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Does the presence of leaves on coleus stem cuttings influence their rooting?

A presença de folhas apicais nas estacas de coleus influencia o enraizamento?

ABSTRACT

Plectranthus scutellarioides (L.) R. Br. is part of the Lamiaceae family and is commonly known as coleus, due to the genus denomination in the past. This ornamental plant species has been gaining importance in the floristic industry due to the great number of exotic cultivars available in the market. Coleus commercial exploitation by seeds is held up by genetic instability, which makes it usually being propagated through semi-hardwood stem cuttings and leaves have great importance in stem cuttings adventitious rooting. The objective of this study was to evaluate the presence or absence of apical leaves and the use of indolebutyric acid (IBA) in coleus stem cuttings rooting. Semi-hardwood coleus stem cuttings with 6 cm long were prepared with or without apical leaves, and then treated with hydroalcoholic solutions (50% v v-1) of 0, 1000 and 2,000 mg L-1 IBA. The propagules were planted in tubes containing vermiculite and kept for 30 days in a greenhouse under intermittent misting (24 ± 2 °C, Relative Humidity 90%) until final evaluation. The experiment was conducted in a completely randomized design, in a 3×2 factorial scheme (3 IBA concentrations x presence and absence of apical leaves). The rooting was evaluated after 30 days. Stem cuttings with and without apical leaves has 100 and 57% rooting, respectively. Stem cuttings rooting did not vary according to IBA concentrations. Roots length and number were higher in stem cuttings with apical leaves in comparison to leafless ones, regardless IBA concentrations. On average, 97.7% sprouting was observed in cuttings with apical leaves, significantly higher rates when compared to the average of 2.2% on leafless cuttings. The presence of apical leaves is fundamental for coleus stem cuttings rooting and the use of IBA is not required for this species propagation.

Keywords:
Plectranthus scutellarioides; auxin; ornamental; rhizogenesis; vegetative propagation

RESUMO

Plectranthus scutellarioides (L.) R. Br. pertence à família Lamiaceae e é comumente conhecida como coleus, devido à antiga denominação do gênero. Essa planta ornamental vem ganhando destaque na indústria florística em função do grande número de cultivares exóticas no mercado. Sua exploração comercial por meio de sementes é dificultada pela instabilidade genética, sendo propagada vegetativamente com estacas semilenhosas e as folhas têm grande importância no enraizamento de estacas. O objetivo deste trabalho foi avaliar a presença e ausência de folhas apicais e do uso de ácido indol butírico (IBA) no enraizamento de estacas caulinares de coleus. Estacas semilenhosas de coleus com 6 cm de comprimento, com e sem de folhas apicais, tratadas em solução hidroalcoólica (50%) de 0, 1000 e 2000 mg L-1 de IBA foram confeccionadas. Os propágulos foram estaqueados em tubetes com vermiculita e mantidos em casa de vegetação (24 ± 2 °C e Umidade Relativa do ar 90%). O enraizamento foi avaliado após 30 dias. As estacas com folhas e sem folhas tiveram 100% e 57% de enraizamento, respectivamente, independente da concentração de IBA. O comprimento e o número de raízes nas estacas com folhas foram maiores que às estacas sem folhas em todas as concentrações de IBA. A brotação das estacas com folhas e sem folhas foi de 97,7% e 2,2%, respectivamente. A presença de folhas apicais aumenta o enraizamento de estacas de coleus e o uso de IBA é desnecessário para a propagação da espécie.

Palavras-chave:
Plectranthus scutellarioides; auxina; ornamental; rizogênese; propagação vegetativa

1. INTRODUCTION

Plectranthus scutellarioides (L.) R. Br., whose scientific synonyms are Coleus blumei Benth. (SYTAR et al., 2015SYTAR, O.; BRUCKOVA, K.; HUNKOVA, E.; ZIVCAK, M.; KONATE, K.; BRESTIC, M. The application of multiplex fluorimetric sensor for the analysis of flavonoids content in the medicinal herbs family Asteraceae, Lamiaceae, Rosaceae. Biological Research, v.48, n.5, 2015. DOI: https://doi.org/10.1186/0717-6287-48-5
https://doi.org/10.1186/0717-6287-48-5...
) and Solenostemon scutellarioides (L.) Codd (YOKO et al., 2015YOKO, I.; TAEKO, T.; YOSUKE, M.; YOSHIAKI, C.; MAMORU, S. Downy mildew of coleus caused by Peronospora belbahrii in Japan. Journal of General Plant Pathology, v.81, n.4, p.328-330, 2015. DOI: http://dx.doi.org/10.1007/s10327-015-0601-3
http://dx.doi.org/10.1007/s10327-015-060...
), is popularly known as coleus (DATILES and ACEVEDO-RODRÍGUEZ, 2014DATILES, M.J.; ACEVEDO-RODRÍGUEZ, P. Plectranthus scutellarioides (coleus). CABI Invasive Species Compendium. 2014. Available in: <https://www.cabi.org/isc/datasheet/118545#470F50C9-6BF2-43C0-9187-941E084D1E7F> Accessed on: 16th March 2018.
https://www.cabi.org/isc/datasheet/11854...
).

This species belongs to the Lamiaceae family, characterized by the abundance of medicinal and ornamental plants. Coleus is mainly used for landscape composition (JUDD et al., 2009JUDD, W.S.; CAMPBELL, C.S.; KELLOGG, E.A.; STEVENS, P.F.; DONOGHUE, M.J. Sistemática vegetal: um enfoque filogenético. 3ed. Porto Alegre: Artmed, 2009. 605p.). In Ouro Preto, Minas Gerais State, Brazil, the plant is popularly known as Christ-blood, and is used for preparation of soothing teas by leaves infusion (MESSIAS et al., 2015MESSIAS, M.C.T.B.; MENEGATTO, M.F.; PRADO, A.C.C.; SANTOS, B.R.; GUIMARÃES, M.F.M. Uso popular de plantas medicinais e perfil socioeconômico dos usuários: um estudo em área urbana em Ouro Preto, MG, Brasil. Revista Brasileira de Plantas Medicinais, v.17, n.1, p.76-104, 2015. DOI: http://dx.doi.org/10.1590/1983-084X/12_139
http://dx.doi.org/10.1590/1983-084X/12_1...
). The species can also be considered a hallucinogenic plant (DUKE, 2002DUKE, J.A. Handbook of Medicinal Herbs, 2ed., Baton Rouge: CRC Press, 2002. 896p.). Another potential application for coleus plant is in phytoremediation of water contaminated by metals, since it showed remarkable capacity for aluminum extraction and accumulation (PANIZZA et al., 2011PANIZZA, D.L.A.; GONZÁLEZ, R.C.; GONZÁLEZ, M.B.; MIER, M.V.; BAZÚA, C.D.D. Exploration of the ability of Coleus blumei to accumulate aluminum, International Journal of Phytoremediation, v.13, n.5, p.421-433, 2011. DOI: https://doi.org/10.1080/15226514.2010.483263
https://doi.org/10.1080/15226514.2010.48...
).

Coleus is characterized as perennial and vigorous foliage with upright, semi-upright, prostrate, semi-trailing, and trailing habits of growth (NGUYEN et al., 2008NGUYEN, P.; QUESENBERRY, K.; LAERK, D. Genetics of growth habit and development of new coleus (Solenostemon scutellarioides (L.) Codd) varieties with trailing habit and bright color. Journal of Heredity, v.99, n.6, p.573-580, 2008. DOI: https://doi.org/10.1093/jhered/esn054
https://doi.org/10.1093/jhered/esn054...
). It has been widely used as flowerbeds in landscaping gardening projects and has gained importance in the floriculture industry with the introduction of a large number of vegetatively propagated varieties with novel foliage colors and shapes (SAHU and DEWANJEE, 2012SAHU, R.; DEWANJEE, S. Differential physiological and biochemical responses under variable culture conditions in micro-propagated Solenostemon scutellarioides: an important ornamental plant. Natural Products and Bioprospecting, v.2, p.160-165, 2012. DOI: https://dx.doi.org/10.1007/s13659-012-0035-y
https://dx.doi.org/10.1007/s13659-012-00...
).

These genotypes are the result of breeding programs based on the genetic instability of this species, which generates unstable colors and shapes through mutations that also obstruct standardized coleus seeds propagation (WEISS, 2002WEISS, D. Introduction of new cut flowers: domestication of new species and introduction of new traits not found in commercial varieties. In: Vainstein, A. Breeding for ornamentals, Dordrecht: Springer, 2002. p.129-137.). Clonal propagation is an alternative to the commercial multiplication of coleus (HAMILTON et al., 2002HAMILTON, C.J.; EMINO, E.R.; BARTUSKA, A.C. The influence of cutting size, leaf area and shipping on coleus cutting quality parameters including rooting. Proceedings Florida State Horticultural Society, v.115, p.134-136, 2002.). Among vegetative propagation methods, the use of stem cuttings has advantages such as homogeneity and the maintenance of stock plants characteristics (HARTMAN et al., 2011HARTMANN, H.T.D.E.; KESTER, F.T.; DAVIES Jr.; GENEVE, R.L. Hartmann and Kester´s Plant propagation: principles and practices. 8ed. Boston: Prentice Hall, 2011. 915p.), such as leaf color pattern. This method is associated with the use of auxin-based plant growth regulator which stimulate adventitious rhizogenesis (OLIVEIRA et al., 2011OLIVEIRA, L.M.; NEPOMUCENO, C.F.; FREITAS, N.P.; PEREIRA, D.M.S.; SILVA, G.C.; LUCCHESE, A.M. Propagação vegetativa de Hyptis leucocephala Mart. ex Benth. e Hyptis platanifolia Mart. ex Benth. (Lamiaceae). Revista Brasileira de Plantas Medicinais, v.13, n.1, p.73-78, 2011. DOI: http://dx.doi.org/10.1590/S1516-05722011000100011
http://dx.doi.org/10.1590/S1516-05722011...
). Indolebutyric acid (IBA) is a synthetic auxin used for rooting. It is recognized to promote homogeneity and speed in stem cuttings rooting, and, consequently, to reduce the time required to commercial plantlets production (HAN et al., 2009HAN, H.; ZHANG, S.; SUN, X. A review on the molecular mechanism of plants rooting modulated by auxin. African Journal of Biotechnology, v.8, n.3, p.348-353, 2009.). However, the required plant growth regulator concentration vary, among other factors, according to the species, the stock plants genetic and physiological quality, and the type of cuttings (PIZZATTO et al., 2011PIZZATTO, M.; WAGNER JÚNIOR, A.; LUCKMAN, D.; PIROLA, K.; CASSOL, D.A.; MAZARO, S.M. Influência do uso de AIB, época de coleta e tamanho de estaca na propagação vegetativa de hibisco por estaquia. Revista Ceres, v.58, n.4, p.487-492, 2011. DOI: https://dx.doi.org/10.1590/S0034-737X2011000400013
https://dx.doi.org/10.1590/S0034-737X201...
).

An additional aspect that may influence stem cuttings rooting is the presence of leaves, since they are sources of auxins and rooting co-factors, that, being transported to cuttings bases, will act synergistically to promote adventitious rooting (FERREIRA et al., 2009FERREIRA, B.G.A.; ZUFFELLATO-RIBAS, K.C.; CARPANEZI, A.A.; TAVARES, F.R.; KOHERLER, H.S. Metodologias de aplicação de AIB no enraizamento de estacas semilenhosas de Sapium glandulatum (Vell.) Pax. Revista Brasileira de Plantas Medicinais, v.11, n.2, p.196-201, 2009. DOI: http://dx.doi.org/10.1590/S1516-05722009000200014
http://dx.doi.org/10.1590/S1516-05722009...
). The present study aimed to evaluate the influence of apical leaves and the use of different IBA concentrations in coleus vegetative propagation through stem cuttings.

2. MATERIAL AND METHODS

Coleus plants (branches with leaves) were collected in a private residence garden in Curitiba, Parana state, Brazil (Coordinates 25°24’58.74 “S and 49°16’35.05” W) on August 7, 2017 (Brazilian winter). Stem cuttings with 6 cm long semi-hardwood were made with a bevel cut at the base and a straight cut at the apex, with or without keeping a pair of apical leaves. Subsequently, the stem cuttings were treated by immersion in 0.5% sodium hypochlorite solution for 10 minutes, and then washed in running water for 5 min.

After phytosanitary procedure, stem cuttings had their bases treated with indolebutyric acid (IBA) at the following concentrations: 0 (control), 1,000 or 2,000 mg L-1. IBA application was done through immersion of cuttings bases in hydroalcoholic solutions (50% v v-1) containing the above-mentioned concentrations during 10 seconds. In the control treatment, cuttings had their bases immersed in hydroalcoholic solution without IBA. After IBA treatments, stem cuttings were planted in plastic tubes filled with previously dampened fine granulometry vermiculite. Cuttings had half their length inserted into substrate. Tubes with cuttings were kept for 30 days in a greenhouse with intermittent misting (24 ± 2 °C and 90% air relative humidity).

After 30 days from planting, the following variables were measured: i) cuttings rooting percentage (living cuttings that emitted roots greater than or equal to 2 mm); ii) number of roots per cutting; iii) average length of the 3 largest roots per cutting; iv) living cuttings percentage (cuttings with living tissues but without rooting or calli formation); v) calli formation percentage (living cuttings without rooting and with undifferentiated mass of cells at the base); vi) cuttings sprouting percentage; and vii) cuttings leaf maintenance percentage (cuttings that kept the original apical pair of leaves).

The study was conducted under a completely randomized experimental design in a 3×2 factorial scheme (3 IBA concentrations x 2 types of cuttings), with 4 replications and 10 stem cuttings per plot, totaling 240 stem cuttings.

Variances were tested for their homogeneity by the Bartlett test and, when homogeneous, data were submitted to variance analysis (ANOVA). Subsequently, the variables with significant differences had their means compared by the Tukey test at 5% probability.

3. RESULTS AND DISCUSSION

The interaction between the types of cuttings and IBA concentrations was only significant for sprouting percentage (p = 0.05). The factors were independent for the other variables.

Rooting in coleus stem cuttings was not affected by IBA. However, the rooting percentages differed significantly according to the type of cuttings. Rooting of coleus cuttings with and without leaves was 100% and 57.3%, respectively (Table 1). These findings corroborate with the study published by Hamilton et al. (2002)HAMILTON, C.J.; EMINO, E.R.; BARTUSKA, A.C. The influence of cutting size, leaf area and shipping on coleus cutting quality parameters including rooting. Proceedings Florida State Horticultural Society, v.115, p.134-136, 2002. on the effects of leaf area on coleus stem cuttings rooting. These authors concluded that leaf area kept on stem cuttings have an expressive effect on rooting quality.

Table 1
Results of variance analysis (ANOVA), rooting percentage, average number of roots per cutting, average length of three largest roots per cutting, living cuttings percentage, and sprouting percentages in coleus (Plectranthus scutellarioides) stem cuttings with presence (PL) or without (WL) apical leaves treated with different indolebutyric acid (IBA) concentrations.

The importance of leaves in stem cuttings rooting is related to the production of carbohydrates by photosynthesis, which will provide a source of energy, necessary for roots emission (VEIERSHOV, 1998).

Considering the average number of roots per cuttings, the presence of leaves also promoted better results. On average, 16 roots per cutting were verified in cuttings with apical leaves, a number almost three times higher than the 5.7 roots in leafless ones (Table 1). This behavior can be attributed to the fact that apical leaves are a major source of endogen auxin, a hormone directly related to adventitious rooting in stem cuttings (HARTMANN et al., 2011HARTMANN, H.T.D.E.; KESTER, F.T.; DAVIES Jr.; GENEVE, R.L. Hartmann and Kester´s Plant propagation: principles and practices. 8ed. Boston: Prentice Hall, 2011. 915p.). Number of roots was affected by IBA concentrations (Table 1). Better results were observed at 2,000 mg L-1 IBA (Table 1). IBA is a plant growth regulator widely used to stimulate stem cuttings rhizogenesis when endogen auxin production is insufficient for rooting and roots development (FRAGOSO et al., 2017FRAGOSO, R.O.F.; STUEPP, C.A.; RICKLI, H.C.; ZUFFELLATO-RIBAS, K.C.; KOEHLER, H.S. Maximum efficiency concentration of indole butyric acid in promoting the rooting of Japanese Flowering Cherry. Ciência Rural, v.47, n.1, p.1-6, 2017. DOI: http://dx.doi.org/10.1590/0103-8478cr20150894
http://dx.doi.org/10.1590/0103-8478cr201...
). In coleus micropropagation, similarly, nodal fragments treated with IBA presented early rooting and an increase in the number of roots per propagule (SAHU and DEWANJEE, 2012SAHU, R.; DEWANJEE, S. Differential physiological and biochemical responses under variable culture conditions in micro-propagated Solenostemon scutellarioides: an important ornamental plant. Natural Products and Bioprospecting, v.2, p.160-165, 2012. DOI: https://dx.doi.org/10.1007/s13659-012-0035-y
https://dx.doi.org/10.1007/s13659-012-00...
).

Stem cuttings with apical leaves also presented longer roots compared to leafless, both not influenced by IBA concentrations (Table 1). This behavior confirms that rooting and root system quality in coleus stem cuttings are highly associated with the presence of leaves, probably because the leaves provide reserves, hormones and co-factors that are not in sufficient concentration solely in coleus stems.

No mortality was observed in any treatment. Similarly, regardless IBA concentrations or presence/absence of leaves, no calli formation occurred on cuttings. This behavior evidences that coleus stem cuttings undergo direct rhizogenesis, i.e., rooting in this species does not require previous formation of undifferentiated mass of cells in cuttings base.

Considering the living cuttings (living tissues with no roots or calli), this was a behavior only observed in leafless cuttings, since all cuttings with leaves have rooted. On average, 42.7% of leafless cuttings remained alive with no roots nor calli formation.

For the leaf maintenance, on average, 45.8% of the cuttings kept the original pair of leaves until the evaluation, regardless IBA concentrations (data not shown).

Regarding the sprouting percentage, cuttings with leaves had the best results, and did not present variations according to IBA concentrations. On average, 97.7% of the cuttings with leaves have sprouted. On leafless cuttings, only a small percentage of sprouting was observed when 2,000 mg L-1 IBA was applied. No sprouting was observed in leafless cuttings submitted to the control treatment or treated with 1000 mg L-1 IBA (Table 1).

The sprouting in cuttings with leaves may also have been a factor influencing better rooting, since, according to Nicoloso et al. (1999)NICOLOSO, F.T.; FORTUNATO, R.P.; FOGAÇA, M.A.F. Influência da posição da estaca no ramo sobre o enraizamento de Pfaffia glomerata (Spreng.) Pedersen em dois substratos. Ciência Rural, v.29, n.2, p.277-83, 1999. DOI: http://dx.doi.org/10.1590/S0103-84781999000200015
http://dx.doi.org/10.1590/S0103-84781999...
, the new leaves produced during the rooting period are an essential feature for roots emission. The importance of leaves and new shoots in cuttings relies on the fact that these organs produce plant hormones, such as indoleacetic acid, and rooting cofactors, both essential for adventitious root formation in vegetative propagules (FACHINELLO et al., 2005FACHINELLO, J.C.; HOFFMAN, A.; NACHTIAGAL, J.C.; KERSTEN, E. Propagação vegetativa por estaquia. In: FACHINELLO, J.C.; HOFFMAN, A.; NACHTIAGAL, J.C. Propagação de plantas frutíferas. Brasília: Embrapa Informações Tecnológicas, 2005. 109p.).

The potential for coleus vegetative propagation is highly associated with the presence of leaves in stem cuttings, since it promoted better results for rooting, roots development and sprouting. IBA had little effect over rooting and sprouting in stem cuttings of this species. Probably the endogenous level of auxins in leaves provide enough stimuli for rooting in P. scutellarioides and, when transported to the cuttings base, promote rooting without the requirement for exogenous auxin application. The lack of IBA effect on leafless cuttings, though, may also suggest that not only auxin is a limiting factor for rooting in coleus cuttings, but also rooting co-factors only produced by the leaves. The nature and quantitative importance of these co-factors, however, are still to be understood for this species.

4. CONCLUSIONS

The use of indolebutyric acid is not necessary to promote coleus stem cuttings rooting and sprouting. The presence of apical leaves on the cuttings is a fundamental feature to promote rooting, roots growth and sprouting in this species.

ACKNOWLEDGEMENTS

The authors are thankful to the Brazilian Federal Agency for Support and Evaluation of Graduate Education – CAPES, by sponsorship of the research and scholarships granted.

REFERENCES

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    » https://www.cabi.org/isc/datasheet/118545#470F50C9-6BF2-43C0-9187-941E084D1E7F
  • DUKE, J.A. Handbook of Medicinal Herbs, 2ed., Baton Rouge: CRC Press, 2002. 896p.
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    » http://dx.doi.org/10.1590/0103-8478cr20150894
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  • HAN, H.; ZHANG, S.; SUN, X. A review on the molecular mechanism of plants rooting modulated by auxin. African Journal of Biotechnology, v.8, n.3, p.348-353, 2009.
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    » https://doi.org/10.1093/jhered/esn054
  • NICOLOSO, F.T.; FORTUNATO, R.P.; FOGAÇA, M.A.F. Influência da posição da estaca no ramo sobre o enraizamento de Pfaffia glomerata (Spreng.) Pedersen em dois substratos. Ciência Rural, v.29, n.2, p.277-83, 1999. DOI: http://dx.doi.org/10.1590/S0103-84781999000200015
    » http://dx.doi.org/10.1590/S0103-84781999000200015
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    » http://dx.doi.org/10.1590/S1516-05722011000100011
  • PANIZZA, D.L.A.; GONZÁLEZ, R.C.; GONZÁLEZ, M.B.; MIER, M.V.; BAZÚA, C.D.D. Exploration of the ability of Coleus blumei to accumulate aluminum, International Journal of Phytoremediation, v.13, n.5, p.421-433, 2011. DOI: https://doi.org/10.1080/15226514.2010.483263
    » https://doi.org/10.1080/15226514.2010.483263
  • PIZZATTO, M.; WAGNER JÚNIOR, A.; LUCKMAN, D.; PIROLA, K.; CASSOL, D.A.; MAZARO, S.M. Influência do uso de AIB, época de coleta e tamanho de estaca na propagação vegetativa de hibisco por estaquia. Revista Ceres, v.58, n.4, p.487-492, 2011. DOI: https://dx.doi.org/10.1590/S0034-737X2011000400013
    » https://dx.doi.org/10.1590/S0034-737X2011000400013
  • SAHU, R.; DEWANJEE, S. Differential physiological and biochemical responses under variable culture conditions in micro-propagated Solenostemon scutellarioides: an important ornamental plant. Natural Products and Bioprospecting, v.2, p.160-165, 2012. DOI: https://dx.doi.org/10.1007/s13659-012-0035-y
    » https://dx.doi.org/10.1007/s13659-012-0035-y
  • SYTAR, O.; BRUCKOVA, K.; HUNKOVA, E.; ZIVCAK, M.; KONATE, K.; BRESTIC, M. The application of multiplex fluorimetric sensor for the analysis of flavonoids content in the medicinal herbs family Asteraceae, Lamiaceae, Rosaceae. Biological Research, v.48, n.5, 2015. DOI: https://doi.org/10.1186/0717-6287-48-5
    » https://doi.org/10.1186/0717-6287-48-5
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    » http://dx.doi.org/10.1007/s10327-015-0601-3
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Publication Dates

  • Publication in this collection
    Jul-Sep 2018

History

  • Received
    14 May 2018
  • Accepted
    11 June 2018
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