Conservation of Schomburgkia crispa Lindl. (Orchidaceae) by reintroduction into a fragment of the Brazilian Cerrado

Journal Pre-proof Conservation of Schomburgkia crispa Lindl. (Orchidaceae) by reintroduction into a fragment of the Brazilian Cerrado Jackeline Schultz Soares, Etenaldo Felipe Santiago, Jose´ Carlos Sorgato

PII:

S1617-1381(19)30084-6

DOI:

https://doi.org/10.1016/j.jnc.2019.125754

Reference:

JNC 125754

To appear in:

Journal for Nature Conservation

Received Date:

27 February 2019

Revised Date:

24 September 2019

Accepted Date:

30 September 2019

Please cite this article as: Soares JS, Santiago EF, Sorgato JC, Conservation of Schomburgkia crispa Lindl. (Orchidaceae) by reintroduction into a fragment of the Brazilian Cerrado, Journal for Nature Conservation (2019), doi: https://doi.org/10.1016/j.jnc.2019.125754

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Conservation of Schomburgkia crispa Lindl. (Orchidaceae) by reintroduction into a fragment of the Brazilian Cerrado Jackeline Schultz Soaresa, Etenaldo Felipe Santiagoa, José Carlos Sorgatob* a

State University of Mato Grosso do Sul, Natural Resources Department, Dourados, 79.804-

970, Mato Grosso do Sul, Brazil. [email protected]; [email protected] b

Federal University of Grande Dourados, College of Agrarian Sciences, Dourados, 79.804-970,

Mato Grosso do Sul, Brazil. [email protected] author

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* Corresponding

ABSTRACT - Asymbiotic germination of native orchids is an important technique for

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producing seedlings for use in species reintroduction programs, as it facilitates the maintenance of genetic variability. The objective of this study was to evaluate the adaptive responses of

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Schomburgkia crispa Lindl. (Orchidaceae) in an area undergoing environmental restoration, and to compare the efficacy of traditional and alternative substrates in order to contribute to the

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management and reintroduction of this species. Plants of S. crispa, obtained from asymbiotic sowing, were reintroduced in an underbrush area at the State University of Mato Grosso do Sul using three planting treatments: without substrate (WS), buriti stem substrate (BU), and coconut

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fiber substrate (CF). Stress was assessed based on chlorophyll a fluorescence measurements and infrared thermography, and the growth environment was evaluated for light conditions and canopy cover. A completely randomized design was used with three treatments, consisting of

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one substrate, with 10 single-plant replicates. At 21 days after reintroduction, the plants showed 100% survival regardless of the substrate used. The largest variations between the temperature

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of the phorophytes and the temperature of the epiphytic orchid plants occurred on the day of reintroduction. There was no significant difference in the potential quantum efficiency of photosystem II (PSII: Fv/Fm) among treatments. Although the different substrates did not significantly affect plant survival, temperature, or the photochemical efficiency of PSII, the variables analyzed allowed us to evaluate the responses of orchids to the transition from the acclimatization phase to the natural environment. Keywords: Orchidaceae; in vitro cultivation; asymbiotic sowing; environmental restoration; plant enrichment

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INTRODUCTION

Epiphytic plants that complete their life cycle without contact with the soil are known as true epiphytes or holoepiphytes. These plants typically grow and develop on arboreal plant hosts, referred to as phorophytes, without parasitizing them (Font‑ Quer, 1953; Duarte & Gandolfi, 2017). Within an ecosystem, epiphytes play a fundamental ecological role, as they can provide micro-habitats and distinct micro-climates, harboring an extensive diversity of organisms and offering resources such as flowers, fruit, and nectar (Cestari, 2009).

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The introduction of epiphytic species, mainly orchids, in ecological restoration projects is not common. Restoration plantings are initially based solely on tree species, as their objective is the reconstruction of vegetation phytophysiognomy (Bellotto, Viani, Gandolfi, & Rodrigues, 2009). However, areas undergoing restoration have typically not been colonized by epiphytes through the natural dispersal of propagules. Even in 50-year-old forests, the richness of non-

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tree species is only approximately half that of natural ecosystems (Garcia et al., 2011).

Asymbiotic germination of native species of the Orchidaceae family is an important

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approach for producing seedlings that can be used in programs aimed at reintroducing species into their original habitats, as this technique facilitates the development of a germplasm bank

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that maintains genetic variability and can contribute to the restoration of natural populations (Schneiders, Pescador, Booz, & Suzuki, 2012). However, in contrast to native tree species, for which there exists an extensive literature describing established techniques and methodologies,

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there have been only a few studies on the reintroduction of orchids and, in general, these either deal with terrestrial species or report investigations of symbiotic associations with mycorrhizal fungi (Yang et al., 2017; Herrera, Valadares, Contreras, Bashan, & Arriagada, 2017).

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As an asymbiotic approach, the reintroduction of material produced in vitro can be even more challenging, since it influences the establishment of individuals based on their complete

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repertoire of morphological and physiological peculiarities. Moreover, these plants must be acclimatized prior to reintroduction and may still need to establish symbiotic associations after reintroduction (Brustulin & Schmitt, 2008). Even so, nursery acclimatization may not be enough to facilitate adaptation the various effects of natural environmental stresses, and thus they may need to develop new adjustment responses. For this reason, there have been relatively few studies on the reintroduction of asymbiotically sown epiphytic orchids into the natural environment (Rubluo, Chávez, Martínez, & Martínez-vázquez, 1993; Decruse, Gangaprasad, Seeni, & Menon, 2003; Dorneles &

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Trevelin, 2011; Endres Júnior, Sasamori, Silveira, Schmitt, & Droste, 2015, Segovia-Rivas, Meave, González, & Pérez-García, 2017), and in none of these has Schomburgkia crispa, an orchid species native to the Brazilian Cerrado, been used (Mendonça et al., 2008; Ostetto, 2015; Barros et al., 2019). Additionally, there is no information regarding the evaluation of reintroductions using different substrates based on the monitoring of temperature or fluorescence analysis of the chlorophyll of reintroduced individuals. Accordingly, in the present study, we evaluated the adaptation of S. crispa plants that had been reintroduced in an area of the Cerrado biome undergoing environmental restoration and compared the efficacy of traditional and alternative substrates in contributing to the

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management and reintroduction of this species in the natural environment.

MATERIAL AND METHODS

In December 2016, plants of Schomburgkia crispa Lindl. were reintroduced in an

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underbrush area at the State University of Mato Grosso do Sul (UEMS), Dourados - MS. This area was the site of a restoration project inaugurated in 2012, in which 32 species of pioneer

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and non-pioneer trees, typical of semi-deciduous tropical forest, were introduced, and at the time of the present study had been undergoing restoration for 5 years. The climate of the region

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is type Am of Köppen (Monsoon Tropical), with average temperatures in the coldest and warmest months of below 18°C and 22°C, respectively (Fietz, Fisch, Comunello, & Flumignan, 2017), and total annual precipitation of between 1,250 and 1,500 mm.

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The S. crispa plants used in this study were derived from asymbiotic germination of seeds obtained from the Orchidarium of the Faculty of Agrarian Sciences - FCA/UFGD, Rio Ivinhema State Park (PEVRI - MS) and acclimatized for 6 months in a nursery covered by the

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overlap of two 50% shading screens [photosynthetically active radiation (PAR) = 235.1 μmol m-2 s-1]. For reintroduction, we randomly selected 30 individuals of S. crispa with a height of

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approximately 4 cm. These plants were affixed to 15 different types of phorophyte, using a nylon mesh, at a height of 1.5 m, all on the eastern side of the stem, such that the incidence of light on plants was not direct during the warmest periods of the day (Figure 1). To evaluate the survival and adaptation of the plants to the new conditions, we assessed the effects of three treatments based on substrate type: treatment 1 - without substrate, treatment 2 - buriti stem substrate, and treatment 3 - coconut fiber substrate. Each phorophyte received up to three individuals, with different treatments, and were identified for later monitoring by an id plate in an aluminum strip.

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On the day of reintroduction (day zero) and at 7, 14, and 21 days after reintroduction, the survival and stress responses of the orchids were evaluated using non-destructive radiation emission techniques. Thermography was performed using a Testo 875-2i thermal imaging camera and fluorescence measurements of chlorophyll a in leaves adapted to the dark for 30 min were obtained using a FluorPen FP 100-Max system, based instantaneous variables and fluorescence emission dynamics (JIP test). The reintroduction environment was evaluated for light conditions and canopy cover (CC). Measurements of PAR, ultraviolet radiation in band B (UVB), and total radiation (RAD) were taken at 21 days at the height of each S. crispa plant, using a radiometer Delta Ohm HD

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2302.0 Light Meter. Canopy cover, canopy opening (CO), and coverage index (CaCo) data were obtained through hemispherical images taken using a smartphone camera incorporating a 180° fisheye lens. The images were processed with the aid of GLAMA software (Tichý, 2016). The experimental design was completely randomized, arranged in a 3 × 4 factorial scheme with three substrates, four times, and 10 replications of one plant each. The fluorescence

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variables were evaluated by analysis of variance and the averages were then compared using Tukey’s test set at 5% probability. The radiation and canopy coverage data were subjected to

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Pearson correlation analysis, followed by a t-test with probabilities set at 1% and 5%.

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RESULTS

At 21 days after reintroduction, S crispa plants showed 100% survival, irrespective of

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the substrate used. Thermographic images of S. crispa individuals on each substrate are shown in Figure 2. In general, the temperature of the S. crispa plants differed from that of the host phorophytes, irrespective of the substrate used.

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The largest variation (DT) between the temperature of the phorophytes and orchid plants was recorded on the day of seedling reintroduction, during which they were removed from the

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protected culture environment and taken to the natural environment. The highest average for this variable was observed in the coconut fiber substrate (CF = 5.37°C), followed by the buriti stem substrate (BU = 2.97°C) and the treatment without substrate (WS = 2.53°C). At 7 and 14 days after reintroduction, we recorded decreases in the DT values, whereas a subsequent increase was recorded after 21 days (Figure 3). During the experiment, no large thermal variations were observed in the environment, as relative humidity (RH) and precipitation varied throughout the experimental period (Figure 4). Moreover, we observed a lower positive correlation between the averages of DT and the

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average temperature of the environment (Pearson correlation = 0.74 and R2 = 0.56) and a higher negative correlation (Pearson correlation = -0.90 and R2 = 0.81) with RH. The higher thermal variations detected at the beginning of the reintroduction of S. crispa reflect a further factor that should be considered as a potential source stress, namely the change from the ex situ acclimatization environment to the natural environment, which is detectable using thermographic techniques. In the forest fragment into which the S. crispa plants were reintroduced, the CO, CC, and CaCo values varied by more than 50% in the environment immediately below each phorophyte (Figure 5 A-C). The radiation conditions showed similar behavior, with the microenvironment of some phorophytes being characterized by high radiation (PAR higher than

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150 μmol m-2 s-1, UVB higher than 70 Wm-2, and RAD higher than 250 Wm-2) and that of other phorophytes by low irradiance (Figure 5 D-F).

A comparison of the curves obtained for the canopy and radiation data revealed a lack of correspondence between these parameters among different phorophytes, which was

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confirmed by Pearson correlation analysis (Table 1). We detected correlations only between the canopy variables or between the radiation variables, with the highest negative correlation

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observed being that between CC and CO (-0.99), and the highest positive correlation being that between CaCo and CC (0.98). The dispersion of data points shown in Figure 6 indicates the

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high linearity of the canopy data, whereas no linearity was observed for the relationship between canopy and radiation variables (Figure 6). Data relating to canopy structure and radiation at the site of S. crispa reintroduction

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reveal a considerable heterogeneity of conditions, characterized by different micro-habitats, even within this generally well-lit environment (mean of PAR 62.09 μmol m-2 s-1; UVB 35.55 Wm-2 and RAD 92.9 Wm-2), considering the reference values for the full-light conditions (PAR

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1.264 μmol m-2 s-1; UVB 638 Wm-2 and RAD 1,825.3 Wm-2). Thus, the development stage of the phorophytes in the fragment provided a cover characteristic of the forest in the initial stages

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of succession, given that sub-forest environments in an already established semi-deciduous forest have a PAR of between 5 and 10 μmol m-2 s-1. With regards to the variables that contribute to the fluorescence emission of chlorophyll

a (FChlo-a), although we detected no significant difference in the potential quantum efficiency of photosystem II (PSII: Fv/Fm), significant differences were observed for initial fluorescence (F0), variable fluorescence (Fv), fluorescence intensity in the J-step (2 μs) (Fj), and maximum fluorescence (Fm), with the highest values being recorded for the coconut fiber substrate (CF),

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which did not differ significantly from those obtained for the buriti stem substrate (BU), and the lowest values being obtained for plants without substrate (WS) (Table 2). Although differences were observed in most of the instantaneous variables of FChlo-a during the experimental period, the different substrates had no apparent effect on the time variables Vj and Vi or quantum efficiency. Similarly, we detected no significant differences among the treatments for the parameters of performance and the index of phenomenological flow (Figure 7), thereby indicating the adequate photosynthetic capacity of reintroduced plants using the available PAR energy in the underbrush. Even though data obtained for the main instantaneous variables or phenomenological

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flow did not indicate environmental stress, and despite the lack any significant differences among the plants grown in different substrates used in the reintroduction of S. crispa, our evaluation of the dynamics of FChlo-a (Figure 8 A-D) revealed variations in the sigmocity of the curves between treatments, with greater uniformity between the averages obtained at 7 days after reintroduction (Figure 8B) and increases in the intensity of fluorescence emission for CF

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DISCUSSION

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over experimental time (Figure 8 C-D).

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Even though the orchid plants used in this study were produced using asymbiotic sowing, the percentage survival was satisfactory. The survival of orchids of this species, even under sub-optimal conditions, may be related to their crassulacean acid metabolism (CAM),

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which is associated with efficient cost/benefit relationships with regards to water consumption and CO2 absorption (Silvera et al., 2010). Other authors have, however, reported that the reintroduction of some species of

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Orchidaceae into their native environments does not ensure survival, and thus it may be necessary to establish symbiotic associations prior to reintroduction, as observed for Vanda

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coerulea Griff. ex Lindl., which showed 80% and 10% survival after symbiotic and asymbiotic propagation, respectively (Aggarwal, Nirmala, Beri, Rastogi, & Adholeya, 2012). On the basis of the findings of the present study, it can be inferred that S. crispa produced in vitro and acclimatized ex situ showed high survival at 21 days after reintroduction, even without symbiotic propagation. The thermographic images obtained in the present study revealed thermal variations between orchid plants and host phorophytes. Thermography has been applied as a tool to evaluate plant–environment interactions, as variations in the temperature of plants are

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associated with the different factors that contribute to environmental stress (Costa, Grant, & Chaves, 2013). Foliar temperature (Tf) is recognized as an indicator of the water status of plants and is therefore considered a potential tool for characterizing functional status under the most diverse cultivation conditions. Tf is directly related to stomatal behavior, with an elevation in Tf values being indicative of stress-related stomatal closure, which is readily measured using infrared thermographic cameras (Chaves et al., 2010; Costa et al., 2013). Thus, thermography can be an effective alternative to conventional evaluations of stress in orchids, given that porometry and other methodologies based on evaluations of gas flow through leaves have certain limitations, owing to the morpho-anatomical characteristics common to those species

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exhibiting crassulacean acid metabolism. Surfaces in the underbrush are associated with the heterogeneity of this environment. An elevation in surface temperature, for example, can be determined by the duration of the incidence of solar radiation that penetrates the vegetation cover, the latter of which can vary in density according to the deciduous behavior of species and seasonality (Larcher, 2006).

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The coloring of a surface, as well as its water content, can also interfere with the way in which it absorbs light energy and dissipates a portion of this in the form of infrared radiation

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(Ponzoni, Pacheco, Santos, & Andrades Filho, 2015). Darker surfaces and those with lower water contents tend to absorb energy across a greater amplitude of the radiation spectrum

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(Kirschbaum et al., 2011), dissipating less of it in the vaporization of the water, and thus being characterized by higher temperatures. Thus, contingent upon the incident light, the darker the tone of phorophyte bark and the lower its water content, the higher will be its temperature,

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consequently influencing the thermal difference between the leaf temperature (Tf) of epiphytic orchids and the host phorophyte.

Given the ideal cultivation temperature of 25ºC for Orchidaceae species (Muller,

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Dewes, Karsten, Schuelter, & Stefanello, 2007), which we maintained throughout the entire experimental period, factors inherent to the bark of the phorophyte, such as color, texture,

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content of water, and temperature, should be taken into consideration when selecting appropriate host phorophytes for the reintroduction of S. crispa. In addition to ambient temperature, stomatal control (and consequently Tf and DT) is

also influenced by light conditions. For plants adapted to underbrush conditions, the canopy coverage provided by the phorophytes is a determinant of the radiation conditions available throughout the day. New tools for evaluating conditions of the underbrush include the use of hemispheric photographs that can be obtained using Android smartphone cameras incorporating “fisheye” lenses (Tichý, 2016).

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Through analysis of such images, it can be determined that the highest correlation between CC and CO obtained in the present study is explained by an inverse relationship between these two variables, since CO reflects the white (open sky) among other pixels of the digital hemispheric image, whereas CC reflects the proportion in the field of centralized analysis (Figure 5B), which is obscured by the canopy. The high positive correlation between CaCo and CC implies a direct relationship between these variables, given that CaCo is also an index of canopy cover, although it is more appropriately used for evaluations of the variation in canopy density; for example, in ecological studies in which photographs are taken at 6-month intervals (Tichý, 2016).

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With regards to photosynthetic variables, an increase in Fo accompanied by a reduction in Fm is typically associated with stress responses (Mathur, Jajoo, Mehta, & Bharti, 2011). However, in the present study, we found that S. crispa plants subjected to the CF treatment, which showed a higher Fo, also showed the highest average Fm, which compensated for the relationship between the variables associated with emission dynamics (Fj and Fi), thereby

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decharacterizing the possibility of damage to the PSII. The Fv/Fm ratio indicates the conditions of the leaf photosynthetic apparatus in terms of the quantum photochemical yield of the primary

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photosynthesis photochemical steps, with values between 0.75 and 0.85 being indicative of efficient transfer of light energy to the PSII complex (Bolhàr-Nordenkampf et al., 1989).

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Under low light intensity (less than 100 μmol m-2 s-1 of radiation), more than 80% of the absorbed quantum energy can be used for photosynthesis (Björkman & Demmig-Adams, 1987), whereas when the light intensity approaches 1000 μm m-2 s-1 (approx. 50% of available

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sunlight), less than 25% of the absorbed quantum energy is used, and under conditions of full sunlight, this rate decreases to 10% (Long, Humphries, & Falkowski, 1994). Endres Júnior et al. (2015), who studied the reintroduction of Cattleya intermedia Graham

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on the border and interior of a fragment of seasonal semi-deciduous forest, reported that a higher percentage of plants reintroduced at the edge of the fragment, where the incidence of solar

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radiation was higher relative to that in the interior, developed roots on the trunk of phorophytes. In the present study, there was a high incidence of light in the underbrush of the reintroduction site and the plants of S. crispa also showed good development, with the formation of new roots and leaves. In general, these observations indicate that when reintroduced under conducive light conditions, orchids are able to develop characteristics important for their survival in the natural environment. Duarte & Gandolfi (2013) have emphasized that, in Brazil, restoration ecology is still very much in its infancy, and accordingly there is currently a lack of empirical data to guide the

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introduction, reintroduction, and transplantation of epiphytes into forests in restoration programs. The authors also point out that only since 2000 has it been recognized that evaluation and monitoring of forests in the process of restoration is fundamental to guarantee the success of the restoration projects already in place. Thus, the need to enrich these environments has become evident, since many are small isolated from fragments. Cardoso, Silva, & Vendrame (2016), in their study on the impact of deforestation on orchids in the state of São Paulo, stated that a change in Brazilian environmental legislation is necessary to enable definition of the success of biodiversity conservation. In addition, it is necessary to define the number of species of orchids that are vulnerable or endangered, their

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richness and diversity, and how these species can be reintroduced and restored in reforested areas.

With regards to the hypothesis that different species of epiphyte may show a preference for certain phorophytes, we found that plants of S. crispa established satisfactorily independent of the species of phorophytes onto which they were transferred. Consistent with these

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observations, Duarte & Gandolfi (2013) suggest that, ultimately, the characteristics of the epiphytes to be transplanted may be more important than those of the host trees, due in part to

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environmental heterogeneity. Thus, with respect to reintroduction, it is necessary to select species of orchids with characteristics that facilitate favorable development within the

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limitations of the epiphytic environment, be they thermal, hydric, or photic. In the context of conservation, objective 15 listed in the Sustainable Development Objectives of Agenda 2030 deals with the conservation of terrestrial life and is aimed at

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protecting, recovering, and promoting the sustainable use of terrestrial ecosystems; managing forests sustainably; combating desertification; halting and reversing land degradation; and halting the loss of biodiversity. In its goals, this objective emphasizes that it is necessary to

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promote the implementation of sustainable management of all types of forests, halt deforestation, restore degraded forests, and substantially increase afforestation and reforestation

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globally by 2020 (ONU, 2019). Studies on the conservation of species of Orchidaceae in the natural environment would thus make an extremely important contribution toward realizing the objectives of this agenda.

CONCLUSION

On the basis of the results obtained in this study, which indicate that, irrespective of the type of substrate used, all the reintroduced plants of S. crispa survived, we can conclude that

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plants of this orchid derived from asymbiotic sowing can be successfully reintroduced in natural environments, thus contributing to the conservation of the species and enhancing the

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biodiversity of degraded areas.

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Segovia-Rivas, A., Meave, J. A., González, E. J., & Pérez-García, E. A. (2018). Experimental reintroduction and host preference of the microendemic and endangered orchid Barkeria whartoniana in a Mexican Tropical Dry Forest. Journal for Nature Conservation, 43, 156-164. Silvera, K., Neubig, K. M., Whitthen, W. M., Williams, M. H., Winter, K. & Cushman, J. C. (2010). Evolution along the crassulacean acid metabolism continuum. Functional Plant Biology, 37, 995-1010.

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Fietz, C. R., Fisch, G. F., Comunello, E. & Flumignan, D. L. (2017). O clima da região de Dourados, MS (2. ed.). Dourados: Embrapa Agropecuária Oeste.

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Tichý, L. (2016). Field test of canopy cover estimation by hemispherical photographs taken with a smartphone. Journal of Vegetation Science, 27, 427-435.

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Yang, F. S., Sun, A. H., Zhu, J., Downing, J., Song, X. Q. & Liu, H. (2017). Impacts of host trees and sowing conditions on germination success and a simple ex situ approach to generate symbiotic seedlings of a rare epiphytic orchid endemic to Hainan Island, China. The Botanical Review, 83, 74-86.

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Table 1. Pearson correlation coefficients (r) between the radiation conditions and canopy cover in sub-forest, in which the Schomburgkia crispa Lindl plants were reintroduced. PAR 1

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PAR

UVB

RAD

AC

CC

0.7951*

1

0.9862**

0.806*

1

AC

0.4127ns

0.3859 ns

0.3534 ns

1

CC

-0.4109 ns

-0.3859 ns

-0.3508 ns

-0.9999**

1

CaCo

-0.3858 ns

-0.3943 ns

-0.3291 ns

-0.9903**

0.9899**

UVB

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RAD

CaCo

1

** significant at 1% probability; * significant at 5% probability; ns not significant, by Student's t test. (PAR = Radiation photosynthetically active; UVB = Ultraviolet B; RAD = Total Visible Radiation; AC = Cup opening; CC = Cup cover and CaCo = Cup cover index).

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Table 2. Fluorescence intensity (Fm), Fluorescence intensity (Fm), Fluorescence intensity (Fm), Fluorescence intensity (Fm), Fluorescence intensity Relative variable fluorescence in step I (30 ms), Maximum efficiency of the photochemical process in FSII (Fv / Fo) and potential

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quantum efficiency of FSII (Fv / Fm) of plants of Schomburgkia crispa Lindl. reintroduced into three different substratum conditions: SS = No substrate, PB = Buriti pau and FC = Coconut

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fiber.

Fo

Fj

Fi

Fm

Fv

BU

276,60 ab

5648, 03 ab

1003,25 ab

1171,28 ab

11585,05 a

FC

307,27 a

6770,50 a

12649,80 a

1465,78 a

8946,78 ab

SS

244,97 b

5227, 96 b

9008,92 b

1041,88 b

7969,17 b

15

Cv (%)

16,05

18,91

18,86

18,37

20,26

Vj

Vi

Fv/Fo

Fv/Fm

BU

0,32 a

0,80 a

38,97 a

0,76 a

FC

0,32 a

0,82 a

30,01 a

0,78 a

SS

0,34 a

0,81 a

32,75 a

0,74 a

Cv (%)

12,72

4,98

75,08

4,87

Means accompanied by the same letter, within the same column, do not differ to 5% of

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probability, by Tukey’s test.

1cm

Figure 1. Plants of Schomburgkia crispa Lindl. at 21 days after reintroduction in a natural environment under different substratum conditions. A = without substrate (WS); B = Buriti palm (BU); and C = Coconut fiber (CF).

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16

A

B

17

D

E

F

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C

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1 cm Figure 2. Thermographic images of plants of Schomburgkia crispa Lindl. at 21 days after reintroduction in a natural environment under different substratum conditions. A and B =

WS

4

1

CF

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3 2

BU

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5

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Temperature Difference ºC

6

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without substrate (WS); C and D = Buriti palm (BU); and E and F = Coconut fiber (CF).

0

0

7 14 Days After Reintroduction

21

Figure 3. Temperature difference between the phorophyte and the plants of Schomburgkia crispa Lindl., during 21 days after the reintroduction of the plants in natural environment, in different substrates. SS = no substrate, BU = Buriti palm and FC = Coconut fiber.

Med. (ºC)

Precipitation (mm)

35

140

30

Precipitation (mm)

25 20

120

100 80

90

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Temperature (ºC)

RH (%)

160

15

40

5

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0

Evaluation Days

60 45 30

20

15

0

0

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10

60

75

Relative Humidity (%)

Min. (ºC)

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Max. (ºC)

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18

Evaluation Days

Figure 4. Climatic data from EMBRAPA - CPAO to Dourados - MS during the experimental

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period of reintroduction of Schomburgkia crispa Lindl plants.

19

100

A

D

200

80

CC

150

-2 -1

PAR (µmol.m .s )

90

70 60

100

50

0

50 50

120

B

40

E

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100 80

AC

-2

UVB (W.m )

30

20

60 40

10 20

0

55

300

C

50

250

45

RAD (W.m )

200 -2

35

150

F

re

CaCo

40

-p

0

30 25

100 50

20

0

2

4

6

8

10

12

Forófitos

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0

15

14

16

18

0

2

4

6

8

10

12

14

Forófitos

Figure 5. Environmental conditions of the understory in a forest fragment in which plants of

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Schomburgkia crispa Lindl. were reintroduced. A = Canopy cover (CC); B = Cup opening (AC); C = crown cover index (CaCo); D = Photosynthetically active radiation (PAR); E =

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Ultraviolet B (UVB) and F = Total Visible Radiation (RAD).

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20

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Figure 6. Scatter matrix (Pearson = r) for canopy and radiation variables in understory in the reintroduction area of Schomburgkia crispa Lindl. (PAR = Radiation photosynthetically active;

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and CaCo = Cup cover index).

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UVB = Ultraviolet B; RAD = Total Visible Radiation; AC = Cup opening; CC = Cup cover

21

3000 WS

2500

BU

CF

2000

1500 1000 500 0 ABS/RC

TRo/RC

ETo/RC

DIo/RC

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PiAbs

Figure 7. The photosynthetic performance index (PIAbs), specific absorption flow per reaction center (ABS / RC), specific capture flux per reaction center (TRo / RC), specific electron transport flux per reaction center ETo / RC) from plants of Schomburgkia crispa Lindl.

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reintroduced into three different substratum conditions: WS = Without substrate, BU = Buriti

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Palm and CF = Coconut fiber.

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22

Figure 8. Chlorophyll a fluorescence emission kinetics in Schomburgkia crispa Lindl plants.

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reintroduced into three different substratum conditions: WS = Without substrate, BU = Buriti palm and CF = Coconut fiber. A: day of reintroduction; B: seven days after reintroduction; C:

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fourteen days after reintroduction and D: twenty-one days after reintroduction.