Implications of Antibiosis and Antixenosis in Four Plant Species Against Oligonychus punicae (Trombidiformes: Tetranychidae)1
Abstract
Oviposition and feeding of herbivorous arthropods influence the plant-arthropod interaction and determine the success in colonization and establishment on their host plant. The avocado brown mite, Oligonychus punicae Hirst (Acari: Tetranychidae), causes severe damage to several crops due to its feeding. This study proposed to evaluate the resistance mechanisms of Moringa oleifera Lamarck (Moringaceae), Phaseolus vulgaris L. (Fabaceae), Persea americana Miller (Lauraceae), and Rosa hybrida L. (Rosaceae) to the attack by avocado brown mite under laboratory conditions. The study was conducted under laboratory conditions at 28 ± 1°C and 70–80% relative humidity (RH), with a photoperiod of 12:12 h (light: dark). Oligonychus punicae females showed no preference to oviposit on M. oleifera (2.10 ± 0.05 eggs/female/day) compared with R. hybrida (2.77 ± 0.06), P. americana (2.73 ± 0.08), and P. vulgaris (3.05 ± 0.08). Females showed a preference to feed on P. vulgaris compared with other host plants. Avocado brown mite recorded the lowest values in r (0.5990 d−1) on M. oleifera foliage square. Moringa oleifera was the most resistant to O. punicae, whereas the most susceptible host plant was P. vulgaris. These results indicate possible resistance of M. oleifera to the attack of O. punicae and that those responses might be due to antibiosis and antixenosis.
The avocado brown mite, Oligonychus punicae Hirst (Acari: Tetranychidae), is found around the world, mainly in countries in the neotropical region, including Brazil, Chile, Colombia, Guatemala, and Mexico (Migeon and Dorkeld 2024, Peña et al. 2013). This mite feeds on 121 species of host plants (Migeon and Dorkeld 2024). In Mexico, O. punicae is an important pest in avocado crops (Peña et al. 2013). Mites feed on the upper surface of the leaf, causing leaf bronzing and a reduction in photosynthetic activity by more than 50%, which can result in losses in avocado yield up to 20% (Castañeda-Cabrera et al. 2022, Maoz et al. 2011). Control of the avocado brown mite mostly depends on the use of chemical pesticides, but tetranychid mites such as O. punicae have a short life cycle, high reproductive rate, and arrenotic reproduction and, thus, can quickly develop resistance to these synthetic compounds (Van Leeuwen et al. 2015).
Avocado brown mite can develop and reproduce on a wide range of host plants and under a variety of climatic factors (Ferraz et al. 2020, 2021; Vásquez et al. 2008). For example, Ferraz et al. (2021) evaluated the behavior of O. punicae on Eucalyptus tereticornis Smith (Myrtaceae) discs at different temperatures ranging from 21 to 37°C. They determined that O. punicae performs better between 25 and 29°C than at other temperatures. Vásquez et al. (2008) reported that the performance of O. punicae was better on the Chenin Blanc cultivar of grapevine, Vitis vinifera L., when compared with other cultivars. Ferraz et al. (2020) found that optimal development for avocado brown mite was on E. tereticornis than on E. pellita F. Mueller, E. brassiana S.T. Blake, E. grandis W. Hill Ex Maiden (Myrtaceae), and Corymbia citriodora L.A.S. Johnson (Myrtaceae). The biology and demographic parameters of O. punicae, such as fecundity, survival rate, and total developmental time (egg-adult), may vary in response to changes in host plant species, temperature, secondary metabolites, and anatomical features of leaves (Ferraz et al. 2020, 2021; Vásquez et al. 2008).
Plant-arthropod interactions are influenced by both the interactions among plants and arthropod feeding and the relationship among plants and arthropods oviposition (Hilker and Meiners 2011). Egg laying by herbivorous arthropods on their host plants is the first and most important event in their interactions because it determines the establishment of a new generation (Hilker and Meiners 2011, Kim et al. 2012, War et al. 2018). Furthermore, the deposition of eggs by herbivorous arthropods on their host plants is a warning sign of future larval herbivory (Hilker and Meiners 2011, Kim et al. 2012). On the other hand, the process of colonization and establishment of a species that inhabits new favorable environments occurs with low population densities, exhibiting exponential growth in a short period (Carey 1993, Smith and Smith 2015).
On the other hand, the response of the plants to the attack of a phytophagous mite can be recorded through various mechanisms such as antixenosis, antibiosis, tolerance, or combinations of these. Antixenosis occurs when a resistant plant is absolutely rejected or accepted by a herbivorous arthropod as a host plant due to the non-preference as a food resource and ovipositional substrate caused by biophysical or allelochemical factors present in the plants. Antibiosis occurs when a plant negatively affects the survival, growth, and fecundity of an herbivorous arthropod, caused by bioactive compounds present in the resistant host plants. Tolerance is a polygenic trait that allows a plant to resist, repair, or recover from damage caused by the herbivorous arthropod (Smith 2005, Smith and Clement 2012). This research aimed to assess antibiosis and antixenosis as resistance mechanisms in four plant species (Moringa oleifera Lamarck [Moringaceae], Persea americana Miller [Lauraceae], Rosa hybrida L. [Rosaceae]), and Phaseolus vulgaris L. [Fabaceae]) to the attack by avocado brown mite under laboratory conditions.
Materials and Methods
Brown avocado mite colony
A mite colony was started with biological material from the Plant Physiology Laboratory, Faculty of Engineering and Sciences, Autonomous University of Tamaulipas. The mite population was increased by placing females and males on bean plants (P. vulgaris) under greenhouse conditions at 29 ± 4°C and 80 ± 10% relative humidity (RH).
Collection and preparation of plant material
Three host plants (M. oleifera, P. americana, and R. hybrida) and P. vulgaris were used to evaluate feeding damage, oviposition, mite mortality, and mite population growth of O. punicae (Migeon and Dorkeld 2024). All plants were grown under field conditions.
Mature leaflets of M. oleifera (30) and R. hybrida (15) and leaves of P. vulgaris (10) and P. americana (2) were collected and transported in resealable plastic bags inside a cooler with gel packs at a temperature of 5 ± 2°C to the Physiology Laboratory of the Faculty of Engineering and Sciences of the Autonomous University of Tamaulipas. The transfer time of the plant material to the laboratory was between 15 and 30 min, depending on the host plant location. The selected leaves and leaflets were clean, that is, without the symptoms of fungi, bacteria, and any damage. However, the plant material was washed for 2 min wash with 1.5% sodium hypochlorite solution. Leaflets and leaves were then cut into 2-cm2 squares.
Experimental design
Oviposition rate and the percentage of damage caused by feeding of O. punicae females were evaluated using a host plant leaf square of 2 cm2. Each square was placed abaxial surface downward on cotton saturated with water and placed in a Petri dish (5 cm diam.). Twenty females in the teleiochrysalis stage (2 d old) and 20 adult males were placed on each foliage square of each host plant. Males were removed 24 h later. Once the females began ovipositing, each was allowed to lay eggs during the first 5 d of the oviposition period, after which we recorded the number of eggs laid (Gotoh et al. 2004). The assay was conducted under laboratory conditions at 28 ± 1°C and 70–80% relative humidity (RH), with a photoperiod of 12:12 h (light: dark). To ensure the freshness of the leaf squares of each tested host plant, individuals were transferred to new leaf squares every 2 d. Six randomly selected foliage squares of each host plant were assigned to one of four groups, one group for each host plant species. One foliage square of each host plant served as a replicate to establish six replicates per group or 24 in total.
The occurrence of antixenosis was based on two criteria: (1) O. punicae did not lay eggs and (2) non-feeding. The number of eggs laid per female on every foliage square of each host plant species was counted using a dissecting microscope (UNICO Stereo and Zoom Microscopes ZM180, Princeton, NJ). Feeding damage was estimated visually on each foliage square using a foliage damage index proposed by Nachman and Zemek (2002), where 0 = 0% damage (no feeding damage) and 5 = 81% to 100% feeding damage (e.g., a dense mark caused by feeding). The number of eggs laid per female and the percentage of feeding damage were recorded at 24 (day 1), 48 (day 2), 72 (day 3), 96 (day 4), and 120 h (day 5).
Antibiosis was determined by mite population growth rate and mite mortality. The growth rate (r, day−1) was calculated using the formula, r = (1/t)×ln(Nt/N0), where Nt is the final number of eggs, larvae, and live adults of O. punicae at time t (days, [equal to 5 d]), N0 is the number of females mites at time 0 (initial cohort = 20). The calculated r provides information on the short-term population growth patterns (Carey 1993). If r = 0, the mite population numbers do not change, whereas, if r > 0 or r < 0, the mite population increases or decreases over time, respectively (Smith and Smith 2015). Mortality was measured by the mean percentage of dead (drowned) individuals outside the foliage square, using the formula, (∑di/n) × 100, where di is the number of individuals drowned and n is the number of individuals on the foliage square. Mite mortality was recorded each day for 5 d.
We also recorded, on the fifth day, the number of O. punicae larvae because the hatch time of a single O. punicae egg is 4.4 to 4.7 d on grapevine leaf discs at 27°C, 80 ± 10% RH, and a photoperiod of 12:12 h (Vásquez et al. 2008). Furthermore, hatching time on discs of 6 species of eucalyptus plants is 5.03 to 5.27 d at 25 ± 2°C, 70 ± 10% RH, and a photoperiod of 12:12 h (Ferraz et al. 2020), while the required time for an O. punicae egg to hatch is 3.70 ± 0.04 d on E. tereticornis at 29°C, 70 ± 10% RH, and 12:12 h of photoperiod (Ferraz et al. 2021).
Statistical analysis
The data (number of laid eggs, dead mites, and the percentage of damage by feeding O. punicae females) was analyzed using analysis of variance of repeated measurements (ANOVArm). The number of larvae and the growth rate (r, day−1) were analyzed using one-way ANOVA and in both cases, the significant differences were analyzed with LSD’s multiple range comparison test. The SAS/STAT software was used for all analyses (SAS Institute, Inc. 2002).
Results
Antixenosis
The number of O. punicae eggs laid per female differed significantly among the host plants tested (F = 127.32; df = 3, 20; P < 0.0001), among the observation times (F = 54.13; df = 4, 80; P < 0.0001), and the host × time interaction (F = 14.37; df = 12, 80; P < 0.0001). The number of eggs laid on P. vulgaris was significantly higher (3.05 eggs/female) and significantly lower on M. oleifera (2.10 eggs/female) (LSD’s test, P < 0.0001) (Table 1). These differences indicate possible M. oleifera resistance to the oviposition of avocado brown mites compared to P. vulgaris, and these responses might be due to antixenosis.
The feeding damage of O. punicae differed significantly among the host plants (F = 338.30; df = 3, 20; P < 0.0001), among the observation times (F = 1281.35; df = 4, 80; P < 0.0001), and the host×time interaction (F = 5.15; df = 12, 80; P < 0.0001). Damage was significantly greater and less on P. vulgaris (28.73%) and M. oleifera (15.20%), respectively (LSD’s test, P < 0.0001) (Table 2). Thus, these discrepancies indicate a possible resistance of M. oleifera to damage caused by female avocado brown mite feeding compared to P. vulgaris and the other host plants, and these responses might be due to antixenosis.
Antibiosis
Growth rate (r, day−1) of O. punicae differed significantly among the host plants (F = 98.60; df = 3,20; P < 0.0001). The highest mean (±SD) r of O. punicae was observed with P. vulgaris (0.6884 ± 0.01), while the lowest was with M. oleifera (0.5990 ± 0.01) (Fig. 1). This indicates that M. oleifera is more resistant to the development of avocado brown mite than P. vulgaris, and these responses might be due to antibiosis.
Citation: Journal of Entomological Science 60, 4; 10.18474/JES24-107
The number of dead O. punicae differed significantly among the host plants (F = 14.89; df = 3, 20; P < 0.0001), among the observation times (F = 24.83; df = 4, 80; P < 0.0001), and the host×time interaction (F = 1.95; df = 12, 80; P = 0.0398). In general, the largest mean percentage of dead mites per day was observed on M. oleifera (8.33%) and the lowest on P. vulgaris (1.67%). Therefore, these results indicate the possible resistance of M. oleifera compared to other host plants to attack by O. punicae (Table 3), this response might be due to antibiosis.
Hatched eggs
The number of O. punicae eggs that hatched was significantly higher on P. vulgaris (36.17 larvae) than on P. americana (31.67 larvae), R. hybrida (28.00 larvae), and M. oleifera (16.00 larvae) (F = 55.28; df = 3, 20; P < 0.0001). The percentage of hatched eggs was significantly higher on P. americana (98.54%) and P. vulgaris (97.31%) than on R. hybrida (90.38%) and M. oleifera (88.07%) (Table 4). This indicates that the O. punicae eggs oviposited during the first 24 h were affected by the M. oleifera host plant. These eggs are expected to hatch on the fifth day after oviposition.
Discussion
The deposition of eggs is the beginning of the attack by herbivorous arthropods. Furthermore, the eggs laid by phytophagous arthropods indicate future damage to the plant since the larvae hatched from those eggs will cause damage due to their feeding. However, the survival of the progeny from those eggs requires a host plant that provides sufficient food for the following stages of development (Hilker and Fatouros 2015). The mean (±SD) number of eggs laid by O. punicae females was greater on P. vulgaris (3.05 ± 0.08) than on P. americana (2.73 ± 0.08), R. hybrida (2.77 ± 0.06), and M. oleifera (2.10 ± 0.05), suggesting that plants influence the ovipositional behavior and biology of the avocado brown mite. Other studies have shown that Oligonychus spp. fecundity is related to the species and variety of the host plant. Ferraz et al. (2020) reported that the total fecundity O. punicae was higher on E. tereticornis (44.75 ± 22.89 eggs/female) than on E. grandis (22.80 ± 11.09), E. brassiana (18.44 ± 8.99), E. pellita (13.35 ± 9.11), E. urophylla (8.45 ± 4.40), and C. citriodora (5.45 ± 3.71). Vásquez et al. (2008) documented the daily egg production of O. punicae on six grape cultivars and found that the daily oviposition rate was higher on Tucupita cultivar (2.82 ± 2.42 eggs/female/day) than on Red Globe (2.72 ± 1.91), Chenin Blanc (2.16 ± 1.99), Sauvignon (2.15 ± 1.41), Villanueva (1.79 ± 0.94), and Sirah (0.94 ± 0.95) cultivars. Yao et al. (2019) reported that litchi cultivars (Litchi chinensis Sonn [Sapindaceae]) affect the number of eggs oviposited per female of Oligonychus litchii Lo & Ho (Prostigmata: Tetranychidae) with O. litchii ovipositing more on the litchi cultivar Nuomici (64.84 ± 3.57 eggs/female) than on Sanyuehong (38.55 ± 2.34), Feizixiao (22.30 ± 1.59), and Baili (14.78 ± 1.66) cultivars. Chaaban et al. (2012) reported the daily fecundity of Oligonychus afrasiaticus (McGregor) (Acari: Tetranychidae) when fed fruits of five date palm (Phoenix dactylifera L. [Arecaceae]) cultivars and sorghum leaves (Sorghum sp.). They found that fecundity of O. afrasiaticus was greater on sorghum leaves (2.00 ± 1.3 eggs/female/day) than on Deglet Noor cultivar (1.5 ± 0.4), Kentichi (0.9 ± 0.5), Bessr (0.9 ± 0.4), Alig (0.7 ± 0.3), and Deglet Noor pinnae (0.7 ± 0.2). Roknuzzaman et al. (2020) found that Oligonychus biharensis (Hirst) (Acari: Tetranychidae) lays more eggs on country bean (Lablab purpureus L.) (1.67 ± 0.06 eggs/female/days) than on mung bean (Vigna radiata L. [Wilczek]) (1.28 ± 0.01 eggs/female/days). It is, therefore, likely that O. punicae females lay a greater number of eggs on P. vulgaris than on P. americana, R. hybrida, and M. oleifera which is likely due to the influence of biotic and abiotic factors. Biotic factors include differences among Oligonychus species, nutritional quality, morphological and chemical features in different host plant species and varieties, and abiotic factors, including adapting specific mite species to the local climate and host plants, handling methods, and observation time intervals (Ferraz et al. 2020, Roknuzzaman et al. 2020, Vásquez et al. 2008).
Feeding damage caused by O. punicae females differed significantly among host plants. Moringa oleifera and P. vulgaris were the host plants that suffered the least and the greatest damage, respectively, caused by feeding of the avocado brown mite. Plants produce various biochemical compounds (alkaloids, flavonoids, terpenes, and phenols) to defend themselves or avoid attack by phytophagous arthropods. These secondary metabolites have acaricidal activity, which generates antixenotic and antibiotic effects on these arthropods. Moringa oleifera may present phytochemicals such as alkaloids, coumarins, flavonoids, phenols, and tannins in its leaves (Heinz-Castro et al. 2021). These phytochemicals are substances with insecticidal and acaricide action (Hamza et al. 2016, Heinz-Castro et al. 2021, Ojo et al. 2013). Koul (2016) and Singh et al. (2021) mentioned that bioactive compounds such as alkaloids, coumarins, terpenes, and phenols adversely affect feeding herbivorous arthropods. Hence, these phytochemicals may explain the low feeding damage rate of O. punicae when fed M. oleifera.
In this study, the number of O. punicae hatched eggs oviposited in the first 24 h was less on M. oleifera than on other host plants. Furthermore, the mortality percentage of female brown avocado mites was higher on the same host plant species, indicating that this plant has a toxic effect on the eggs and females of O. punicae females and, hence, a negative effect on the growth rate (r) of the avocado brown mite. Hilker and Meiners (2011) mentioned that low humidity due to dry air and closed plant stomata and gas concentrations in the host plant leaf boundary layer can cause desiccation of eggs. Hence, this may explain the low growth rate of O. punicae when fed M. oleifera compared to other host plants. In addition, many factors influencing the population growth of spider mites, including the nutritional value of the host plant, environmental factors, and feeding inhibitions caused by the different morphological and physiological characters of host plants (Roknuzzaman et al. 2020). The growth rate estimated the process of colonization and establishment in new environments for O. punicae, which had exponential growth (r > 0), characteristic of populations inhabiting favorable environments at low population densities (Smith and Smith 2015).
Female mite feeding and oviposition on beans in comparison to avocado, rose, and moringa. Using the same host plant in the test as for the mite rearing could induce changes in the responses of the adult females of O. punicae on the other host plants in comparison to the mites that experienced in the pre-adult stages on the plants of beans, suggesting memory conditioning in O. punicae. Thorpe (1939) described this as preimaginal conditioning. More studies are required to determine if this behavior is due to preimaginal conditioning. Arthropods with preimaginal experiences can learn the characteristics of a high-quality food resource and, as adults, females with retention of this learning quickly locate an oviposition site suitable for larval development (Barron and Corbet 1999).
In conclusion, the results of these tests show that the host plant M. oleifera presents possible resistance to the attack of O. punicae females and that those responses might be due to antibiosis and antixenosis, causing a lower oviposition, lower feeding damage, a low population growth rate, higher mortality, and lower number of hatched eggs as compared to P. vulgaris, P. americana, and R. hybrida. While promising, further research is required, including the morphological and biochemical characters of plants to correlate them with their observed antibiotic and antixenotic effects on O. punicae. Furthermore, to have a clear and systematic image of the specific age at birth (mx) and survival (lx) of the O. punicae population on tested plants, both for females and males must be analyzed in detail, the development time from egg to adult, the pre- and post-oviposition, oviposition periods; total and daily fecundity and its demographic parameters.
Population growth rate of Oligonychus punicae on four tested plants.
Contributor Notes
Faculty of Agronomy and Veterinary, Autonomous University of San Luis Potosí, Soledad de Graciano Sánchez, 78321, San Luis Potosí, Mexico.
Faculty of Engineering and Business San Quintín, Autonomous University of Baja California, San Quintín, 22930, México.
Parasitology Department, Antonio Narro Agrarian Autonomous University, Saltillo, 25315, Coahuila, Mexico