Distribution and redistribution of phosphorus forms in grapevines

Scientia Horticulturae 218 (2017) 125–131

Contents lists available at ScienceDirect

Scientia Horticulturae journal homepage: www.elsevier.com/locate/scihorti

Distribution and redistribution of phosphorus forms in grapevines Rogério Piccin a , João Kaminski b , Carlos Alberto Ceretta b , Tales Tiecher c , Luciano Colpo Gatiboni d , Roque Junior Sartori Bellinaso b , Carina Marchezan b , Rodrigo Otávio Schneider de Souza b , Gustavo Brunetto b,∗ a

Graduate Program in Soil Science (PPGCS), Federal University of Santa Maria (UFSM), Santa Maria, Rio Grande do Sul, Brazil Department of Soil Science at UFSM, Santa Maria, Rio Grande do Sul, Brazil Universidade Federal do Rio Grande do Sul (UFGRS), Porto Alegre, Rio Grande do Sul, Brazil d State University of Santa Catarina (UDESC), Agroveterinary Science Center (CAV), Lages, Santa Catarina, Brazil b c

a r t i c l e

i n f o

Article history: Received 13 September 2016 Received in revised form 30 January 2017 Accepted 10 February 2017 Keywords: Phosphate fertilizer Chemical fractionation of P in the tissue Vitis vinifera L.

a b s t r a c t Increased phosphorus (P) available in soil can modify the partitioning of P forms in annual and perennial organs of grapevines throughout the growing season. This study was to evaluate the distribution and redistribution of P forms in organs of grapevines grown in soils with different contents of available P. The study was conducted in two vineyards installed in the city of Santana do Livramento, state of Rio Grande do Sul (RS), in southern Brazil. The treatments were vineyard 1 (V1) with 11.8 mg kg−1 of available P in soil and vineyard 2 (V2) with 34.6 mg kg−1 . The cultivar of both vineyards is Tannat (Vitis vinifera L.) grafted on SO4 (Vitis berlandieri × Vitis riparia) rootstock. Plant density per hectare was 2525 (1.2 m × 3.2 m) on a spur pruned cordon system. The grapevines were uprooted and partitioned into roots, trunks, arms, spurs, new-year shoots, leaves and clusters (when present) at flowering (F), veraison (V), harvest (H) and dormancy (D). The organs were dried, prepared and subjected to chemical fractionation of P, to estimate fractions of total acid-soluble P (PST ), acid-soluble inorganic P (PSI ), acid-soluble organic P (PSO ) (by difference between TSP and PSI ), phospholipids P (PLIP ), P associated with RNA (PRNA ), P associated with DNA (PDNA ) and residual P (PRES ). P in grapevines of V1 and V2 accumulated mainly in the PSI fraction in leaves and clusters, which was collected at F, V and H, and in PSO fraction in roots, collected at D. Part of the root PSI was redistributed at F to the leaves and clusters in vines of V1. Vines grown in V2 accumulated more P in PSO form in roots and tended to redistribute less PSI to the leaves and clusters after F. Grapevines accumulated P in roots, both in soils with low and high available P contents, and P was subsequently redistributed and accumulated in leaves and clusters in inorganic form. © 2017 Elsevier B.V. All rights reserved.

1. Introduction Phosphorus is one of the elements that most often limits agricultural productivity in the world. In the soil-plant system, P is characterized by having very low mobility in soil, because it is adsorbed strongly by iron and aluminum oxides of highly weathered soils and by forming complexes with Ca in calcareous soils (Bortoluzzi et al., 2015; Fink et al., 2014, 2016). However, it has high mobility within the plant and it is redistributed from one

∗ Corresponding author. E-mail addresses: [email protected] (R. Piccin), [email protected] (J. Kaminski), [email protected] (C.A. Ceretta), [email protected], [email protected] (T. Tiecher), [email protected] (L.C. Gatiboni), roquejunior [email protected] (R.J.S. Bellinaso), [email protected] (R.O.S.d. Souza), [email protected] (G. Brunetto). http://dx.doi.org/10.1016/j.scienta.2017.02.023 0304-4238/© 2017 Elsevier B.V. All rights reserved.

organ to another according to demand and availability of P in soil (Schachtman et al., 1998). In the face of a predicted imminent shortage of phosphate reserves in the middle of this century (Cordell et al., 2009), it is essential to improve understanding of the physiological and metabolic aspects of the use of P by plants in order to design strategies to improve the efficiency of P-fertilization. Within plants such as grapevines (Vitis vinifera L.), P can be accumulated in organic forms such as metabolically active organic P in the cytoplasm (PSO ), phospholipids (PLIP ), P associated with RNA and DNA (PRNA and PDNA ), phosphoproteins (PRES ) and in inorganic phosphorus (PSI ) form (Bieleski, 1973; Veneklaas et al., 2012). It is believed that plants grown in soils with greater P availability tend to accumulate higher levels of P in organs in PSI form, especially in annual organs, such as leaves, and in annual crops, in grains (Martinez et al., 2005; Lambers et al., 2011). The PLIP fraction represents P contained primarily in cell membranes (Bieleski, 1973) and its increase typically happens because of the increase in com-

126

R. Piccin et al. / Scientia Horticulturae 218 (2017) 125–131

plexes of cell membranes, especially thylakoid at plant flowering (Thomas and Sadras, 2001), in response to increase in PSI content (Reef et al., 2010; Veneklaas et al., 2012). P content in ribonucleic acids (PRNA and PDNA ) usually differs among organs, tissues and cells, and is higher especially in expanding leaves, lower in adult leaves, and very low in senescent leaves (Suzuki et al., 2001; Niklas, 2006). Generally, PSO content in annual organs such as leaves and fruit varies slightly with increasing availability of soil P (Lee and Ratcliffe, 1983). However, when cell division in these organs ceases, P may be redistributed and accumulated in perennial organs such as roots, the main nutrient reserve organ in fruit trees (Lima et al., 2011). In adult grapevines, it is not sufficiently known if the increase in P content available in soil can actually change the distribution of P forms, such as PSI, PSO, PRNA and PLIP, in annual organs (leaves, new year shoots and clusters) and perennial organs (roots, trunks, arms and spurs) throughout the phenological stages. For example, if the amount of P forms in annual organs is high, P depletion within the plant is expected, especially if the soil does not possess P in sufficient quantities to meet plant demand. On the other hand, if P forms are primarily accumulated in reserve organs such as roots, they are expected to contribute to the growth of annual organs in the following cycle, thus decreasing the dependence of the plant on soil P. However, this dynamic redistribution of P forms from perennial to annual organs throughout phenological stages of adult grapevines at field level has rarely been studied. Therefore, this study aimed to evaluate the distribution and redistribution of P forms in grapevines grown in soil with different contents of available P. 2. Material and methods 2.1. Description of the experimental area The study was conducted from August 2014 to March 2015 in two vineyards (V1 and V2) of the city of Santana do Livramento (Latitude 30◦ 49 8” S, Longitude 55◦ 27 3” W and altitude of 320 m), located in the Campanha Gaúcha region of Rio Grande do Sul (RS), southern Brazil. The soil of both vineyards is a Typic Hapludalf (Soil Survey Staff, 2006). The relief in both vineyards is slightly undulated with a 12% declivity. The physical and chemical characteristics of the both vineyard soils for the 0 − 20 cm layer are shown in Supplementary material 1. The climate is humid subtropical Cfa according to Köppen classification, which is characterized by mild temperatures and rain with little variation throughout the year. The average annual rainfall is approximately 1600 mm. The average temperature of the hottest month of the year (January) is 23.8◦ C and the average temperature of the coldest month (July) is 12.4◦ C. The annual sunshine is approximately 2500 h. Phenological stages, monthly average values of rainfall, temperature, humidity and sunshine observed during the study are shown in Supplementary material 2. 2.2. Treatments The treatments were two vineyards with two contents of available P in soil (extracted by Mehlich-1). Soil in V1 had 11.8 mg P kg−1 (considered low in a soil with 15% clay) and V2 34.6 mg P kg−1 (considered high in a soil with 10% clay) (Committee on Soil Chemistry and Fertility, 2004). V1 was established in 2004 and V2 in 2003. The cultivar of both vineyards is Tannat (Vitis vinifera L.) grafted on SO4 (Vitis berlandieri × Vitis riparia) rootstock. Plant density per hectare was 2525 (1.2 m × 3.2 m) on a spur pruned cordon system. The experimental design was randomized blocks with three replications. Each replication was formed by five plants and the three central grapevines were evaluated. During the experiment, the

grapevines were subjected to applications of 40 kg N ha−1 year−1 (urea) and 20 kg K2 O ha−1 year−1 (potassium chloride), according to the recommendation established by CQFS-RS/SC (2004). Phosphorus was never applied to the soil of the vineyards in the evaluated growth season.

2.3. Collection of grapevines and fractionation in organ The grapevines were uprooted and partitioned at four phenological stages: flowering (F) (October 12, 2014), when 50% of flowers were open; veraison (V) (December 20, 2014), when 50% of berries changed color; harvest (H) (January 27, 2015) at which point 100% of the grapes showed intense color development; and dormancy (D) (April 11, 2015) (Baillod and Baggiolini, 1993). The grapevines were cut close to the soil surface, separated into roots, trunks, arms, spurs, new year shoots, leaves and clusters (when present in some phenological stages). The roots were collected in a 1.5 m3 trench with a 0.7 m radius (the trunks was the central point) and 1.0 m depth. The roots were collected manually and stored. The trunks was cut close to the soil surface. The shoots were separated into new year shoots, spurs and arms. The leaves and clusters were cut at the intersection of the shoots and collected. All the organs were weighed in the field with the use of a hook digital precision scale. A subsample of each organ was collected, weighed and stored. The subsamples of the organs were dried in an oven with forced air at 65 ◦ C until constant weight. Then the subsamples of dried organs were ground in a Wiley mill and passed through a 2 mm mesh sieve. Dry matter of different vine organs are presented in Supplementary material 3. Grape yield was 5.4 (±1.3) and 9.8 (±0.9) Mg ha−1 in V1 and V2, respectively (data not shown). The tissue of the grapevine organs previously stored was subjected to chemical fractionation of P according to the methodology proposed by Casali et al. (2011). P forms obtained were: Total soluble P in acid (PST ), inorganic soluble P in acid (PSI ), organic soluble P in acid (PSO ) (by the difference between PST and PSI ), lipid P (PLIP ), P associated with RNA (PRNA ), P associated with DNA (PDNA ) and residual P (PRES ). The determination and quantification of all P forms was done according to Murphy and Riley, 1962 in a UV–vis spectrophotometer.

2.4. Statistical analyses The content of P forms (PST , PSI , PLIP, PRNA , PDNA , PRES ) in grapevine organs were subjected to analysis of variance using SISVAR (Ferreira, 1998). When the effects were significant by analysis of variance, the results obtained were subjected to a mean comparison test, based on levels of significance lower than 5% (p < 0.05) by Scott-Knott test (1974). To compare the effect of the P content available in both vineyards, we made orthogonal contrasts by comparing the values of the P content of each fraction (PST , PSI , PLIP, PRNA , PDNA , PRES ) within each phenological stage, between grapevines grown in soil with low (n = 3) and high (n = 3) levels of available P. To evaluate the distribution of P in vines during the production cycle, we used only PSI and PSO contents, as they contribute most to total P content and presented greater variation throughout the production period. We used Eq. (1) to calculate the percentages: F (%) = PFraction (1)where: F (%) is the percentage of P in each TotalP fraction; P fraction is the P content of the fraction in mg kg−1 and total P is the P content in each organ in mg kg−1 .

R. Piccin et al. / Scientia Horticulturae 218 (2017) 125–131

127

Table 1 P forms (mg kg−1 ) in grapevine organs of Vineyard 1 (V1), grown in soil with low available P content and collected during flowering (F), veraison (V), harvest (H) and dormancy (D). Vineyard

Stage

PSI

PSO

PLIP

PRNA

PDNA

PRES

Total

CV (%)

V1

Flowering Roots Trunks Arms Spurs New year shoots Leaves Clusters CV (%) Roots Veraison Trunks Arms Spurs New year shoots Leaves Clusters CV (%) Harvest Roots Trunks Arms Spurs New year shoots Leaves Clusters CV (%) Dormancy Roots Trunks Arms Spurs New year shoots CV (%)

729.2 aB 320.1 aC 310.0 aC 666.5 aB 2093.8 aA 2050.6 aA 1997.1 aA 8.32 582.3 aB 408.5 aC 419.0 aC 648.4 aB 806.9 aA 917.5 aA 1046.8 aA 12.6 432.3 bB 258.2 aB 298.8 aB 391.4 aB 662.1 aA 768.2 aA 761.6 aA 18.34 299.1 bB 235.1 bB 302.9 bB 387.3 aB 618.9 aA 17.2

633.0 aA 80.1 bD 65.1 cD 47.0 cC 151.8 bD 252.2 dB 212.5 cB 13.69 495.7 aA 316.8 bB 96.2 bC 131.0 bC 298.0 bB 350.6 cB 521.0 bA 13.96 574.8 aA 152.4 bD 167.4 bD 247.2 bC 318.0 bB 193.5 dD 300.2 cB 13.23 605.8 aA 398.0 aB 403.4 aB 332.0 aB 512.2 bA 9.54

60.9 cC 57.7 cC 57.7 cC 57.7 cC 191.3 bB 358.9 cA 298.9 cA 29.16 82.1 cC 83.8 cC 90.6 bC 127.9 bB 176.6 bA 206.0 aA 207.2 cA 18.33 151.7 cB 104.2 dB 114.3 cB 126.8 cB 157.2 cB 378.1 cA 227 cB 14.21 79.3 cA 70.5 cA 67.5 cA 90.0 bA 101.8 cA 18.62

266.2 bC 105.1 bE 121.7 bE 194.7 bD 337.9 bB 625.8 bA 657.6 bA 9.88 241.7 bB 129.1 cD 127.5 bD 135.4 bD 192.1 bC 459.7 bA 223.2 cB 12.96 238.8 cC 303.9 aC 226.5 bC 303.7 bC 381.3 bB 554.9 bA 443.8 bB 6.96 147.2 cA 62.0 cB 59.9 cB 83.7 bB 81.3 cB 15.03

23.8 cB 25.1 cB 23.6 dB 22.0 cB 22.8 cB 28.6 eA 31.2 dA 12.61 17.7 cB 14.9 cB 16.2 cB 16.7 bB 17.0 cB 49.7 eA 20.6 dB 14.42 32.4 dB 31.9 dB 32.4 dB 35.0 cB 36.2 dB 54.0 dA 53.4 dA 8.52 31.0 cA 28.3 cA 21.1 cA 26.6 bA 27.2 cA 10.82

37.6 cB 40.9 cB 26.0 dB 42.7 cB 33.7 cB 59.7 eA 51.7 dA 11.76 109.2 cB 82.9 cC 74.3 bC 108.5 bB 181.5 bA 165.0 dA 124.9 cB 12.33 53.8 dC 31.7 dC 36.9 dC 36.9 cC 25.2 dB 98.3 dA 48.7 dB 13.53 68.9 cA 58.3 cA 57.8 cA 67.7 bA 69.8 cA 11.35

1750.7C 636.0 E 604.2 E 1030.8 D 2831.3 B 3375.7A 3249.0A 6.83 1528.7 B 1036.1C 823.8 D 1168.0C 1672.2 B 2148.7A 2143.6A 6.64 1483.9C 882.2 E 876.3 E 1140.9 D 1580.0C 2047.1A 1835.2 B 8.74 1231.3A 852.1 B 912.6 B 987.4 B 1411.1A 8.97

7.38 8.68 9.24 19.08 9.45 11.04 11.24

Flowering Roots Trunks Arms Spurs New year shoots Leaves Clusters CV (%) Roots Veraison Trunks Arms Spurs New year shoots Leaves Clusters CV (%) Harvest Roots Trunks Arms Spurs New year shoots Leaves Clusters CV (%) Dormancy Roots Trunks Arms Spurs New year shoots CV (%)

675.4 aB(2) 486.3 aC 433.8 aC 754.5 aB 2129.5 aA 2002.2 aA 2036.6 aA 8.32 397.8 bC 312.3 aC 312.8 aC 477.5 aC 706.6 aB 935.6 aA 999.7 aA 12.6 352.9 bC 356.2 aC 315.3 aC 383.2 aC 879.3 aB 1719.4 aA 876.7 aB 18.34 515.3 bB 360.6 bC 322.3 bC 556.1 aB 706.9 aA 17.2

550.8 bA 394.1 bB 215.7 bC 151.6 bC 189.5 bC 184.2 cC 500.4 bA 13.69 624.3 aA 153.1 bD 170.8 bD 141.0 bD 350.3 bC 334.0 cC 455.2 bB 13.96 630.8 aA 281.3 bB 151.1 cC 215.9 bC 431.8 bB 274.6 bB 438.2 bB 13.23 1139.9 aA 651.7 aB 501.2 aC 542.4 aC 622.8 aB 9.54

70,5 dC 74.7 cC 70.2 cC 98.3 bC 301.6 bB 405.3 bA 437.5 bA 29.16 87.0 dC 85.0 cC 101.6cC 86.0 cB 151.7 cB 236.6 cA 205.2 cA 18.33 144.9 cD 144.9 cD 141.5 cD 135.9 cD 208.3 cC 461.9 bA 296.6 cB 14.21 106.7 cA 76.3 cA 84.2 cA 99.8 bA 132.1 bA 18.62

147.6 cC 105.0 cC 108.1 cC 111.1 bC 210.2 bB 506.8 bA 543.3 bA 9.88 255.9 cB 147.5 bC 146.5 bC 181.5 bB 202.0 cB 580.8 bA 246.4 cB 12.96 169.9 cD 185.5 cD 228.2 bC 247.7 bC 252.6 cC 471.9 bA 325.1 cB 6.96 196.8 cA 85.9 cB 67.4 cB 96.8 bB 120.7 bB 15.03

24.0 dA 27.4 dA 27.1 dA 30.9 bA 24.3 cA 25.8 cA 24.7 cA 12.61 34.2 dC 33.4 dC 44.6 dB 36.0 cC 40.3 dB 62.6 dA 46.9 dB 14.42 35.7 dB 32.2 dB 32.2 dB 33.9 cB 31.9 dB 51.2 cA 47.9 dA 8.52 39.8 cA 28.8 cB 23.4 cB 28.3 bB 31.5 bB 10.82

35.6 dB 24.9 dC 26.0 dC 35.9 dB 23.8 cC 61.7 cA 61.0 cA 11.76 104.1 dB 45.8 dC 55.8 dC 71.2 cC 100.7 dB 159.6 dA 112.0 dB 12.33 56.5 dB 44.1 dC 30.0 dD 39.1 cC 29.3 dD 77.7 cA 26.5 dD 13.53 105.9 cA 87.4 cB 78.0 cB 75.6 bB 69.3 bB 11.35

1503.9 D 1112.4 E 880.9 F 1182.4 E 2878.9C 3186.0 B 3603.6A 6.83 1503.3C 777.1 E 832.1 E 993.2 D 1551.6C 2309.1A 2065.4 B 6.64 1390.7C 1044.2 D 898.2 D 1055.6 D 1833.3 B 3056.6A 2011.0 B 8.74 2104.3A 1290.7C 1076.5 D 1399.0C 1683.3 B 8.97

7.38 8.68 9.24 19.08 9.45 11.04 11.24

V2

Organs

10.03 10.12 7.06 11.42 8.11 11.26 9.04 16.92 10.7 4.99 26.24 10.45 16.17 12.06 12.44 12.44 13.81 15.04 14.31

10.03 10.12 7.06 11.42 8.11 11.26 9.04 16.92 10.7 4.99 26.24 10.45 16.17 12.06 12.44 21.19 13.81 15.04 14.31

PSI = soluble inorganic P; PSO = soluble organic P; PLIP = phospholipids; PRNA = P associated with RNA; PDNA = P associated with DNA and Pres = residual P. Means followed by the same letter, lower case on the line and capitals on the column at the same phenological stage did not statistically differ by the Scott-Knott test at 5% probability of error. Values in bold highlight the highest values of each fraction within each organ and phenological stage. CV% = coefficient of variation.

3. Results 3.1. Distribution of P fractions in grapevine organs grown in soil with low available P content at different phenological stages In vineyard 1 (V1), where soil has low available P content, the highest content of total P was found in leaves and clusters at pheno-

logical stages of flowering (F) and veraison (V); in leaves at harvest (H) and in roots and new year shoots at dormancy (D) (Table 1). The highest content of PSI was observed in the new year shoots, leaves and clusters at F, V and H; and only in new year shoots at D. The highest content of PSO was found in the roots at F and H; in roots and clusters in V, and in roots and new year shoots at D. The high-

128

R. Piccin et al. / Scientia Horticulturae 218 (2017) 125–131

est content of PLIP was observed in the leaves and clusters at F; in new year shoots, leaves and clusters at V; in leaves at H. Also, there was no significant difference in the fraction PLIP between organs at D. Similar to the PLIP fraction, the highest contents of PRNA , PDNA and PRES were observed in the leaves and clusters at F. The highest content of PRNA was in the leaves at V and H, and in roots at D. The PDNA fraction was highest in the leaves at V and in leaves and clusters at H. The PRES fraction was highest in the new year shoots and leaves at V and only in leaves at H. The contents of the PDNA and PRES fractions did not differ significantly between organs at D. In the grapevines roots of V1, contents of total P, PSI and PRNA decreased from F to D (Table 1). PSO content of roots decreased at V and increased at D. In the trunks and arms, total P content increased from F to V and PSO content increased from H to D. PRNA content of trunks and arms increased from V to H and decreased from H to D. In the spurs, PSI content decreased from V to H. PSO content increased from F to D. PSI and Total P contents decreased in the new year shoots from F to D; and in the leaves and clusters from F to H. PSO content increased in the new year shoots from F to D. In the leaves and clusters PSO content increased from F to V and decreased from V to H. PRNA content decreased in the new year shoots, leaves and clusters from F to V, and increased from V to H, reducing again from H to D. Fractions of PLIP , PDNA and PRES did not change between phenological stages (Table 1). 3.2. Distribution of P fractions in grapevine organs grown in soil with high available P content at different phenological stages In grapevines of vineyard 2 (V2), grown in soil with high available P content, the highest content of total P was observed in the clusters at F; in leaves at V and in roots at D (Table 1). The highest content of PSI was found in the new year shoots, leaves and clusters and clusters at F; in leaves and clusters at V, in leaves at H and in new year shoots at D. The highest content of PSO was observed in the roots and clusters at F and only in roots at V, H and D. The highest contents of PLIP and PRNA were found in the leaves and clusters at F and in leaves at H. In V, the highest content of PLIP was observed in the leaves and clusters; and PLIP content did not differ significantly between the organs at D. However, PRNA fraction was the highest in the leaves at V and in the roots at D. The highest contents of PDNA and PRES were found in the leaves at V and in the roots at D. In V2, the roots of the vines had the highest P content in the PSI fraction at F and PSO at V, H and D (Table 1). In the trunks, arms, spurs, shoots, leaves and clusters the highest P content was found in PSI fraction at F, V and H; and in PSO fraction at D. New year shoots was an exception, because P content in PSI and PSO fractions did not differ significantly at D. In the roots of grapevines in V2, contents of total P and PSI decreased from F to H, and increased from H to D (Table 1). In the trunks, arms and spurs contents of total P and PSI decreased from F to V. Contents of total P and PSO increased from V to D. In the new year shoots, leaves and clusters total P, PSI and PLIP contents decreased from F to V. Total P and PSI contents increased only in the leaves from V to H. PSO content increased only in the new year shoots from F to D. On the other hand, PRNA content decreased only in the clusters from F to V. PLIP , PRES and PDNA fractions did not change between phenological stages (Table 1). 3.3. Influence of available P content in soil on the distribution of P forms in grapevine organs throughout the phenological stages The available P content in soil changed total P content in the trunks and arms at F; in the trunks at V, in the leaves at H; and in the roots, trunks, arms and spurs at D (Table 2). P content in the soil increased PSI content in the trunks and arms at F; and in the leaves and trunks at D. P content in soil increased PSO content in

the trunks, arms, spurs and clusters at F; in the roots and trunks at V; in the trunks at H; and in the roots, trunks and spurs at D. The highest contents of total P, PSI and PSO were observed in V2, with the exception of PSO content in the trunks at V of vines grown in V1. P content in soil increased PLIP content only in the new year shoots at F (Table 2). PRNA fraction increased in the roots, spurs, new year shoots, leaves and clusters at F; in the leaves at V; in the roots, trunks, new year shoots and clusters at H of grapevines in V1, with the exception of the trunks that decreased at V. PDNA fraction was the highest in the roots, trunks, arms, spurs, new year shoots and clusters at F; and in the roots at D of grapevines grown in V2. PRES content was the increased in the trunks, arms, spurs and new year shoots at V of vines grown in V2; and highest in the roots, trunks and arms at D of grapevines in V1. 3.4. Redistribution of PSI and PSO among grapevine organs throughout the phenological stages The PSI form at F in grapevines of both vineyards (V1 and V2) was greater than 40% of the total P content in the roots, trunks and arms; greater than 55% in the clusters; and greater than 60% in the spurs, new year shoots and leaves (Fig. 1). However, PSI content at D in grapevines of V1 and V2 decreased to less than 25% in the roots, less than 30% in the trunks, less than 35% in the arms, and less than 45% in the spurs and new year shoots. On the other hand, PSO content at F in grapevines of V1 and V2 was less than 15% in the clusters and spurs; less than 10% in the leaves and new year shoots and less than 40% in the roots. But it increased to over 35% in the new year shoots and spurs, and over 45% in the arms at D. In grapevines of V1 and V2, PSO content in the trunks and arms at F was less than 15%, but increased to over 40% at D. In vines of V2 PSO content in the arms went from 24% at F to 50% at D, and in the trunks from 35% at F to 54% at D (Fig. 1). 4. Discussion The highest contents of total P and PSI in the leaves and clusters of grapevines in vineyards V1 and V2 collected at phenological stages of F and V can be attributed to intense cell division in the tissue of these two growing organs, which promotes the increase of dry matter (Supplementary material 3) and consequently become P sinks (Williams, 1987; Borém and Ramos, 2002; Tagliavini et al., 2005; Lambers et al., 2010). However, the increase of PSI in the leaves and clusters also results from to its allocation to the vacuole of the tissue cells, because plants absorb amounts of P above their metabolic needs, as a result of consumption (Chapin et al., 1982; Pereira et al., 2008; Lambers et al., 2011; Veneklaas et al., 2012). On the other hand, the highest content of total P and PSO in the roots of grapevines in V1 and V2 collected at D is probably because P demand by leaves ceases, as cell division and increase of dry matter production cease. Thus, the leaves change from being a sink to being a source of P in F and V. P is redistributed to perennial organs, such as to the roots, typically the main reserve organ of fruit trees (Lima et al., 2011), because P has great mobility in the phloem (Marschner, 2012). The highest content of PTOTAL in the leaves of V2 at H shows that the redistribution of P from the leaves to the clusters decreases after V in vines grown with high available P content. This probably occurs because the soil is meeting cluster demand for P and so P begins to accumulate in leaves in the PSI form at H, which is redistributed at D to the roots in PSO form. The PLIP fraction is contained mainly in cell membranes (Bieleski, 1973). Therefore, the highest content of PLIP in the leaves of grapevines in V1 and V2 collected at F, V and H happens because of the increase in cell membranes complexes, especially thylakoid

R. Piccin et al. / Scientia Horticulturae 218 (2017) 125–131

129

Table 2 Orthogonal contrasts of P forms of grapevine organs in Vineyard 1 (V1) and Vineyard 2 (V2), respectively, grown in soil with low and high available P content in the soil and collected during flowering (F), veraison (V), harvest (H) and dormancy (D). Stage

Organs

PSI

PSO

PLIP

PRNA

PDNA

PRES

Total

Flowering

Roots Trunks Arms Spurs New year shoots Leaves Clusters

ns ** * ns ns ns ns

ns * ** *** ns ns *

ns ns ns ns ** ns ns

* ns ns ** * *** ***

ns ns ns ns ns ns ns

ns ns ns ns ns ns ns

ns * * ns ns ns ns

Veraison

Roots Trunks Arms Spurs New year shoots Leaves Clusters

ns ns ns ns ns ns ns

*** ** ns ns ns ns ns

ns ns ns ns ns ns ns

ns ns ns ns ns *** ns

* * * * * ns *

ns ** *** ** * ns ns

ns *** ns ns ns ns ns

Harvest

Roots Trunks Arms Spurs New year shoots Leaves Clusters

ns ns ns ns ns * ns

ns *** ns ns ns ns ns

ns ns ns ns ns ns ns

*** * ns ns * ns ***

ns ns ns ns ns ns ns

ns ns ns ns ns ns ns

ns ns ns ns ns * ns

Dormancy

Roots Trunks Arms Spurs New year shoots

ns *** ns ns ns

* * ns * ns

ns ns ns ns ns

ns ns ns ns ns

*** ns ns ns ns

** *** ** ns ns

* * *** ** ns

PSI = soluble inorganic P; PSO = soluble organic P; PLIP = phospholipids; PRNA = P associated with RNA; PDNA = P associated with DNA and Pres = residual P. * = significant at 5%; ** = significant at 1%; *** = significant at 0.1%.

(Thomas and Sadras, 2001), in response to increase in PSI content (Reef et al., 2010; Veneklaas et al., 2012). However, excess complexes of thylakoid membranes in leaves can be degraded if there is a reduction in the availability of soil P and may be substituted by glycolipids and phospholipids (Van Mooy et al., 2006, 2009). The proportion of P in ribonucleic acids differs between tissues and cells. It is higher particularly in expanding leaves, lower in mature leaves and very low in senescent leaves (Suzuki et al., 2001). The highest content of PRNA in the leaves of grapevines in V1 and V2 collected at F, V and H, and in the roots at D happened because part of the P that was absorbed, between 5–15% on average, was allocated for the production of ribosomal RNA, which is used for rapid protein synthesis (Niklas, 2006; Suzuki et al., 2001). Typically the increase in PRNA content is correlated with the growth rate of the tissues. This is due to increased protein production by virtue of higher availability of soil P, as well as a higher growth rate (Niklas, 2006; Reef et al., 2010). A larger amount of proteins may also be regarded as a form of P storage in cells (Bieleski, 1972). It should be noted that the highest accumulation of PRNA of grapevines in V1 at F, compared to those in V2 and different than expected, may be due to increased transcription of P carrier proteins (Vance et al., 2003; Veneklaas et al., 2012), thus meeting nutrient requirements of plants (Hammond et al., 2004). The lowest contents of PDNA and PRES compared to contents of PRNA in all organs and phenological stages of grapevines in V1 and V2, may be because PDNA and PRES fractions are more recalcitrant and less susceptible to changes during the phenological stage or soil conditions. In addition, changes in PDNA content are undesirable (Holford 1997; Veneklaas et al., 2012). The response of the PDNA fraction to P availability in soil is linked to the size of the genome, especially of the number of noncoding regions, which are regions within the genome where genes are controlled (Veneklaas et al., 2012). These results are concurrent with those obtained by Bieleski (1972), who found that in Spirodela oligorriza, the PRNA fraction was 7.5 times higher than the PDNA fraction.

ns

= not significant;

The decrease in total P, PSI and PRNA in the new year shoots, leaves and clusters of vines in V1 and V2 from F to H may occur because of the increase in green mass and dry matter production of these organs, which promotes dilution of P fractions (Raghothama, 2000; Marschner, 2012; Camargos and Muraoka, 2007). On the other hand, the increase in PSO content in the new year shoots of vines in V1 and V2 collected at D was probably these shoots started to accumulate P after harvest as documented for N (Neilsen et al., 2010; Millard, 2005). The redistribution of P among the organs of vines in V1 was verified by the reduction of PSI and PRNA contents and increase of PSO in the roots collected from phenological stages F to D. This may be because of the export of root P to the leaves and clusters, and its accumulation mainly in the vacuole of the cells (Marschner, 2012; Veneklaas et al., 2012). Thus, the PSI content in the vacuole acts as a P reserve capable of being mobilized in order to maintain growth rate, mainly of leaves and fruits (Bieleski and Ferguson, 1983). On the other hand, the lowest redistribution of root PSO to the leaves and clusters of vines in V2 from phenological stage H to D is due to greater availability of soil P. The increase in PSI content in the roots of vines in V2 at D can be attributed to the lack of P sinks, such as leaves and fruit (Zambrosi et al., 2012). The redistribution of leaf P to other organs during senescence is an important physiological process for P and N retention in the plant (Tagliavini et al., 1997; Resende et al., 2005; Millard and Grelet, 2010; Zambrosi et al., 2012). P that was redistributed and accumulated in reserve organs may be redistributed to growing organs in the following growing seasons, thus contributing to maintenance of grape productivity (Pommer, 2003).

5. Conclusion P in grapevines is primarily accumulated in soluble inorganic fractions (PSI ) in the leaves and clusters collected during flowering,

130

R. Piccin et al. / Scientia Horticulturae 218 (2017) 125–131

PSO

PSI Clusters

61 56

Leaves

61 53

New year shoots

6 6

V2 - High available P V1 - Low available P

14

7

5 7

74 74 5

65 64

Spurs Arms

49

Trunks

13

11

24

51 13

44

35

42 45

Roots

36 37

24 22

49 48

Clusters

16 14

40 43

Leaves

48

New year shoots Spurs

18 23

45 55

12 14

48

Arms

57

Trunks

39 38

Roots

Clusters

44

Leaves

12 20

48

30

20

32

27

16

41 9

37

56

New year shoots

Flowering

Arms

19 17

29

17

34

Roots

29 25

Harvest

24

22 20

34 35

Trunks

39 45

22

20

Spurs

Veraison

9

42 34 36

41

27

Clusters Leaves New year shoots

42

Spurs

44

36 40

39

44

33 30

Arms Trunks

28

Roots 2500

39

24

2000

1500

1000

28

34 47

47

50 49

24

500

37

0

P (mg kg-1)

500

Dormancy

54

1000

1500

2000

2500

Fig. 1. Redistribution of inorganic P (PSI ) and organic (PSO ) in vine organs of Vineyard 1 (V1) and Vineyard 2 (V2) collected at four phenological stages, flowering (F), veraison (V), harvest (H) and dormancy (D). Values next to of each vertical bar are in percentages.

veraison and harvesting, and in soluble organic fraction (PSO ) in the roots at dormancy. Part of the soluble inorganic P (PSI ) of the roots is redistributed at flowering to the leaves and clusters of grapevines grown in soil with low available P content. Grapevines grown in soils with high available P content accumulate higher amounts of soluble organic P (PSO ) in the roots and tend to redistribute less soluble inorganic P (PSI ) to the leaves and clusters after flowering.

Appendix A. Supplementary data Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.scienta.2017. 02.023.

References Baillod, M., Baggiolini, M., 1993. Lês stades repères de la vigne. Rev. Suisse Vitic. Hortic. 25, 7–9. Bieleski, R.L., Ferguson, J.B., 1983. Physiology and metabolism of phosphate and its compounds. In: Pirson, A., Zimmermann, M.H. (Eds.), Encyclopedia of Plant

R. Piccin et al. / Scientia Horticulturae 218 (2017) 125–131 Physiology: Inorganic Plant Nutrition. Spring, Berlin, pp. 422–449, http://dx. doi.org/10.1007/978-3-642-68885-0 15. Bieleski, R.L., 1972. Turnover of phospholipids in normal and phosphorus-deficient spirodela. Plant Physiol. 49 (5), 740–745. Bieleski, R.L., 1973. Phosphate pools, phosphate transport, and phosphate availability. Annual Review Plant Physiol. 24, 225–252, http://dx.doi.org/10. 1146/annurev.pp.24.060173.001301. Borém, R.A.T., Ramos, D.P., 2002. Variac¸ão estacional e topográfica de nutrientes na serapilheira de um fragmento de Mata Atlântica. Revista Cerne. 8, 42–59. Bortoluzzi, E.C., Péreza, C.A.S., Ardissonb, J.D., Tiecher, T., Canerd, L., 2015. Occurrence of iron and aluminum sesquioxides and their implications for the P sorption in subtropical soils. Appl. Clay Sci. 104, 196–204, http://dx.doi.org/10. 1016/j.clay.2014.11.032. Camargos, S.L., Muraoka, T., 2007. Teores, acúmulo e redistribuic¸ão de macronutrientes em castanheira-do-brasil. Rev. Agric. Tropical. 10, 72–83. Casali, C.A., Kaminski, J., Arbugeri, F.E., Piccin, R., Doneda, A., 2011. Mineralizac¸ão das formas de fósforo do tecido de plantas de cobertura. Informac¸ões Agronômicas − IPNI. 135, 21–24. Chapin, F.S., Follett, J.M., O’connor, K.F., 1982. Growth, phosphate absorption, and phosphorus chemical fractions in two Chionochloa species. J. Ecol. 70, 305–321, http://dx.doi.org/10.2307/2259881. Comissão de Química e Fertilidade do Solo -CQFS-RS/SC., 2004. Manual de adubac¸ão e de calagem para os Estados do Rio Grande do Sul e Santa Catarina. Brazilian Society of Soil Science, Porto Alegre. Cordell, D., Drangert, J.O., White, S., 2009. The story of phosphorus: global food security and food for thought. Global Environ. Change 19, 292–305, http://dx. doi.org/10.1016/j.gloenvcha.2008.10.009. Ferreira, D.F., 1998. SISVAR − Sistema De Análise De Variância Para Dados Balanceados. Universidade Federal de Lavras, Lavras (19 p). Fink, J.R., Inda, A.V., Bayer, C., Torrent, J., Barrón, V., 2014. Mineralogy and phosphorus adsorption in soils of south and central-west Brazil under conventional and no-tillage systems. Acta Scientiarum. 36 (3), 379–387, http:// dx.doi.org/10.4025/actasciagron.v36i3.17937. Fink, J.R., Inda, A.V., Bavaresco, J., Bayer, C., Torrent, J., Barrón, V., 2016. Adsorption and desorption of phosphorus in subtropical soils as affected by management system and mineralogy. Soil Tillage Res. 155, 62–68, http://dx.doi.org/10.1016/ j.still.2015.07.017. Hammond, J.P., Broadley, M.R., White, P.J., 2004. Genetic responses to phosphorus deficiency. Ann. Bot. 94 (3), 323–332, http://dx.doi.org/10.1093/aob/mch156. Holford, I.C.R., 1997. Soil phosphorus: its measurement and its uptake by plants. Aust. J. Soil Res. 35, 227–239, http://dx.doi.org/10.1071/S96047. Lambers, H., Brundrett, M.C., Raven, J.A., Hopper, S.D., 2010. Plant mineral nutrition in ancient landscapes: high plant species diversity on infertile soils is linked to functional diversity for nutritional strategies. Plant Soil 334, 11–30, http://dx. doi.org/10.1007/s11104-010-0444-9. Lambers, H., Finnegan, P.M., Lalibert, E., Pearse, S.J., Ryan, M.H., Shane, M.W., Veneklaas, E.J., 2011. Phosphorus nutrition of proteaceae in severely phosphorus-impoverished soils: are there lessons to be learned for future crops? Plant Physiol. 156, 1058–1066 (10. 1104/ pp. 111. 174318). Lee, R.B., Ratcliffe, R.G., 1983. Phosphorus nutrition and the intracellular distribution of inorganic phosphate in pea root tips: a quantitative study using 31P-NMR. J. Exp. Botany. 34, 1222–1244, http://dx.doi.org/10.1093/jxb/34.9. 1222. Lima, R.L.S., Severino, L.S., Cazetta, J.O., Azevedo, C.A.V., Sofiatti, V., Arriel, N.H.C., 2011. De: redistribuic¸ão de nutrientes em folhas de pinhão-manso entre estádios fenológicos. Revista brasileira de engenharia agrícola e ambiental 11, 1175–1179. Marschner, H., 2012. Mineral Nutrition of Higher Plants, 3. ed (London). Martinez, H.E.P., Novais, R.F., Rodrigues, L.A., Sacramento, L.V.S., 2005. Phosphate forms in plant and their internal buffering in five soybean cultivars. Revista Brasileira de Ciência do Solo. 29, 249–257, http://dx.doi.org/10.1590/S010006832005000200010. Millard, P., Grelet, G.A., 2010. Nitrogen storage and remobilization by trees: ecophysiological relevance in a changing world. Tree Physiol. 30, 1083–1095, http://dx.doi.org/10.1093/treephys/tpq042. Millard, P., 2005. Ecophysiology of the internal cycling of nitrogen in deciduous fruit trees. Italus Hostus 12 (1), 3–10.

131

Murphy, J., Riley, J.P., 1962. A modified single solution methods for the determination of phosphate in natural waters. Anal. Chim. Acta v.27, 31–36. Neilsen, G.H., Neilsen, D., Bowen, P., Bogdanoff, C., Usher, K., 2010. Effect of timing, rate, and form of n fertilization on nutrition vigor, yield, and berry yeast-Assimilable N of grape. Am. J. Enol. Vitic. 61 (3), 327–336. Niklas, J.K., 2006. Plant allometry, leaf nitrogen and phosphorus stoichiometry, and interspecific trends in annual growth rates. Ann. Bot. 97, 155–163, http://dx. doi.org/10.1093/aob/mcj021. Pereira, J.M., Cambraia, J., Júnior, E.M.F., Ribeiro, C., 2008. Efeito do alumínio sobre a absorc¸ão, o acúmulo e o fracionamento do fósforo em sorgo. Bragantia 67 (4), 961–967, http://dx.doi.org/10.1590/S0006-87052008000400019. Pommer, C.V., 2003. Uva tecnologia de produc¸ão, pós-colheita, mercado. Cinco Continentes. Porto alegre. Raghothama, K.G., 2000. Phosphate transport and signaling. Curr. Opin. Plant Biol. 3, 182–187. Reef, R., Ball, M.C., Feller, I.C., Lovelock, C.E., 2010. Relationship between RNA: DNA ratio, growth and elemental stoichiometry in mangrove trees. Funct. Ecol. 24, 1064–1072, http://dx.doi.org/10.1111/j.1365-2435.2010.01722.x. Resende, A.V., Neto, A.E.F., Curi, N., 2005. Mineral nutrition and fertilization of native tree species in brazil: research progress and suggestions for management. J. Sustain. For. 20, 45–81, http://dx.doi.org/10.1300/J091v20n02 03. Schachtman, D.P., Reid, R.J., Ayling, S.M., 1998. Phosphorus uptake by plants: from soil to cell. Plant Physiol. 16 (2), 447–453, http://dx.doi.org/10.1104/pp.116.2. 447. Scott, A., Knott, M., 1974. Cluster-analysis method for grouping means in analysis of variance. Biometrics 30 (3), 507–512, Washington D.C. Soil Survey Staff, 2006. Soil Taxonomy: A Basic System of Soil Classification for Making and Interpreting Soil Surveys, 2nd ed. US Government Printing Office, Washington DC. Suzuki, Y., Makino, A., Mae, T., 2001. An efficient method for extraction of rna from rice leaves at different ages using benzyl chloride. J. Exp. Bot. 52, 1575–1579. Tagliavini, M., Quartieri, M., Millard, P., 1997. Remobilised nitrogen and root uptake of nitrate for spring leaf growth, flowers and developing fruits of pear (pyrus communis l.) Trees. Plant Soil 195, 37–142, http://dx.doi.org/10.1023/ A:1004207918453. Tagliavini, M., Baldia, E., Lucchic, P., Antonellia, M., Sorrentia, G., Baruzzib, G., Faedibet, W., 2005. Dynamics of nutrients uptake by strawberry plants (fragaria x ananassa dutch.) Grown in soil and soilless culture. Eur. J. Agron. 23, 15–25, http://dx.doi.org/10.1016/j.eja.2004.09.002. Thomas, H., Sadras, V., 2001. The capture and gratuitous disposal of resources by plants. Funct. Ecol. 15, 3–12, http://dx.doi.org/10.1046/j.1365-2435.2001. 00488.x. Van Mooy, B.A.S., Rocap, G., Fredricks, H.F., Evans, C.T., Devol, A.H., 2006. Sulfolipids dramatically decrease phosphorus demand by picocyanobacteria in oligotrophic marine environments. Proc. Natl. Acad. Sci. 103, 8607–8612, http://dx.doi.org/10.1073/pnas.0600540103. Van Mooy, B.A.S., Fredricks, H.F., Pedler, B.E., Dyhrman, S.T., Karl, D.M., Koblozec, M.K., Lomas, M.W., Mincer, T.J., Moore, L.R., Moutin, T., Rappé, M.S., Webb, E.A., 2009. Phytoplankton in the ocean use non-phosphorus lipids in response to phosphorus scarcity. Nature 458, 69–72, http://dx.doi.org/10.1038/ nature07659. Vance, C.P., Uhde-stone, C., Allan, D.L., 2003. Phosphorus acquisition and use: critical adaptations by plants for securing a nonrenewable resource. New Phytol. 157 (3), 423–447, http://dx.doi.org/10.1046/j.1469-8137.2003.00695.x. Veneklaas, E.J., Lambers, H., Bragg, J., Finnegan, P.M., Lovelock, C.E., Plaxton, W.C., Price, C.A., Scheible, W.R., Shane, M.W., White, P.J., Raven, J.A., 2012. Opportunities for improving phosphorus-use efficiency in crop plants. New Phytol. 195 (2), 306–320, http://dx.doi.org/10.1111/j.1469-8137.2012.04190.x. Williams, L.E., 1987. Growth of ‘Thompson seedless’ grapevines: ii. nitrogen distribution. J. Am. Soc. Hortic. Sci. 112, 330–333. Zambrosi, F.C.B., Junior, D.M., Boaretto, R.M., Quaggio, J.A., Muraoka, T., Syvertsen, J.P., 2012. Contribution of phosphorus (32P) absorption and remobilization for citrus growth. Plant Soil 355 (1–2), 353–362, http://dx.doi.org/10.1007/ s11104-011-1107-1.