**1. Introduction**

10 Agricultural Science

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Heliconia cultivation has intensified in Brazil as a cut flower, especially in the Northeast region. This ornamental rhizomatous herbaceous plant from the *Heliconia* genus, belongs to the Musaceae family, now constitutes the Heliconiaceae family in the Zingiberales order. The various species of *Heliconia* are subdivided into five subgenera: *Heliconia, Taeniostrobus, Stenochlamys, Heliconiopsis* and *Griggisia*; and 28 sections (Kress *et al*., 1993). In *Heliconia* genus, the number of species ranges from 120 to 257 and there are also a great number of cultivars and 23 natural hybrids (Berry and Kress, 1991; Castro *et al*., 2007), these plants can be found either in shaded places, such as forests or at full Sun areas, such as forest edges and roadsides (Castro and Graziano, 1997). They are native from Tropical America (Berry and Kress, 1991), found at different altitudes, from sea level up to 2.000 meters in Central and South America, and up to 500 meters in the South Pacific Islands (Criley and Broschat, 1992).

Heliconia hybrids comprise many of the major cultivars as cut flowers, like *H. psittacorum* x *H. spathocircinata* cv. Golden Torch, cv. Golden Torch Adrian, cv. Alan Carle and *H. caribaea* x *H. bihai* cv. Carib Flame, cv. Jacquinii, cv. Richmond Red (Berry and Kress, 1991). Many heliconia species are identified through their morphological differences, such as the size and color of its flowers and bracts. These characteristics can be influenced either by geographic isolation or by environmental factors, such as light and nutrients (Kumar *et al*., 1998). *H. psittacorum* clones, even when closely grown, can vary in blooming, size and color of bracts, as well as post harvest durability (Donselman and Broschat, 1986).

<sup>\*</sup> Corresponding author

Genetic Diversity Analysis of *Heliconia psittacorum* Cultivars

Genotypes*<sup>a</sup>* Location Subgenus

*H. psittacorum* L.f. cv. Sassy Paulista - PE *Stenochlamys*

*H. psittacorum* L.f. cv. Lady Di Ubatuba - SP *Stenochlamys*

*H. psittacorum* L.f. cv. St. Vincent Red Ubatuba - SP *Stenochlamys*

*H. psittacorum* L.f. cv. Red Gold Paulista - PE *Stenochlamys*

bract color; OV: ovary; PD: pedicel; SE: sepals. *d*Ploidy (Costa *et al*., 2008).

at a single plan (Berry and Kress, 1991).

*H.* x *nickeriensis Maas and de Rooij* (*H. marginata* x *H. psittacorum*)

*H. psittacorum* L.f x *H. spathorcircinata*  Aristeguieta cv. Golden Torch

*H. psittacorum* L.f x *H. spathorcircinata*

*H. psittacorum* L.f. x *H. spathocircinata* 

*Heliconia* sp. (suposed to be *H.* 

*H. psittacorum* L.f *.*cv. Strawberries

*psittacorum* cv. Sassy)

*Suposed triploid*

and Cream

*Hybrids* 

Adrian

cv. Golden Torch

*H. psittacorum* L.f. cv. Suriname Sassy

cv. Red Opal

*Triploid<sup>d</sup>*

and Interspecific Hybrids Using Nuclear and Chloroplast DNA Regions 13

average annual temperature is 25.1ºC and monthly rainfall of 176 mm, with maximum of 377 mm and minimum of 37 mm (ITEP, 2008). This study evaluated 11 *Heliconia psittacorum* cultivars and interspecific hybrids (Table 1) obtained by exchange with research institutions and farmers from the states of Pernambuco (PE), Alagoas (AL) and Sao Paulo (SP) in Brazil. The analyzed genotypes presented short size, musoid habit and erect inflorescence disposed

Paulista - PE *Heliconia*

Paulista - PE *Stenochlamys*

Maceió - AL *Stenochlamys*

Paulista - PE *Stenochlamys*

*<sup>a</sup>*Identification based on Berry and Kress (1991) and Castro *et al*. (2007); *b*Based on Kress *et al*. (1993); *c*BC:

Table 1. Genotypes, location, classification and description for 11 *Heliconia psittacorum* cultivars and interspecific hybrids of the UFRPE Heliconia Germplasm Collection used in this study

and Section*<sup>b</sup>*

*Pendulae* 

*Stenochlamys*

*Stenochlamys*

*Stenochlamys*

*Stenochlamys*

*Stenochlamys*

*Stenochlamys*

*Stenochlamys*

Description*<sup>c</sup>*

BC- yellow-orange; OV- dark yellow distally, light yellow proximally, PD-

yellow with indistinct blackish green area distally, SE- dark yellow

with indistinct blackish green area

BC- pink-green; OV- orange distally, yellow proximally, PD- orange to cream, SE- orange with indistinct blackish green area distally

BC- pink-green; OV- orange distally, yellow proximally, PD- yellow green, SE- orange with indistinct blackish green area distally

BC- pink-lilac; OV- green distally and yellow green proximally, PDyellow green, SE- orange with indistinct blackish green area distally

BC- pink-yellow; OV- yellow to cream, PD- cream, SE- pale yellow with green spot on distally corner

BC- red; OV- yellow, PD- light yellow to cream, SE- light yellow with distally dark green band and white tip

BC- red-orange; OV-orange distally, orange to cream proximally, PDorange, SE- orange with indistinct blackish green area distally

BC- red-orange; OV- yellow, PDyellow, SE- dark yellow with indistinct blackish green area distally

SE- dark yellow with indistinct blackish green area distally

yellow, SE- dark yellow

distally, SE- dark yellow

Paulista - PE *uncertain* BC- yellow-Red; OV- yellow, PD-

Paulista - PE *uncertain* BC- yellow; OV- yellow, PD- yellow

Paulista - PE *uncertain* BC- orange; OV- yellow, PD- yellow,

The natural variation among heliconia individuals or populations has led to taxonomic identification doubts among farmers and researchers. Thus, genebanks have played an important role in genetic diversity conservation, providing raw material for crop breeding, including landraces and their wild relatives. DNA markers, which allow the access to variability at DNA level, emerge as an efficient alternative for plant species characterization by quantifying diversity and determining its genetic structure (Bruns *et al*., 1991).

The choice on which molecular marker technique shall be used depends on its reproducibility and simplicity. Kumar *et al*. (1998) distinguished three cultivars of the hybrid *H. psittacorum* x *H. spathocircinata* cvs. Golden Torch, Red Torch and Alan Carle which showed only slight differences in RAPD markers profile from *H.* x *nickeriensis* Maas and de Rooij (*H. psittacorum* x *H. marginata*), they also observed similarities in RAPD profiles and morphology. The authors concluded that two triploid *H. psittacorum*: cv. Iris and Petra, are supposed to be the same genotype.

Genetic diversity studies grew up in interest during the last years (Jatoi *et al*., 2008; Kladmook *et al*., 2010). As a result, nucleotide sequences of ribosomal genes (rDNA) and chloroplast genes (cpDNA) have been exploited to investigate several individuals of the Zingiberales order (Kress, 1990, 1995; Kress *et al*., 2001) once they are not capable of lateral transfers and are not subject to the same functional limitations, they allow greater confidence in the results (Camara, 2008). The unit of ribosomal eukaryotic organisms consists of three genic and three non-genic regions. On one hand, the genic regions (18S, 5,8S and 26S) are conserved and evolve slowly. On the other hand, non-genic regions, known as ITS - Internal Transcribed Spacer (ITS-1 and ITS-2), evolve rapidly, showing high polymorphism and, therefore, allowing its use at higher hierarchical levels. The variability found in these regions could be the result of mutations in these areas, since they suffer less selection pressure and may be well used to study genetic diversity in plants (Bruns *et al*., 1991).

This molecular marker is important from the genetic variability assessment point of view, because the rDNA mutltigenic family once subjected to a rapid evolution in concert event, allows greater precision in the reconstruction process of the relationship between species based on sequencing, since this phenomenon increases the intragenomic uniformity (Baldwin *et al*., 1995). These authors also affirm that due to the biparental inheritance of the nuclear genome it is possible to study the origin of hybrids and their parents. Moreover, chloroplast genes (cpDNA), such as the leucyn and fenilalanyn of RNA transporter (*trnLtrnF*), the treonyn and leucyn of RNA transporter (*trnT-trnL*) and the protein small 4 (*rps4*), have been used successfully to solve genetic diversity doubts in taxonomic lower levels. Johansen (2005), for example, studying the genetic diversity in Zingiberales order, using cpDNA, has positioned all Heliconiaceae and Musaceae within a same clade.

The aim of this study was to evaluate genetic diversity involving *Heliconia psittacorum* cultivars and interspecific hybrids of the Federal Rural University of Pernambuco *Heliconia* Germplasm Collection (UFRPE-HCG), using nuclear and chloroplast DNA regions.
