Phytotaxa 208 (2): 175–181 ISSN 1179-3155 (print edition)

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Correspondence PHYTOTAXA
Copyright © 2015 Magnolia Press ISSN 1179-3163 (online edition)

http://dx.doi.org/10.11646/phytotaxa.208.2.7

First occurrence of Gracilaria chilensis, and distribution of Gracilariopsis

lemaneiformis (Gracilariaceae, Gracilariales) in Peru on the basis of rbcL sequence
analysis
NATALIA ARAKAKI1*, WILLIAM E. SCHMIDT2, PATRICIA CARBAJAL3 & SUZANNE FREDERICQ2
1
Instituto del Mar del Perú, Banco de Germoplasma de Organismos Acuáticos. Esquina Gamarra y General Valle s/n, Chucuito, Callao,
Perú.
2
University of Louisiana at Lafayette, Lafayette, LA 70504-2451, U.S.A.
3
Instituto del Mar del Perú, Area Funcional de Investigaciones en Biodiversidad. Esquina Gamarra y General Valle s/n, Chucuito, Cal-
lao, Perú.
*corresponding author

The terete taxa Gracilariopsis (hereafter Gp.) lemaneiformis (Bory) Dawson, Acleto & Foldvik and Gracilaria (hereafter
G.) chilensis Bird, McLachlan & Oliveira resemble each other in branching pattern. This study aims to analyze samples
attributed to G. chilensis from a southern locality in Peru using molecular tools, in order to differentiate it from Gp.
lemaneiformis samples collected along the Peruvian coast. Species identification of the Gracilariaceae, especially of
the terete forms, is notoriously difficult due to morphological plasticity and convergence of their thalli, and the correct
taxonomic identification of non-reproductive, cylindrical specimens remains a difficult task (Gurgel & Fredericq
2004). Utilizing traditional morphometric techniques, Edding et al. (2006) concluded that G. chilensis, comprising a
single morphotype, is the main species of Gracilaria present along the Chilean coast, in which natural populations are
distributed between 30°S and 45°S, from Coquimbo to Southern Chiloe (Bird et al. 1986), with farms extending further
north to Antofagasta (24°S) (Santelices & Ugarte 1990). Hoffmann & Santelices (1997) found the establishment of
G. chilensis on the coast of Coihaique (45°S) mainly due to aquaculture introduction. This species also occurs in the
western Pacific Ocean, ranging from southeastern Australia to New Zealand (Nelson 1987). Gracilariopsis Dawson
was long merged into Gracilaria until Fredericq & Hommersand (1989) resurrected Gracilariopsis on the basis of lack
of nutritive cells in the cystocarp, and with chorda-type surface spermatangia. Gp. lemaneiformis has a geographical
range from Paita, Peru to Antofagasta in northern Chile (Ramírez & Tapia 1991). DNA sequence analysis has been
the most reliable and widely used molecular technique to infer phylogenic relationships at the species level within
the Gracilariaceae (Gurgel & Fredericq 2004), and recognition of Gracilariopsis as a genus distinct from Gracilaria
received strong support from the molecular studies of Bird et al. (1994), and Gurgel et al. (2003), among others.
According to Gurgel et al. (2003), Gp. lemaneiformis is shown not to have a worldwide distribution but to be restricted
to the vicinity of Peru, whereas the species going under the name Gp. “lemaneiformis” from North and South Carolina
is now referred to as Gp. carolinensis Liao et Hommersand, and Gp. “lemaneiformis” from China and Japan constitutes
an undescribed species that is related to Gp. heteroclada Zhang et Xia.

TABLE 1. Silica gel samples for molecular analyses.

GenBank
Taxon Locality Collection Date, Collector
Accession Numbers
G. chilensis Morro Sama, Sama, Tacna 13.v.2014, Coll. L. Mina & V. Chili KP857578
Gp. lemaneiformis Eten, Lambayeque Dec. 2014, Coll. P. Ramírez KP857574
Gp. lemaneiformis San Andres, Pisco, Ica 21.x.2014, Coll. N. Arakaki KP857575
Gp. lemaneiformis Ancon, Lima 30.xii.2014, Coll. N. Arakaki & M. Arakaki KP857576
Gp. lemaneiformis Chorrillos, Lima 30.xii.2014, Coll. N. Arakaki & M. Arakaki KP857577
Gp. lemaneiformis Yacila, Paita, Piura 03.iii.1994, Coll. C. Acleto & R. Zúñiga AY049415

DNA extraction was performed and specific gene regions were amplified by PCR and prepared for sequencing
following the protocols described in Gurgel et al. (2003). The rbcL primers used for amplification and sequencing
reactions were as follows: F15 (5’-GTAATHCCNTAHGCNAAAATGGG-3’)-R916 (5’-CCWGCCATDCKCATCCA-

Accepted by Rafael Riosmena-Rodriguez: 21 Apr. 2015; published: 19 May 2015 175

3’) designed in-house at UL Lafayette by T. Sauvage, and F753-RrbcS-start (Freshwater et al. 1994), for a total
alignment of 1359 bp (93%). Phylogenetic analyses were conducted as described in Camacho et al. (2015). Species
examined, collection localities, and GenBank accession numbers for generated sequences are listed in Table 1. Based
on the phylogenetic analysis of chloroplast-encoded rbcL sequences of terete members of Gracilariaceae from Peru,
we report on the occurrence of G. chilensis in the southernmost part of the country and establish the distribution of
Gp. lemaneiformis as being continuous along Peru’s central and northern part. Previous molecular analyses of nuclear
and plastid DNA verified that the different morphotypes of Gracilaria on the Chilean coast correspond to G. chilensis
(González et al. 1996, Meneses 1996, Candia et al. 1999).

FIGURE 1. Distribution map of terete Gracilariaceae from Peru. Occurrence of Gracilariopsis lemaneiformis along the northern and
central coast: Paita (Piura), Eten (Lambayeque), Ancon and Chorrillos (Lima), and San Andres (Ica). Also, Gracilaria chilensis from the
southern coast (Sama, Tacna).

This study provides molecular evidence that an unidentified species from Morro Sama (Sama, Peru) (Fig. 1)
is G. chilensis, representing the first report of the species for Peru (Fig. 4). This species has robust axes with many
branches (Fig. 2), contrary to Gp. lemaneiformis (Fig. 3) that has narrower, shorter axes. We also provide molecular
confirmation for five area collections of Gp. lemaneiformis (Fig. 1). The rbcL dataset included in the analyses consists
of 19 taxa of Gracilariaceae, 2 of which represent the outgroup. The Maximum Likelihood (ML) analysis resulted in a
tree indicating that samples of Gp. lemaneiformis from Paita (type locality), Eten, Ancon and Chorrillos group together
in a well-supported clade (Fig. 4). There is a 5 bp (0.37%) rbcL difference between the San Andres specimen and all
other specimens, and a 2 bp (0.15%) rbcL difference between the Paita, and Chorrillos, Ancon and Eten specimens.
The uncorrected genetic distance between Gp. lemaneiformis and G. chilensis is 12.6–12.8%.

176 • Phytotaxa 208 (2) © 2015 Magnolia Press ARAKAKI ET AL.

FIGURE 2. Habit of non-reproductive specimens of Gracilaria chilensis from Morro Sama, Peru. Fig. 2A. IMARPE 05-000326-1. Fig.
2B. IMARPE 05-000326-2.

G. chilensis is a species so far known only from Chile and New Zealand (Cohen et al. 2004). Since this is the
first convincing report of this species out of its center of distribution in Chile, we are extending the distribution of G.
chilensis from Chile to southern Peru. We also confirm the distribution of Gp. lemaneiformis in Peru from Paita, Piura
to San Andres, Ica (Fig. 1). Even if its geographical range extends to Antofagasta, Chile (Ramírez & Tapia 1991), it is
unclear at this point whether the plant known in Peru and Chile as Gp. lemaneiformis comprises one or more species.
The reliability of morphological and molecular characters for separating all the taxonomic entities in the complex
will require an extensive phylogeographic investigation of samples throughout their geographic range (Gurgel et al.
2003). Cross-sections through thalli from Gp. lemaneiformis from Peru have been illustrated earlier by Fredericq &
Hommersand (1989), and from G. chilensis from Chile by Bird et al. (1986), among others.
Extended beds of the agarophyte G. chilensis growing on soft and unstable substrates have been intensively
harvested along the Chilean coast for about 20 years. Subsequently, planted and farmed within their area of origin
(30°S and 45°S according to Bird et al. 1986), some populations were introduced outside of their natural range of
distribution in northern Chile (Guillemin et al. 2008). The success of traditional methods used in the commercial
harvesting of G. chilensis is due to four attributes, namely 1) terete branching allowing for rapid regrowth even
though the thalli are covered by sediment; 2) tolerance of a broad range of both salinity (ca. 7–37%) and temperature
(7–28°C); 3) tolerance of mild desiccation; and 4) a very rapid growth rate (Alveal et al. 1997). Farming success of G.
chilensis in Chile depends on vegetative propagation of thallus fragments, planted in soft bottoms, that remain free-
floating, grow indefinitely and propagate at a rapid rate (Guillemin et al. 2008). The ability to alternate between sexual
reproduction and vegetative propagation allows G. chilensis to colonize different environments of bays and estuaries
(Guillemin et al. 2013) and to survive changes characteristic of shallow waters as temperature, water turbidity, light
intensity and salinity (Gómez et al. 2005).
G. chilensis has not been detected along the Peruvian coast prior to this study. We do not know the events responsible
for these introductions, but as Saunders (2009) indicated for G. vermiculophylla (Ohmi) Papenfuss, “its introduction
at one site with secondary transfers along the coast by local shipping or movement of species for aquaculture, is a
possibility”. G. vermiculophylla originated from Japan and has only recently been recognized as an invasive species
in Europe (Brittany, France) and on the North American Atlantic coast (Virginia, North Carolina) (see references in
Saunders 2009). The presence of natural beds of G. chilensis in southern Peru opens the possibility to investigate the
economic potential this available resource could have in the future for the country.

occurrence of Gracilaria chilensis in Peru Phytotaxa 208 (2) © 2015 Magnolia Press • 177
FIGURE 3. Habit of a female gametophyte of Gracilariopsis lemaneiformis from San Andres, Ica, Peru (IMARPE 05-000325).

178 • Phytotaxa 208 (2) © 2015 Magnolia Press ARAKAKI ET AL.

FIGURE 4. Phylogenetic tree based on rbcL sequences inferred from a maximum-likelihood analysis as implemented by RAxML. Taxa
highlighted in red were newly sequenced for this study. GenBank numbers are listed after each taxon. Bootstrap proportions (>50%) for
ML (1000 replicates, Left) and Bayesian Posterior Probability (Right) are shown at the nodes. Full support denoted as *.

occurrence of Gracilaria chilensis in Peru Phytotaxa 208 (2) © 2015 Magnolia Press • 179
Acknowledgements

Thanks to the several individuals (listed in Table 1) that made their collections available for this study.

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