Characterization of phenology, physiology, morphology and biomass traits across a broad Euro-Mediterranean ecotypic panel of the lignocellulosic feedstock Arundo donax

Abstract

Giant reed (Arundo donax L.) is a perennial rhizomatous grass, which has attracted great attention as a potential lignocellulosic feedstock for bioethanol production due to high biomass yield in marginal land areas, high polysaccharide content and low inhibitor levels in microbial fermentations. However, little is known about the trait variation that is available across a broad ecotypic panel of A. donax nor the traits that contribute most significantly to yield and growth in drought prone environments. A collection of 82 ecotypes of A. donax sampled across the Mediterranean basin was planted in a common garden experimental field in Savigliano, Italy. We analysed the collection using 367 clumps representing replicate plantings of 82 ecotypes for variation in 21 traits important for biomass accumulation and to identify the particular set of ecotypes with the most promising potential for biomass production. We measured morpho-physiological, phenological and biomass traits and analysed causal relationships between traits and productivity characteristics assessed at leaf and canopy levels. The results identified differences among the 82 ecotypes for all studied traits: those showing the highest level of variability included stomatal resistance, stem density (StN), stem dry mass (StDM) and total biomass production (TotDM). Multiple regression analysis revealed that leaf area index, StDM, StN, number of nodes per stem, stem height and diameter were the most significant predictors of TotDM and the most important early selection criteria for bioenergy production from A. donax. These traits were used in a hierarchical cluster analysis to identify groups of similar ecotypes, and a selection was made of promising ecotypes for multiyear and multisite testing for biomass production. Heritability estimates were significant for all traits. The potential of this ecotype collection as a resource for studies of germplasm diversity and for the analysis of traits underpinning high productivity of A. donax is highlighted.

REFERENCES Ackerly, D. D., Dudley, S. A., Sultan, S. E., Schmitt, J., Coleman, J. S., Linder, C. R., … Lechowicz, M. J. (2000). The evolution of plant ecophysiological traits: Recent advances and future directions. BioScience, 11, 979–995.https://doi.org/10.1641/0006-3568(2000)050[0979:TEOPET]2.0.CO;2 Adiredjo, A. L., Navaud, O., Muños, S., Langlade, N. B., Lamaze, T., & Grieu, P. (2014). Genetic control of water use efficiency and leaf carbon isotope discrimination in sunflower (Helianthus annuus L.) subjected to two drought scenarios. PLoS ONE, 9, e101218. https://doi.org/10.1371/journal.pone.0101218 Ahmad, R., Liow, P. S., Spencer, D. F., & Jasieniuk, M. (2008). Molecular evidence for a single genetic clone of invasive Arundo donax in the United States. Aquatic Botany, 88, 113–120. https://doi.org/10.1016/j.aquabot.2007.08.015 Amaducci, S., & Perego, A. (2015). Field evaluation of Arundo donax clones for bioenergy production. Industrial Crops and Products, 75, 122–128. https://doi.org/10.1016/j.indcrop.2015.04.044 Angelini, L. G., Ceccarini, L., & Bonari, E. (2005). Biomass yield and energy balance of giant reed (Arundo donax L.) cropped in central Italy as related to different management practices. European Journal of Agronomy, 22, 375–389. https://doi.org/10.1016/j.eja.2004.05.004 Angelini, L. G., Ceccarini, L., Nassi o Di Nasso, N., & Bonari, E. (2009). Comparison of Arundo donax L. and Miscanthus × giganteus in a long-term field experiment in central Italy: Analysis of productive characteristics and energy balance. Biomass and Bioenergy, 33, 635–643. https://doi.org/10.1016/j.biombioe.2008.10.005 Aravanopoulos, F. A. (2010). Contribution of leaf morphometrics in the study of genetic entries in Salix L. Electronic Journal of Plant Breeding, 1, 1320–1328. Barney, J. N., & Ditomaso, J. M. (2008). Nonnative species and bioenergy: Are we cultivating the next invader? BioScience, 58, 64–70. https://doi.org/10.1641/B580111 Bogeat-Triboulot, M. B., Brosche, M., Renaut, J., Jouve, L., Le Thiec, D., Fayyaz, P., … Dreyer, E. (2007). Gradual soil water depletion results in reversible changes of gene expression, protein profiles, ecophysiology, and growth performance in Populus euphratica, a poplar growing in arid regions. Plant Physiology, 143, 876–892. Bolmgren, K., & Cowan, P. D. (2008). Time-size tradeoffs: A phylogenetic comparative study of flowering time, plant height and seed mass in a north-temperate flora. Oikos, 117, 424–429. https://doi.org/10.1111/j.2007.0030-1299.16142.x Borin, M., Barbera, A. C., Milani, M., Molari, G., Zimbone, S. M., & Toscano, A. (2013). Biomass production and N balance of giant reed (Arundo donax L.) under high water and N input in Mediterranean environments. European Journal of Agronomy, 51, 117–119. https://doi.org/10.1016/j.eja.2013.07.005 Bylesjö, M., Segura, V., Soolanayakanahally, R. Y., Rae, A. M., Trygg, J., Gustafsson, P., … Street, N. R. (2008). LAMINA: A tool for rapid quantification of leaf size and shape parameters. BMC Plant Biology, 8, 82. https://doi.org/10.1186/1471-2229-8-82 Ceotto, E., & Di Candilo, M. (2010). Shoot cuttings propagation of giant reed (Arundo donax L.) in water and moist soil: The path forward? Biomass and Bioenergy, 34, 1614–1623. https://doi.org/10.1016/j.biombioe.2010.06.002 Ceotto, E., Di Candilo, M., Castelli, F., Badeck, F.-W., Rizza, F., Soave, C., … Marletto, V. (2013). Comparing solar radiation interception and use efficiency for the energy crops giant reed (Arundo donax L.) and sweet sorghum (Sorghum bicolor L. Moench). Field Crops Research, 149, 159–166. https://doi.org/10.1016/j.fcr.2013.05.002 Charrad, M., Ghazzali, N., Boiteau, V., & Niknafs, A. (2013). NbClust: An examination of indices for determining the Number of Clusters. R package version 1.4. Chaves, M. M., Flexas, J., & Pinheiro, C. (2009). Photosynthesis under drought and salt stress: Regulation mechanisms from whole plant to cell. Annals of Botany, 103, 551–560. https://doi.org/10.1093/aob/mcn125 Clifton-Brown, J. C., Robson, P. R., & Allison, G. G. (2008). Miscanthus: Breeding our way to a better future. Aspects of Applied Biology, 90, 199–206. Condon, A. G., Richards, R. A., Rebetzke, G. J., & Farquhar, G. D. (2004). Breeding for high water-use efficiency. Journal of Experimental Botany, 55, 2447–2460. https://doi.org/10.1093/jxb/erh277 Cornic, G., & Fresneau, C. (2002). Photosynthetic carbon reduction and carbon oxidation are the main electron sinks for photosystem II activity during a mild drought. Annals of Botany, 89, 887–894. https://doi.org/10.1093/aob/mcf064 Corno, L., Pilu, R., & Adani, F. (2014). Arundo donax L.: A non-food crop for bioenergy and bio-compound production. Biotechnology Advances, 32, 1535–1549. https://doi.org/10.1016/j.biotechadv.2014.10.006 Cosentino, S. L., Copani, V., D'Agosta, G. M., Sanzone, E., & Mantineo, M. (2006). First results on evaluation of Arundo donax L. clones collected in Southern Italy. Industrial Crops and Products, 23, 212–222. https://doi.org/10.1016/j.indcrop.2005.06.004 Cosentino, S. L., Copani, V., Patanè, C., Mantineo, M., & D'Agosta, G. M. (2008). Agronomic, energetic and environmental aspects of biomass energy crops suitable for Italian environments. Italian Journal of Agronomy, 2, 81–95. https://doi.org/10.4081/ija.2008.81 Cosentino, S. L., Copani, V., Scalici, G., Scordia, D., & Testa, G. (2015). Soil erosion mitigation by perennial species under Mediterranean environment. Bioenergy Research, 8, 1538–1547. https://doi.org/10.1007/s12155-015-9690-2 Cosentino, S. L., Scordia, D., Sanzone, E., Testa, G., & Copani, V. (2014). Response of giant reed (Arundo donax L.) to nitrogen fertilization and soil water availability in semi-arid Mediterranean environment. European Journal of Agronomy, 60, 22–32. https://doi.org/10.1016/j.eja.2014.07.003 Dillen, S. Y., Marron, N., Sabatti, M., Ceulemans, R., & Bastien, C. (2009). Relationships among productivity determinants in two hybrid poplar families grown during three years at two contrasting sites. Tree Physiology, 29, 975–987. https://doi.org/10.1093/treephys/tpp036 Donald, C. M. (1968). The breeding of crop ideotypes. Euphytica, 17, 385–403. https://doi.org/10.1007/BF00056241 Fagnano, M., Impagliazzo, A., Mori, M., & Fiorentino, N. (2015). Agronomic and environmental impacts of giant reed (Arundo donax L.): Results from a long-term field experiment in hilly areas subject to soil erosion. BioEnergy Research, 8, 415–422. https://doi.org/10.1007/s12155-014-9532-7 Farquhar, G. D., Ehleringer, J. R., & Hubick, K. T. (1989). Carbon isotope discrimination and photosynthesis. Annual Review of Plant Physiology and Plant Molecular Biology, 40, 503–537. https://doi.org/10.1146/annurev.pp.40.060189.002443 Farrar, K., Bryant, D. N., Turner, L., Gallagher, J. A., Thomas, A., Farrell, M., … Donnison, I. S. (2012). Breeding for bio-ethanol production in Lolium perenne L.: Association of allelic variation with high water-soluble carbohydrate content. Bioenergy Research, 5, 149–157. https://doi.org/10.1007/s12155-011-9156-0 Giessow, J., Casanova, J., Lecler, R., MacArthur, R., & Fleming, G. (2011). Arundo donax – distribution and impact report. Berkeley, CA: California Invasive Plant Council, 240 pp. Gifford, J. M., Chae, W. B., Swaminathan, K., Moose, S. P., & Juvik, J. A. (2015). Mapping the genome of Miscanthus sinensis for QTL associated with biomass productivity. Global Change Biology Bioenergy, 7, 797–810. https://doi.org/10.1111/gcbb.12201 Granier, A., Loustau, D., & Bréda, N. (2000). A generic model of forest canopy conductance dependent on climate, soil water availability and leaf area index. Annals of Forest Science, 57, 755–765. https://doi.org/10.1051/forest:2000158 Haddadchi, A., Gross, C. L., & Fatemi, M. (2013). The expansion of sterile Arundo donax (Poaceae) in southeastern Australia is accompanied by genotypic variation. Aquatic Botany, 104, 153–161. https://doi.org/10.1016/j.aquabot.2012.07.006 Hardion, L., Verlaque, R., Baumel, A., Juin, M., & Vila, B. (2012). Revised systematics of Mediterranean Arundo (Poaceae) based on AFLP fingerprints and morphology. Taxon, 61, 1217–1226. Hardion, L., Verlaque, R., Saltonstall, K., Leriche, A., & Vila, B. (2014). Origin of the invasive Arundo donax (Poaceae): A trans-Asian expedition in herbaria. Annals of Botany, 114, 455–462. https://doi.org/10.1093/aob/mcu143 Harfouche, A., Meilan, R., & Altman, A. (2014). Molecular and physiological responses to abiotic stress in forest trees and their relevance to tree improvement. Tree Physiology, 34, 1181–1198. https://doi.org/10.1093/treephys/tpu012 Holland, J. B., Nyquist, W. E., & Cervantes-Martinez, C. T. (2003). Estimating and interpreting heritability for plant breeding: An update. Plant Breeding Reviews, 22, 9–112. Jakob, K., Zhou, F. S., & Paterson, A. (2009). Genetic improvement of C4 grasses as cellulosic biofuel feedstocks. In Vitro Cellular and Developmental Biology-Plant, 45, 291–305. https://doi.org/10.1007/s11627-009-9214-x Jiang, Q., Webb, S. L., Yesudas, C. R., Bhandari, H. S., Narasimhamoorthy, B., Bouton, J. H., & Saha, M. C. (2014). Variance components and heritability of biomass yield in switchgrass(Panicum virgatum L.) grown in the Southern Great Plains. Field Crops Research, 168, 148–155. https://doi.org/10.1016/j.fcr.2014.07.016 Kooyers, N. J. (2015). The evolution of drought escape and avoidance in natural herbaceous populations. Plant Science, 234, 155–162. https://doi.org/10.1016/j.plantsci.2015.02.012 Kottek, M., Grieser, J., Beck, C., Rudolf, B., & Rubel, F. (2006). World map of the Köppen-Geiger climate classification updated. Meteorologische Zeitschrift, 15, 259–263. https://doi.org/10.1127/0941-2948/2006/0130 Kou, L., Song, Y., Zhang, X., & Tan, T. (2017). Comparison of four types of energy grasses as lignocellulosic feedstock for the production of bio-ethanol. Bioresource Technology, 241, 424–429. https://doi.org/10.1016/j.biortech.2017.04.078 Krishnamurthy, L., Kashiwagi, J., Tobita, S., Ito, O., Upadhyaya, H. D., Gowda, C. L. L., … Varshney, R. K. (2013). Variation in carbon isotope discrimination and its relationship with harvest index in the reference collection of chickpea germplasm. Functional Plant Biology, 40, 1350–1361. https://doi.org/10.1071/FP13088 Lebrija-Trejos, E., Pérez-García, E. A., Meave, J. A., Bongers, F., & Poorter, L. (2010). Functional traits and environmental filtering drive community assembly in a species-rich tropical system. Ecology, 91, 386–398. https://doi.org/10.1890/08-1449.1 Lewandowski, I., Scurlock, J. M. O., Lindvall, E., & Christou, M. (2003). The development and current status of perennial rhizomatous grasses as energy crops in the US and Europe. Biomass and Bioenergy, 25, 335–361. https://doi.org/10.1016/S0961-9534(03)00030-8 Mariani, C., Cabrini, R., Danin, A., Piffanelli, P., Fricano, A., Gomarasca, S., … Soave, C. (2010). Origin, diffusion and reproduction of the giant reed (Arundo donax L.): A promising weedy energy crop. Annals of Applied Biology, 157, 191–202. https://doi.org/10.1111/j.1744-7348.2010.00419.x Mårtensson, L.-M., Carlsson, G., Prade, T., Kørup, K., Lærke, P. E., & Jensen, E. S. (2017). Water use efficiency and shoot biomass production under water limitation is negatively correlated to ¹³C discrimination in the C₃ grasses Dactylis glomerata, Festuca arundinacea and Phalaris arundinacea. Plant Physiology and Biochemistry, 113, 1–5. https://doi.org/10.1016/j.plaphy.2017.01.021 Mommer, L., Lenssen, J. P. M., Huber, H., Visser, E. J. W., & De Kroon, H. (2006). Ecophysiological determinants of plant performance under flooding: A comparative study of seven plant families. Journal of Ecology, 94, 1117–1129. https://doi.org/10.1111/j.1365-2745.2006.01175.x Nackley, L. L., Vogt, K. A., & Kim, S. H. (2014). Arundo donax water use and photosynthetic responses to drought and elevated CO₂. Agricultural Water Management, 136, 13–22. https://doi.org/10.1016/j.agwat.2014.01.004 Nassi o Di Nasso, N., Roncucci, N., & Bonari, E. (2013). Seasonal dynamics of aboveground and belowground biomass and nutrient accumulation and remobilization in giant reed (Arundo donax L.): A three-year study on marginal land. Bioenergy Research, 6, 725–736. https://doi.org/10.1007/s12155-012-9289-9 Nassi o Di Nasso, N., Roncucci, N., Triana, F., Tozzini, C., & Bonari, E. (2011). Productivity of giant reed (Arundo donax L.) and miscanthus (Miscanthus × giganteus Greef et Deuter) as energy crops: Growth analysis. Italian Journal of Agronomy, 6, e22. https://doi.org/10.4081/ija.2011.e22 Nsanganwimana, F., Marchand, L., Douay, F., & Mench, M. (2014). Arundo donax L. a candidate for phytomanaging water and soils contaminated by trace elements and producing plant-based feedstock. International Journal of Phytoremediation, 16, 982–1017. https://doi.org/10.1080/15226514.2013.810580 O'Leary, M. H. (1988). Carbon isotopes in photosynthesis. BioScience, 38, 328–336. https://doi.org/10.2307/1310735 Pilu, R., Bucci, A., Badone, F. C., & Landoni, M. (2012). Giant reed (Arundo donax L.): A weed plant or a promising energy crop? African Journal of Biotechnology, 11, 9163–9174. Pilu, R., Cassani, E., Landoni, M., Cerino Badone, F., Passera, A., Cantaluppi, E., … Adani, F. (2014). Genetic characterization of Italian giant reed (Arundo donax L.) clones collection: Exploiting clonal selection. Euphytica, 196, 169–181. https://doi.org/10.1007/s10681-013-1022-z Pilu, R., Manca, A., & Landoni, M. (2013). Arundo donax as an energy crop: Pros and cons of the utilization of this perennial plant. Maydica, 58, 1–6. Ping, H., Bransby, D. I., & van Santen, E. (2014). Long-term biomass yields of giant reed, mimosa and switchgrass in Alabama. Biofuels, Bioproducts & Biorefining, 8, 59–66. https://doi.org/10.1002/bbb.1437 Pison, G., Struyf, A., & Rousseeuw, P. J. (1999). Displaying a clustering with CLUSPLOT. Computational Statistics & Data Analysis, 30, 381–392. https://doi.org/10.1016/S0167-9473(98)00102-9 Plomion, C., Lalanne, C., Claverol, S., Meddour, H., Kohler, A., Bogeat-Triboulot, M. B., … Bonneu, M. (2006). Mapping the proteome of poplar and application to the discovery of drought-stress responsive proteins. Proteomics, 6, 6509–6527. https://doi.org/10.1002/(ISSN)1615-9861 Robson, P., Jensen, E., Hawkins, S., White, S. R., Kenobi, K., Clifton-Brown, J., … Farrar, K. (2013). Accelerating the domestication of a bioenergy crop: Identifying and modelling morphological targets for sustainable yield increase in Miscanthus. Journal of Experimental Botany, 64, 4143–4155. https://doi.org/10.1093/jxb/ert225 Rose, L. W. IV, Das, M. K., & Taliaferro, C. M. (2008). Estimation of genetic variability and heritability for biofuel feedstock yield in several populations of switchgrass. Annals of Applied Biology, 152, 11–17. https://doi.org/10.1111/j.1744-7348.2007.00186.x Rossa, B., Tüffers, A. V., Naidoo, G., & von Willert, D. J. (1998). Arundo donax L. (Poaceae) – a C₃ species with unusually high photosynthetic capacity. Plant Biology, 111, 216–221. Saikia, R., Chutia, R. S., Kataki, R., & Pant, K. K. (2015). Perennial grass (Arundo donax L.) as a feedstock for thermo-chemical conversion to energy and materials. Bioresource Technology, 188, 265–272. https://doi.org/10.1016/j.biortech.2015.01.089 Sánchez, E., Scordia, D., Lino, G., Arias, C., Cosentino, S. L., & Nogués, S. (2015). Salinity and water stress effects on biomass production in different Arundo donax L. clones. BioEnergy Research, 8, 1461–1479. https://doi.org/10.1007/s12155-015-9652-8 Scordia, D., Cosentino, S. L., Lee, J., & Jeffries, T. W. (2011). Dilute oxalic acid pretreatment for biorefining giant reed (Arundo donax L.). Biomass and Bioenergy, 35, 3018–3024. https://doi.org/10.1016/j.biombioe.2011.03.046 Scordia, D., Cosentino, S. L., Lee, J., & Jeffries, T. W. (2012). Bioconversion of giant reed (Arundo donax L.) hemicellulose hydrolysate to ethanol by Scheffersomyces stipitis CBS6054. Biomass and Bioenergy, 39, 296–305. https://doi.org/10.1016/j.biombioe.2012.01.023 Sharma, K. P., Kushwaha, S. P., & Gopal, B. (1998). A comparative study of stand structure and standing crops of two wetland species, Arundo donax and Phragmites karka, and primary production in Arundo donax with observations on the effect of clipping. Journal of Tropical Ecology, 39, 3–14. Silva, C. F. L., Schirmer, M. A., Maeda, R. N., Barcelos, C. A., & Pereira, N. Jr (2015). Potential of giant reed (Arundo donax L.) for second generation ethanol production. Electronic Journal of Biotechnology, 18, 10–15. https://doi.org/10.1016/j.ejbt.2014.11.002 Slavov, G., Robson, P., Jensen, E., Hodgson, E., Farrar, K., Allison, G., … Donnison, I. (2013). Contrasting geographic patterns of genetic variation for molecular markers vs. phenotypic traits in the energy grass Miscanthus sinensis. Global Change Biology Bioenergy, 5, 562–571. https://doi.org/10.1111/gcbb.12025 Spencer, D. F., Liow, P. S., Chan, W. K., Ksander, G. G., & Getsinger, K. D. (2006). Estimating Arundo donax shoot biomass. Aquatic Botany, 84, 272–276. https://doi.org/10.1016/j.aquabot.2005.11.004 Sun, S., & Frelich, L. E. (2011). Flowering phenology and height grow pattern are associated with maximum plant height, relative growth rate and stem tissue mass density in herbaceous grassland species. Journal of Ecology, 99, 991–1000. https://doi.org/10.1111/j.1365-2745.2011.01830.x Sun, L., Wang, Y., Yan, X., Cheng, T., Ma, K., Yang, W., … Zhang, Q. (2014). Genetic control of juvenile growth and botanical architecture in an ornamental woody plant, Prunus mume Sieb. et Zucc. as revealed by a high-density linkage map. BMC Genetics, 15(Suppl. 1), S1. https://doi.org/10.1186/1471-2156-15-S1-S1 Venables, W. N., & Ripley, B. D. (2002). Modern applied statistics with S (4th ed., p. 495). New York, NY: Springer-Verlag. https://doi.org/10.1007/978-0-387-21706-2 Verhulst, N., Cox, R., & Govaerts, B. (2013). Yield and yield components: A practical guide for comparing crop management practices. Mexico, DF: CIMMYT International Maize and Wheat Improvement Center, 23 pp. Watts, D. A., & Moore, G. W. (2011). Water-Use Dynamics of an Invasive Reed, Arundo donax, from Leaf to Stand. Wetlands, 31, 725–734. https://doi.org/10.1007/s13157-011-0188-1 Zucaro, A., Forte, A., Fagnano, M., Bastianoni, S., Basosi, R., & Fierro, A. (2015). Comparative attributional life cycle assessment of annual and perennial lignocellulosic feedstocks production under Mediterranean climate for biorefinery framework. Integrated Environmental Assessment Management, 11, 397–403. https://doi.org/10.1002/ieam.1604