEE) (Table 3). The agaves of the different varieties sorted by management category, with wild agaves found on the extreme left of the arrangement and cultivated in the right, while the transplanted and tolerated individuals are located in the center (Fig. 3A).
Table 3
Vegetative morphological characteristics measured in the traditional varieties of agaves recorded in the localities Rancho La Coyotera and El Cubo studied in the state of Hidalgo. The columns show eigenvectors of the first (PC1) and second (PC2) principal components according to PCA and eigenvectors of the first (DF1) and second (DF2) factors of the DFA. Bold letters indicate the characteristic with greatest contribution to ordination.
Vegetative character | PC1 | PC2 | DF1 | DF2 |
General plant lenght (GPL) | 0.350 | 0.055 | 0.302 | 0.339 |
Stem lenght (SL) | 0.308 | -0.027 | 0.016 | 0.096 |
Mean diameter of the plant (D) | 0.274 | -0.005 | -0.135 | 0.165 |
Leaf lenght (LL) | 0.355 | 0.061 | 1.492 | 0.598 |
Leaf number (LN) | 0.057 | -0.125 | -0.425 | -0.303 |
Thickness leaf (TS) | -0.020 | 0.216 | -0.290 | -0.295 |
Suckers (S) | not included in the analysis | not included in the analysis | -0.127 | -0.220 |
Leaf width at middle (LW) | 0.186 | -0.219 | 0.416 | -0.039 |
Leaf width at base (LB) | 0.266 | -0.037 | 0.160 | 0.338 |
LL/LW | 0.179 | 0.264 | 1.054 | 0.131 |
LL/SL | -0.143 | 0.078 | -0.721 | 0.768 |
Terminal thorn length (TTL) | 0.078 | -0.303 | -0.054 | 0.038 |
Terminal thorn width at the base (TTW) | 0.235 | 0.013 | 0.554 | -0.349 |
Terminal thorn distance a first tooth (TTL-TEE) | not included in the analysis | not included in the analysis | not included in the analysis | not included in the analysis |
TTL/TTW | -0.155 | -0.182 | 0.406 | 0.109 |
TTL/LL | -0.132 | -0.120 | -0.368 | 0.053 |
Total number of teeth (TEET) | 0.183 | 0.262 | -0.583 | -0.070 |
TEET/LL | -0.184 | 0.227 | 0.454 | -0.076 |
Number of teeth in 10 cm2 (TEE10) | -0.198 | 0.311 | 0.167 | -0.138 |
Teeth length (LTEE) | -0.154 | -0.312 | -0.118 | -1.805 |
LTEE/LL | -0.280 | -0.214 | 0.678 | 2.174 |
Teeth width (WTEE) | 0.131 | -0.264 | -0.402 | -0.186 |
LTEE/WTEE | -0.211 | -0.184 | -0.560 | 0.761 |
Distance between teeth (DTEE) | 0.205 | -0.299 | 0.956 | 0.835 |
DTEE/LL | 0.072 | -0.337 | -0.150 | -0.241 |
Color (C) | not included in the analysis | not included in the analysis | not included in the analysis | not included in the analysis |
The heatmap showed that the agaves were grouped according to the management category. The cultivated agaves had larger size and were less thorny, regardless of their variety. They had smaller lateral teeth and a teeth in a smaller proportion with respect to the leaf. Wild individuals were grouped in the middle part of the heatmap, with smaller size than agaves of the other categories, but increased thorniness. The bottom of the heatmap had a mixture of individuals from all categories, especially the agaves of the tolerated and transplanted categories, which had intermediate characteristics between the other extremes of the management classification (Fig. 3B).
Regarding the characteristics associated with gigantism, the wild variety Corriente (A. salmiana ssp. crassispina) was the smallest in terms of GPL, SL, D, and LL. The cultivated variety Penca larga (A. mapisaga) was the largest in terms of GPL, D and LL, while the cultivated variety Poblano (A. salmiana var. salmiana) had the highest average SL. In terms of thorniness, Penca larga had more teeth and more closely spaced teeth than the other varieties, but the size of the teeth was smaller than the wild varieties. The variety Manso de Zoqui also had a smaller tooth size. This confirms the domestication syndrome of lesser dentition for easier manipulation (Table 4). Carrying out the morphological comparisons among the management categories, we found that the wild category was the smallest (GPL, SL, D, LL) and cultivated the largest. Wild agaves had larger lateral tooth sizes, but not a larger Terminal thorn (TTL), which was similar among all management categories (Table 5).
Table 4
Vegetative morphological characteristics of eight traditional varieties of pulque agave, identified by producers in the localities of La Coyotera and El Cubo, in the state of Hidalgo.
Vegetative character | Traditional varieties of agave |
Corriente | Corriente- Penca ancha | Corriente-Penca Larga | Manso de Zoqui | Poblano | Guanté | Mutá | Penca larga |
General plant length (GPL) | 172.955 ± 4.185 | 194.000 ± 7.625 | 218.714 ± 6.578 | 256.286 ± 8.050 | 252.308 ± 8.209 | 225.333 ± 6.227 | 240.333 ± 10.813 | 260.000 ± 10.206 |
Stem length (SL) | 42.898 ± 2.520 | 50.000 ± 5.882 | 80.243 ± 5.096 | 68.829 ± 3.233 | 81.692 ± 4.532 | 71.667 ± 3.333 g | 78.444 ± 3.271 | 66.750 ± 3.728 |
Mean diameter of the plant (D) | 253.523 ± 6.498 | 271.417 ± 13.679 | 303.000 ± 10.341 | 321.024 ± 9.249 | 368.692 ± 14.644 | 331.500 ± 4.770 | 296.944 ± 45.123 | 369.500 ± 8.663 |
Leaf length (LL) | 116.561 ± 2.801 | 132.167 ± 5.089 | 143.857 ± 4.748 | 188.438 ± 6.138 | 185.154 ± 6.052 | 148.000 ± 2.000 | 180.000 ± 5.336 | 209.750 ± 11.213 |
Terminal thorn length (TTL) | 5.051 ± 0.165 | 5.554 ± 0.319 | 4.138 ± 0.611 | 5.408 ± 0.224 | 6.069 ± 0.339 | 3.713 ± 1.408 | 6.479 ± 0.462 | 3.225 ± 0.183 |
Number of teeth in 10 cm2 (TEE10) | 3.636 ± 0.187 | 2.667 ± 0.333 | 3.143 ± 0.340 | 2.429 ± 0.177 | 2.000 ± 0.000 | 2.000 ± 0.000 | 2.000 ± 0.167 | 4.750 ± 0.629 |
Teeth length (LTEE) | 0.769 ± 0.043 | 0.750 ± 0.042 | 0.716 ± 0.057 | 0.469 ± 0.027 | 0.794 ± 0.060 | 0.699 ± 0.121 | 0.768 ± 0.066 | 0.301 ± 0.029 |
LTEE/LL | 0.007 ± 0.000 | 0.006 ± 0.000 | 0.005 ± 0.000 | 0.003 ± 0.000 | 0.004 ± 0.000 | 0.005 ± 0.001 | 0.004 ± 0.000 | 0.001 ± 0.000 |
Distance between teeth (DTEE) | 1.687 ± 0.015 | 2.595 ± 0.597 | 2.020 ± 0.304 | 3.058 ± 0.254 | 4.509 ± 0.277 | 4.389 ± 0.618 | 4.892 ± 0.698 | 0.778 ± 0.066 |
DTEE/LL | 0.015 ± 0.0 | 0.020 ± 0.0 | 0.014 ± 0.0 | 0.017 ± 0.0 | 0.025 ± 0.002 | 0.030 ± 0.004 | 0.028 ± 0.004 | 0.004 ± 0.001 |
Allele richness (A) | 6.000 ± 0.428 | 3.313 ± 0.285 | 2.813 ± 0.306 | 3.500 ± 0.316 | 3.313 ± 0.435 | 1.563 ± 0.182 | 3.063 ± 0.452 | 2.313 ± 0.237 |
Allele effective (Ae) | 4.073 ± 0.303 | 2.590 ± 0.221 | 2.226 ± 0.238 | 2.390 ± 0.235 | 2.495 ± 0.345 | 1.459 ± 0.146 | 2.294 ± 0.347 | 1.975 ± 0.193 |
Observed Heterocigozity (Ho) | 0.190 ± 0.052 | 0.167 ± 0.050 | 0.161 ± 0.070 | 0.143 ± 0.056 | 0.214 ± 0.086 | 0.146 ± 0.074 | 0.203 ± 0.083 | 0.125 ± 0.065 |
Expected Heterocigozity (He) | 0.730 ± 0.023 | 0.562 ± 0.046 | 0.493 ± 0.040 | 0.517 ± 0.048 | 0.487 ± 0.059 | 0.219 ± 0.065 | 0.435 ± 0.066 | 0.410 ± 0.061 |
Inbreeding coefficient (FIS) (CI) | 0.716 (0.680–0.719) | 0.702 (0.565–0.709) | 0.562 (0.378–0.575) | 0.643 (0.523–0.647) | 0.159 (0.150–0.275) | 0.194 (0.003–0.228) | 0.324 (0.077–0.326) | 0.650 (0.353–0.656) |
Table 5
Vegetative morphological characteristics according to the management categories of the eight traditional varieties of pulque agaves, identified by producers in the localities of La Coyotera and El Cubo, in the state of Hidalgo.
Vegetative character | Management category |
Wild (W) | Tolerated (T) | Transplanted (TR) | Cultivated (C) |
General plant length (GPL) | 170.850 ± 5.085 | 202.455 ± 9.646 | 207.875 ± 11.311 | 246.615 ± 4.436 |
Stem length (SL) | 40.025 ± 2.459 | 64.909 ± 4.689 | 68.963 ± 5.498 | 73.460 ± 2.156 |
Mean diameter of the plant (D) | 249.900 ± 9.275 | 289.091 ± 17.687 | 294.938 ± 20.720 | 328.603 ± 8.135 |
Leaf length (LL) | 115.200 ± 3.792 | 135.273 ± 7.232 | 142.500 ± 8.480 | 181.998 ± 3.326 |
Terminal thorn length (TTL) | 5.049 ± 0.210 | 4.777 ± 0.400 | 4.473 ± 0.469 | 5.479 ± 0.184 |
Number of teeth in 10 cm2 (TEE10) | 3.450 ± 0.178 | 3.091 ± 0.339 | 3.750 ± 0.398 | 2.500 ± 0.156 |
Teeth length (LTEE) | 0.800 ± 0.041 | 0.713 ± 0.077 | 0.660 ± 0.091 | 0.622 ± 0.036 |
LTEE/LL | 0.007 ± 0.000 | 0.005 ± 0.001 | 0.005 ± 0.00 | 0.004 ± 0.000 |
Distance between teeth (DTEE) | 1.851 ± 0.216 | 2.099 ± 0.411 | 1.567 ± 0.482 | 3.556 ± 0.189 |
DTEE/LL | 0.016 ± 0.001 | 0.015 ± 0.003 | 0.012 ± 0.003 | 0.020 ± 0.001 |
Allele richness (A) | 5.587 ± 0.407 | 2.250 ± 0.348 | 3.063 ± 0.347 | 5.188 ± 0.572 |
Allele effective (Ae) | 3.562 ± 0.221 | 2.386 ± 0.281 | 2.389 ± 0.255 | 3.418 ± 0.572 |
Observed Heterocigozity (HO) | 0.187 ± 0.048 | 0.204 ± 0.079 | 0.135 ± 0.064 | 0.167 ± 0.058 |
Expected Heterocigozity (He) | 0.701 ± 0.020 | 0.495 ± 0.055 | 0.504 ± 0.053 | 0.646 ± 0.040 |
The DFA explained 96.03% of the variance (DF1 83.46%; DF2 12.58%). Three large groups were differentiated; one contained the wild traditional varieties (at the left in Fig. 3C), a second was composed of cultivated traditional varieties (at the right in Fig. 3C), and a third group contained the tolerated and transplanted traditional varieties (center bottom). The variables with the highest eigenvalues in DF1 were LL, LL/LW, LL/SL, TTW, TEET, LTEE/LL, LTEE/WTEE and DTEE. In the case of DF2, they were LL, LL/SL, LTEE, LTEE/WLEE, and DTEE. These characteristics are associated with the length of the leaves and the dentition. The first group, in the upper left it is made up of wild individuals of the traditional varieties Corriente, Corriente-Cenizo and Corrriente-Penca ancha. The second group (top right) is made up of individuals cultivated from traditional varieties Penca larga, Manso de Zoqui, Mutá and Poblano. The third group (bottom center) corresponds to tolerated and transplanted individuals of different varieties (their centroids did not differ), even wild and cultivated individuals are distinguished. The Wilk’s Lambda had a value close to zero (0.05; p < 0.001), indicating that the information provided by the variable is statistically significant, allowing the discrimination of groups whose centroids are not same (Wild and cultivated) and have little overlap. Only 7% of the individuals were not correctly classified into the category management assigned a priori. 90% of the individuals in the management category W were classified correctly; 7.5% were classified T and 2.5% to TR. 85% of the individuals in the T category were classified correctly and 15% classified as TR. All individuals in the TR category were correctly classified. In the case of management category C, 96% were classified correctly and 4% to category T.
Genetic diversity and structure
Null alleles were found at 16 loci, suggesting homozygote excess. Only one locus (BYU4463) did not exhibit null alleles. The estimated null allele frequencies over traditional varieties and managed category varied from − 0.0738 (7%) at BYU4463 (this suggest heterozygote excess) to 1 (monomorphic loci) at APAR3-11, APARLC28, APARLC34 and BYU 4012. Sixteen loci showed departures from HWE (p < 0.05). LD was observed between four pairs of loci (p < 0.05: APAR2-12 x APAR3-11, APARLC-21 x APARLC-28, APARLC-34 x BYU3674 and APARLC-35 x BYU4012. The percentage of polymorphic loci (PPL) in traditional varieties ranged from 43.75% (Guanté) to 100% (Corriente, Corriente-Penca Larga and Manso de Zoqui). In the managed categories, this was 100% for wild (W) and cultivated (C) and 93.75% for tolerated (T) and transplanted (TR). The allele richness (A) in the traditional varieties ranged from 1.563 (Guanté) to 6 (Corriente), and the effective number of alleles (Ae) varied from 1.459 to 4.076 in the same traditional varieties (Guanté and Corriente respectively; Table 4). In the managed categories, the allele richness ranged from 3.063 (TR) to 5.875 (W), and the effective number of alleles (Ae) was 2.386 to 3.562 in the same managed categories (TR, T and W, Table 5). The observed heterozygosity (Ho) in traditional varieties ranged from 0.125 (Penca larga) to 0.214 (Poblano) and the expected heterozygosity (He) varied from 0.219 (Guanté) to 0.730 (Corriente) (Table 4). In managed categories, the observed heterozygosity (HO) ranged from 0.135 (TR) to 0.204 (T) and the expected heterozygosity (He) varied from 0.495 (T) to 0.701 (W, Table 5). Average levels of genetic variation at species level for Agave salmiana was (Hs = 0.564, N = 7), for A. salmiana ssp. crassispina (Hs = 0.738; N = 73), for Agave salmiana var. salmiana (Hs = 0.540, N = 21); for A. mapisaga (Hs = 0.526, N = 4) and for A. americana (HS = 0.291, N = 3). FST with ENA correction was 0.127, indicating a moderate genetic differentiation between managed categories. FIS ranged from 0.194 (Guanté) to 0.720 (Corriente), indicating strong endogamy. The neighbor joining tree based on Nei’s distances showed three groups. The first included the traditional varieties: Corriente, Corriente-Cenizo and Corriente-Penca ancha, the second group was formed by the Corriente-Colorado, Corriente-Espina china, Corriente-Penca larga and Manso de Zoqui varieties, and the third contained the Poblano, Mutá, Guanté, Penca Larga and Xaminí varieties (Fig. 4A). The Bayesian cluster analysis indicated that the most likely number of genetics group was three (K = 3, Fig. 4B). The blue genetic group corresponds to the category of wild management and the traditional varieties Corriente, Corriente-Cenizo and Corriente-Penca ancha. The orange group is made up of individuals that are tolerated, transplanted, and cultivated, with the traditional varieties Corriente, Corriente penca larga, Corriente-Colorado, Corriente-Espina China, and Manso de Zoqui. The purple group corresponds to the category of cultivated management and the traditional varieties Xaminí, Poblano, Guanté, Mutá and Penca larga. This analysis corresponds to the grouping of the dendrogram. The DAPC also grouped the plants according to the management categories, in the upper part in the center the wild traditional varieties (Fig. 4C) matched the dendrogram and Bayesian clustering. In the lower left, the cultivated traditional varieties (Poblano, Guanté, Mutá, Corriente-Penca Larga and Xaminí) and in the lower right a mixed group between the tolerated and transplanted traditional varieties together with a cultivated traditional variety Manso de Zoqui.