Introduction

Forest restoration with native species has led to discussions about the ideal size of seedlings for planting (FERRAZ and ENGEL, 2011). Research by Cunha et al. (2005) suggested that one of the difficulties faced by those working in native seedlings production is that many show slow growth, particularly those classified as late or climax species.

It is expected that larger seedlings, provided with well-formed roots, have a greater chance of survival in the field, mainly due to their greater competitiveness with grasses. On the other hand, larger containers imply higher implantation costs, both due to higher substrate consumption and greater need of space in the nursery, higher transport costs and lower yields in the planting (FERRAZ and ENGEL, 2011).

In view of this, it is of fundamental importance to define protocols and strategies that favor the production of seedlings of high quality that take less time to produce and in conditions accessible to small and medium rural producers, who are the most interested public for this type of input ( CUNHA et al., 2005).

The in-nursery vegetative propagation of the Cerrado species is still under studied, however, and it is beginning to arouse interest. Moreover, several methods of asexual propagation, including cutting, have already been tested with the Cerrado species and recent studies have achieved promising results (SANO et al., 2008).

For several species, cutting is one of the main processes of producing good quality seedlings. Despite this type of propagation tending to decrease biodiversity in the environment, the method is very promising for the recovery of degraded areas, especially for species with seed production difficulty (OLIVEIRA and RIBEIRO, 2013).

However, with respect to native species, little is known about the need for germination and emergence of seedlings of each species, as well as what the substrate characteristics are for the production of high quality seedlings (BAO et al., 2014). Thus, the choice of a substrate should consider the technical aspects, but also the local availability of the container to be used (CUNHA et al., 2005).

The proper choice of native species Miconia albicans, Croton urucurana and Solanum paniculatum for recovery of degraded areas is essential as they are suitable to the local ecological conditions and food of the local fauna. Of note, the species were classified as zoocoric dispersion (dispersed by animals) and autocore (self-dispersion), according to Van der Pijl (2012).

Croton urucurana Baill. 1864, from the Euphorbiaceae family, known as Sangra d'agua, has a height of 7 to 14 m, acts as a pioneer, grows in very humid and swampy terrain, mainly of semideciduous broadleaf forest and is excellent for mixed plantations in degraded riparian areas (LORENZI, 2002). Croton urucurana, on the other hand, presents fruits of spontaneous opening (explosive dehiscence), being classified as autocoric (CALEGARI et al., 2013).

Miconia albicans Steud. 1841, of the family Melastomataceae, commonly called White Leaf, Lares-fake or Maria-branca is characteristic of Cerrado (SIQUEIRA, 1988), being common in forest edges (NERI et al., 2005). It is shrub that grows up to 2 , 5 meters high, has coriaceous leaves with densely tomentosa lower face (CARREIRA, 2004). Miconia is one of the major genera of Melastomataceae in Brazil and its species are common in forest and cerrado areas, where Miconia stenostachya and Miconia albicans (SOUZA and LORENZI, 2012) stand out. Miconia albicans also presents zoocoric dispersion by birds and small mammals of Cerrado (APPROBATO and GODOY, 2006).

Solanum paniculatum L. 1762, of the family Solanaceae (GARCIA et al., 2008), commonly known as jurubeba, is a shrub reaching roughly 2 m in height, displays a branched structure with alternate leaves, petiolate and having darker shade on the upper face, which is covered by hairs and thorns. It is native to almost all of Brazil, growing spontaneously in degraded lands, mainly Cerrado, pastures, vacant lots and roadside (LORENZI, 2000; NUNES and ARAUJO, 2003). It too shows predominant dispersion via zoocoria, mainly by bats and birds (SAMPAIO, 2013).Considering all the arguments mentioned above, the objective of this work was to produce seedlings of native species by induction of rooting and foliar sprouting in stakes of Miconia albicans, Croton urucurana and Solanum paniculatum.

Material and methods

Study area

The study was carried out at Instituto São Vicente (20 ° 23'16.80 ''S and 54° 36'36.30''W), at 661 m altitude, approximately 10 km from the city center, located in the urban perimeter of the region of Lagoa da Cruz in Campo Grande, MS.

The total area is 191 hectares, of which 20 hectares are allocated to the legal reserve area. Approximately 30 hectares of the region are destined to for agriculture, with planting of corn, beans, soybean and araruta among others. There are also pastures for farming of sheep, goats, horses, cattle and pigs as well as a pisciculture tank.

Planting Stakes

Apical cuttings of five Miconia albicans, Croton urucurana and Solanum paniculatum of roughly 15 cm long were collected onto which a bevel cut was made at the basal end of the cutting followed by leaf removal. For species identification, exsicates were sent to the herbarium of the Federal University of Mato Grosso do Sul (UFMS), where the material was deposited in the herbarium CGMS with the following discrimination CGMS 51397, CGMS 51398 and CGMS 51399.

The apical stakes were placed in 20x30 cm polyethylene bag containers containing 180 mL of water (mini greenhouse) closed with a knot at the end (Figure 1).

Figure 1.  Apical cuttings of Solanum paniculatum. (A) no use of regulator (B), 15 days after treatment (C) with phytoregulator (IBA), (D) after 15 days of treatment with IBA and (E) container used for treatments.

The treatments were: T1) with the application of indolbutyric acid phytoregulator (AIB) with a concentration of 0.5%, on the basis thereof; T2) the control (without IBA), and in two types of substrates: I) Cerrado Land and II) Sand. In total, 120 stakes were used, 10 replicates in each treatment.

The cuttings were kept in the deactivated physical structure used for the wormhole. Before the test installation, chemical analyses of both soils were carried out (Table 1).

Table 1. Chemical characteristics of the soil used in the production of seedlings.

* Soil chemical analysis performed by the Laboratory of Soils and Nutrition of Plants of Fazenda Escola / UCDB. P and K: Mehlichl; MO: Colorimetry; Ca, Mg and Al: 1M KCl; H + Al: Calcium Acetate pH 7.

The evaluation parameters were the numbers of rooted cuttings and number of leaves in each treatment. The observation of the emission of roots, buds and leaves were counted every 15 days in order to minimize damage to the roots at the moment of handling the polyethylene bag. The observation period was over 208 days.

Statistical Analyzes

  The percentage of difference between treatments was analyzed using the statistical software SISVAR (FERREIRA, 2000). Comparison of non-parametric Chi-square means (ZAR, 1999) between different treatments of the same species and for each species was performed to verify the difference between the observed periods (15-day interval). When the analysis indicated that there was at least one difference, the means were then compared by the Tukey test at 95%.

Results

The substrates were not significant and did not affect the establishment between M. albicans, C. urucurana and S. paniculatum (Table 1).

Regardless of the indolbutyric acid in the propagation of cuttings of M. Albicans, C. Urucurana and S. Paniculatum, all of them presented buds, where the highest percentage of budding occurred in S. paniculatum. Tukey test analysis indicated that the number of leaves was significant since on average 2.3 leaves were observed for S. paniculatum while none were observed for cuttings of M. albicans, C. urucurana. (Figure 2).

Figure 2. Average number of buds (A) and average number of leaves (B) of Miconia albicans, Croton urucurana and Solanum paniculatum cuttings. Test Tukey, p <0.05.

The rooting was significant between M. albicans, C. urucurana and S. paniculatum with or without the use of the AIB phytoregulator (Table 2). However, Solanum paniculatum showed shoot, leaf and root emission, independent of substrates and IBA treatment.

Table 2. Root length cuttings for Miconia albicans, Croton urucurana and Solanum paniculatum, with or without IBA phytoregulator.

* Means followed by the same letter in the column do not differ statistically from each other, by the Tukey test P <0.05. CV (%) = 30.70

Discussion

Bao et al. (2014) verified that sand, earth and APR (earth, sand and manure) in containers such as black polyethylene bag (15 x 25 cm), tubes (2.60 cm in diameter x 13 cm in height) and polystyrene tray (128 cells in dimensions of 680 mm x 340 mm x 60  mm in depth) were not suitable for seedling emergence. As for the number of leaves per plant in apical papaya cuttings (Carica papaya L.), no significant differences were observed between the substrates used (NEGREIROS et al., 2005). Our study also did show significant differences for the substrates, suggesting that land of Cerrado and sand could be appropriate for the production of seedlings of the studied species.

 The reduction of expenses with water and containers used is advantageous for rural producers and nursery workers, with a seedling acquisition in a shorter period of time.

According to the propagation technique adopted in this study, the use of transparent polythene bags in the form of a mini greenhouse using only 180 ml of water can be used for the same period of time during the whole seedling period. Some species can then use different containers at each stage of development, until the formation of a molt (DA SILVA et al., 2009).

On average, roughly R$ 4000 is spent irrigation roughly 400 000 coffee seedlings in nurseries per month in a medium sized nursery (CARVALHO et al., 2013). The price of a molt is $ 1.40, if it is silent clonal. However, if seeds are purchased, the price decreases to R$ 0.35.

Different types of containers are used for the production of seedlings (plastic tubing, trays, plastic bags) and these are made of different materials (including expanded polystyrene, polypropylene) and range in sizes, such as specially expanded polystyrene trays which are most commonly used for the production of seedlings (FILGUEIRA, 2008).

The method with closed polyethylene bag containing only 180 mL of water provided production of asexual propagation seedlings, and it can be stated that it decreased the time and cost of several methods to obtain S. paniculatum seedlings. However for jurubeba, the plastic bag was a container that, in general, allowed better development conditions for seedlings (GUIMARAES et al., 2012).

Indeed, following the first 15 days in the treatment without IBA, sprouts were observed in M. albicans, C. urucurana and S. paniculatum stakes. Leaves and roots were only observed in S. paniculatum, suggesting that eliminating phytoregulators could decrease costs.

According to Dias (2012), the supply of exogenous auxin can promote hormonal alteration, favoring or not rooting. Similar results of rooting without application of IBA were recorded in several studies, such as Hernandez et al. (2013), where they indicated that IBA had little influence on the vegetative propagation of the jequitibá-rosa (Cariniana estrellensis) by cuttings. The species Cestrum laevigatum and Salix humbolditiana studied by Santos et al. (2011) can also be easily propagated by wood cuttings independently of IBA. Oliveira and Ribeiro (2013) in their study, with different application methods, IBA concentrations and Benlate 500 (systemic fungicide) did not allow for rooting of Euplassa inaequalis.

Ohland et al. (2009) observed greater rooting for Ficus carica cuttings collected in August without treatment with IBA (42.5%). However, when measuring rooting ability of cuttings taken in different months subjected to the basal treatment of 2000 mg/L of IBA, only cuttings collected in June showed a 70% increase in rooting, with no differences between for cuttings collected in other collection seasons.

Importantly, the stakes of M. albicans, C. urucurana and S. paniculatum without IBA treatment were collected in February, and those with 0.5% IBA treatment were collected in June.

The main factors affecting the rooting of cuttings are the physiological conditions of the matrix plant, such as the presence of carbohydrates, nitrogenous substances, amino acids, auxins, phenolic compounds and other substances, the period and position of collection of the stakes, youthfulness of the plant, oiling, the presence of leaves and gems, the age of the matrix plant and environmental factors, such as water availability, light incidence and substrate (HARTMANN et al., 2002; HESS, 1969). This may explain the absence of rooting in M. albicans and C. urucurana, as the collection of their cuttings in the vegetative period did not favor the rooting. Since auxin requires carbohydrates for the production of nucleic acids and proteins, thus requiring energy for the production of roots (FACHINELLO et al., 1994, HARTMAN et al., 2002).

Therefore, it is important to collect apical cuttings at the beginning or end of the dormancy due to the presence of photoassimilates in the branches, as observed by Ohland et al. (2009) and Hartmann et al. (2002), among others. However, this information does not corroborate the results found for S. paniculatum, which, sprouted, emitted leaves and rooted despite being an suboptimal collection period. The same was observed for two bamboo species (Bambusa vulgaris and B. vulgaris var. Vitatta) (GASPARETTO et al., 2013).

According to Hartmann et al. (2002), it is of paramount importance to use the correct phytoregulator concentrations at the base of the cuttings and this ideal concentration varies within species. In contrast, Stuepp et al. (2013) suggested that the rooting of cuttings of Melaleuca alternifolia was not influenced by the position of the branches in which the cuttings are removed nor by the AIB concentrations tested.

A number of successful and unsuccessful attempts to propagate Baru (Dypterix alata), Jatobá (Hymenaea stigonocarpa), White Ipê (Tabebuia roseoalba), Ingá banana (Inga laurina), Water bleed (Croton urucurana), Pororoca guianensis), Grapevine (Serjania hebecarpa), Astrapéia (Dombeya wallichii) and Cambotã (Matayba guianensis) performed by the authors of this work, asexually, by cuttings with phytoregulators and stimulants (GONDIM, unpublished data).

Santos et al. (2011) in their study showed that species such as C. laevigatum and S. humboldtiana can be easily propagated by wood cuttings for all classes of diameters tested and independently of IBA application. Furthermore, their study was carried out in order to evaluate the effect of rooting on Crotalus urucurana, Sebastiana scottiana, Ficus adathodigifolia, Ficus citrifolia, Ficus citrifolia, Nectandra nitidula, Schinus terebintifolius and Siparuna guianensis and the results were promising.

The time of year is directly related to the consistency of the stake, wherein stakes collected in the period of intense vegetative growth (spring / summer) are more herbaceous and, generally speaking, show higher rooting capacity when collected at that time, while cuttings collected in winter have a higher degree of lignification and tend to root less (FACHINELLO et al., 2005). Also, depending on the species, in the case of wood cuttings, there is a higher concentration of phenolic compounds, which inhibit or delay any manifestation of budding.

Conclusion

Our results indicate that rooting and sprouting of M. Albicans, C. Urucurana and S. Paniculatum could occur from wood cuttings independently of IBA with a high percentage of rooting for S. Paniculatum. Furthermore, the vegetative propagation via cutting without the use of regulators is feasible for S. Paniculatum, independent of the time of the stake collection.

Acknowledgement

We would like to thank the São Vicente Institute for the availability of the study area and the herbarium of the Federal University of Mato Grosso do Sul (UFMS) for the aid in botanical identification.

References

Approbato, A. U., & Godoy, S. A. P. (2006). Survey of diaspores in Cerrado areas in the Municipality of Luiz Antônio, SP. Hoehnea, 33(3), 385–401.

Bao, F., Lima, L. B., & Luz, P. B. (2014). Morphological characterization of branch, seeds and seedlings of Matayba guianensis Aubl. and seedling production in different containers and substrates. Tree Review, 38(1), 63–71.

Calegari, L., Martins, S. V., Campos, L. C., Silva, E., & Gleriani, J. M. (2013). Evaluation of soil seed bank for forest restoration in Carandaí, MG. Revista Árvore, 37(5), 871–880.

Carreira, R. C. (2004). Germination in seeds of Miconia albicans (Sw.) Triana and Miconia rubiginosa (Bonpl.) DC. Melastomataceae, from the cerrado of Mogi Guaçu, SP. Master’s dissertation.

Carvalho, C. H. S., Paiva, A. C. R. S., Silva, E. Q., & Custodio, A. A. (2013). Cost of production of clonal Arabica coffee seedlings produced by somatic embryogenesis. Technical Circular, 3. Brasília, DF.

Cunha, A. O., Andrade, L. A., Bruno, R. L. A., Silva, J. A. L., & Souza, V. C. (2005). Effects of substrates and vessel sizes on seedling quality of Tabebuia impetiginosa (Mart. Ex D.C.) Standl. Revista Árvore, 29(4), 507–516.

Da Silva, E. A., Maruyama, W. I., de Oliveira, A. C., & Dógenes, M. B. (2009). Effect of different substrates on the production of mangabeira seedlings (Hancornia speciosa). Revista Brasileira de Fruticultura, 31(3).

Dos Santos, J. P., Davide, A. C., Teixeira, L. A. F., Melo, A. J. S., & Melo, L. A. (2011). Rooting of woody cuttings of forest species. Hoehnea, 17(3), 293–301.

Facchinello, J. C., Hoffmann, A., & Santos, A. M. (1994). Amoreira-preta, raspberry, and blueberry: Small fruits for the south of Brazil. In Brazilian Fruit Congress, 13, Salvador (Vol. 3, pp. 989–990). Brazilian Society of Fruticulture.

Fachinello, J. C., Hoffmann, A., & Nachtigal, J. C. (2005). Propagation of fruit plants. Brasília, DF: Embrapa.

Ferraz, A. V., & Engel, V. L. (2011). Effect of tubet size on seedling quality of Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang., Tabebuia chrysotricha (Mart. Ex DC.) Sandl., and Parapiptadenia rigida (Benth.) Brenan. Tree Review, 35(3), 413–423.

Ferreira, D. F. (2000). Statistical analysis through SISVAR (System for Analysis of Variance) for Windows version 4.0. In Annual Meeting of the Brazilian Region of the International Society of Biometry, 45, São Carlos (pp. 255–258). UFSCar.

Filgueira, F. A. R. (2000). New manual of olericultura: Modern agrotechnology in the production and commercialization of vegetables. Minas Gerais: UFV.

Garcia, J., Jacobson, T. K. B., Farias, J. G., & Boaventura, R. F. (2008). Effectiveness of methods to increase the germination rate of Solanum paniculatum L. seeds. Pesquisa Agropecuária Tropical, 38, 223–226.

Gasparetto, G. A., Davalo, M. J., & Rondon, J. N. (2013). Decreased production and acclimatization time of two bamboo species under greenhouse conditions. Biotemas, 26(1), 17–23.

Guimarães, M. A., Garcia, M. F. N., Damasceno, L. A., & Viana, C. S. (2012). Production of cocona and jurubeba seedlings in different types of containers. Horticultura Brasileira, 30(4), 720–725.

Hartmann, H. T., Kester, D. E., Davies, F. T. Jr., & Geneve, R. L. (2002). Plant propagation: Principles and practices (7th ed.). New Jersey: Prentice Hall.

Hernandez, W., Xavier, A., Paiva, H. N., & Wendling, I. (2013). Vegetative propagation of the pink jequitibá (Cariniana estrellensis (Raddi) Kuntze) by cuttings. Tree Review, 37(5), 955–967.

Hess, C. E. (1969). Internal and external factors regulating root initiation: Root growth. London: Butterworths.

Lorenzi, H. (2000). Weeds of Brazil - Terrestrial, aquatic, parasitic and toxic (3rd ed.). Nova Odessa: Plantarum Institute.

Lorenzi, H. (2002). Brazilian trees: A manual for identification and cultivation of native tree plants in Brazil (4th ed., Vol. 1). Nova Odessa: Plantarum Institute.

Neri, A. V., Campos, E. P., Duarte, T. M., Neto, J. A. A. M., Silva, A. F., & Valente, G. E. (2005). Regeneration of native woody species under Eucalyptus plantation in the Cerrado area of the Paraopeba National Forest, MG, Brazil. Acta Botanica Brasilica, 19(2), 369–376.

Nunes, A. P. M., & Araujo, A. C. (2003). No genotoxicity of stevioside in E. coli. In X Week of Scientific Initiation of UERJ, Rio de Janeiro (p. 15).

Ohland, T., Worse, E., Chagas, E. A., Barbosa, W., Kotz, T. E., & Daneluz, E. (2009). Rooting of apical cuttings of 'Roxo de Valinhos' fig tree as a function of the collection time and IBA. Ciência e Agrotecnologia, 33(1), 74–78.

Oliveira, M. C., & Ribeiro, J. F. (2009). Rooting of cuttings of Euplassa inaequalis (Pohl) Engl. of gallery forest in different seasons of the year. Bioscience Journal, 25(2).