The piquin pepper is a wild species of great importance from an economic and nutritional point of view. Its preference is due to its pleasant taste and spiciness, comparable to other peppers such as the serrano and jalapeño (Villalón-Mendoza, Medina-Martinez, Ramírez-Meraz, Solis-Urbina, & Maiti, 2014). It is a wild plant distributed in Mexico and the southern United States, which demonstrates its wide environmental adaptability, probably associated with its genetic diversity. According to their nature, the fruits are obtained by harvesting in wild populations, and not from commercial plantations, which diminishes the natural populations and threatens the genetic diversity of this species (Rueda-Puente et al., 2010).
On the other hand, environmental conditions modify the phenology of plants; specifically, the wild populations of piquin pepper are perennial and commonly associated with partially shaded conditions in the understory, but when planted in the open field they can be considered annual crops due to frost, disease and susceptibility to pests (Rodríguez-del Bosque et al., 2003). In addition, in recent years, global warming has caused changes in environmental conditions, such as temperature and precipitation (Conde-Álvarez & Saldaña-Zorrilla, 2007). To mitigate these changes, shading nets or plastic covers can be used that, in addition to generating a certain amount of shade, simulate their wild habitat and promote the normal development of the plant, which increases the yield and quality of the fruit in comparison with open field production (Ayala-Tafoya et al., 2011).
This is of utmost importance, as plants that are not adapted to intense sunlight can develop heat stress. Most negative effects can be avoided with the protection of a greenhouse or with shading structures (Castronuovo, Statuto, Muro, Picuno, & Candido, 2017).
In order to preserve this species and increase its commercial cultivation, protected production systems are used as a viable alternative for the planting of Capsicum annuum var. Glabriusculum (Kitta, Katsoulas, Kandila, González-Real, & Baille, 2014). This is because plants cultivated under shade undergo morphological changes as mechanisms of adaptation to the low availability of light. This adaptation includes changes in the leaf area index, temperature and relative humidity, as well as better leaf area distribution in height, all of which can have effects on the plant’s productive and physiological responses (Li, Chen, & Li, 2012).
Photoselective nets for agricultural use filter the intercepted solar radiation (Shahak, Gal, Offir, & Ben-Yakir, 2008), as well as being designed to detect several spectral bands of solar radiation and transform direct light to diffuse light. In crops such as cucumber (Cucumis sativus) and pepper (Capsicum annuum), using white, blue, green and white-polyethylene covers increased temperature, relative humidity and evapotranspiration, resulting in greater growth, yield and biochemical compound content per plant (Casierra, Matallana, & Zapata, 2014; Hashem, Medany, Abd, & Abdallah, 2011).
Changes in spectral light promote different morphogenetic and photosynthetic responses (Fu, Li, & Wu, 2012). Each plastic net modifies the solar radiation that reaches the crop, reducing the flow of light and varying the available radiant spectrum. Apart from the net structure, the transmittance spectrum is also influenced by the diameter of the thread, color and thickness of the net, and the absorbance, transmittance and reflectance properties of the plastic material (Sica & Picuno, 2008). Based on the above, this study aimed to evaluate the effect of different photoselective covers (nets and greenhouse) on agronomic variables in wild piquin pepper ecotypes. It is hypothesized that the use of different covers will increase yield and agronomic traits, at least in one of the six ecotypes evaluated.
Materials and methods
The research was carried out during the spring-summer 2016 agricultural cycle in the Horticulture Department of the Universidad Autónoma Agraria Antonio Narro in Saltillo, Coahuila (25° 35’ 63” NL and 101° 03’ 49” WL, at 1 581 m of elevation). Wild piquin pepper seeds collected in the states of Zacatecas, Nuevo León and Coahuila, Mexico, during October and November 2015, were used (Table 1).
|Abbreviation||Location-State||Elevation (masl)||Collection coordinates|
|RTZ||Río Tuxpan, Zacatecas||2 060||22° 39’ 10.5’’ NL, 102° 42’ 16.2’’ WL|
|PTZ||Puente Tepetatilla, Zacatecas||1 888||22° 47’ 14.1’’ NL, 103° 35’ 11.4’’ WL|
|MZC||Múzquiz, Coahuila||499||28° 00’ 02.9’’ NL, 101° 44’ 45.8’’ WL|
|SAC||San Alberto, Coahuila||365||27° 42’ 83.1’’ NL, 101° 38’ 16.1’’ WL|
|SNL||Santiago, Nuevo León||480||25° 23’ 53.7’’ NL, 100° 06’ 22.8’’ WL|
|LNL||Linares, Nuevo León||356||24° 50’ 14.5’’ NL, 99° 35’ 21.8’’ WL|
The seeds, previously treated with 500 ppm of gibberellic acid (Biogib®), were sown in 200-cavity polystyrene trays in order to break the seed coat and obtain greater germination. Sphagnum peat moss (Pro Mix®) and perlite (Hortiperl®) were used at a 2:1 (v/v) ratio. The plants began to emerge 25 and 38 days after sowing. When the seedlings reached 15 cm in height, they were transplanted into 10-L polyethylene bags and placed at a distance of 40 cm between plants and 1 m between furrows (at a density of 20 750 plants·ha-1).
The crop was grown in macro-tunnels (4 m wide, 6 m long and 2.30 m high) with raschel netting with 30 % shading and hole size of 6 x 8 mm, and a macro-tunnel covered with milky white polyethylene with 20 % shading, while open field production had 100 % light transmission. Steiner’s nutrient solution (1976) was used at 25 % in seedling stage, at 50 % during vegetative development, at 75 % in flowering and at 100 % in fructification. The water supply was from 0.50 to 2.50 L·plant-1·day-1 through a stake-based fertigation system.
An experimental design was used with a split-plot arrangement, where the large plot includes covers and the small plot the ecotypes (Table 2), giving a total of 36 treatments. Analysis of variance and Tukey’s means comparison (P ≤ 0.05) of the evaluated variables were carried out with Statistical Analysis System software (SAS Institute Inc., 2004). On the other hand, principal component analysis and biplot graphs were carried out with the prcomp function in R version 3.4.3.
|Small (Ecotypes)||RTZ: Rio Tuxpan, Zacatecas||PTZ: Puente Tepetatilla, Zacatecas||MZC: Múzquiz, Coahuila||SAC: San Alberto, Coahuila||LNL: Linares, Nuevo León||SNL: Santiago, Nuevo León|
|Large (Covers)||OF: Open field||GRE: Greenhouse||BLAN: Black netting||RN: Red netting||BLUN: Blue netting||WN: White netting|
The microclimatic variables recorded were ambient temperature and relative humidity with a digital thermohygrometer (1452, Taylor®, China), and photosynthetically active radiation (PAR) was recorded with a portable Quantum sensor (SM-700, Apogee®, USA). All measurements were made between 7:00 and 19:00 h in the center of each macro-tunnel and under clear sky conditions. The morphological variables evaluated were: plant height (PH) and internode length (IL) with a tape measure, basal stem diameter (BSD) and fruit diameter (FD) with a digital Vernier caliper (HER-411, Digital Caliper®, China), and average fruit yield (AFY). The last was estimated by the weight of the fruits per plant, using an electronic scale (BABOL-100G, Rhino®, China) with a maximum capacity of 100 g and minimum resolution of 0.01 g. In addition, the number of fruits per plant (NFP), number of seeds (NS), number of leaves (NL) and days to flowering (DF) were quantified.
Results and discussion
Effect of nets on the crop environment
PAR, ambient temperature and relative humidity were different in each cover with respect to the field. Only the field and under-greenhouse measurements exceeded 1 000 μmol·m-2·s-1 (Figure 1), and there were reductions of between 43 and 50 % PAR with the different covers compared to that presented in the field (Table 3). The maximum PAR indices consistently occurred between 13:00 and 15:00 h, while in the colored nets these indices were between 500 and 700 μmol·m-2·s-1, with the red netting recording the highest indices and the blue the lowest. PAR above 1 000 μmol·m-2·s-1 has a direct effect on the photosynthesis rate, which negatively affects productivity, growth and yield (Kitta et al., 2014). The results of the decline in PAR were notable because the shade netting had a buffering effect on the changes in this parameter over the course of the day (Figure 1).
|Environment||Reduction of PAR1 with respect to OF (%)||Temperature (°C)||RH (%)|
|Red netting||43.59||31.23 bz||31.84 c|
|Blue netting||50.10||30.18 d||33.36 a|
|White netting||49.14||30.01 d||32.83 b|
|Black netting||48.86||29.07 e||32.96 b|
|Greenhouse||7.16||31.68 b||31.63 d|
|Open field||0||33.94 a||32.72 b|
According to Retamales, Montecino, Lobos, and Rojas (2008), in highbush blueberry cultivation it has been observed that PAR, transmitted by white and red nets with 35 % shading, and grey netting with 50 % shading, was reduced by up to 29 %, while red netting with 50 % shading and black netting with 35 % shading decreased PAR by 41 and 47 %, respectively, compared to the field. Therefore, the use of shading nets is relevant for reducing the impact of high light intensity in open field conditions and transforming direct radiation into dispersed radiation. This allows light to penetrate the inner canopy of the plant, which prevents burns and gives a moderate cooling effect (Ilic et al., 2017).
Shahak (2014) points out that with any cover placed on plants, in addition to the effects on light intensity and quality, the climatic elements (solar radiation, wind speed, temperature and relative humidity) can be modified (Arthurs, Stamps, & Giglia, 2013), altering the metabolism and water consumption of plants, which has a positive impact on fruit yield and quality (Tanny, 2012). Temperature and relative humidity play an important role in foliar stomatal conductance and, therefore, in the rate of transpiration and photosynthesis of the plant (Righi, Buriol, Angelocci, Heldwein, & Tazzo, 2012).
Effect of the nets on the crop
Highly significant (P ≤ 0.01) differences were found among covers in all the growth variables studied (Table 4). Plants grown under white netting showed the greatest statistical difference in seven of the nine variables studied (PH, BSD, AFY, NFP, FD, NS and NL), and plants grown under blue netting had the best values in four of the variables (DF, FD, NS and IL); therefore, the latter can be considered as the second-best cover for piquin pepper cultivation (Table 5).
|Source of variation||DF1||PH||BSD||DF||NFP||AFY||FD||NS||IL||NL|
|Cover||5||1 192.4**||12.3**||1 001.7**||720.6**||103.8**||154.5**||548.4**||432.6**||162 629.7**|
|Ecotype||5||1 333.4**||14.9**||633.0**||5 328.7**||312.4**||58.0**||125.2**||40.0**||23 717.3**|
|Cover x ecotype||25||104.1**||2.1**||68.2**||383.0*||84.3**||4.4**||6.1**||4.1**||5 035.7**|
|Ecotype||PH1 (cm)||BSD (mm)||DF (days)||AFY (g)||NFP||FD (mm)||NS||IL (cm)||NL|
|RTZ||36.95 az||6.15 a||43.66 a||9.29 a||40.04 a||11.11 a||14.79 a||12.45 a||193.58 c|
|PTZ||34.70 b||6.01 b||53.54 b||0.98 b||8.79 b||6.46 b||11.50 c||9.75 c||152.16 e|
|LNL||25.75 c||5.11 c||58.62 d||0.16 b||1.95 d||6.29 b||10.79 c||9.66 cd||219.95 a|
|MZC||23.54 d||4.92 d||54.12 b||0.51 b||4.62 d||8.30 ab||14.66 ab||10.91 b||206.66 b|
|SNL||23.25 d||4.71 e||56.29 c||0.35 b||2.79 d||7.02 b||12.04 bc||9.25 cd||166.50 d|
|SAC||17.12 e||4.09 f||54.37 b||0.26 b||2.20 d||6.60 b||7.91 d||9.04 d||141.37 f|
|WN||36.41 a||6.29 a||47.29 b||5.68 a||19.33 a||10.77 a||18.87 a||13.66 b||303.08 a|
|BLUN||32.58 b||5.39 b||45.12 a||3.10 b||13.95 b||12.17 a||17.66 a||15.67 a||175.12 c|
|GRE||27.83 c||5.37 b||55.95 d||0.86 b||8.58 c||6.02 b||7.37 cd||6.62 e||139.12 d|
|RN||24.45 d||4.76 d||55.62 d||0.70 c||6.50 d||6.86 b||12.83 b||7.87 d||248.79 b|
|BLAN||23.04 e||4.15 e||53.66 c||0.69 c||6.87 cd||5.39 b||8.95 c||12.16 c||135.91 d|
|OF||17.00 f||5.03 c||62.95 e||0.44 c||5.16 d||4.56 b||6.00 d||5.08 f||78.20 e|
On the other hand, plants grown in the open field showed the worst results in eight of the nine variables studied (except BSD) (Table 5). Likewise, plants grown under black netting showed the lowest values in four of the analyzed variables (BSD, AFY, NFP and FD), which supports the need to look for the best conditions for the development of wild piquin pepper, in this case through the use of nets that provide the optimal ranges in the microclimatic variables.
The white netting produced a higher NFP and AFY (374.61 and 1 290.90 %, respectively), compared to open field production (Table 5). Likewise, fruits grown under white and blue nets produced larger FD (236.18 and 266.88 %, respectively) and NS (314.50 and 294.33 %, respectively), both with respect to open field production.
The decreased PAR availability may induce morphogenetic responses such as increased leaf area and lengthening of the stem and internodes, because cells expand more with low solar radiation intensities to capture light and carry out photosynthesis. This is associated with the phenomenon known as "shade avoidance", since it is influenced by the phytochromatic activity that regulates stem lengthening, by transforming non-photosynthetic plastids (etioplasts) into fully developed chloroplasts with photosynthetic function, and the leaf area, with the consequent effects on photoassimilation distribution and fruit growth (Bastías, Manfrini, & Grappadelli, 2012).
In this study, the nets and greenhouse caused an increase in the final height of the piquin pepper plants (Table 5), probably in response to the reduced light (Salisbury & Ross, 2000). It has been reported that when using nets the PH changes depending on the color of the net; in bell pepper the highest PH was obtained with blue nets (Ayala-Tafoya et al., 2015), and in basil with black ones (Martínez-Gutiérrez, Nicolás-Santana, Ortiz-Hernández, Morales, & Gutiérrez-Hernández, 2016).
Radiation is one of the factors capable of producing greater photosynthetic activity in plants; in addition, the quality and distribution of the light spectrum can influence the length of the internode and the development of flowering (Runkle & Heins, 2006). In this case, this effect stood out in the white and blue nets, where an increase in PH of 114.05 and 91.53 %, respectively, was recorded compared to the open field. Additionally, the white netting increased BSD by 34.1% in contrast to the black netting, where the lowest result was obtained. This coincides with the findings of Ayala-Tafoya et al. (2011), who report that colored shading nets increased BSD in tomato.
Principal component analysis (PCA) revealed a grouping of plants in the second quadrant (Figure 2), which indicates that they showed lower PH, NFP and FD, and higher DF (CP1), as well as greater BSD and fruit weight and lower NS (PC2), this considering the relevance of these variables for each of the components (Table 6). However, in general, the materials behaved in a dispersed manner in the analyzed conditions, which reveals that the ecotype played a determining role in the variables studied. It is important to note that a larger BSD allows the plant to have a better-developed vascular system, through which it conducts water and nutrients, which improves physiological processes (Bahena-Delgado, Bustos-Rangel, Broa-Rojas, & Jaime-Hernández, 2012).
|Basal stem diameter||0.26704|
|Days to flowering||-0.35263||0.05962|
|Number of fruits per plant||0.32774|
|Average fruit yield||0.31251||0.30546|
|Number of seeds||0.31634|
|Number of leaves||0.22385||-0.4196|
Díaz-Pérez (2014) found an increase in the number and size of bell pepper fruit under shade conditions. On the other hand, Shahak et al. (2008) when evaluating "raschel" colored nets (red, yellow and pearl, with 30 to 40 % shading) in bell pepper obtained 115 to 135 % higher AFY compared to results obtained with black netting and in the open field. In contrast, Ayala-Tafoya et al. (2015) found no significant differences in the NFP in bell pepper when using different colored nets and production without nets.
Effect of ecotypes
All growth variables showed highly significant (P ≤ 0.01) differences among the different ecotypes analyzed (Table 4). The RTZ ecotype had the best performance in eight of the nine variables (PH, BSD, DF, AFY, NFP, FD, NS and IL), while the SAC ecotype had the worst performance in almost all variables. The community of origin of the latter is at an elevation of 365 masl, which is much lower than that of the place where the experiment was established (1 580 masl). On the other hand, the place of origin of the RTZ ecotype is at an elevation of 2 060 m, which could have influenced the adaptation to the conditions of the experiment. However, this does not coincide with what was reported by Martínez-Sánchez, Pérez-Grajales, Rodríguez-Pérez, and Moreno-Pérez (2010), who collected wild species of Capsicum annuum at elevations of 940 to 1 600 m, and evaluated them at 2 247 m without finding significant statistical differences among species in the variables PH, BSD, NFP and fruit size.
PCA did not reveal distinct ecotype groupings, which supports the idea that the environment also plays an important role in the dispersion of data. However, some trends were observed; for example, the RTZ ecotype was grouped to a greater extent in quadrant 1 (Figure 3), indicating that these plants performed better in terms of FD, NFP, PH and BSD. In addition, this ecotype showed greater PH (46.5 %) and BSD (34.5 %) compared to the SAC ecotype (Table 5), and exhibited greater precocity (DF), coming 14 days earlier than the latest-flowering ecotype (LNL), and had 95 % more fruits per plant compared to the same ecotype (Table 5).
On the other hand, blue netting increased the precocity to flowering by almost 18 days with respect to the open field, and, of all the covers, it also produced the largest IL, which was 62.81 % longer than in the production without cover. In this same variable, the RTZ ecotype was higher (27.38 %) than the SAC ecotype (Table 5).
The yield, size and quality in Capsicum, caused by the shading effect, depend largely on the geographical area and production technology (Zhu, Peng, Liang, Wu, & Hao, 2012). In general, the RTZ and MZC ecotypes had the largest FD (40.59 and 20.48 %, respectively) and NS (46.52 and 46.04 %, respectively), compared to the SAC ecotype (Table 5). Salinas-Hernández, Liévano-Liévano, Ulín-Montejo, Mercado, and Petit-Jiménez (2010) also obtained greater length, FD and NS in two types of wild amashito pepper ("Pico de Paloma" and "Garbanzo"), when compared with other types of pepper evaluated.
Santos and Salame-Donoso (2012) found that the use of 35 % shading in southern highbush blueberry (Vaccinium oxycoccus) cultivation increases flowering, fruit weight and yield, in contrast to open field production. On the other hand, Nooryazdan, Serieys, Baciliéri, David, and Bervillé (2010) report that differences in morphological traits, when evaluating 77 populations of wild Helianthus annuus collected in the United States, were correlated with the climatic variables of the sites of origin of the populations, which suggests adaptation to the local conditions of the evaluated species.
Effect of the interaction
Two-way analysis of variance revealed that the interaction between the covers used and the ecotypes had a significant effect on all variables. This coincides with the findings of Hernández-Verdugo et al. (2015), who report significant differences in all phenotypic characteristics evaluated in wild pepper under different shade levels. In general terms, it can be pointed out that the use of white shading nets provides better results in the variables analyzed in all the ecotypes (Table 5), and that the RTZ ecotype presents the best values in all the covers used.
Analysis of the vectors revealed that a certain group of variables had a high correlation with each other (BSD, AFY, PH and NFP) (Figures 2 and 3). These results are similar to those reported by Meena-Prakash and Bahadur (2015) for PH and NFP, and by Ogwulumba and Ugwuoke (2013) for NL and NFP. On the other hand, the variables FD, IL, NS and NL formed another group, which showed little correlation with the previous group.
Meena-Prakash and Bahadur (2015) indicate that PH showed a significant negative correlation with respect to fruit weight and polar diameter, which reveals that at a higher PH, fruit weight and diameter decrease, while the DF variable presented a negative correlation with all variables, which indicates that the higher the number of DF, the lower the performance of the other variables. In this study, biplot analysis revealed that all variables, except DF, are positively related to crops under white or blue nets (Figure 2); that is, crops under those covers had the best results in almost all variables. In terms of ecotypes, RTZ was positively related to the variables BSD, AFY, PH and NFP (Figure 3).
The DF variable in open field and greenhouse (Figure 2) reveals that plants grown under these conditions take longer to present flowers and have a lower performance in the rest of the variables. The same happened in all ecotypes when they were grown in the open field (Figure 3).
The use of covers reduced photosynthetically active radiation, which had a positive effect on plant morphology. White and blue nets increased yield, number of fruits and their size, and the RTZ ecotype had the best results in all variables studied.
The combination of the RTZ ecotype with white netting was the one that improved the agronomic traits and yield in piquin pepper, so it can be considered as an alternative for the production of Capsicum annuum var. Glabriusculum, under similar conditions to those of this study.