Morphological Study of Lemna Minuta Kunth, an Alien Species Often
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Aquatic Botany 131 (2016) 51–56 Contents lists available at ScienceDirect Aquatic Botany jou rnal homepage: www.elsevier.com/locate/aquabot Short communication Morphological study of Lemna minuta Kunth, an alien species often mistaken for the native L. minor L. (Araceae) ∗ Simona Ceschin , Ilaria Leacche, Stefano Pascucci, Silverio Abati Roma Tre University, Department of Science, Viale G. Marconi 446, Rome 00146, Italy a r t i c l e i n f o a b s t r a c t Article history: Lemna minuta Kunth and Lemna minor L. are two small aquatic floating plants easily mistakable for similar Received 16 April 2015 morphology. The need to distinguish with certainty these two species is a relevant issue, not only from a Received in revised form 20 January 2016 floristic viewpoint, but also for establishing management plans in wetlands where L. minuta is an invasive Accepted 30 January 2016 alien species as in Europe. The vein number and frond length are considered by most authors as the main Available online 28 February 2016 morphological characters discriminating the two species. However, in this study the use of these two characters has not been effective for the determination of 43 specimens out of 248 living specimens of Keywords: Lemna collected from 15 different wetlands of the Central Italy. Statistical analyses of the dataset made it Alien species possible to better define the variability of the morphological characters of these two species and to identify Floating plant additional diagnostic characters to use for improving the morphological discrimination between them. Congeneric species Decision Tree Among these characters, width, apex and shape of the frond, were the most helpful. Decision Trees were Duckweed elaborated for differentiating L. minor from L. minuta with greater efficiency both in the laboratory and in Morphological characterization the field. The increase in percentage of correct determination of Lemna specimens through measurement and parallel utilization of the emerged morphological characters will facilitate the botanists’ activities, but also will have practical implications, such as the ability to contribute better to the decision-making system in drawing up plans for plant biodiversity protection and/or eradication of the alien species. © 2016 Elsevier B.V. All rights reserved. 1. Introduction five aliens: L. minuta, L. aequinoctialis Welw., L. perpusilla Torrey, L. turionifera Landolt and L. valdiviana Phil. (DAISIE 2008). Aquatic ecosystems are among the most threatened habitats The Lemna’s vegetative body (frond) is a complex of tissues worldwide (Millenium Ecosystem Assessment, 2005) and among with few differentiations, reduced morphological features, and the most vulnerable to biological invasions (Vitousek et al., 1997; frequently lacking of reproductive structures. This scarcity of char- Capers et al., 2007). For a better conservation of their native com- acters for distinguishing the species of Lemna, makes the taxonomic munities, it becomes necessary to correctly verify the presence of discrimination between them difficult (Landolt 1986; Donald et al., invasive alien species and then to undertake management actions 2002; Wang et al., 2010), and in particular between L. minor and L. that can limit the spread and potential impact of these species in minuta, which exhibit very similar morphological characters (Lucey the invaded ecosystem. 2003; Mifsud 2010). Thus, these two Lemna species are easily mis- One invasive plant species widespread in European standing takable between them. inland waters is Lemna minuta Kunth (Verloove 2006; Branquart In literature, for the morphological discrimination between et al. 2007; Thiébaut 2007; Celesti-Grapow et al., 2009; Lukács et al., these two species there is no complete concordance between 2014), which is among the smallest and fastest growing flower- authors due to different and sometimes conflicting set of pheno- ing plants (Landolt 1986). In Europe, the Lemna genus (duckweeds; typic characters to use (e.g., Landolt 1992; Wolff and Landolt, 1994; Araceae) is represented by three native species: Lemna minor L., the Stace 2010; Eckehart 2011). For example the number of fronds most common European duckweed, L. gibba L. and L. trisulca L., and is not considered by Stace (2010) and Wolff and Landolt (1994) as a character for discriminating L. minuta from L. minor, while other authors recognize this character as diagnostic although they disagree among themselves about the numerical range associated ∗ Corresponding author. with L. minor and L. minuta. However, most authors agree that E-mail addresses: [email protected] (S. Ceschin), [email protected] primarily vein number and secondarily frond length are the best (I. Leacche), [email protected] (S. Pascucci), [email protected] (S. Abati). characters for differentiating the two species (Landolt 1992; Wolff http://dx.doi.org/10.1016/j.aquabot.2016.01.005 0304-3770/© 2016 Elsevier B.V. All rights reserved. 52 S. Ceschin et al. / Aquatic Botany 131 (2016) 51–56 Table 1 and Landolt, 1994; Galán and Castroviejo 2008; Stace 2010). Par- Repartition of the Lemna specimens based on vein number and length frond. ticularly, L. minor is usually described by 3–5 veins, frond 1–8 (10) mm long, while L. minuta by 0–1 vein and frond 0.8–2.0(4.0) mm Length frond (mm) long. <2 2–3 >3 However, both characters, although necessary to distinguish L. Vein n 0–1 58 (23%) (A) 33 (13%) (B) 10 (4%) (C) minor from L. minuta, show some limits. Vein number is a character >1 11 (4%) (D) 38 (15%) (E) 98 (40%) (F) often scarcely observable either in the field or in the laboratory, even on samples treated properly (Landolt 1992); frond length U-CMAD3). To better observe some qualitative characters, such is a character partially overlapping between the two species (e.g. as vein number and aerenchyma extension, the fronds were dis- Landolt 1992; Wolff and Landolt, 1994; Stace 2010). ◦ coloured with ethanol 95 for at least 24 h following Landolt (1992) Therefore it is necessary to identify other morphological char- and subsequently observed under a stereoscope with a light source acters, which combined with those mentioned above, can better from below. distinguish L. minor from L. minuta. This need is even more relevant For the analysis of quantitative characters, digital photos were in Europe since L. minor is a native species while L. minuta is an processed with the program ImageJ vers. 1.47. The measurements invasive alien species which often shares the same habitats with L. were carried out only on the major frond, if the specimen consisted minor, threatening its presence (Landolt 1986; Iberite et al., 2011; of contiguous fronds. Mazzini et al., 2014). The present study aimed to better define the ranges of the mor- phological characters usually used for discriminating L. minor from 2.2. Statistical analysis L. minuta, and to identify additional morphological characters for better differentiating these two species. Decision Trees were devel- Firstly, the Lemna specimens were characterized on the basis oped for differentiating the two species directly in the field or in the of the vein number and frond length, the two main mor- laboratory with the minimum number of morphological characters phological characters recognized in literature as discriminating and the lowest degree of error. between the two species. In particular, the specimens were assigned to one of two Lemna species on the basis of non- overlapping ranges of these two characters to allow a certain 2. Materials and methods assignment. On these specimens certainly assigned to L. minuta or L. minor 2.1. Collection and analysis of Lemna specimens and belonging to populations sampled at different sites, the mor- phological variability was tested (inter-population variability) by A morphological comparison between L. minuta and L. minor was applying the non-parametric Kruskal–Wallis test (H) on the medi- carried out. This analysis is based on observations and measure- ans of the measured values for the frond length. ments of morphological characters made on 248 living specimens Subsequently, the whole dataset was processed through of Lemna collected in 2013–2014 from 15 wetlands of the Central Non-Metric Dimensional Scaling (NMDS), for evaluating the con- Italy, between May and September. tribution of each variable (character) for discriminating the two Only the vegetative characters were chosen for the investigation species. Gower index for quantitative variables and Hamming index because no specimen was observed with flowering structures or for qualitative variables were used as measures of similarity dis- fruits, as indeed happens in most of specimens occurring in Europe, tance and a biplot was obtained correlating the scores of the two especially as regards L. minuta (Landolt 1980; Bramley 1997). In first ordination axes with each morphological character (Kendall addition, the characters related to the roots (total length, root cap) concordance coefficient, ). were not considered because their correct measurement would Exclusively for quantitative characters, a Spearman correlation not have been possible in some specimens, since the roots of both was performed for identifying any redundant characters and for species are very fragile and break easily. highlighting potential differences in the morphological plastic- A total of fourteen morphological characters, including both ity of the two species. To check significant differences between qualitative (*) and quantitative ones,