SANTA-ROSA S; SOUZA FVD; VIDAL AM; LEDO CAS; SANTANA JRF. 2013. Micropropagation of the ornamental vulnerable bromeliads Aechmea blanchetiana and Aechmea distichantha. Horticultura Brasileira 31: 112-118. Micropropagation of the ornamental vulnerable bromeliads Aechmea blanchetiana and Aechmea distichantha Sandra Santa-Rosa1; Fernanda VD Souza2; Ádila M Vidal3; Carlos Alberto da S Ledo2; José RF de Santana1 1UEFS-Depto. Ciências Biológicas, Av. Universitária km 03 BR116, 44031-460 Feira de Santana-BA; 2Embrapa Mandioca e Fruticultura, C. Postal 007, 44380-000 Cruz das Almas-BA; 3UFRB-Depto. Ciências Agrárias, 44380-000 Cruz das Almas-BA; sandra_santarosa@ yahoo.com.br; [email protected]; [email protected]; [email protected]; [email protected] ABSTRACT RESUMO Aechmea blanchetiana and Aechmea distichantha are Brazilian Micropropagação das bromélias ornamentais vulneráveis native bromeliads with great ornamental value, although none of the Aechmea blanchetiana e Aechmea distichantha species are produced commercially. Therefore, market demands are Aechmea blanchetiana e Aechmea distichantha são bromélias met through nature predatory exploitation. This study intended to nativas do Brasil, de grande valor ornamental. Não sendo produzidas (1) evaluate the in vitro seed germination of A. blanchetiana and A. comercialmente, ambas sofrem exploração predatória. Este trabalho distichantha and (2) establish micropropagation protocols for both teve como objetivos (1) avaliar a germinação de sementes in vitro species, aiming at producing plantlets for the market, as well as to de A. blanchetiana e A. distichantha e (2) estabelecer protocolos de germplasm in vitro conservation. Germination experiments were micropropagação destas bromélias, visando a produção de mudas carried out in completely randomized designs with two treatments para atender ao mercado e à conservação in vitro. Os experimentos (presence and absence of light) and seven and eight replications de germinação foram conduzidos em delineamento inteiramente respectively for A. blanchetiana and A. distichantha. In vitro casualizado, com dois tratamentos (presença e ausência de luz) e seedlings, produced under light conditions, were used as explants sete e oito repetições respectivamente para A. blanchetiana e A. in the multiplication experiment (MS medium). The experimental distichantha. Plantas in vitro, obtidas em presença de luz, foram uti- design was again completely randomized, with treatments in lizadas como explantes no experimento de multiplicação (meio MS). factorial 2 (NAA concentrations; 0.05 and 0.5 μM) x 2 (cytokines: O delineamento utilizado foi inteiramente casualizado, em fatorial BAP and KIN) x 2 (cytokine levels: 2.2 and 4.4 μM) + 1 (control 2 (concentrações de ANA: 0,05 e 0,5 μM) x 2 (citocininas: BAP e with basal MS, without growth regulators), with ten replications per CIN) x 2 (concentrações de citocininas: 2,2 e 4,4 μM) + 1 (controle treatment. We evaluated fungal and bacterial contamination, total and com meio MS básico, sem reguladores de crescimento), com 10 re- cumulative seed germination and number of shoots per treatment in petições por tratamento. Foram avaliadas as contaminações fúngica each subculture. We observed only fungal contamination and only e bacteriana, germinação total e cumulativa e número de brotos por in A. blanchetiana (14.0%). Light favored in vitro seed germination tratamento em cada subcultivo. Observou-se apenas contaminação in both species (99 and 62% respectively for A. blanchetiana and A. fúngica e somente em A. blanchetiana (14,0%). A presença de luz distichantha). MS medium + 0.5 μM NAA + 2.2 μM BAP resulted favoreceu a germinação de sementes in vitro de ambas as espécies in the highest multiplication rate for both species, with significant (99 e 62% para respectivamente A. blanchetiana e A. distichantha). contrasts between this treatment and the control in both species Meio MS + 0,5 μM ANA + 2,2 μM BAP promoveu a maior taxa de (105.62 more shoots in A. blanchetiana; 223.80 in A. distichantha). multiplicação para ambas as espécies, com contrastes significativos BAP promoted the formation of rootless shoots, while kinetin favored entre este tratamento e o controle em ambas as espécies (105,62 brotos the formation of roots rather than shoots. A. blanchetiana and A. a mais em A. blanchetiana; 223,80 em A. distichantha). BAP promo- distichantha showed 90 and 97% of survival respectively in plantlet veu a formação de brotos sem raízes, enquanto cinetina favoreceu a acclimatization. formação de raízes em detrimento de brotos. Na aclimatização das plântulas, A. blanchetiana e A. distichantha apresentaram 90 e 97% de sobrevivência respectivamente. Keywords: Bromeliaceae, tissue culture, in vitro multiplication rate, Palavras-chave: Bromeliaceae, cultura de tecidos, taxa de growth regulators. multiplicação, reguladores de crescimento. (Recebido para publicação em 13 de abril de 2012; aceito em 5 de novembro de 2012) (Received on April 13, 2012; accepted on November 5, 2012) enus Aechmea is one of the in the Atlantic Rainy Forest (Smith Brazilian native bromeliad, terrestrial, Glargest in family Bromeliaceae. & Downs, 1979). Most bromeliads occasionally epiphytic, perennial (Bert Aechmea belongs to the subfamily within the genus have high ornamental & Luther, 2005) and frequent among Bromelioideae and comprises about potential, such as Aechmea blanchetiana sand coastal plants and in the Atlantic 240 species (Luther, 2006), from which and Aechmea distichantha, and thus are Rainy Forest in states of Bahia and 70% are estimated to be distributed in target of predatory exploitation. Espírito Santo (Martinelli et al., 2008). Brazil, having its center of diversity A. blanchetiana (Figure 1A) is The inflorescence consists of a set of red 112 Hortic. bras., v. 31, n. 1, jan. - mar. 2013 Micropropagation of the ornamental vulnerable bromeliads Aechmea blanchetiana and Aechmea distichantha various concentrations, according to A B the particularities of each step of the micropropagation process (Souza et al., 2003; Droste et al., 2005; Bellintani et al., 2008; Silveira et al., 2009). As in other species, the need to develop specific in vitro protocols for Aechmea blanchetiana and A. distichantha comes mainly from the possibility that a fortuitous genotype-specific dependency appears, which would require optimization procedures to avoid compromising the results. On the other hand, in spite of all the benefits ofin vitro propagation, it is important to bring into the spot that the cloning of poorly studied species may jeopardize the preservation of genetic Figure 1. Flowering plants of Aechmea blanchetiana (A) and Aechmea distichantha (B) variability in natural populations and, in (plantas em florescimento deAechmea blanchetiana (A) e Aechemea distichantha (B)). Cruz the long run, germplasm conservation, das Almas, Embrapa Mandioca e Fruticultura, 2009. resulting in irreversible genetic erosion. Given this, seed production, seems to suit best as propagation strategy for bracts that can last for several months. Micropropagation can guarantee not species like Aechmea at this point. This characteristic adds to the large only plantlet production in large scale, Sexual reproduction results in offspring ornamental potential the species already but also the means for establishing in that ensures the preservation of genetic has, a remarkably long postharvest life vitro banks for both species, expanding variability and avoids gene loss (Mercier (Lorenzi & Souza, 1998). The growing the conservation strategies of this & Nievola, 2003; Carneiro & Mansur, employment of A. blanchetiana in germplasm, currently under threat. 2004; Silveira et al., 2009). landscaping and gardens using plants In vitro propagation has been Taking into account what was supplied by predatory exploitation, showing great potential when compared exposed so far, the objective of this places the species in constant threat. to conventional propagation methods study was to develop protocols for Kanashiro et al. (2007) stated that the in bromeliads. In vitro propagation micropropagation of the ornamental lack of plant commercial production to allows for a quick genetically safe bromeliads Aechmea blanchetiana meet market demands resulted in the multiplication of a high number of and A. distichantha using plantlets presence of A. blanchetiana in the list plants, which matches with the large originated from in vitro germination of endangered species. market demand. Studies involving of seeds. A. distichantha (Figure 1B), typical in vitro propagation resulted in the from the Brazilian savannah, has development of multiplication protocols MATERIAL AND METHODS epiphytic, lithophytic or terrestrial habit, for several species (Mendes et al., 2007; arched leaves reaching up to 90 cm, Silva et al., 2008; Silveira et al., 2009; In vitro seed germination and an erect rosette which lasts up to a Santos et al., 2010; Huang et al., 2011), of Aechmea blanchetiana and A. month (Bert & Luther, 2005). Similarly including A. blanchetiana (Kanashiro et distichantha - Seeds from both species to the A. blanchetiana, the species is not al., 2007; Galvanesi et al., 2007; Chu et were taken from ripe fruits; but while grown commercially and has been used al., 2010). for A. blanchetiana fruits were collected as an ornamental plant due to predatory Several factors can influence the at the farm Flor de Brotas, in the supply, which led A. distichantha to efficiency of a micropropagation city of Irará,