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Performance of C4 versus C3 grasses in calcareous soil irrigated by slightly saline water in terms of the S:Fe index

Conference Paper · June 2014

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Performance of C4 versus C3 grasses in calcareous soil irrigated by slightly saline water in terms of the S:Fe index

D.L. Bouranis1,*, S.N. Chorianopoulou1, S. Nikologiannis2, D. Gasparatos3 1Agricultural University of Athens, Crop Science Department, Plant Physiology Laboratory, Iera Odos 75, 115885 Athens, Greece 2Nikologiannis Agricultural Store, Vassilissis Sofias and 4 Ioannou Metaxa Str., Peania, Greece 3Aristotle University of Thessaloniki, School of Agriculture, Laboratory of Soil Science, Thessaloniki 54124, Greece

*Corresponding author: email: [email protected]; tel: 00302105294287; www.aua.gr/pnpg

Abstract In a medium-textured calcareous soil (pH 8.04, CaCO3 41.2%) the following graminaceous species were sown in lanes among others: the warm season C4 dactylon, and Pennisetum clandestinum, as well as the cold season C3 Festuca arundinacea, and Lolium perenne. Sowing took place in December 15th 2012, and the lanes were irrigated regularly with slightly saline water (pH 7.5, Cl- 56.8 mg L-1, Na+ 90 mg L-1, 1080 μS cm-1). The situation of the was evaluated during July 2013. At the biodiversity level in the C3 lanes appeared mostly broadleaf weeds, whilst in the C4 lanes the prevalent weeds were the C4 Digitaria and Setaria. Analysis of each ’s nutrient status in total iron and sulfur revealed differential results. The examined warm season grasses presented in their aerial part concentrations of iron between 1.47-1.86 μmol g-1DM and of sulfur between 116-196 μmol g-1DM. In contrast the examined cold season grasses presented concentration ranges between 2.41-2.56 μmol g-1DM for iron and between 27-125 μmol g-1DM for total sulfur respectively. These values provided a S:Fe index of 79-105 for the examined warm season and of 14-52 for the cold season grasses. This index (1) indicates more acquisition of sulfur by the warm season grasses grown in this adverse system, and (2) it seems to be of diagnostic value; therefore it is proposed for further evaluation in more graminaceous species under the specific conditions.

Keywords: sulphur, iron, cold/warm season grasses, calcarerous soil, nutrition status index

1. INTRODUCTION

The material of the present study was the outcome of an internship project at the Agricultural University of Athens. The Hassioti Building atrium was not in its best shape, so to make a virtue of necessity for its restoration constituted as the primary aim. The establishment and maintenance of lawn, became one of the main goals in this study. An in depth analysis of the atrium’s current infrastructure revealed the existence of a calcareous type soil, and a slightly saline irrigation water derived from drilling. Under the circumstances a selection of cold season grasses was used and their performance was compared to that of the selected warm season grasses. Moreover, a decision for the non-grass repens and Trifolium repens to be included in this research was considered as necessary, due to their broad usage in lawns. How would the selected species react under the prevailing environmental conditions? Were these conditions adverse? What weeds would appear? The comparative performance of the selected cool and warm season grasses under the atrium’s environmental conditions, could be described as a very challenging attempt, indeed. An important finding to emerge in this study is that weeds can reveal a lot about the condition of the established lawn. Weeds thrive in unbalanced pH levels, poorly fertilized, and/or compacted soils, as well as in wrongly seeded, mowed and/or watered lawns. Trifolium repens (white clover) grows among turfgrass. It can tolerate close mowing, and can grow on many different types of soil and Proceedings of the 12th International Conference on Protection and Restoration of the Environment 653 Editors: A. Liakopoulos, A. Kungolos, C. Christodoulatos, A. Koutsopsyros ISBN 978-960-88490-6-8 levels of pH. It is considered to be a beneficial component of natural or organic lawn care, due to its ability to fix nitrogen and out-compete lawn weeds. Natural nitrogen fixing reduces leaching from the soil and can reduce the incidence of some lawn diseases that are enhanced by the availability of synthetic fertilizer. (kidney weed) can be used as a lawn substitute or groundcover in gardens. In general, cool season grasses start growing at 5 °C and are best adapted to grow at their fastest rate in temperatures ranging between 10 °C and 25 °C, in climates with relatively mild summers. This type is defined by two periods of rapid growth in the spring and autumn. Their typical growth is very dense, known as carpetlike lawns, with a thin layer of thatch. On the other hand, warm season grasses start growing at temperatures above 10 °C, and are best adapted to growth in the temperature range of 25 °C and 35 °C. This group is characterized by one long growth period over the spring and summer, however in colder months they enter dormancy and slightly tanned or brown leaves appear. Many warm season grasses are quite drought tolerant, and can survive drought conditions. The third aim of the project was the assessment of the nutritional status of the used species after a period of cultivation. Our focus was especially on sulfur and iron nutrition status, due to the intimate relationship of these nutrients and their ecological significance.

Table 1. Characteristics of the atrium soil at Hassioti Building, Agricultural University of Athens .

Sand (%) Silt (%) Clay (%) Texture CaCO3 (%) 47.1 26.3 26.6 SCL* 41.2 pH Organic matter (%) N (%) P (mg kg-1) K (mg kg-1) 8.04 1.21 0.13 46.9 704 * Sandy Clay Loam

2. MATERIALS AND METHODS

The cultivated area of the Hassioti Building atrium consisted of 360 m2 was divided in 10 lanes 15 m × 2.31 m each (in total 346.5 m2), separated by narrow lanes 15 m × 0.10 m each (in total 13.5 m2). Soil preparation prior to sowing included: soil and water evaluation, weed recognition, weed elimination (250 mL in 15 L of water were used), removal of stones, gravels, and existing vegetation. The existing irrigation system was repaired. Ground preparation required tilling at 30-40 cm deep which was conducted by a rotavator, along with a peripheral digging with the use of a hand tool. Soil samples were taken from topsoil for analysis. Afterwards, it was applied a 10- 18-18 (Anderson) fertilizer together with an 8-6-12+0.3B+25% organic material (Biofil olea) that consists of perlite and peat as well. At a later date, sowing took place in December 15th 2012. The irrigation schedule differed among the seasons with 3 minutes of water to be applied twice daily at 08:00 and 12:00 during winter, contrary to 12 minutes at 08:00 and 14:00 daily, during the summer. Finally, the results were evaluated in July 2013.

Fresh weight per sample (aerial part) was recorded and samples were oven-dried at 80 0C; dry weight was recorded, and the samples were ground to pass a 40-mesh screen using an analytical mill prior to chemical analysis. Sulfur (S) content was determined after dry ashing at 600 0C. The ash was dissolved in 2% (v/v) acetic acid aqueous solution, filtered through Whatman No. 42 paper, 2- and total sulfate (SO4 ) was determined turbidimetrically. Iron (Fe), potassium (K) and phosphorus (P) were determined following a wet acid digestion procedure based on the combination of nitric acid and 30% hydrogen peroxide. Phosphorus quantitative analysis in the diluted digests was carried out colorimetrically by determining the absorption of the blue phosphomolybdate complex at 660 nm, using the ammonium molybdate and stannus chloride procedure. The concentrations of Fe and K were determined in the diluted digests by atomic absorption spectrophotometry. Kjeldahl nitrogen (N) was determined by micro-Kjeldahl digestion followed by distillation [1].

654 Table 2. Characteristics of irrigation water vs tap water. The irrigation water comes from drilling in the campus of Agricultural University of Athens. Irrigation Tap water water pH 7.5 7.3 EC (μS cm-1) 1080 310 Total hardness (gd) 2.1 8.54 Cl- (mg L-1) 56.8 17.75 Na+ (mg L-1) 90 6 - -1 NO3 (mg L ) - 0.12 + -1 NH4 (mg L ) - -

Table 3. The mixtures or monocultures of plant species cultivated in Hassioti atrium were arranged in lanes. The cold or warm season type of each case is given.

Lane Species type 1 Wembley mixture cold season C3 2 Dichondra repens warm season C4 3 Pennisetum clandestinum (Κikuyu) warm season C4 4 Trifolium repens Barbian cold season C3 5 warm season C4 6 Festuca arundinacea Rebel cold season C3 7 Agrostis stolonifera Bengal cold season C3 8 Lollium perenne cold season C3 9 Festuca arundinaceae Tomanhank cold season C3 10 Mixture suitable for shadow cold season C3

3. RESULTS

The soil analysis of the atrium prior to fertilization revealed a medium-textured calcareous soil with pH 8.04, due to the presence of carbonates (41.2%, Table 1). Analysis of irrigation water showed a slightly saline water with pH 7.5, and an electric conductivity of 1080 μS cm-1 (Table 2). As seen in Tables 3 and 4, the selected warm season graminaceous species (lanes 2 and 5, Table 3) and the cold season ones (lanes 6-9, Table 3) were used as monocultures, along with two mixtures of graminaceous species (lanes 1 and 10, Table 4). The non-graminaceous species Dichondra repens (lane 2) and Trifolium repens (lane 4) were included for comparative reasons (both of them are used in lawns). No significant observations in Agrostis stolonifera was found since its establishment (lane 7) failed and it was not analyzed further. The initial situation included the weeds: Convolvulus arvensis, Taraxacum officinale, Oryzopsis hymenoides, Cynodon dactylon, Malva sylvestris, Parietaria judaica, Trifolium sp, Sonchus olevaceus, Lactuca seriolla, Veronica sp, Poa annua, Urtica sp, Gallium aparine, Capsella bursa-pastoris, and Ranunculus sp.

Table 4. The composition (species and percentage contribution) of each mixture of lanes 1 and 10.

Wembley mixture Mixture suitable for shadow 10% Festuca rubra Polka 40% Festuca rubra Barustic 15% Festuca arundinaceae Borneo 35% Festuca arundinaceae Palladio 15% Festuca arundinaceae Guardian 21 10% Festuca ovina Hardtop 40% Lollium perenne Caddieshack 10% Lolium perenne Pinacle II 20% Poa pratensis Geronimo 5% Poa pratensis Barimpala

655 Table 5. The distribution of weeds within the various lanes. Asterisk indicates the presence of the corresponding weed species, whilst a double asterisk denotes dominant presence.

Weed species type Lane 1 2 3 4 5 6 7 8 9 10 Taraxacum officinale C3 * * * * Plantago major * * * * * * Euphorbia chamaesyce * * Digitaria verticilate * Parietaria judaica * * * * Oxalis per-carpae * * * C4 ** ** * * * * * Setaria verticilata C4 ** ** * * * Picris echioides * * * ** * * Polycarpon tetraphyllum * * * * Convolvulus arvensis * * ** * Poa annua C3 * Malva sylvestris * Kickxia commutata * *

In July 2013, the following weeds were recognized: Taraxacum officinale, Plantago major, Euphorbia chamaesyce, Digitaria verticilate, Parietaria judaica, Oxalis per-carpae, Digitaria sanguinalis, Setaria verticilata, Picris echioides, Polycarpon tetraphyllum, Convolvulus arvensis, Poa annua, Malva sylvestris, Kickxia commutata, and their distribution within the lanes is depicted in Table 5. The underlined weeds were common in both examinations. Among the tested species, water content ranged between 2.7-8.6 g g-1 DM, and the concentration ranges were for N: 430-1180, P: 19.6-99.1, K: 6.7-112.4, Fe: 0.2-7.8, total S: 26.5-195.9, sulfate: 23.6-85.7, and organic S: 2.9- 110.2 μmol g-1 DM, respectively. Festuca arundinacea Rebel presented significantly lower sulfate and sulfur contens, and significantly high N:S ratio, compared to Festuca arundinacea Tomahawk. Cynodon dactylon presented the lowest water, N and K contents, and the higher total S, organic S and sulfate contents (Table 6); N:S value was the lowest one, too. The S:Fe ratios ranged between 9-450, and the N:S ratios between 2.2-28.3 (Table 7).

Table 6. Water content (W; g g-1Dry Mass) of the aerial part of each lane, along with its content in 2- nitrogen (N), phosphorus (P), potassium (K), iron (Fe), total sulfur (S), sulfate (SO4 ) and organic -1 sulfur (Sorg) (expressed as μmol g DM). In bold the higher and lower values.

W N P K Fe S SO4 Sorg Lane g g-1DM μmol g-1DM 1 4.3 660 68.7 8.2 7.8 74.0 49.7 24.3 2 7.1 790 71.2 18.7 1.3 138.6 38.5 100.1 3 8.6 920 64.7 48.1 1.5 116.1 36.7 79.4 4 7.3 820 70.0 112.4 0.2 89.9 51.4 38.5 5 2.7 430 59.4 6.7 1.9 195.9 85.7 110.2 6 4.4 750 82.1 21.1 2.6 26.5 23.6 2.9 7 – – – – – – – – 8 3.1 760 19.6 20.4 2.4 97.4 49.2 48.2 9 4.0 710 56.1 39.6 2.4 125.2 42.2 83.0 10 4.3 1180 99.1 47.7 4.9 49.1 35.5 13.6

656 Table 7. The S:Fe ratio of the aerial part of the species or mixture of each lane, along with the ratios 2- SO4 :S and N:S.

Lane S:Fe SO4:S N:S Lane S:Fe SO4:S N:S 1 9 0.672 8.9 6 10 0.891 28.3 2 110 0.278 5.7 7 – – – 3 79 0.316 7.9 8 40 0.505 7.8 4 450 0.572 9.1 9 52 0.337 5.7 5 105 0.437 2.2 10 10 0.723 24.0

4. DISCUSSION

In the time course of this project, we did have indeed the opportunity and the pleasure to restore the atrium’s picture (1st goal), by establishing and maintaining lawns of several species for educational reasons (2nd goal). Both aims were achieved successfully. Weed diversity within a lane and between lanes revealed various preferences and obstructions. The most prominent preference was that of the C4 weeds and grasses Setaria verticilata and Digitaria sanguinalis in C4 warm season grasses Pennisetum clandestinum and Cynodon dactylon. Digitaria sanguinalis was also found in some of the lanes of C3 cold season grasses, however not in dominance, whilst it did not appear at all in the rest of these lanes.

The results of the nutritional status of the lawns (3rd goal) revealed the S:Fe ratio as the most significant finding of this project so far. Plant requirements for sulfur are closely linked to nitrogen availability and growth rate. Intensive crop production requires sulfur inputs, especially as the anthropogenic S inputs have decreased in many regions, over the last years [2]. Establishing and maintaining a lawn is an intensive agronomic activity. Furthermore, S is intimately related to Fe [3,4]. The S:Fe ratio of warm season grasses (values 79 and 105; Table 7) presented a remarkable deviation to that of cold season grasses (values ranging between 9-52). Interestingly, the S:Fe ratio was 110 and 450 for the non-grasses Dichondra repens and Trifolium repens. However, a number of limitations need to be considered, as knowledge on sulfur biology and ecology of these systems is rather fragmentary and further research is needed towards confirmation and understanding of this finding.

5. CONCLUSIONS

The S:Fe ratio perhaps indicates more acquisition of sulfur relative to iron, by the warm season grasses grown in this adverse system. Moreover, the study adds to the body of knowledge on this field, because this ratio seems to be of diagnostic value; therefore the S:Fe ratio is proposed as an index of performance for further evaluation in the graminaceous species under the specific conditions.

Acknowledgements The authors gratefully acknowledge the technical contribution of the students Elma Bali, Anastasia Chatzopoulou, Natassa El Trouk, Ioanna Kanaki, Nikos Katris, Konstantina Kounougeri, Danai Martoni, Katerina Ntoutsouli, Yiorgos Paraskevaidis, Petros Sigalas, the colleagues Philippa Maniou, Milena Nikolopoulou, Panagiotis Trigas, and Barrenbruck company for providing the seeds used in this project. The Traineeship Programme for undergraduate Students of the Agricultural University of Athens has been funded by the National Strategic Reference Framework 2007–2013 and the European Union. The authors are also grateful to the reviewers for their input.

657 References

1. Bouranis D.L., S.N. Chorianopoulou, V.F. Siyiannis, V.E. Protonotarios, C. Koufos, P. Maniou (2012) Changes in nutrient allocation between roots and shoots of young maize plants during sulfate deprivation. Journal of Plant Nutrition and Soil Science, 175, 499-510. doi: 10.1002/jpln.201100154 2. Hawkesford M. J. (2007) Sulfur and plant ecology: A central role of sulfate transporters in responses to sulfur availability, pp.1-55, In: M.J. Hawkesford, L.J. De Kok (eds) Sulfur in Plants. An ecological perspective, Springer. 3. Bouranis D.L., P. Buchner, S.N. Chorianopoulou, L. Hopkins, V.E. Protonotarios, V.F.Siyiannis, M.J. Hawkesford (2008). Chapter 1 - Responses to sulfur limitation in maize. In: Sulfur assimilation and abiotic stress in plants (N.A. Khan, S. Singh, S. Umar Eds.), Springer, pp. 1-19.doi: 10.1007/978-3-540-76326-0_1 4. S. Astol, S. Zuchi, G. Neumann, S. Cesco, L. Sanita` di Toppi and R. Pinton (2012) Response of barley plants to Fe deciency and Cd contamination as affected by S starvation. Journal of Experimental Botany, Vol. 63, No. 3, pp. 1241–1250. doi:10.1093/jxb/err344

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