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Analyses of Phylogenetics, Starch Granule Morphology and Consumer Preference of Canna Indica L. Grown in Sri Lanka

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Analyses of Phylogenetics, Starch Granule Morphology and Consumer Preference of Canna Indica L. Grown in Sri Lanka RESEARCH ARTICLE Analyses of phylogenetics, starch granule morphology and consumer preference of Canna indica L. grown in Sri Lanka R.W.K.M. Senevirathna, L.T. Ranaweera, N.D.U.S. Nakandala, H.M.T.N. Senavirathna, W.M.D.A. Wijesundara, H.S.M. Jayarathne, C.K. Weebadde and S.D.S.S. Sooriyapathirana Highlights • The sequence polymorphism of the atpB gene differentiates Canna indica (Arrowroot/Buthsarana) from ornamental Canna spp. • Canna indica has the significantly large starch granules compared to those of Xanthosoma sagittifolium, Manihot esculenta, Solanum tuberosum, and Ipomoea batatas. • Canna indica has unique scallop-seashell shaped starch granules. • The cooked Canna indica tubers are accepted, better than those of Xanthosoma sagittifolium, and rated equally to Solanum tuberosum, and Manihot esculenta. Ceylon Journal of Science 49(3) 2020: 261-273 DOI: http://doi.org/10.4038/cjs.v49i3.7777 RESEARCH ARTICLE Analyses of phylogenetics, starch granule morphology and consumer preference of Canna indica L. grown in Sri Lanka R.W.K.M. Senevirathna1, L.T. Ranaweera1, N.D.U.S. Nakandala1, H.M.T.N. Senavirathna1, W.M.D.A. Wijesundara1, H.S.M. Jayarathne1, C.K. Weebadde2 and S.D.S.S. Sooriyapathirana1,3,* 1Department of Molecular Biology and Biotechnology, Faculty of Science, University of Peradeniya, Peradeniya, Sri Lanka. 2Department of Plant, Soil and Microbial Sciences, College of Agriculture and Natural Resources, Michigan State University, East Lansing, 48824, USA. 3Postgraduate Institute of Science, University of Peradeniya, Peradeniya, Sri Lanka. Received: 22/05/2019; Accepted: 29/06/2020 Abstract: Canna indica is a tuber crop which has many Devkali in India, Buthsarana in Sinhala, and Ampurut Alai in medicinal values. In Sri Lanka, C. indica tubers are consumed Tamil (Bachheti et al., 2013). It is believed that C. indica was in rural areas and mainly available in street-markets of Nuwara- originated in Latin America (Andrade-Mahecha et al., 2012) Eliya and Kandy Districts. In the present study, we assessed and later naturalized in many countries including Sri Lanka the phylogenetics of C. indica, starch granule morphology and (Olango et al., 2013; Gunarathna et al., 2016). The rhizome consumer preference of C. indica tubers in comparison to the is the edible part of the plant and all parts of the plant contain popular tuber crops. The phylogenetic analysis was conducted medicinal values including antioxidant property (Mahesh based on the sequence polymorphism at rbcL, atpB gene, trnL- et al., 2014; Joshi et al., 2009a), HIV reverse transcription trnF and trnH-psbA marker-loci with respect to the ornamental inhibitory activity (Woradulayapinij et al., 2005), anti- Canna spp. in Sri Lanka and the previously published sequences diarrheal, hemostatic, hepatoprotective, antibacterial and anti- of Canna spp. The starch granules were isolated and observed worming effects (Rahmatullah et al., 2010; Josephine et al., under optical and scanning electron microscopes. The diameter 2013; Lin et al., 2011; Joshi et al., 2009b; Anisuzzaman et and the surface area of the starch granules were measured under al., 2007; Indrayan et al., 2011; Thepouyporn et al., 2012; the optical microscope and subjected to analysis of variance. Abdullah et al., 2012; Nirmal et al., 2007). As C. indica tubers are consumed as boiled tuber pieces in Sri The tubers of C. indica are a common commodity in Lanka, the consumer preference analysis was conducted using the ayurvedic herbal medical preparations (Kankanamalage boiled tuber pieces C. indica, Xanthosoma sagittifolium, Manihot et al., 2014). Although C. indica possesses significant esculenta, Solanum tuberosum, and Ipomoea batatas. The medicinal and food values, it is currently an underutilized phylogenetic tree based on rbcL marker revealed that C. indica tuber crop in Sri Lanka (Malkanthi, 2017). The tubers of in Sri Lanka is slightly divergent from the other Canna spp. Only C. indica rarely appear in the country-wide market. The the polymorphism of the atpB gene can be used to differentiate younger generation who generally appreciate fast food and C. indica from the ornamental Canna sp. in Sri Lanka. The limited array of choices, even do not know that C. indica morphological analysis of starch granules revealed that C. indica tuber is a human food. However, C. indica is fast growing has the biggest scallop-seashell shaped starch granules compared (Zhang et al., 2008) with an excellent ornamental value to other tuber species. The boiled C. indica tubers were accepted (Zhang et al., 2008) and recently being identified as one better than that of X. sagittifolium, rated equally to the tubers of of the best species for the phytoremediation of the polluted S. tuberosum and M. esculenta, and rated less than I. batatas. The water released from reverse osmosis process of the water hardy and fibrous nature of C. indica tubers must be the major purification units in Sri Lanka (Gunarathna et al., 2016). limiting factors for achieving the highest consumer preference It is believed that C. indica contains the largest starch highlighting the need of breeding for better texture in tubers. granules ever known (Hermann et al., 1997) thus, can be Keywords: Devkali, Indian-shot, Large starch granules in plants, used as a candidate species to broaden our knowledge on Ornamental Canna spp., Underutilized tuber crops. starch science and technology. In Sri Lanka, C. indica is grown in lowland plains (30 – INTRODUCTION 200 m above sea level) of Sri Lanka without much attention. Canna indica of family Cannaceae is considered as one However, C. indica plants thrive well in the central highlands of the traditionally used tuber crops in the world (Prince, of Sri Lanka (500 m above mean sea level, 2000 mm annual 0 2010). The vernacular names of C. indica are Indian-Shot rainfall and 23-26 C of mean temperature) and grow as in English, Arrowroot in Africa, Achira in Latin America, large dense bushes where suckers keep coming from the mother plants. The tubers can be harvested for human *Corresponding Author’s Email: [email protected] http://orcid.org/0000-0002-5592-1742 This article is published under the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 262 Ceylon Journal of Science 49(3) 2020: 261-273 consumption after six months of planting. There are Phylogenetic analysis slight variations in leaf size, leaf margin and plant height exist among the C. indica populations found in different The initial and end noises of the sequencing data obtained habitats. However, when suckers from one location is from four DNA markers (Table 1) were observed planted in a different habitat, these variations get changed independently using MEGA 7 (Kumar et al., 2016). Then based on the soil fertility and moisture content. However, the identities of the trimmed sequences were confirmed the genetic diversity or the exact species delimits of the C. independently by subjecting to a search in Basic Local indica have not been studied in Sri Lanka. In addition to the Alignment Search Tool (BLAST). We created separate edible C. indica, there are ornamental Canna plants which alignments for each marker in MEGA 7. In order to relax the can produce attractive flowers and foliage grown in many family Cannaceae topology, we did not use phylogenetically places of the country. The species differences between distant outgroup, or a too closely related one. Thus, we edible C. indica (i.e. Arrowroot) and the ornamental used two species from family Commelinaceae as outgroups Canna spp. have not been studied to date. The extensive which can also be found in the Commelinids clade where multidisciplinary research attempts are needed to uplift C. family Cannaceae is located. To compare the Canna spp. Sri indica from its current underutilized status. Also, the taste Lanka, the sequences generated were aligned for the marker, and other sensory attributes of C. indica in comparison to rbcL, with the dataset generated in Prince, (2010) (Table 2). the popular tuber crops must be studied. Therefore, the A model selection analysis in Jmodel test (Posada, 2008) present study was conducted to assess the species delimits was implemented to infer the best nucleotide substitution and phylogenetic relationships, starch granule morphology, process of the dataset. A phylogeny was constructed in and consumer preference of C. indica in Sri Lanka. Maximum Likelihood framework in RAxML (Stamatakis, 2006). We implemented rapid+boostrap algorithm with MATERIALS AND METHODS 1000 iterations and GTRGAMMA as the nucleotide substitution model. The analysis was implemented in the DNA extraction, PCR and DNA sequencing CIPRES supercomputer (Miller et al., 2010). We also The DNA was extracted from the immature leaves of carried out a tree search in the Bayesian framework in C. indica, plants collected from Kundasale, Matale, MrBays (Huelsenbeck and Ronquist, 2001) in CIPRES Nuwara-Eliya, Peradeniya (7°16’42.2”N 80°40’33.7”E, science gateway. Two hot and cold chains of Markov Chain 7°27’29.7”N 80°36’31.3”E, 6°52’53.7”N 80°48’55.5”E, Monte Carlo (MCMC) running for 30 million generations 7°16’01.1”N 80°36’23.2”E) and ornamental Canna spp. were used to probe trees in the tree-space. The 10% of the plants in Kandy, (7°15’26.8”N 80°35’08.9”E) using initial trees were removed as burn-in, and the rest of the modified cetyltrimethylammonium bromide (CTAB) probed trees were used to draw the final 50% majority rule method (Porebski et al., 1997). The extracted DNA consensus tree. The Effective Sample Size (ESS) of all the samples were stored at -20 °C. The PCR was performed priors used were checked in TRACER v1.
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