Scientia Horticulturae 257 (2019) 108710

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

Root growth dynamic and plant performance of nectarine trees amended T

with biochar and compost
Giovambattista Sorrenti , Enrico Muzzi, Moreno Toselli
⁎

Department of Agricultural and Food Sciences, University of Bologna, viale G. Fanin 46, 40127, Bologna, Italy

ARTICLE INFO ABSTRACT

Keywords: A 2-year experiment was carried out outdoor on 1-year-old nectarine trees (Big Top grafted on Adesoto 101
Cumulative root length Puebla (P. insititia) grown in large (˜0.5 m3) pots filled with a sandy and poorly fertile soil in which, with 4
Root survivorship replicates, the following soil-applied amendment strategies were compared: a) unamended control; b) biochar
C allocation (16.4 g kg−1 (d.w.)); c) compost (40.0 g kg−1 (d.w.)) and d) biochar mixed with compost, at the same rates of the
Synergism
previous two strategies. Only the first two strategies received mineral fertilizers. We monitored root growth
Root: shoot
dynamic and assessed tree growth, nutritional status, leaf photosynthetic rate, yield, fruit quality, tree archi-
tecture and tree biomass partitioning. Our findings demonstrate that benefits on soil properties and tree growth
were mainly attributable to the addition of compost and, to a less extent, by biochar alone (mostly without
significant effects), indicating that perennial agro-ecosystems may not immediately respond to biochar appli-
cation. Strategies influenced root physiology rather than morphology or biomass, as a possible adaptation to the
changed growing conditions. However, root growth pattern did not reflect the aboveground tree performance.
Compost extended root lifespan and increased photosynthetic leaf efficiency, while biochar reduced root sur-
vivorship and increased root turnover, signifying that amendment strategies modify C fixation and alter C budget
partitioning within tree organs. The PCA analysis suggests that biochar-induced benefits on tree yield could
presumably occur over time. Tree physiology and performance were not outperformed by the mixture of the two
amendments, likely because compost hindered potential synergism with biochar. While we conclude that the
contemporaneous addition of biochar and compost may not always reflect additive responses on plant perfor-
mance, we speculate that these matrices can be conveniently combined in the sustainable management of
nectarine trees with the aim to replace mineral inputs, preserve soil fertility and fight climate changes.

1. Introduction growing conditions and counteract anthropogenic-derived C emissions

Long-term overuse of mineral-based fertilizers in intensive culti-
vated agroecosystems negatively impact soil biota and biogeochemical
processes, posing environmental and agronomical risks (Lal, 2009).
Organic amendments, including compost and biochar, are identified as
economically and ecologically sustainable inputs able to restore and
enhance organic matter pools in soils, with agronomical, ecological and
social benefits (Woolf et al., 2010; Hargreaves et al., 2008).
Biochar is the porous and carbon (C)-rich material resulting from
the thermo-chemical decomposition under limited oxygen availability
(pyrolysis) of a range of organic residues (Spokas et al., 2012), whilst
compost is the stabilized outcome of the aerobic microbial biode-
gradation of biomasses (Hargreaves et al., 2008). Serving as soil con-
ditioners, both organic matrices are deliberately added to soils (Sorrenti
et al., 2012, 2016) with the aims to ameliorate soil properties, plant
(Ros et al., 2006; Diacono and Montemurro, 2010; Woolf et al., 2010;
Spokas et al., 2012).
A growing body of literature reports mostly agronomical and en-
vironmental benefits upon biochar and/or compost inputs in a wide
range of agroecosystems, proposing peculiar mechanisms to explain
such improvements (Verheijen et al., 2010; Hargreaves et al., 2008;
Sorrenti et al., 2012), although some of them are yet unclear (Abiven
et al., 2015).
Evidences frequently investigated the repercussions of the two
amendments (generally applied separately) on soils properties, growth
and yield of annual crops (Jeffery et al., 2011), while long-term out-
comes on perennial species are limited.
Given that once in soil biochar and compost are likely to interact
(Fischer and Glaser, 2012; Sorrenti and Toselli, 2016; Sorrenti et al.,
2017), different responses in terms of soil properties and tree

⁎
Corresponding author at: Department of Agricultural and Food Sciences, University of Bologna, viale G. Fanin 44, 40127, Bologna, Italy.
E-mail addresses: g.sorrenti@unibo.it (G. Sorrenti), enrico.muzzi@unibo.it (E. Muzzi), moreno.toselli@unibo.it (M. Toselli).

https://doi.org/10.1016/j.scienta.2019.108710
Received 7 December 2018; Received in revised form 19 July 2019; Accepted 22 July 2019
Available online 19 August 2019
0304-4238/ © 2019 Elsevier B.V. All rights reserved.
G. Sorrenti, et al.
performance may be expected if compared to the effects of the two
amendments added separately. Indeed, biochar usually promotes sy-
nergistic effects when incorporated into soils either with stabilized (i.e.
composted) organic residues or used as a co-composting agent of or-
ganic matrices (e.g. increasing compost stability and nutrient avail-
ability, altering soil water retention potential and stimulating biomass
biodiversity), leading to improved soil growing environment and plant
health (Fisher and Glaser 2012, Liu et al., 2012; Schulz et al., 2013;
Sorrenti et al., 2017). On the other hand, a recent and comprehensive
study reported not prevalent additive interactions (antagonistic re-
sponses were more frequent than synergistic interactions) when bio-
chars were mixed with organic amendments (Bonanomi et al., 2017),
highlighting that synergistic interactions occurred mainly when N rich
and lignin-poor organic amendments were mixed with biochars. Mostly
confirming results of the latter study, Seehausen et al. (2017) concluded
that the synergisms on plant performance, either on annual or perennial
species, did not occur. Definitively, evidences of combined soil-applied
biochar with organic amendments in temperate regions are scarce and
those documented propose often contradictory results (Kammann et al.
2016).
Therefore, direction and extent of the effects on plant performance
as a consequence of the mix of biochars with organic amendments
seems not always granted (Bonanomi et al., 2017), requiring full fac-
torial experiments (Seehausen et al., 2017).
As far we know, very few studies assessed the influence of these
strategies on the whole-plant system (i.e. growth, nutritional status,
physiology, biomass organ partitioning, allometric trend, yield and fruit
quality) in perennial fruit trees (Eyles et al., 2015) and even less studies
included the root growth dynamic in the attempt to link potential
above- and belowground changes of resource allocation, thereby
adopting a holistic approach. In fact, it is widely accepted that root
system architecture and development greatly controls aboveground
biomass growth (Benjamin and Nielsen, 2006) and that the pattern of
the tree root development often reflects the adaptation to the changed
soil properties as well as in terms of resources abundance or restriction
that plant experiences (Körner, 2011; Poorter and Sack, 2012). Thus,
representing the point of intimate contact, roots may be associated as
the “bridge” between the non-native inputs in soil (e.g. biochar and/or
compost particles) with the growing plants. Therefore, scientific ac-
quisitions on the tree root dynamic may contribute to increase our
understanding of the mechanisms that govern the aboveground plant
responses once soil is enriched with organic amendments (Prendergast-
Miller et al., 2014; Razaq et al., 2017). Both biochar and compost were
shown to affect plant root systems. For instance, shifting soil properties
and nutrient availability, compost stimulated root production and
lifespan of young nectarine trees (Baldi et al., 2010). Abiven et al.
(2015) recorded a greater developed root system and extensively
modified root architecture on maize plants while Amendola et al.
(2017) documented benefits in the fine root seasonal pattern and life-
span in biochar-treated plots of field-grown grapevines. In line with
previous authors, the conclusions of Bruun et al. (2014) on barley,
Reyes-Cabrera et al. (2017) on soybean and Ventura et al. (2014) who
reported a promoted root length intensity in mature biochar-amended
apple trees. Finally, Lehmann et al. (2011) reviewed an increase in root
and shoot biomass upon biochar additions while in other cases also an
increased shoot-to-root ratio, suggesting improved root efficiency.
In this study, we hypothesized that benefits in the short-term, if any,
would be induced mainly by compost then by the amendment with
biochar. Additionally, we hypothesized that the improved soil condi-
tions induced by the amendments would positively affect root growth
first with subsequent repercussion to the aboveground tree perfor-
mance. Finally, we tested the hypothesis that the mix of biochar and
compost, due to synergistic actions, would positively affect plant re-
sponse to a greater extent than the mere addition of the two amend-
ments separately. In this context, we assessed the impact of soil-applied
biochar, compost or their mixture on soil pH and water content (SWC),
2
Scientia Horticulturae 257 (2019) 108710
root growth dynamic and survivorship, tree physiology, growth, yield,
fruit quality and organ biomass partitioning of young bearing nectarine
trees. Specific objectives of this study were to elucidate potential me-
chanisms behind responses (positive, neutral or negative) of perennial
trees subjected to different amendment strategies.
2. Materials and methods
2.1. Experimental and growing conditions
We performed a 2-year (2012–2013) experiment outdoors at the
University of Bologna (44°54′ N, 11°41′ E; 33 m a.s.l.) on nectarine trees
(Prunus persica (L.) Batsch), variety Big Top grafted on Adesoto (Puebla
de Soto 101 - Prunus insititia (L.) Bullace). At the end of winter (March
2012), 1-year old leafless grafted trees (∼0.60 m height, provided with
4 ± 1 lateral branches) were transplanted in large (∼0.5 m3) pots (one
tree per pot), filled with ∼610 kg soil. The soil was sandy (88% sand)
and poorly fertile (O.M. < 0.6 g 100 g−1); this was collected from an
uncultivated and unfertilized field corner specifically selected to avoid
potential carry over effects induced by previous cultural practices (e.g.
fertilization). The physico-chemical properties of the soil are reported
in table S1 (Supplementary Material). Trees were trained as slender
spindle, protected with anti-hail shade netting (in summer) and wa-
tered by microirrigation (4 drippers of 2 L h-1 ea. per plant) to return
the evapotranspiration (ETc) rate (from May through September).
Weeds were mechanically removed and from petal fall until the end of
the vegetative season, trees grown on the unamended or amended with
the mere biochar soils received 41.7, 9.3 and 6.9 g tree−1 of N, P and K,
respectively in the first season and 62.4, 12.0 and 22.9 g tree−1 of N, P
and K, respectively in the second season, supplied under mineral form
(commercial fertilizers) through fertigation. Contrarily, compost-
amended soils did not receive chemical fertilization. Climate of the area
is classified as temperate sub-continental with humid, warm summers
and cold winters. Daily temperatures ranged in average from 2.1 °C to
28.9 °C (annual average temperature was 14.1 °C), while annual pre-
cipitation fluctuated between 680 and 821 mm, with an average of 81
rainy days per year, mostly occurred in spring and autumn.
2.2. Amendment strategies, rates and experimental design
We compared the effect of the following soil-applied amendment
strategies: I) unamended control; II) biochar (16.4 g kg−1 (d.w.)); III)
compost (40.0 g kg−1 (d.w.)) and IV) biochar mixed with compost
(combi from now on) at the same rates of the treatments II and III.
Before filling the pots, amendments were carefully homogenized into
the soil. Pots were N–S oriented and arranged in a single row, dis-
tributed in a randomized block design with four replicates (single pot).
Main physico-chemical properties of the biochar and compost are re-
ported in tables S2 and S3. Compost quality was approved by the Italian
Composting and Biogas Association (C.I.C.) and biochar meet the
European Biochar Certificate (European Biochar Certificate (EBC, 2012)
requirements.
2.3. Measurements
2.3.1. Soil parameters
2.3.1.1. pH and soil water content. Soil samples were obtained by
homogenizing 4 cores per pot collected between 0.05-0.30 m depth at
2-month intervals. A subsample was oven-dried, then 10 g were added
to 25 mL of deionized water (D-H2O) and shaken 60 min at 95 rpm
through an orbital shaker. The pH was measured on the filtered
supernatant under continuous stirring by a pH-meter (BasiC 20,
Crison, Barcelona, Spain). SWC (w w−1) was gravimetrically
measured by oven drying representative subsamples at 105 °C.
Additional soil cores were collected at two-week intervals to monitor
the top-soil (0-0.05 m depth) SWC, as mentioned above.
G. Sorrenti, et al.
2.3.2. Tree growth and performance
2.3.2.1. Root growth dynamic and survivorship. At transplanting, one
rectangular (0.55 m x 0.30 m × 0.12 m, HxWxD, respectively)
parallelepiped-shaped (∼0.020 m3) root box made by transparent
polycarbonate screens was vertically buried into the soil of each pot,
according to the method proposed by Dannoura et al. (2008). Root
boxes were N-E oriented, placed at 0.45 m from the tree trunk and
maintained permanently closed by a dark waterproof cap to prevent
light and water infiltration. Digital images (212.5 × 292.5 mm at 300
dpi) of the soil profile (towards the tree) were taken at about 2-week
intervals, from mid-July (2012) through mid-September of the
following year by inserting a charge-coupled device (CCD) sensor
scanner (HP Scanjet G2710, Hewlett-Packard, CA, USA). Images were
then analyzed by WinRHIZO Tron MF software (Regent Instrument,
Quebec, Canada). We measured root number, length and diameter at
each sampling. Additionally, we recorded the number of root white tips
(new root apex). Root length density (RLD) was calculated by dividing
the total root length by the image area. Root lifespan was estimated by
the difference between birth and death time of individual root,
assuming its disappearance from the scanned images as the death
date, not the point at which the root simply turned brown. Roots that
were still alive after the end of the experiment were excluded from
lifetime estimation.
2.3.2.2. Leaf chlorophyll content, net photosynthetic rate and stomata
conductance. We monitored leaf chlorophyll (Chl) content by the
nondestructive estimation of the leaf Chl concentration made by a
handheld Chl meter (SPAD 502, Minolta Co. Ltd., Osaka, Japan) on 10
young, fully expanded leaves (2 readings per leaf) per tree, from May
through August at about 10-day interval of both seasons. We also
measured leaf net photosynthesis and stomata conductance (between
11:00 am and 1:00 pm) on 4 fully expanded leaves per tree using an
open circuit infra-red gas exchange system (Li−COR 6400, LI−COR
Inc., Lincoln, Nebraska, USA) equipped with a light emitting diode
(LED) source and an external photosynthetic photon flux density
(PPFD) sensor. Light intensity inside the cuvette was constant during
measurements.
2.3.2.3. Leaf weight, leaf area, specific leaf weight and tree nutritional
status. In summer, we randomly sampled 20 fully expanded leaves per
tree from annual shoots. Petioles were removed, then fresh weight and
leaf area (LI 3000, Li-COR Inc., Lincoln, Nebraska, USA) were recorded.
Subsequently, leaf laminas were washed, oven dried (65 °C), weighted,
milled (0.2 mm mesh) and analyzed for nitrogen (N), phosphorus (P),
potassium (K), calcium (Ca), magnesium (Mg), Fe (Iron), manganese
(Mn), zinc (Zn) and copper (Cu) concentration as described in Sorrenti
et al. (2016a). Specific leaf weight (SLW) was calculated dividing leaf
dry weight by leaf area.
2.3.2.4. Trunk girth, pruned wood and thinned fruits. We measured trunk
girth at 0.10 m above the grafting union at 6-month interval. As per
common commercial practices, trees were pruned and thinned the
second growing season, in March and May, respectively. Pruned wood
was oven dried (65 °C) until constant weight, then weighted. Thinned
fruitlets were counted then oven dried (65 °C) and weighted.
2.3.2.5. Yield and fruit quality assessments. Ripen fruits (picked only in
the second season) were individually collected through two consecutive
pickings at 7-day interval. Picked fruits were counted, weighted and
classified, according to their equatorial diameter, in 8 commercial fruit
size categories (AAA Ø > 80 mm; AA Ø 73–80 mm; A Ø 67–72 mm; B Ø
61–66 mm; C Ø 56-60 mm; D Ø 51-47 mm; E Ø < 47 mm). Yield was
calculated by adding the individual fruit weight per tree.
At both pickings, we selected 15 representative fruits per tree for the
determination of the fruit skin color (L*a*b coordinates) (Chroma
meter CR-200, Minolta Co. Ltd., Osaka, Japan), whilst flesh firmness
3
Scientia Horticulturae 257 (2019) 108710
was measured by a pressure tester (Effe.Gi, Ravenna, Italy) fitted with
an 8-mm-diamenter plunger on two sides of the fruits previously
peeled. Two slices per fruit were cut and homogenized to obtain the
juice for the determination of the soluble solids content (SSC) by a di-
gital refractometer (PR-1, Atago, Tokio, Japan), titratable acidity and
pH (Compact Tritator I, Crison, Barcelona, Spain). Additionally, flesh
slices (only from the last fruit picking) were lyophilized, milled and
analyzed for macro and micronutrients concentration, according to the
methods described in Sorrenti et al. (2012). Fruit dry matter con-
centration was also recorded.
2.3.2.6. Tree architecture and biomass partitioning. At the end of the
second growing season (September 2013) we measured the tree height
(from grafting point) and the canopy diameter by 2 orthogonal
measures per plant. Afterwards, we individually removed and
counted the standing leaves and the number of 1-year old shoots per
tree. Then, the length of each 1-year old shoot was measured. These
were oven dried (65 °C) until constant weight and weighted.
Subsamples of 50 representative leaves per tree were used to estimate
the total leaf area and biomass (65 °C). Leaf density was calculated
dividing the number of leaves counted only on the 1-year-old shoots
and the length of the same shoots. We also determined SLW (as
previously described), total leaf area (multiplying the average leaf
area by the total leaf number) and leaf area index (LAI), calculated by
the ratio of the total leaf area per unit ground surface area.
The aboveground canopy (above the grafting point union) was di-
vided in 2-year old wood (skeleton) and trunk, and then organs were
individually weighted. Representative wood rings per each organ were
oven dried (65 °C) until constant weight and used to calculate the organ
dry biomass. Rootstock was accurately removed from soil by a self-
propelled hydraulic excavator, carefully washed to remove adhering
soil then entirely oven dried (65 °C). Roots remained in the soil mixture
after rootstock removal were also recovered. Total plant biomass was
partitioned in fruits, leaves, shoots, skeleton, trunk and roots dry
weight.
2.4. Statistical procedures
Measurements recurrent along the seasons (i.e. soil pH, SWC, leaf
Chl content) were submitted to the repeated measures analysis of var-
iance with 4 replicates using PROC MIXED (Littell et al., 1998) with a
compound symmetry covariance structure, according to a randomized
block experimental design. Data collected only once in the season (i.e.,
yield, leaf number, organ biomass) were submitted to the analysis of
variance using PROC MIXED according to a randomized block design
with 4 replicates. When the interaction between amendment strategy
and season (e.g. leaf nutritional status) was significant, 3 times standard
error of means (SEM) was used as the minimum difference between two
statistically different means (Saville and Rowarth, 2008). Homogeneity
of variance was checked using Levene’s test before analysis. When
analysis of variance showed statistical effects, means were separated by
using Tukey's (HSD) test (at p ≤ 0.05). Statistical analyses were per-
formed by using SAS 9.0 software (SAS Institute Inc., Cary, NC, USA).
We adopted the Principal component analysis (PCA) approach to re-
present the relative effect of the amendments on the tree yield perfor-
mance. PCA was performed by using STATISTICA v. 12 (StatSoft, Inc.,
Tulsa, OK, USA). Data of root dynamic and survivorship were submitted
to the survival, partykit and survfit packages of R Project for Statistical
Computing software (R Core Team, 2013).
3. Results
3.1. Soil pH and water content (SWC)
Soil pH fluctuated in the sub-alkaline range, between 7.52 and 8.06.
Compost and combi, except for the sampling of July (2012),
G. Sorrenti, et al. Scientia Horticulturae 257 (2019) 108710

Fig. 1. Effect of soil-applied biochar and compost on soil water content (SWC) trend at 0-0.05 m soil depth.
ns, *, ** and ***: effect of the amendment not significant or significant at p ≤ 0.05, p ≤ 0.01 and p ≤ 0.001, respectively. Within dates, means with the same letter
are not statistically different (p ≤ 0.05, Tukey's HSD Test). Values represent the mean of 4 replicates.

significantly decreased soil pH compared to the unamended (by 0.18 Table 1

units in average at each sampling time) and biochar-enriched soils, Main effect (ANOVA) of the amendment (TRT), sampling time (TIME) and their
while values between the latter strategy and the control (unamended) interaction on root morphological parameters and proliferation (Avg. ± S.E.;
were always comparable (Tab. S4). n = 4).
In the 0.05-0.30 m depth soil profile, SWC was often increased by Root parameter Degrees of freedom (DF) F ratio P value
compost and, to a greater extent, by the mix of the two amendments
(Tab. S4). Compared to the unamended soils, the mere addition of Total root number
TRT 2 0.07 0.9374
biochar and compost statistically increased the SWC in two and four
GROUP 1 1.72 0.2805
sampling (out of 9), respectively (Tab. S4). Similarly, the highest SWC TIME 29 15.5 < 0.0001
in the top soil layer (0-0.05 m) were mostly achieved by combi, then by TRT*TIME 58 1.44 0.0263
compost, while values of the biochar-amended soil did not statistically Cumulative root length
differ from the control (Fig. 1). TRT 3 4.27 0.0391
TIME 29 4.28 0.0008
TRT*TIME 87 1.02 0.4457
3.2. Tree growth and performance
Root diameter
TRT 3 3.79 0.0524
3.2.1. Root growth dynamic and survivorship TIME 29 2.88 < 0.0001
For an easier understanding, Table 1 summarizes the output of the TRT*TIME 87 2.16 < 0.0001
analyses of variance for the main effects induced by amendment stra- Root length density (RLD)
tegies (TRT), sampling time (TIME) and their interactions on root TRT 3 1.98 0.1954
morphological traits and proliferation throughout the experiment, TIME 29 7.85 < 0.0001
TRT*TIME 87 1.15 0.2000
whereas Table 2 shows trends observed in the root growth dynamic on
selected dates, specifically 109, 159, 227, 337, 435, 484 and 535 days New root apex
TRT 3 2.89 0.1024
after planting (DAP). Nonetheless, full results are detailed in tables S5
TIME 29 10.09 < 0.0001
and S6 of the supplementary materials. TRT*TIME 87 1.20 0.1341
As expected, independently of the strategy, time mostly influenced
root growth parameters and values increased over time, with higher
rates recorded during the vegetative seasons (Table 2). Amendments second season, with an even faster increase from mid-June until the end
affected only cumulative root length, while the two factors (time and of the experiment (September) (Tab. S4 and S5). We observed a similar
amendment strategy) significantly interacted for the total root number course with regard to the cumulative root length (Fig. 2) and the RLD
and average root diameter (Table 1). In details, root diameter increased (Table 2), while the appearance of new root apex (white tips) followed
over time, reaching at the end of the experiment about 36.5 μm root−1, an irregular trend (Table 2).
whereas total root number was separated according to a two by two The root survivorship analyses estimated that over 70% of the roots
groups: the first group composed by unamended and combi strategies in the soil amended with biochar survived less than 100 days (S100 =
(with often overlapping trends), which values were higher than the a root survivorship percentage within 100 days), significantly shorter
second group represented by biochar and compost (Table 2). We ob- than other strategies (Fig. 3) which instead showed a S100 of about 37%
served, independently of the strategy, a fast increase in the root number of the total roots. S200 was over 80% for biochar, close to 50% for combi
(with values over than doubled) until the natural leaf drop (mid-Oc- and unamended soils and less than 40% for compost (figure not shown).
tober of the first season), then such rate was much slower or even Considering the entire experimental period (all the observed roots
suspended, during winter and spring (Tab. S4 and S5). The resumption survived less than 500 days), no roots survived over than 350 days in
in root number was again appreciable from the beginning of May of the

4
G. Sorrenti, et al. Scientia Horticulturae 257 (2019) 108710

Table 2
Effect of soil-applied biochar, compost and combi on root morphological parameters and proliferation as measured 109, 159, 227, 337, 435, 484 and 535 days after
planting (DAP). Data (e.g. n tree−1) refer to the area (0.06215 m2) of the image (soil profile) that was periodically analyzed, rather than the whole tree (Avg. ± S.E.;
n = 4).
Measurement 1 5 10 16 23 27 30
Date Jul-19 Sep-07 Nov-14 Mar-04 Jun-10 Jul-29 Sep-18
DAP 109 159 227 337 435 484 535

Total root number (n tree−1)

Unamended 31.5 ± 13.9 59.5 ± 27.0 72.2 ± 26.4 74.1 ± 27.5 77.2 ± 26.9 82.5 ± 27.1 128.2 ± 24.2
Biochar 9.50 ± 1.7 18.2 ± 3.3 24.2 ± 4.7 32.1 ± 3.7 36.7 ± 3.4 58.7 ± 6.9 76.5 ± 5.6
Compost 18.3 ± 11.6 38.0 ± 14.5 41.3 ± 13.3 42.0 ± 22.0 43.3 ± 12.8 50.3 ± 13.7 69.7 ± 12.2
Combi 37.2 ± 17.5 52.0 ± 19.0 68.0 ± 19.4 71.2 ± 18.7 77.2 ± 19.3 83.0 ± 18.6 111.2 ± 24.2

Cumulative root length (mm tree−1)

Unamended 373 ± 19 684 ± 308 791 ± 306 804 ± 315 854 ± 302 917 ± 303 1519 ± 301
Biochar 141 ± 46 292 ± 82 356 ± 76 434 ± 55 48.9 ± 40 709 ± 48 949 ± 42
Compost 230 ± 162 484 ± 249 525 ± 256 565 ± 434 555 ± 250 616 ± 268 798 ± 238
Combi 523 ± 259 784 ± 252 1024 ± 252 1047 ± 247 1160 ± 248 1228 ± 235 1600 ± 309

Root diameter (μm root−1)

Unamended 29 ± 8 23 ± 3 23 ± 3 23 ± 3 24 ± 3 25 ± 2 35 ± 4
Biochar 26 ± 3 24 ± 1 24 ± 2 24 ± 2 26 ± 3 36 ± 6 37 ± 3
Compost 24 ± 6 29 ± 9 26 ± 5 28 ± 10 27 ± 6 35 ± 8 38 ± 7
Combi 26 ± 2 31 ± 4 31 ± 3 30 ± 3 33 ± 2 34 ± 1 37 ± 2

Root length density (RLD) (m m−2)

Unamended 6.08 ± 3.05 11.0 ± 4.96 12.7 ± 4.92 12.9 ± 5.08 13.7 ± 4.87 14.7 ± 4.87 24.4 ± 4.84
Biochar 2.27 ± 0.74 4.70 ± 1.32 5.72 ± 1.22 6.988 ± 0.89 7.87 ± 0.65 11.4 ± 0.77 15.3 ± 6.68
Compost 3.71 ± 2.61 7.79 ± 4.00 8.44 ± 4.12 9.10 ± 6.99 8.93 ± 4.03 9.91 ± 4.31 12.8 ± 3.82
Combi 8.41 ± 4.17 12.6 ± 4.06 16.5 ± 4.05 16.8 ± 3.97 18.7 ± 3.99 19.8 ± 3.78 25.7 ± 4.97

New root apex (n tree−1)

Unamended 31.0 ± 13.9 5.2 ± 4.6 0.5 ± 0.5 0.2 ± 0.2 1.2 ± 1.2 0.0 ± 0.0 1.7 ± 0.7
Biochar 9.2 ± 1.6 0.2 ± 0.2 0.7 ± 0.5 3.0 ± 2.4 0.0 ± 0.0 11.2 ± 3.9 0.0 ± 0.0
Compost 18.3 ± 10.0 2.7 ± 1.1 0.7 ± 0.6 0.3 ± 0.3 0.3 ± 0.3 6.3 ± 1.0 0.3 ± 0.3
Combi 37.0 ± 17.6 1.7 ± 0.5 2.2 ± 0.2 0.2 ± 0.2 0.0 ± 0.0 1.7 ± 1.4 0.5 ± 0.3

(Table 3).
Amendment strategies did not affect leaf gas exchange in the first
growing season, with leaf net photosynthetic rates ranging between 8.6
and 12.6 μmol CO2 m2 s−1 (data not shown). Leaf photosynthesis and
stomata conductance were consistently higher in the second season
and, compared to the unamended soil, significantly promoted by
compost and combi in the two measurements of July, with intermediate
values recorded in biochar treated trees (Table 4).

Fig. 2. Effect of the amendment strategies on the cumulative root length dy-
namic throughout the experiment. Data represent the root length measured
periodically and limited to the root box images, not to the whole root system.
the soil treated with biochar, significantly shorter than the unamended
soil, which however did not exceed 400 days (Fig. 3). Finally, in-
dependently of the mixture with biochar, compost statistically pro-
longed the root survivorship (Fig. 3) and the oldest roots survived in
average 426 days, observed only in compost-amended soils. Finally,
50% of the observed roots survived in average 60, 190, 202 and 270
days in the biochar, unamended, combi and compost-amended soils,
respectively (Fig. 3).
3.2.2. Leaf Chl content and net photosynthetic rate
Until late spring of the second season, plants fertilized with mineral
inputs (unamended and only biochar-amended soils) showed the
highest SPAD values, without differences between unamended control
and only biochar-amended trees (Table 3). Later on, values were similar
among strategies, with the exception of the measurement of July 30 in
which the highest value was recorded in biochar-amended trees
3.2.3. Leaf weight, area, specific leaf weight (SLW) and tree nutritional
status
Leaf fresh and dry weight, leaf area and SLW were 733 ± 10.6 mg
leaf−1, 287 ± 7.8 mg leaf−1, 37.0 ± 0.47 cm2 leaf−1 and
7.77 ± 0.29 mg cm-2 leaf-1 (average ( ± SE) of both seasons; n = 20),
respectively, without changes ascribed to the soil conditioning strate-
gies (data not shown).
Amendment and sampling time affected tree nutritional status with
the two factors interacting on leaf N and Mn concentration (Table 5).
Leaf N concentration was statistically higher in the first season, in
which values were increased in trees amended with compost, without
synergic effect when mixed with biochar (Table 5). In the same year, a
significantly reduced N concentration emerged in leaves sampled from
trees amended with biochar, even compared with the unamended trees
(Table 5). Nonetheless, this trend was not confirmed the following year,
when leaf N values were comparable among strategies (Table 5).
Similarly to N, leaf Mn concentration was higher in the first season
and depressed by biochar (Table 5). In the second season, leaf Mn
concentration was significantly reduced also in plants amended with
compost (Table 5). Independently of the amendment strategies, the
concentration of leaf P, Mg, Fe and Zn was higher in the first season,
whereas an opposite trend emerged for Ca (Table 5). Season did not
alter leaf Cu and K concentrations. The latter metal was promoted by
the three amendments compared to the untreated control, with a higher
extent induced by compost (as recorded also for P) and combi (Table 5).

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G. Sorrenti, et al. Scientia Horticulturae 257 (2019) 108710

Fig. 3. Effect of the amendment strategies on root survivorship after 100 and 500 days from transplanting.

Leaf Mg concentration decreased in amended trees and significantly
lower leaf Ca values were recorded in compost-amended trees, with
intermediate concentrations induced by biochar (Table 5). Finally, the
addition of compost to soils increased leaf Zn concentration, without
synergic effect with biochar (Table 5).
3.2.4. Trunk girth, pruned wood and thinned fruits
Amendments did not alter trunk girth throughout the experiment
resulting, after 18 months since transplanting, equal to 190.8 mm
tree−1, with a net annual increase of 45.9% (data not shown).
Winter pruned wood was heavier in plants amended with the
combination of biochar and compost (+30% compared to untreated
trees), while the mere addition of biochar showed similar wood re-
sidues than the unamended trees (Table 6). Compost induced inter-
mediate values (Table 6).
Although the number of thinned fruitlets was unaffected by strate-
gies, their weight was statistically higher (without synergic effect) in
compost-amended trees (Table 6), where fruitlet fresh and dry weight
resulted 72.9 and 79.5% higher than trees that did not receive compost,
respectively (Table 6).
3.2.5. Yield and fruit quality assessments
A significantly higher total fruit number was picked from compost-
and combi-amended trees compared with the unamended soil, with

Table 3
Effect of the amendment strategies on the leaf chlorophyll content in the second season.
Amendment Leaf Chlorophyll Content (Spad Units)

May-8 May-14 May-28 Jun-5 Jun-10 Jun-20 Jun-27 Jul-5 Jul-12 Jul-24 Jul-30 Aug-26

Unamended 31.9 31.0 a 31.5 a 29.5 a 29.4 ab 30.3 29.9 32.6 32.4 31.2 33.7 b 34.8
Biochar 33.3 28.5 ab 30.7 a 29.1 a 30.5 a 29.9 32 33.3 34.2 32.7 36.3 a 36.3
Compost 30.8 27.1 b 27.2 b 26.4 b 27.0 b 30 31.4 32.3 32.8 32 33.3 b 35.6
Combi 29.5 25.6 b 27.3 b 25.7 b 27.5 b 31.6 32.1 32.3 33.6 31 33.8 b 35.1
Significance ns ** ** ** ** ns ns ns ns ns * ns

ns, * and **: effect of the amendment not significant or significant at p ≤ 0.05 and p ≤ 0.01, respectively. In the same column, means followed by the same letter are
not statistically different (p ≤ 0.05, Tukey's HSD Test).

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G. Sorrenti, et al. Scientia Horticulturae 257 (2019) 108710

Table 4
Effect of the amendment strategies on the leaf photosynthetic rate (μmol CO2 m2 s−1) and stomata conductance (μmol H2O m2 s−1) in the second growing season.
Amendment Leaf photosynthetic rate (μmol CO2 m2 s−1) and stomata conductance (μmol H2O m2 s−1)

Jun-5 Jul-5 Jul-31 Aug-30

Photosyntesis Stom. Cond. Photosyntesis Stom. Cond. Photosyntesis Stom. Cond. Photosyntesis Stom. Cond.
Unamended 7.05 0.12 12.2 b 0.20 b 14.7 b 0.21 b 19.3 0.23
Biochar 5.62 0.10 15.9 ab 0.25ab 18.8 ab 0.29 ab 18.9 0.22
Compost 7.55 014 18.2 a 0.31 a 23.6 a 0.44 a 21.3 026
Combi 6.15 0.13 16.4 a 0.25 a 20.4 a 0.34 a 19.0 0.22
Significance ns ns ** ** ** ** ns ns

ns
and **: effect of the amendment not significant or significant at p ≤ 0.01, respectively. In the same column, means followed by the same letter are not statistically
different (p ≤ 0.05, Tukey's HSD Test).

biochar showing values not different from untreated control (Table 7). Table 6
This trend mirrored the response of the cumulative yield (Table 7). In Effect of the amendment strategies on winter pruned wood (March 2013) and
fact, without synergic effects between amendments, the addition of thinned fruits (May 2013).
compost yielded over than 150% compared to unamended trees Amendment Pruned Wood Thinned fruits
(Table 7). However, although limited to the first picking, we picked a
higher number of fruits from biochar-amended trees (but statistically Fresh Dry weight Thinned Fresh Dry weight
weight fruits weight
similar to compost), even though this increase was not enough to dif-
g tree−1 fruitlets g fruit −1 g fruit −1
ferentiate cumulative yield (Table 7). tree−1
Independently of the amendment strategies and picking, fruits
showed similar juice pH and flesh firmness, with softer values in the Unamended 874.8 b 421.4 b 42.0 1.69 b 0.20 b
second picking (Table 8). On the contrary, the fruit SSC from trees Biochar 772.3 b 398.5 b 61.5 1.78 b 0.24 b
Compost 1046.0 ab 535.7 ab 33.7 3.07 a 0.40 a
amended with compost (alone or in combination with biochar) was
Combi 1178.7 a 605.6 a 48.0 2.93 a 0.39 a
statistically higher than biochar alone and, limited to the first picking, Significance * * ns *** **
also compared to the control (Table 8). In particular, fruits grown in
plants amended with compost showed an increase (average of both ns, *, ** and ***: effect of the amendment not significant or significant at
pickings) equal to 2.07°Bx (Table 8). p ≤ 0.05, p ≤ 0.01 and p ≤ 0.001, respectively. In the same column, means
Juice titratable acidity, as well as fruit skin color, exhibited only followed by the same letter are not statistically different (p ≤ 0.05, Tukey's HSD
marginal differences attributable to amendments, without a clear trend Test).
(Table 8).
3.2.7. Principal component analyses
The multivariate analyses (PCA) shows that the cumulative variance
3.2.6. Fruit mineral concentration explained by the first two components was over than 78% (58.9% axis 1
Potassium was the most concentrated mineral in the fruit flesh, and 19.8% axis 2) (Fig. 4). This approach clearly separated the effect of
followed by N, P, Mg and Ca (Table 9). Among micronutrients, Fe was the different strategies on tree yield with the unamended soils opposed
the most abundant, with Zn, Cu and Mn in decreasing order (Table 9). to the effect induced by compost and biochar, positioned inter-
However, amendments did not alter fruit N, Ca and micronutrient mediately. Finally, the cluster area representing the combi strategy
concentration (Table 9) while, similarly to leaves, compost increased fully overlapped that of compost, indicating no synergic effects between
flesh P and K concentration. The latter element was promoted (but amendments (Fig. 4).
lesser than compost) by biochar, which also increased Mg concentration
in fruits compared to the control (Table 9). 3.2.8. Tree architecture and biomass partitioning
After 18 months, tree height, trunk cross sectional area (TCSA),
canopy width, shoot length, leaf number, total leaf area, SLW and LAI

Table 5
Effect of the amendment strategies and season on the leaf macro and micro nutrient concentration.
Amendment N P K Ca Mg Fe Mn Cu Zn

2012 2013 2012 2013

−1
g kg dw mg kg−1 dw

Unamended 26.6 19.4 1.73b 15.2c 19.0a 3.93a 66.7 40.4 33.3 9.71ab 21.1b
Biochar 22.2 19.7 1.92b 21.0b 17.0ab 3.00b 65.5 30.2 25.2 8.75b 20.2b
Compost 33.4 19.1 2.39a 24.1a 15.0b 2.96b 60.7 38.3 22.1 10.7a 27.0a
Combi 34.0 18.9 2.06ab 24.8a 15.9b 3.17b 71.4 40.6 20.0 10.2ab 28.5a
Significance 3SEM=3.52 * *** ** ns 3SEM=7.21 * **
Season
2012 2.42 21.3 14.6 3.52 84.8 9.78 29.5
2013 1.62 21.4 18.9 3.02 47.4 9.91 18.8
Significance *** ns *** *** *** ns ***
Interaction *** ns ns ns ns ns ** ns ns

ns, *, ** and ***: effect of the amendment and season not significant or significant at p ≤ 0.05, p ≤ 0.01 and p ≤ 0.001, respectively. In the same column, means
followed by the same letter are not statistically different (p ≤ 0.05, Tukey's HSD Test). ** and ***: interaction between amendment and season significant at
p ≤ 0.05. Values differing by ≥3 standard error of means (SEM) are statistically different.

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G. Sorrenti, et al. Scientia Horticulturae 257 (2019) 108710

Table 7
Effect of the amendment strategies on the number of harvested fruits, fruit weight and yield on the two commercial pickings (July 2013).
Amendment Number of fruits Fruit Weight Yield

st nd st nd
I picking II picking Total I picking II picking Average Ist picking IInd picking Cumulative
fruit tree−1 g fruit−1 kg tree−1

Unamended 3.75 b 8.25 b 12.0 b 152.8 135.0 142.0 0.57 b 1.12 b 1.69 b
Biochar 12.0 a 14.2 ab 26.2 ab 166.4 129.4 146.1 1.98 a 1.83 b 3.82 ab
Compost 13.3 a 18.5 a 30.7 a 173.4 124.4 144.2 2.12 a 2.33 a 4.45 a
Combi 9.5 ab 23.0 a 32.5 a 159.8 137.2 143.7 1.52 a 3.15 a 4.67 a
Significance ** * * ns ns ns ** * *

ns, * and **: effect of the amendment not significant or significant at p ≤ 0.05 and p ≤ 0.01, respectively. In the same column, means followed by the same letter are
not statistically different (p ≤ 0.05, Tukey's HSD Test).

resulted analogous among strategies which values were 2.81 m tree−1, Table 9
31.7 cm2 trunk−1, 1.20 m tree−1, 0.221 m shoot−1, 6701.1 leaf tree−1, Effect of the amendment strategies on the fruit flesh macro and micro nutrient
24.1 m2 tree−1, 7.90 mg cm-2 and 5.36 m2 m-2 in average, respectively concentration at harvest.
(Table 10). However, shoot number and consequently leaf density Amendment N P K Ca Mg Fe Mn Cu Zn
(number of leaves per unit of shoot length) were promoted by compost g kg−1 mg kg−1
(without synergic effects). Finally, cumulative shoot length was in-
creased in unamended trees compared to compost (Table 10). Unamended 6.12 1.43 b 8.9 b 0.16 0.42 b 18.9 1.54 5.15 7.70
Biochar 6.65 1.48 b 11.7 a 0.16 0.49 a 17.4 2.01 5.46 7.88
At the end of the experiment, leaf, trunk and root dry biomass was Compost 6.24 1.72 a 12.8 a 0.15 0.44 ab 16.3 1.42 6.73 8.46
comparable among experimental trees. Although slight differences were Combi 6.10 1.81 a 12.4 a 0.17 0.46 ab 14.5 1.85 5.72 7.94
recorded in fruit, 1-year old shoot and 2-year old branch biomass, the Significance ns * ** ns * ns ns ns ns
cumulative dry biomass of the whole tree was unaffected by the
amendment strategies (Table 11). In fact, even though fruit (as a con- ns, * and **: effect of the amendment not significant or significant at p ≤ 0.05
and p ≤ 0.01, respectively. In the same column, means followed by the same
sequence of a higher yield) biomass was significantly increased by
letter are not statistically different (p ≤ 0.05, Tukey's HSD Test).
compost, such increase was counterbalanced by an overall lower 1- and
2-year-old woods biomass in the compost-amended trees (irrespective
2010; Sorrenti et al., 2016b). Although liming effects have been often
of the addition of biochar) (Table 11). Independently of the amend-
reported as a consequence of biochar incorporation in acid and
ment, the whole tree biomass was partitioned as follow: roots (30.5%),
weathered soils, neutral responses are common, as observed in a 3-year
leaves (24.5%), 2-year old wood (21.8%), trunk (9.83%), fruits (7.74%)
field experiment in which biochar was distributed up to 30 Mg ha−1
and 1-year old shoots (5.66%) (Table 11).
(Sorrenti et al., 2016a).
Given that total above- and belowground tree biomass was similar,
The lower soil pH values measured in compost-amended soils (0.2-
the root:shoot ratio originated comparable values, ranging from 0.416
0.4 units lower than the unamended soil) suggests that the composted
(compost) and 0.468 (unamended; Table 11).
organic residues were rich in humic substances and validate the buffer
property of the organic matter, preserving the soil reaction next to
4. Discussion neutrality. This was confirmed also in the combi strategy, indicating
that the compost buffer ability was predominant.
4.1. Compost, not biochar, altered soil pH
4.2. Soil water content was improved synergically by compost and biochar
Despite the high rate and the alkaline pH of the carbonaceous ma-
trix, the mere incorporation of biochar did not change soil pH. This While biochar alone poorly contributed to increase SWC, a more
response may be due to the alkaline pH (8.0) and to the abundance of pronounced effect was observed in the compost-amended soils, even
carbonates of the unamended soil, which likely contributed to buffer greater (always statistically increased) in the combi-amended soils, ei-
the soil reaction. Unlikely the pH of the biochar-enriched soil is ex- ther on the top-soil or in the deeper soil profiles, indicating an additive
pected to increase afterwards, as the environmental weathering pro- effect of two matrices. The beneficial effect of the OM sources (in-
cesses lead to a development of carboxylic acids functional groups on cluding biochar and compost) in increasing SWC is well documented
the exposed biochar surfaces (Cheng and Lehmann, 2009; Yao et al., (Baronti et al., 2014; Evanylo et al., 2008; Liu et al., 2007; Reicosky,

Table 8
Effect of the amendment strategies on fruit firmness, solid soluble content (SSC), titratable acidity, juice pH and skin color (L, a, b components) at the two commercial
picking.
Amendment Fruit firmness SSC Titratable acidity Juice pH Fruit skin color

st nd st nd st nd st nd
I picking II picking I picking II picking I picking II picking I picking II picking Ist picking IInd picking

kg °Bx g L−1 malic acid L a b L a b

Unamended 4.73 3.56 17.8 b 18.5 ab 3.91 b 3.69 3.75 3.89 33.6 ab 33.4 8.3 33.8 33.2 8.7
Biochar 5.11 3.95 17.1 b 16.5 b 5.56 ab 4.44 3.71 3.89 33.3 b 32.3 7.9 33.7 33.7 8.8
Compost 4.49 3.36 18.6 a 19.2 a 6.18 a 4.42 3.68 3.87 34.2 ab 33.5 8.7 34.5 35.5 9.8
Combi 4.72 3.10 20.0 a 20.4 a 5.55 ab 4.13 3.93 4.03 34.9 a 34.5 9.6 34.0 33.7 9.0
Significance ns ns * * * ns ns ns ** ns ns ns ns ns

ns, * and **: effect of the amendment not significant or significant at p ≤ 0.05 and p ≤ 0.01, respectively. In the same column, means followed by the same letter are
not statistically different (p ≤ 0.05, Tukey's HSD Test).

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G. Sorrenti, et al. Scientia Horticulturae 257 (2019) 108710

positive responses on root growth parameters upon the addition of

Fig. 4. Principal component analyses of the effect of the amendment strategies
on individual tree yield. Values along X and Y axes express the variance (%) of
the two main factors. CK, B, C and BC indicate Unamended and amended with
biochar, compost and combi, respectively.
2005), while less evidences are reported about synergisms of these two
matrices. Our evidences indicate that the effect of biochar on SWC
could be not as persistent as expected (likely related to the biochar-soil
interaction), but that the mix of biochar and compost may result an
effective strategy in increasing the SWC to a greater extent than the
mere addition of the two amendments, with potential and positive
implications in the irrigation management of commercial agricultural
crops.
4.3. Root growth dynamic and survivorship
Soil conditioners poorly altered the main root morphological traits
and growth dynamic, with mostly and apparently not beneficial neither
synergic effects. Amendments were homogeneously incorporated into
the soil prior transplanting in large pots (˜ 610 kg soil per tree), thereby
the root growing environment was certainly conditioned by amend-
ments and roots were in intimate contact with either biochar fragments
and/or compost residues.
Digital images scanned the full soil profile available for root de-
velopment (0.35 m soil depth), hence we exclude that roots were
growing in uninvestigated (e.g. deeper soil depth) soil volumes. As we
did not observe differences in the root distribution pattern along the
soil profile, we assume that roots were not escaping either from biochar
particles or compost residues. This is also corroborated by results of
Prendergast-Miller et al. (2014), who describe that sycamore roots were
attracted towards biochar and that the rhizosphere was richer in bio-
char fragments than the bulk soil. Similarly, Olmo et al. (2014) con-
cluded that roots may be attracted by biochar particles with the aim to
access to the biochar-provided resources. Literature reports mostly
biochar or compost to both tropical and temperate soils (Ishii and
Kadoya, 1994, Donn et al. 2014, Lehmann et al., 2011; Jindo et al.,
2012). However, in our study, benefits on root growth were limited.
This lack of response was not likely induced by the growing environ-
ment as it was not limiting for root growth (i.e. sandy textured soil,
irrigated and drained, free of weeds). In fact, in our conditions, roots
were free to colonize the soil volume. Thus, although beneficial effects
on root morphology were negligible, it is worth to mentions that rates
of biochar (˜15.5 Mg dw ha−1) and compost (˜35.5 Mg dw ha-1, as-
suming a tree density of 1481 trees ha-1 (4.5 m * 1.5 m))adopted in this
study were not harmful for root growth (e.g. toxicity).. This is sup-
ported by the fact that amendments did not proportionate detrimental
symptoms on the root morphological traits (e.g. root color and hydra-
tion, root branching and architecture, presence of root rot symptoms
etc.) and root dry biomass, as well.
Interestingly, biochar markedly reduced root survivorship. Indeed,
almost 40% of the biochar-conditioned roots disappeared within 50
days (data not shown) and over than 70% of the total observed roots
survived less than 100 days whilst in the other strategies root mortality,
after 50 and 100 days, resulted below than 20% and 40%, respectively.
The oldest roots survived 426 days and a considerable number of these
were retrieved in the combi treatment and, sporadically, in the com-
post-treaded soil. Conversely, when biochar was soil-incorporated
alone, most of the roots (98%, out of 163 monitored roots) survived
significantly shorter than the other strategies. Our data (shorter root
survivorship, reduced cumulative root length, equal cumulative root
number) suggest that biochar promoted a faster root turnover and
changes in root morphology (shorter roots). Reasons for changes in root
growth and physiology have been explained by different mechanisms,
mainly including biochar properties and relative conditions that restrict
root or shoot growth in diverse soil environments (Lehmann et al.,
2011). It has been reported that biochar could potentially generate
negative effects on root growth due to modifications of the physico-
chemical interface where soil, roots, and biochar converge, hence af-
fecting root growth and distribution patterns (Lehmann et al., 2009,
2015). However, we believe that this was not our case. We speculate
that biochar may have interfered at physiological level (e.g. releasing of
hormone-like molecules), stimulating root turnover. Biochar may have
affected root growth through the releasing of stimulatory signals
(Graber et al., 2015) or by altering signaling between microbes and
plant roots at the rhizo-char sphere (Warnock et al., 2007; Masiello
et al., 2013). On the contrary, the extended root lifespan induced by
compost (previously observed also by Baldi et al. (2010) on field-grown
nectarine trees) is likely related to improved soil properties (e.g. por-
osity, soil moisture, stable aggregates) and higher resources availability
(i.e. mineral N) occurred in the compost-amended soils which in turn
may improve the root environment and stimulate plant growth. This is
supported by the considerable higher soil nitrate-N and ammonium-N
availability (as well as in the soil leachate) measured in the compost-
amended soils in the same experiment (Sorrenti et al., 2017; Sorrenti

Table 10
Effect of the amendment strategies on the tree height, trunk cross section area (TCSA), canopy radius, shoot number and length, leaf number, leaf density, total leaf
area, specific leaf weight (SLW) and leaf area index (LAI) at the end of the experiment.
Amendment Tree height TCSA Canopy radius Shoot number Shoot length Leaf number Leaf density Total leaf area SLW LAI

m tree−1 cm2 trunk−1 m tree−1 n tree−1 m shoot−1 m tree−1 leaf tree−1 leaf cm−1 m2 tree mg cm−2 m2 m−2

Unamended 2.830 33.8 1.18 282 b 0.263 74.2 a 6797.0 0.84 b 23.1 7.93 5.26
Biochar 2.811 29.8 1.17 279 b 0.238 66.4 ab 6887.7 0.88 b 25.5 8.10 5.93
Compost 2.813 31.6 1.20 307 ab 0.195 59.9 b 6238.0 0.96 a 22.2 7.91 5.06
Combi 2.802 31.6 1.26 343 a 0.189 64.8 ab 6881.7 0.98 a 25.6 7.68 5.19
Significance ns ns ns * ns * ns * ns ns ns

ns
and *: effect of the amendment not significant or significant at p ≤ 0.05, respectively. In the same column, means followed by the same letter are not statistically
different (p ≤ 0.05, Tukey's HSD Test).

9
G. Sorrenti, et al.
Table 11
Effect of the amendment strategies on the tree biomass (dw) partitioning and the root:shoot ratio at the end of the experiment.
Amendment Fruits Leaves 1-year-old shoots 2-year-old wood
g (d.w.) tree1
Unamended 261.6 b 1835.6 566.1 a 1822.8
Biochar 620.3 ab 2056.9 457.7 ab 1685.5
Compost 742.7 a 1780.3 353.9 b 1542.5
Combi 761.2 a 1965.2 362.5 b 1747.8
Significance * ns * *
ns
and *: effect of the amendment not significant or significant at p ≤ 0.05, respectively. In the same column, means followed by the same letter are not statistically
different (p ≤ 0.05, Tukey's HSD Test).
and Toselli, 2016) compared to soils fertilized with mineral inputs. The
mineral N availability is among factors controlling root efficiency and
lifespan, based on resource optimization in organ deployment
(Eissenstat and Yanai, 1997; Baldi et al., 2010). Our results indicate
that: a) the predominant effects on root physiology and morphology
were driven by the addition to the soil of compost, either alone or
mixed with biochar, while changes induced by the mere addition of
biochar were less pronounced, b) Changes observed in the combi
treatment resulted primarily induced by compost, without synergic
effect with biochar. In this strategy, the effect of compost likely hin-
dered that of biochar. and c) the extended root lifespan induced by
compost implies shifting in the tree resource allocation, with potential
benefits in the aboveground biomass. However, interactions at root
level are expected to vary depending on species (e.g. annual vs. per-
ennial), soil type (e.g. sandy vs. textured), biochar and compost prop-
erties (e.g. feedstock, pyrolysis conditions, pH, C/N), rates and place-
ment (surface application vs. soil incorporation).
4.4. Tree growth, nutritional status and physiology
Despite leaf traits (e.g. leaf weight, area, SLW) and the main tree
morphological parameters (i.e. tree height, TCSA, canopy radius) re-
sulted unchanged, amendment strategies influenced both aboveground
growth and physiology, with the most evident effects proportioned by
compost (without synergic effects). The higher Spad values recorded
throughout the first season (data not shown) in trees amended ex-
clusively with organic inputs (i.e. compost and combi) are likely a
consequence of the higher nutrients (i.e. easily-available N) availability
supplied by compost, as previously mentioned (Sorrenti et al., 2017).
Then, Spad values mirrored the trend of the leaf N concentration
(Rubio-Covarrubias et al., 2009) as a likely consequence of the com-
bination between compost and soil properties. Indeed, compost was
fairly reach in N (2.3%) with a well-balanced C/N ratio (17.0), in-
dicating a mature and stabilized organic matter source with adequate
N-nutritional value. Thereby, the mineralization of compost was pro-
moted in the sandy (88% sand) soil we used, rapidly increasing N mi-
neral forms in soil contributing to fulfil the limited nutrient require-
ments of the young trees in the first season. The opposite behavior
observed in the second season (lower Spad values) in compost-amended
trees may be partially due to a dilution-like effect in response of a an
increased plant growth (see pruned wood), a higher fruit set (see
thinned fruits) and consequent higher crop load (see cumulative yield).
However, most importantly, soil mineral N availability (N-NO3 and N-
NH4) was lower compared to the previous season, often below
7 mg kg−1, indicating that the residual nutritional potential provided
by compost was less pronounced (even after only one season) or that
most of the compost-derived N was leached out during the first season
(Sorrenti and Toselli, 2016). We suggest that, to support nutrient re-
quirements of perennial crops, high compost quality should be yearly
applied, even at a lower rate compared to the one adopted in this study
in order to avoid excessive nutrient availability (creating imbalances)
and consequent losses (e.g. N) through leaching, especially in coarse-
textured soils. In the second season, although acceptable for this
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Scientia Horticulturae 257 (2019) 108710
Trunk Roots Total biomass Root:Shoot
a 857.3 2513.5 7857 0.468
ab 641.3 2461.7 7923 0.451
b 771.6 2212.8 7496 0.426
ab 754.6 2323.9 7915 0.416
ns ns ns ns
species, leaf Spad values of the compost-amended trees were lower
during the early phenological stages of fruit cell division and sub-
sequent enlargement. Given that fruits are a major sink for nutrients
and photoassimilates and that crop load was higher in these trees, fruits
likely drained resources to sustain their growth at the expense of the
vegetative organs. This is corroborated by the fact that, once fruits were
picked (early July), Spad values were increased and returned in line
with remaining strategies.
On the other hand, in the first season tough, while biochar did not
affect leaf Spad values, it significantly reduced leaf N concentration,
confirming that, despite trees amended with only biochar received
additive mineral N inputs, this was not enough to prevent N imbalance.
We suppose that biochar temporarily reduced N availability for plant
uptake due to biochar-induced N adsorption or immobilization in soils,
as reported also in other experiences (Rondon et al., 2007; O’Toole
et al., 2013). To avoid N imbalance in cultivated plants, we recommend
increasing N supply, at least during the first season after wood-derived
N-poor biochar incorporation.
Noteworthy, the increased leaf and fruit K concentration of nec-
tarine trees grown in biochar-amended soil confirms that the high
concentration of exchangeable K present in the biochar may represent a
valuable plant-available K source (Sorrenti et al., 2016a). An even
greater increase in leaf K concentration was measured in compost-
amended trees. Similarly, the same strategy increased fruit flesh P and K
concentration. However, in the latter case, this implicated a reduced
leaf Ca, Mg and Mn concentrations (leaf Mg and Mn concentrations
were also reduced in biochar-amended trees), as a further evidence of
the antagonistic interaction between K with other cations and as a proof
that fruits are a major sink for translocatable nutrients in higher plants.
4.5. Yield, fruit quality and biomass organ partitioning
Benefits on soil fertility induced either directly (increased SWC,
exchangeable cations and nutrient availability, organic C) or indirectly
(reduced soil pH, physical properties), by compost may be the main
factors for an improved growing environment. Although such benefits
were of poor importance for the root system development, clear ad-
vantages were measured on plant growth and performance, with special
emphasis on the tree reproductive potential. In fact, despite a com-
parable number of thinned fruits, these were bigger in size, with higher
DM accumulation. Moreover, a higher fruit number was picked from
these trees at commercial harvest, as evidence of an increased fruit set.
Benefits clearly emerged on the leaf physiological performance in term
of photosynthetic rates, which benefited C fixation potential. This is
likely the main aspect responsible for the increased aboveground tree
growth and crop response, in term of yield and C allocation to fruits (i.e.
SSC) in compost-amended trees.
Despite leaf photosynthetic rates of biochar-amended trees were
similar to the unamended ones, biochar benefited the crop load in-
creasing the number of picked fruits (of similar fruit weight) in the first
picking though, suggesting that biochar anticipated fruit ripening. On
the other hand, improved C fixation in biochar-amended trees grown in
temperate and not limiting soil conditions was not taken for granted,
G. Sorrenti, et al.
especially in young trees. However, interestingly, the PCA approach
clearly grouped the effect of the amendments on the tree yield in which
biochar-amended trees performed evidently better than unamended
control, arranged in an intermediate spot between unamended and
compost-amended trees, as evidence of the biochar potential in im-
proving plant performance. In this context we conclude that pyrolized
biomass was i) not detrimental to tree physiology and performance and
ii) that benefits in a longer term may be supposed. On the other hand,
immediate positive effects of the combined use of biochar with organics
amendments in the field have mostly predominantly been observed in
the tropics in highly weathered soils (Cornelissen et al., 2013; Steiner
et al., 2007). Similarly, the mere addition of biochar was shown to
benefit poor tropical soils, in particular when they were sandy or acidic,
while results were negligible or absent in already fertile soils
(Cornelissen et al., 2013; Haefele et al., 2011; Nguyen et al., 2008;
Oguntunde et al., 2004; Sorrenti et al., 2016a).
The amendment with wood-derived biochar (at a rate of 10 t ha−1),
compost (72 m3 ha−1) and the mix of biochar and compost (same rates)
demonstrates that biochar compost treatments induced only small and
mostly non- significant effects over the two years in field grown
grapevine (Smith et al. 2014), with limited effects on grape quality
parameters only in the driest year in the biochar compost treatment
compared to pure biochar. Authors concluded that the top soil in-
corporation of biochar was of insignificant immediate economic values
for vine growing in alkaline-temperate soils. Similarly, Trupiano et al.
(2017) stressed that combined application of biochar and compost did
not outperform compost amendment in terms of lettuce biomass yield
and soil fertility.
C allocation was altered within trees as a function of the amendment
strategy. C stored in fruits was increased in compost-amended trees
shown by the higher crop load, supported by an increased leaf photo-
synthetic efficiency. Based on this, a decrease in the below:above
ground biomass ratio was expected as an indication of improved re-
source supplied by compost which requires fewer roots to sustain the
same above-ground biomass production. Conversely, lower shoot-to-
root ratio would have indicated a lower growth rate and lower resource
supply. However, in our conditions, differences were not enough to
differentiate aboveground tree biomass. Contrarily to our observation,
in most biochar-amended crops, Lehmann et al. (2011) reported a de-
creased root-to-shoot ratio as a consequence of an increased above-
ground biomass, while an opposite trend was described by Abiven et al.
(2015) who systematically measured an increased ratio. However such
changes were reported mostly in annual crops species which are faster
responsive to the altered growing conditions than perennial trees for
which results reported in literature are yet limited. However, while we
demonstrated that compost can successfully replace mineral inputs in
the sustainable fertilization management of fruit tree crops, we suppose
that differences induced by biochar and combi compared to unamended
control and compost, respectively would become significantly evident
in the longer term.
Probably, the synergic effect between amendments could faster
emerge if biochar is co-composted with organic amendments before
their incorporation in soils. This would allow to exploit the stability of
biochar and the nutrient-rich compost (Kammann et al. 2016), given
that the interaction of biochar with organic amendments (i.e. compost)
during composting, positively affect each other’s properties (Wu et al.,
2017), resulting in different soil and tree responses.
5. Conclusions
Our findings indicate that benefits on soil properties and tree
growth were primarily induced by the addition of compost then, but
mostly negligible, by biochar alone, confirming our first hypothesis.
Strategies influenced root physiology rather than morphology or
biomass, as a possible adaptation to the changed growing conditions
(e.g. resources availability). However, root growth pattern did not
11
Scientia Horticulturae 257 (2019) 108710
reflect the aboveground tree performance, hence the second hypothesis
cannot be accepted.
Compost extended root lifespan and increased photosynthetic leaf
efficiency, signifying that this amendment strategy improves C fixation
and alters C budget partitioning within tree organs.
Conversely, the application of biochar in temperate or fertile soils
should focus on potential non-yield benefits such as C sequestration and
other ecosystem services. Additionally, we conclude that perennial
systems characterized by high inputs may not immediately respond to
biochar application.
This is the first study reporting the effect of biochar on root survi-
vorship of perennial species. The reduced root survivorship and in-
creased root turnover, indicates that shifting in C allocation in biochar-
amended trees are likely to occur. On the other hand, rather than
detrimental effects, biochar improved tree nutritional status.
Furthermore, the benefits of charred biomasses on the yield potential
highlighted by the PCA analyses confirms that biochar-induced benefits
would presumably occur over time and/or in limiting soil conditions.
Synergic effects induced by amendments were scarcely evident.
Indeed, responses of tree physiology and performance were not sy-
nergistically outperformed, thereby our last hypothesis should be re-
jected. This because the outcome of compost appeared predominant,
likely hindering the potential synergism with biochar. While we con-
clude that the contemporaneous addition of biochar with compost may
not always reflect additive responses on plant performance, we spec-
ulate that biochar and compost could be conveniently combined in the
sustainable management of nectarine trees with the aim to replace
mineral inputs, preserve soil fertility and fight climate changes.
Acknowledgements
The authors thank Dr. A. Cipriano and Dr. E. Baldi for their assis-
tance with root image analyses.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.scienta.2019.108710.
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