Introduction
Moringa oleifera is a plant with ecologic plasticity, that allow it to adapt to various climates and soil conditions (Pérez et al. 2010). According to the goodness that show as protein plant is considered ideal as animal food, being attractive for ruminants (García-López et al. 2017 and Alvarado-Ramírez et al. 2018) as to monogastric (Valdivié et al. 2017). It is also attributed several beneficial effects for human health (Hernández e Iglesias 2022 and Príncipe and Soto 2023) and as plants bio-stimulant (Pérez et al. 2023). However, the edaphoclimatic conditions in which it develops has a marked influence on its yields. In any regions of Cuba, the susceptibility it has showed to the attack of harmful organisms, mainly insect-pest and pathogens, involve even more it use when great biomass volume is required, as it happens when is about ruminants feeding.
To guarantee healthy plants and of good growing in the first development stages is important for the plant rooted and future performance in the adult age, aspect that are essentials to achieve satisfactory establishments. This research was develop with the objective of monitoring and control the organisms caused of plagues, associated to different Moringa oleifera origins under nursery and field conditions.
Materials and methods
Experimental area and climate
Two tests, one in nursery phase and the other in field phase were developed, at Instituto de Ciencia Animal (ICA) from San José de las Lajas municipality, Mayabeque province, Cuba. The Institute is located at 22º 55 NL and 82º 0 WL, at 92 m. o.s.l. The climate according to the climatic classification of Köppen is Aw type, tropical grassland climate, with a dry season (November - April) and a rainy season (May -October). The annual rainfall is of 1244 mm and mean temperature of 24.9 ºC (Anon 2016).
Treatments
The treatments were constituted by six Moringa oleifera origins collected in three provinces of the country (table 1). The collections start from adult plants, apparently healthy, with good growing, in fruiting phase.
Experimental design
A completely random design, with 66 repetitions for each of the treatments was used.
Experimental procedure
A total of 396 polyethylene bags were used. The substratum used in each bag was the mixture of three parts of the red ferrallitic soil (Hernández et al. 2015), and a part of organic matter (decomposed bovine manure). At the moment of sowing, October 13, 2016, two seeds apparently healthy were place at 2 cm deep, according to Cardoso et al. (2006), to keep only the most healthy and vigorous plant.
The nursery was place separated from the soil surface, at a 1cm height approximately. The irrigation was manually, with the help of a watering can in alternate days, until cover their filed capacity. The bags were perforated to guarantee the drainage and flooding. It is important to highlight that the selected place was at opencast, so the plantlets received directly the sun, as the rainfalls too. The manual control of the arvenses in each bag was carried out.
Once the germination of more than 70 % of the plants was guarantee, the record of agronomic and phytosanitary measures were started:
Emergence percentage, %
Plantlets survival, %
Plants height, cm
Stem diameter, mm
Presence and quantification of arthropods and pathogens
Determination of the caused damage, %
The emergence percentage (% E) was determined counting the emerged plantlets with more than 1 cm height, in accordance to the formula described by Ede et al. (2015):
The plantlets survival (% S) was determined by the counting of emerged plantlets:
The methodology described by Centeno et al. (1994) was considered, who establish a classification according to the obtained value: a) 80 -100 % very good; b) 60-79 % good; c) 40-59 % so- so and d) less than 40 % bad.
The plants height (cm) was determined measuring with a tape measure, from the soil to the apex of the apical branch.
The diameter or stem thickness (cm) was taken in the interception of the base of the first branch and the stem with a vernier.
The growing dynamics (GD) was determined by the following equation:
where:
GD= |
Growing dynamics |
IH = |
Initial height (The measurement of the height taking after the germination and the previous to the data taking) |
FH = |
Final height (height taking in each sampling) |
Number of sampling days |
(at 10, 30 and 40 d after sowing) |
The nursery phase lasted 45 days. The samplings were carried out in the morning, taking into account three samplings in the period. The arthropods and pathogens were evaluated through visual observations in a sampling system presence-absence. The present organisms were noted and quantified, classifying in pollinators, phytophagous and bioregulators, in accordance to the assignation of the functional groups, according to Metcalf and Flint (1965), Triplehorn and Johnson (2005), Mancina and Cruz (2017) and World Spider Catalog (2020).
The damage was evaluated according to Calderón (1982) methodology for stinging-sucking or biting insects and for both. If to be necessary, the capture of arthropods and phytophatogen microorganisms unknown for their identification in the laboratory was made. The damages by pathogen agents were evaluated according to Lenné (1982) methodology.
The plantlets were in nursery phase until reached heights higher than 25cm, moment in which their transplant was made. To counteract the risk of plant loss by plague attacks the use of control measures (chemical or biological, and both) was foresee, is to be necessary. All the agronomic and phytosanitary measurements were performed in fixed plantlets with the optimums sample size (22).
Field phase
The test was conducted in a red ferrallitic soil, in accordance with Hernández et al. (2015) in areas from the experimental center “Miguel Sistachs Naya” attached to Grasses and Forages department from ICA.
On December 5, 2016 a total of 378 plantlets of M. oleifera were transplanted to the field, with approximately height from 25 to 30 cm, previously cultivated under nursery conditions in an opencast place. The experiment lasted two years.
Experimental design
The plantation was carried out under a random block design with three replications in which the six origins were randomized in each replication.
Experimental procedure
A preparation of conventional soil was initially carried out, leaving the bed loose and free of weeds. The area included a total of 23 furrows, established at 0.70 m for a dimension of 16.10m wide and 70 m length. Each block was separated 3m among them.
At a week of transplant fertilization to the whole area was carried out, with complete formula of N-P-K (9-13-17) to cover the crop requirements, according to that reported by Pérez et al. (2010). The used dose was at a rate of 10 g per bunch, which leads to 200 g per plot.
The area was protected of weeds invasion. The manual cleaning was with spud and hoe, mainly around the plant.
The area was maintained in the dry season, sprayed with mobile irrigation machine (OCMIS brand, model 82 with sprinkling bar) operated with a pressure of 500 kPa and the volume of sprinkler used was of 33600 L/h. During the irrigation time (2.5 h) a water lamina of 35 mm was applied. The irrigation was applied at the beginning of the transplant, with weekly frequency which was interval from 10 to 15 d until the first rains arrives.
The area was also under phytosanitary alertness against possible attacks of leaf-cutting ants (Atta insularis) or other harmful agents. The application of chemical or biological products was prevented.
The sampling of the arthropods and pathogen incidence were monthly carried out through visual observations in the internal furrows, leaving those of the external as border effect. A total of 10 plants of each origin/replication were sampled trying to visualized, counted and identified the organisms associated in each honological stage of the crop. When it was necessary, samples were taken and transfers to the laboratory for their processing through the help of similar taxonomic keys and with the use of the stereoscopic microscopic. The classification of the organisms in accordance with the assignation of functional groups and the damages was performed as it is described in the nursery phase. In the fruiting stage, five pods from each origin/replication were random taking to determine, under laboratory conditions, the phytosanitary state and the damages caused by the organisms associated to the crop.
The area was for seed production. There were measured as morphologic indicators the survival (%), plants height (cm), the number of branches and the stem diameter (cm), following the methodology described for nursery phase.
Statistical analysis
All the collected data in nursery and field phase were included in Microsoft Excel database. They were statistically processed and analyzed according to statistical package InfoStat version 2012 (Di Rienzo et al. 2012). The theoretical assumptions of the analysis of variance were verified for the analyzed variables from the Shapiro and Wilk (1965) test for the normality of errors and Levene (1960) test for the variance homogeneity. It was taking into account that in case that the analyzed variables do not fulfill with the theoretical assumptions of ANAVA, the arcosen√% transformation will be used. If they even will not improve the fulfillment of those assumptions it was determined to carry out non-parametric analysis of variance of one-way classification (Kruskal and Wallis 1952) and to apply the Conover (1999) test for the comparison of average ranges. The height and growth dynamic, in nursery stage were analyzed according to a completely random statistically model and Duncan (1955) test was applied for the differentiation between means. The variables height and stem thickness, in field phase, were analyzed according to random block design. Tukey (1958) test was applied in necessary cases.
Results and discussion
Nursery phase
Regarding the presence of insects and pathogens, there were only visible some phytophagous (table 2) as the leaf miner Liriomyza sp. (Dipteran: Agromyzidae) in three isolated plantlets from the origin 3, 5 and 6.
Class | Order | Family | Scientific name | Common name | Origin |
---|---|---|---|---|---|
Insecta | Diptera | Agromyzidae |
|
Leaf miner | 3, 5 and 6 (one plant from each origin) |
Hemiptera | Aphididae |
|
Aphid or plant louse | 1 (two plants) | |
- | - | - | Not identified causal agent | Leaf spot by fungus | 1 (some plants) |
- | - | - | Not identified causal agent | Whitish spots | 2 (some plants) |
It was also found a source of aphids from Aphis sp. (Hemiptera: Aphididae) genus with higher intensity in a plant and, lower in another one, from origin 1. It was visible in some plants leaf spots by fungus, whose causal agent could not being identified. There were five plants in the origin 2 with whitish discoloration in the leaves whose origin is unknown. However, the harmful organisms associated did not caused economic damages to the plants under nursery conditions, so the damages did not exceeded the slight degree, in accordance with the Calderón (1982) scale, in this case for insects with stinging- sucking habits.
Results from Reyes (2005) and Alfaro and Martínez (2008) reported damages in this specie in nursery phase, due to the attack of cutting ants from Atta genus. To keep the plants under nursery conditions at 1.5m separated from the soil prevent the attack of leaf-cutting ants to moringa plantlets, which constitutes an effective resource to minimize the damages that these organisms could caused.
Regarding the morphologic performance of the plantlets, in table 3 are show the average ranges of the emergency and survival (%) of the different M. oleifera origins at 30 days after sowing. The origin 6 (Matanzas, Boca Camarioca) showed the lower average ranges (53.50 and 57.00 for emergency and survival, respectively), although without statistical differences with relation to the rest of the evaluated origins.
Indicator | Average ranges | Sign. | |||||
---|---|---|---|---|---|---|---|
Origins | |||||||
1 | 2 | 3 | 4 | 5 | 6 | ||
Survival | 69.00 (95) SD=0.21 | 69.00 (95) SD=0.21 | 69.00 (95) SD=0.21 | 66.00 (91) SD=0.29 | 69.00 (95) SD=0.21 | 57.00 (77) SD=0.43 | p=0.1810 |
Emergency | 71.50 (95) SD=0.21 | 68.50 (91) SD=0.29 | 71.50 (95) SD=0.21 | 65.50 (86) SD=0.35 | 68.50 (91) SD=0.29 | 53.50 (68) SD=0.48 | p=0.0570 |
( ): True data, %
SD: Standard deviation
Comparison of average ranges according to Conover (1999) test
When considering the mortality as the survival loss, coincide that is the origin 6 which has more plantlets loss (33 %). However, even with these values, the survival is classifying as good, according to Centeno et al. (1994). In nursery phase, all the origins showed acceptable values as to the evaluated morphologic indicators. Researchers from García and Mora (2018) assure that when the emergency and the initial development of M. oleífera is very good in nursery phase, this constitutes an indicator of the good vigor of the used seeds.
Figure 1 show the result of the plants diameter at 40 days after sowing. The diameter was in the range of 0.23 to 0.28, without statistical differences among the origins. Slightly high values, between 0.33 and 0.40, Toral et al. (2013) stated, when studying eight origins of moringa under nursery conditions, some collected in the country and other imported.
The plantlets height (figure 2) in the first sampling, 10 days after sowing, had a high performance in the origin 5 with 11.41 cm. There were not statistical differences with the origin 1 and 2, which showed values of 10.80 and 10.20 cm, respectively. In the second sampling, 30 d after sowing, the origin 1 had the highest value with 30.55 cm, without statistical differences with the origin 2 (28.91 cm), that in turn did not differ from the origin 5 with 28.23 cm. The origin 6 showed the lower height (25.32 cm) without statistical differences with the 3 and 4, whose heights were around 25 cm. At 40d, the origin 1 showed the higher size (p < 0.0001) (34.41 cm), followed by the origin 2 (31.95 cm) that, in turn, did not differ from the origin 5 (30.32 cm) neither this one from the rest, that ranges between 27 and 28 cm. The origin 6, similar to that occur in other agronomic indicators, showed the lowest height (27.82 cm), although in this time did not statistically differ from the origins 3, 4 and 5. These results were superior to those obtained by García and Mora (2018), whose did not reached sizes similar to those recorded in this test until after 15 weeks. Even, there were very superiors to that reported by Cadillo Rojas (2022), which used biostimulants to accelerate the germination, emergency and plants height.
Researchers from Martínez et al. (2013) stated that the height is an indicator of the development of the aerial part, which determines the photosynthetic process and the plant transpiration. Miranda et al. (2014) studies refers heights of 7.6 cm in the first week with the use of compost and manure as substrate and low values with soil only (5.1 cm). In the tenth week heights of 104.9 cm were recorded with compost, 59.7 cm with soil and 32.7 cm with manure. In this study, the plant height is similar to the treatment with compost, maybe the result of the plants development will be conditional by the substrate used in the bags under nursery conditions, which consisted in organic (decomposed cattle manure), the quality of the used soil (red ferrallitic soil), water availability (irrigation) and other agroclimatic factors, whose performance caused variations from a study to other.
Table 4 shows the growth dynamic of the origins in each of the samplings. The origin 6 had a low growth dynamic and differing from the rest of origins at 10 d with 0.78 cm/d. At 30 d, the origin 6 was similar to 4 and 3, with values of 0.84, 0.85 and 0.86 cm/d, respectively. At 40 d, had a low growth dynamic too, without differ from the origins 3, 4 and 5 with values that ranges between 0.71 and 0.78. These results were, even, higher to those obtained by Toral et al. (2013). These authors reported higher increases of 0.41 a 0.43 cm/d in origins imported in the country, as Supergenius and Plain, as well as in a collection in Holguín-Mayarí. These results show the fast growth the plant species Moringa oleifera have. In addition, they confirm antecedents that distinguish the plant because of their great growth speed (Anon 2002). In this indicator is more reliable show that the plant has a fast growth in the first weeks, what coincides with Medina et al. (2007), whose state that the moringa has a fast growth from the beginning, due to it develops a root system very deep, which makes higher the use of soil nutrients and of the available water.
Treatments Indicator | Origins | SE± and sign. | |||||
---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | ||
Sampling 1 | |||||||
Growth, cm/d | 1.08ab | 1.02ab | 0.93b | 0.96b | 1.14ab | 0.78c | 0.05 p<0.0001 |
Sampling 2 | |||||||
Growth, cm/d | 1.02a | 0.96ab | 0.86c | 0.85c | 0.94b | 0.84c | 0.02 p<0.0001 |
Sampling 3 | |||||||
Growth, cm/d | 0.86a | 0.80b | 0.71c | 0.71c | 0.76bc | 0.78c | 0.02 p<0.0001 |
a,b,c Means with a common letter in each sampling there are not significantly different (p<0.05) (Duncan 1955)
Field phase
Regarding the incidence of insects-plague a total of 45 species from four classes of arthropods were collected, as is show in table 5, being the Insecta class the most represented with 39 morph species, included in 8 orders and 18 families. From them, 87 %, a total of 34 morph species, corresponding to phytophagous insects, coinciding with Foidl et al. (1999) and Reyes (2005) researchers, when reporting some organisms which causes plagues in the establishment stage, associated to M. oleifera. Medina et al. (2007) assure that the moringa is resistant or immune to plagues and diseases. According to the results of this test, under field conditions, the criteria of the cited authors is not share, so in the course of the study it was evident the presence and incidence of harmful organisms in the different phenological stages of the crop. This caused, in some occasions, economic damages to the plants.
Class | Order | Family | Scientific name | Common name |
---|---|---|---|---|
Insecta | Hemiptera | Cicadellidae |
|
Leafhopper |
|
Leafhopper | |||
|
Leafhopper | |||
Flatidae |
|
Palm flatid planthopper | ||
Aphididae |
|
Aphid | ||
Coleoptera | Chrysomelidae |
|
Flea beetle | |
|
Leaf beetle | |||
|
Leaf beetle | |||
|
Leaf beetle | |||
Coccinellidae |
|
Ladybird | ||
Curculionidae |
|
Citrus root weevil | ||
Not identified species | True weevils | |||
Lycidae |
|
Coleoptera | ||
Diptera | Syrphidae | Not identified species1 | Syrphid fly | |
Drosophilidae |
|
Fruit fly | ||
- | 5 morph-species not identified | Fly | ||
Dolichopodidae |
|
Fly | ||
Chamaemyiidae |
|
Fly | ||
Culicidae | 1 morph-specie not identified | Mosquito | ||
Lepidoptera | Crambidae |
|
Bean-leaf webworm moth | |
- | 4 morph-species not identified | Moth | ||
Orthoptera | Tettigonidae |
|
Long-horned grasshoppers | |
Gryllidae |
|
Crickets | ||
Hymenoptera | Formicidae |
|
Fire ants | |
|
Crazy ants | |||
|
Little fire ants | |||
|
Leaf-cutting ants | |||
Apididae |
|
Bee | ||
Ichneumonidae |
|
Wasp | ||
- | Not identified species | Black wasp | ||
Dermaptera | Forficulidae |
|
Earwig | |
Odonata | - | Not identified species | Dragonfly | |
Arachnida | Araneae | 4 morph-species not identified1 | Spider | |
Gastropoda | Not identified species | Snails | ||
Malacostraca | Isopoda | Armadillidae |
|
Common pill woodlouse |
1 Bioregulators
The M. oleifera plantlets, just transplanted, had a strong attack of leaf-cutting ants (Atta insularis Guérin) (Hymenoptera: Formicidae), which caused defoliation and loss of plantlets, equivalent to average damage according to the established by Calderón (1982) for biting insects. This led to the immediate application of management strategies to minimize the damages. The application of Blitz (i.a. Fipronil) granulated, near to the active nests found in the area control the initiated forage in the plantlets of the six moringa origins. These results coincide with Padilla et al. (2017) studies, which affirm that at present the A. insularis constitutes one of the organisms that most affects the survival of M. oleifera plantlets under Cuba conditions.
Researchers in other regions from the American tropic about M. oleifera coincide that the plagues that has most affect the plants, immediately after germination, are the cutting ants (Atta sp.) and also mention the small mocis moth (Mocis latipes Guén.) (Lepidoptera: Noctuidae). These organisms normally make an attack and did not come again to the crop, although they emphasize that even so they should be controlled to reduce the damages (Foild et al. 1999).
In the vegetative phase, with the emission of leaves and branches, the attack of coleoptera from Curculionidae (Pachneus litus and True weevils) family was proved. These last, not identified, with high population levels, caused significant reduction of the active photosynthetic area, when defoliate some plants from different origins. The degree of damage was moderate according to Calderón (1982) scale for biting insects.
Subsequently, in the fruiting phase, it was proved in all origins, damages in the pods caused by the fruit fly Gitona sp. which allow the secondary presence of fungus, which caused, even more, severe damages to the fruits and their seeds, equivalent to an intense damage, according to those described by Lenné (1982), preventing the healthy seeds harvest.
Gitona sp. is considered most important plague of the fruit in almost all the region of the world. The female infests the fruits, causing their putrefaction. Also, the opening makes possible the presence of other harmful agents that invade the inside of the fruit, as grubs and mature coleopteran, lepidopterous grubs, mites and phytopathogen microorganisms, which accelerate the damage and fruit loss (Anon 2023). The above completely coincide with the damages proved in this test in the inside of the fruits.
Every 15 d a control variant to counteract the leaf cutting ants attacks was systematically applied. Leaves of Tithonia or Marigold (Tithonia diversifolia), protein plant from Asteraceae family were spread, near to the nets according to Montoya (2020) and chemical application of Blitz (i.a. Fipronil). Any other insecticide or biopesticide was not used to counteract the attack of flying phytophagous insects, so the plants size exceeded 2cm height, and it was impossible to made any correct spraying that cover the stem and the affected branches.
The survival, measure at 90 d, it was not significant (Table 6), since the average ranges were similar in all the evaluated origins, and oscillate from 3.40 to 3.60, which means values above 90 % of survival, classified as very good, according to Centeno et al. (1994). The average ranges, corresponded to the count of the number of branches, showed with the best results the origin 4 (4.40), which in turn did not differed from the origin 6 (4.18). The origins 3 and 2 had low number of branches with the lowest average ranges (2.50 and 2.92, respectively). It is important to highlight that, despite the incidence of harmful organism showed in the field phase, the evaluated origins showed acceptable morphologic indexes for the M. oleifera species too. Maybe these optimums result of the crop growth were conditional by the cultural practices made in the field phase. In this phase, immediately after the transplant, fertilization with complete formula N-P-K was carried out. The arvenses competition was reducing, during the first years, when maintaining the manual cleaning by means of spud and hoe in the area. The irrigation in the dry season was guarantee and a constant protection against the attack of leaf cutting ants was maintained. Therefore, the applied agronomic management obviously improves the establishment, in terms of survival rates with a faster growing. Similar criteria showed researchers by Holguín et al. (2018). Other authors give more important to the good species performance, favored by climatic and edaphic variables that generates the agroecological potential in which the M. oleifera is developed (Carrión Delgado et al. 2022).
Indicator | Average ranges | Sign. | |||||
---|---|---|---|---|---|---|---|
Origins | |||||||
1 | 2 | 3 | 4 | 5 | 6 | ||
Number of branches | 3.60bcd (12.33) SD=4.62 | 2.92de (10.50) SD=2.80 | 2.50e (9.27) SD=4.12 | 4.40a (15.80) SD=6.76 | 3.50cd (11.80) SD=4.71 | 4.18abc (14.00) SD=5.00 | P=0.0002 |
Survival at 90 d | 3.60 (1.00) SD=0.00 | 3.60 (1.00) SD=0.00 | 3.40 (0.93) SD=0.25 | 3.50 (0.97) SD=0.18 | 3.40 (0.93) SD=0.25 | 3.50 (0.97) SD=0.18 | P=0.5553 |
( ) - True data
SD- Standard deviation
Average ranges comparison according to Conover (1999) test
The height and stem thickness (figure 3) were measured at 120 d after the transplant. The plantlets from the origin 6 and 2 were highlighted by their high size (148.1 cm and 146.3 cm, respectively). The higher stem thickness (31.2 mm) was confirmed in the plantlets from origin 2, which differ from the rest. It was proved that the morphological performance of the plants can varied depending on the biotic and abiotics factors. In this case, the palatability showed to some plants in the nursery phase, from origin 1, caused of the attack of aphids from the Aphis genus, caused that when they were transplanted to the field with the incidence of this phytophagous in the affected plants occur a growth lateness, with respect to others from the same origin that there were not affected. This could be possible because this insects, located with preference in the reverse of leaves, suck up the plants sap causing roll, chlorosis and necrosis, which prevent the normal development of the host plant. Hence the importance of having healthy seeds and plants, free of harmful organisms, in the moment of sowing or crop planting, and in both.
Conclusions
It is concluded that the evaluated origins, given the performance of their morphologic indicators, has potentialities for their use. It is recommended to maintain the phytosanitary alertness of the crop and to regulate the forage activity of A. insularis until achieving the moringa establishment. It should be alert in the fruiting phase to the incidence of Gitona sp. fly to minimize the damage to the seeds, to avoid the secondary presence of pathogens and to allow the harvest of healthy seeds.