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1
Agrociencia
1
Universidad Juárez del Estado de Durango. Facultad de Ciencias Forestales y Ambientales.
Avenida Papalopan y Boulevard Durango s/n, Durango, Durango, Mexico. C. P. 34120.
2
Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias. Campo Experimental
Valle del Guadiana. Carretera Durango-El Mezquital km 4.5, Durango, Durango, Mexico. C. P.
43000.
* Author for correspondence: jprieto@ujed.mx
ABSTRACT
Substrate and mycorrhization are important aspects when producing plants. The objective
of the study was to determine the eect of three substrates inoculated with ectomycorrhizal
fungal spores on the morphological characteristics of nursery-grown Pinus durangensis as well
as the inherent costs of the substrate. A randomized block experimental design with a 3 x 3
factorial arrangement was used. The substrates evaluated were: 1) peat, vermiculite, and perlite
(50:25:25) [PVP]; 2) peat, composted bark, and raw sawdust (50:25:25) [PBS]; and 3) peat and
bark (50:50) [PB]. The mycorrhizal inoculants were: 1) Pisolithus tinctorius; 2) Laccaria laccata;
and 3) control. The plant was produced in black polyethylene tubes (160 mL) and evaluated
at 13 months. The substrate factor presented statistical dierences (p ≤ 0.05) in the variables
evaluated, with higher values in PVP, followed by PBS and PB, the laer being similar to
each other. The mycorrhizal inoculation factor showed dierences (p ≤ 0.05) in root and total
biomass, Dickson’s quality index, and mycorrhizal colonization, obtaining higher values when
P. tinctorius was inoculated, except in the last variable, where the application of L. laccata stood
out. In the substrate-mycorrhizal inoculation interaction, the most favorable combination was
PVP and P. tinctorius. The PVP substrate was 28.4 and 34.6 % more expensive than PB and PBS,
respectively. The combined eect of PVP and P. tinctorius produced the best growth, although
the substrate was more expensive.
Keywords: growth media, controlled mycorrhization, plant quality, morphological
characteristics, reforestation.
INTRODUCTION
Deforestation is directly related to the loss of biodiversity and the decrease in the
eciency of environmental services provided by forests and jungles, which has
contributed to global warming. These ecosystems, in addition to being reservoirs of
Citation: Martínez-Casas R,
García-Rodríguez JL, Salcido-
Ruiz S, Goche-Télles JR, Prieto-
Ruíz JA. 2024. Production
of Pinus durangensis Mart.
under dierent substrate
and mycorrhizal inoculation
conditions in nursery.
Agrociencia. hps://doi.
org/ 10.47163/agrociencia.
v58i4.2960
Editor in Chief:
Dr. Fernando C. Gómez Merino
Received: February 02, 2023.
Approved: January 31, 2024.
Published in Agrociencia:
June 18, 2024.
This work is licensed
under a Creative Commons
Aribution-Non- Commercial
4.0 International license.
PRODUCTION OF Pinus durangensis Mart. UNDER DIFFERENT
SUBSTRATE AND MYCORRHIZAL INOCULATION
CONDITIONS IN NURSERY
Ricardo Martínez-Casas1, José Leonardo García-Rodríguez2, Silvia Salcido-Ruiz1,
José Rodolfo Goche-Télles1, José Ángel Prieto-Ruíz1*
1
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 2
biodiversity, are a source of consumer goods such as timber, fuelwood, and other
non-timber forest products (FAO, 2018). In Mexico, due to the deterioration and
decrease of forest resources, with an average annual rate of gross deforestation of 212
834 ha between 2001 and 2019, there is a need to implement strategies that favor the
restoration of forest ecosystems (CONAFOR, 2021).
Using nursery-produced plants allows the generation of new forest stands, which
in the future will contribute to re-establish forests and obtain economic gains in a
sustainable way (Buamscha et al., 2012; Scholz and Morera, 2016); however, the
propagation material must have quality, which is fundamental in reforestation
programs (Grossnickle and MacDonald, 2018). Plant quality is the result of nursery
production practices and the management of propagated material until planting
(Villar-Salvador et al., 2021). After the nursery culture process, the initial survival and
growth potential of individuals is related to their morphological and physiological
aributes, as well as their eco-physiological response to site environmental conditions
(Grossnickle, 2018).
The main cultural procedures that plants receive in nursery include irrigation,
fertilizer, mycorrhization, pest and disease control, and environmental management
(Sanchún and Obando, 2016; Grossnickle, 2018). Other important aspects include the
correct selection of substrates and the proportions for mixing them, as well as the type
of container. These factors are related to the formation of the root ball and the growing
space for the root system of the plants (Landis, 1990). The importance of the substrate
lies in the functions it performs in anchoring the root system to support the plants as a
temporary store and diuser of water and nutrient solutions that are absorbed by the
root hairs. The physical and chemical characteristics of substrates are directly related
to the morphology and functional performance of plants and, therefore, inuence
their growth and survival in the eld (Villar-Salvador et al., 2021).
There is a search for materials to replace the substrate based on peat (50–60 %), perlite
(20–25 %), and vermiculite (20–25 %), known as a base mix, whose main disadvantages
are its high cost and immediate unavailability. These reasons limit their use for most
nurserymen (Aguilera-Rodríguez et al., 2016). Therefore, it is necessary to look for
alternative materials with an ecological approach (Mateo-Sánchez et al., 2011; Gayosso-
Rodríguez et al., 2016). Pine bark and sawdust have favorable physical and chemical
characteristics for the development of seedlings of the genus Pinus (Hernández-
Zárate et al., 2014); in addition, they are by-products of forestry activity with relative
abundance (Fregoso-Madueño et al., 2017), and it is desirable to nd a potential use
for them.
An important component in plant production is mycorrhizal inoculation through the
application of spores of wild ectomycorrhizal fungi (HSECM) (Escobar-Alonso and
Rodríguez-Trejo, 2019), which promotes plant-fungus symbiosis. This condition is
desirable in species of the genus Pinus in nursery and eld to favor plant survival and
growth (García-Rodríguez et al., 2017).
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The objective of this study was to determine the eect of three substrates and inoculation
with spores of two species of ectomycorrhizal fungi and a control on the morphological
characteristics of nursery-grown Pinus durangensis Mart. Similarly, it was sought to
determine the costs inherent to the type of substrate. It was hypothesized that at least
a combination of substrate and ectomycorrhizal fungus favors the survival, growth,
and mycorrhizal colonization of the plant in a nursery at the lowest cost due to the
components of the substrate.
MATERIALS AND METHODS
Location
The experiment was carried out in the forest nursery of the Experimental Field Valle del
Guadiana, of the National Institute of Forestry, Agriculture, and Livestock Research
(INIFAP), located at Carretera Durango-El Mezquital km 4.5, Durango, Durango,
Mexico (24° 01’ N and 104° 44’ W), with an altitude of 1860 m and average annual
rainfall of 504 mm.
Plant production
The plant was produced under three environmental conditions, with the following
sequence: a) from October 17, 2019 (sowing) until seven months later, a saw-type
greenhouse was used, 50 m long and 30 m wide, with plastic lm cover caliber 720
µm, milky white color, treated against UV rays, and 50 % black color shade mesh, with
an average temperature of 22 °C, maximum of 46 °C, and minimum of -2 °C; b) from
May 8, 2020, for four months, using 50 % shade mesh, with an average temperature
of 23 °C, maximum of 39 °C and minimum of 9 °C; and c) from September 3, 2020,
for two months, under outdoor conditions, without sun protection, with an average
temperature of 20 °C, maximum of 42 °C, and minimum of 2 °C. The production cycle
lasted 13 months. Temperature was monitored with an Elitech data logger (Elitech
Technology, San Jose, CA, USA).
The plants were produced in black rigid containers mounted on 98-cavity racks. Each
tube was 3.8 cm in top diameter, 19 cm long, 160 mL in capacity, and had internal
root guides. Prior to sowing, the seed was soaked in water for 24 hours to activate
germination; subsequently, it was dried. To avoid possible damage by pathogenic
damping-o fungi, it was impregnated with Tecto 60® fungicide (thiabendazole,
2-(4-thiazolyl) benzimidazole).
Experimental design and treatments
Nine treatments were evaluated, considering three substrate mixtures based on
peat, composted pine bark, raw pine sawdust, vermiculite, and perlite, and two
spore inocula from wild ectomycorrhizal fungi (HSECM): a) Laccaria laccata (Scop.)
Cooke, native inoculum made from HSECM fruiting bodies collected in mid-2019
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Scientic article 4
from cold temperate forest stands in the Iztaccíhuatl-Popocatépetl National Park
and surroundings in Central Mexico, dehydrated fruiting bodies were ground in a
Willye type mill, sieved through a 1 mm mesh, and stored at 5 °C until inoculation; b)
Pisolithus tinctorius (Pers.) Coker et Couch, commercial spore inoculum Ecto-Rhyza®;
and c) control (no inoculum) (Table 1). A randomized, complete block experimental
design with a 3 x 3 factorial arrangement was used. Each treatment consisted of four
replicates and 49 plants per experimental unit.
Table 1. Substrate composition and inoculum of ectomycorrhizal fungi in Pinus durangensis
Mart. plant production in nursery conditions.
Treatment Substrate Mycorrhizal inoculant
1 Peat + vermiculite + perlite (50:25:25) [PVP]. No inoculum [NI].
2 Peat + bark + sawdust (50:25:25) [PBS]. No inoculum [NI].
3 Peat + bark (50:50) [PB] No inoculum [NI].
4 Peat + vermiculite + perlite (50:25:25) [PVP]. Laccaria laccata [Ll]
5 Peat + bark + sawdust (50:25:25) [PBS]. Laccaria laccata [Ll]
6 Peat + bark (50:50) [PB] Laccaria laccata [Ll]
7 Peat + vermiculite + perlite (50:25:25) [PVP]. Pisolithus tinctorius [Pt]
8 Peat + bark + sawdust (50:25:25) [PBS]. Pisolithus tinctorius [Pt]
9 Peat + bark (50:50) [PB] Pisolithus tinctorius [Pt]
After preparing the substrate mixtures for disinfection, they were impregnated
with Anibac® PLUS bactericide at a dose of 6.3 mL L-1 of water and covered for 15
d with black rubber. During mixing, eight-month-old Multicote® (12N-25P5O-12K2O)
controlled-release fertilizer was added to each substrate at a rate of 3 g L-1. Seven
months after sowing, a second application of fertilizer of the same brand was made,
but with a 17N-17P5O-17K2O ratio and with four months of release, at a dose of 2 g L-1.
The second application was made in each tube, burying the fertilizer 1 cm deep.
To complement the nutrition, fertilization was carried out through irrigation: a) one
month after germination and up to seven months after sowing, the soluble fertilizer
Master® Finalizer (4N-25P5O5-35K2O) was added at a dose of 1 g L-1 of water (except
in the period from 3 to 5.5 months of plant age, due to inoculation) (40-109-290 ppm of
N-P-K); and b) after seven months of age, Master® Development (20N-7P2O5-19K2O)
was used at a dose of 0.5 g L-1 of water (100-15-79 ppm of N-P-K). During the fall and
winter, water-soluble fertilizers were applied twice a week, and in the spring and
summer, fertilizer was applied three times a week.
For each substrate mixture, total porosity, aeration porosity, and moisture holding
capacity were estimated (Table 2) based on the methodology developed by Landis
(1990). For mycorrhizal inoculants, the spore concentration of each inoculum was
determined in a Neubauer chamber. On each plant, 5 mL of spore suspension were
applied, equivalent to 1 643 750 spores on average. Two inoculations were performed,
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 5
the rst at 3.5 months, and 30 days later (4.5 months), the second application was
performed using the same dose.
Table 2. Total porosity, aeration porosity, and moisture holding capacity of the substrate
mixtures used in the production of Pinus durangensis Mart. plants in nursery conditions.
Substrate
mixture
Total
porosity (%)
Aeration
porosity (%)
Humidity retention
capacity (%)
PVP 58.3 31.7 26.6
PBS 57.2 28.9 28.3
PB 63.4 31.4 32.0
Recommended value+60 a 80 25 a 35 25 a 55
+Landis (1990). PVP: peat + vermiculite + perlite (50:25:25); PBS: peat + bark + sawdust
(50:25:25); PB: peat + bark (50:50).
Variable evaluation and statistical analysis
The percentage of survival was evaluated thirteen months after sowing. To analyze
morphological growth, 10 plants per experimental unit were randomly taken, and the
height (measured using a PILOT® model graduated ruler in millimeters), root collar
diameter (using a CALDI-6MP model digital vernier; Truper, Mexico), and wet and
dry biomass of the aerial part and the root were recorded. Dry biomass was obtained
by placing the plants at 70 °C in a forced ventilation oven (model FE-291D; Felisa,
Mexico) for 72 hours. The samples were weighed on an analytical balance (model
PA1502, accurate to 0.001 g; Ohaus, NJ, USA). Similarly, the dry biomass ratio of the
aerial part, the dry biomass of the root, and the lignication index (Equation 1) were
calculated (Villalón-Mendoza et al., 2016), as well as the Dickson quality index (DQI)
(Equation 2) (Dickson et al., 1960).
Total dry biomass (g)
Lignication index = × 100
Total wet biomass (g) (1)
Total dry biomass (g)
DQI =
Height (cm) + Aerial dry biomass (g)
Diameter (mm) Root dry biomass (g) (2)
To determine mycorrhizal colonization (MC) (Equation 3), the methodology described
by Salcido-Ruiz et al. (2020) was used by randomly selecting three plants per
experimental unit and extracting 100 cm of secondary roots, which were preserved
in FAA xative (formaldehyde, glacial acetic acid, 96° alcohol, and distilled water) in
a 10:5:50:35 ratio, respectively. In each sample, direct observations were made on the
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 6
roots with an optical stereoscope (Leica S9i, Swierland), recording the number of
apices with and without mycorrhizae.
Mycorrhizal apices
MC = × 100
Mycorrhizal apices + Non-mycorrhizal apices (3)
The percentage results of MC and survival, being binomial variables, were transformed
with the arcsine and square root functions. Furthermore, for the variables evaluated,
analyses of variance with Tukey’s multiple comparison of means tests (p ≤ 0.05)
were performed using SAS software (SAS Institute, 2009). To calculate the costs per
plant, a volume of 160 mL per cavity was considered, according to the inputs used
per substrate mix. The evaluation was carried out based on three quotations for the
products in Durango, Durango, Mexico, and the proportions used per mixture.
RESULTS AND DISCUSSION
Substrate factor
The substrate factor showed dierences (p ≤ 0.05) in the variables evaluated; in all
cases, the PVP-based treatment stood out, except in nursery survival, where PBS and
PB were superiors. Likewise, there was a similarity of results between PBS and PB. For
the aerial/root ratio (< 2.5) and robustness index (< 6), the values are in agreement with
those suggested by Ritchie et al. (2010) (Tables 3 and 4).
Regarding total porosity, aeration porosity, and moisture holding capacity, the
percentages are in the ranges suggested by Landis (1990) (Table 2), except for total
porosity, where only the mixture composed of peat and bark was in the recommended
range, which may have inuenced the plants to reach the morphological characteristics
stipulated by the Mexican Standard for the Certication of Forest Nursery Operation
NMX-AA-170-SCFI-2016 (DOF, 2016).
Mycorrhizal inoculum factor
In the mycorrhizal inoculum factor, only the variables root and total biomass
production, Dickson’s quality index, and percentage of mycorrhizal colonization
presented statistical dierences (p ≤ 0.05); the best results were obtained when Pt or
Ll was applied for mycorrhizal colonization, but there were no dierences between Ll
and T for Dickson’s quality index. Regarding the hardiness index (< 6) and aerial/root
ratio (≤ 2.5), all treatments presented values in the range recommended by Ritchie et
al. (2010) (Tables 3 and 4).
Vicente-Arbona et al. (2019) reported a similar trend when growing Pinus greggii Engelm.
ex Parl. on ve substrates, generated from combinations with sawdust as the main
component (60 to 80 % in each substrate), mixed with bark, peat, compost, vermiculite,
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 7
Table 4. Quality indices in nursery-grown Pinus durangensis Mart. plants 13 months after sowing.
T I S Aerial/
root ratio
Robustness
index
Lignication
index (%) DQI MC (%)
S
All PVP 1.5 a 2.8 a 30.1 a 1.0 a 27.1 a
All PBS 1.6 a 2.9 a 28.1 b 0.7 b 21.6 b
All PB 1.5 a 2.7 a 28.7 ab 0.8 b 21.7 b
NI All 1.5 a 2.9 a 28.0 a 0.7 b 15.4 b
I Ll All 1.4 a 2.7 a 29.4 a 0.8 ab 28.4 a
Pt All 1.5 a 2.7 a 29.5 a 0.9 a 26.6 a
1 NI PVP 1.6 a 3.1 a 28.8 a 0.9 abc 16.9 bc
2 NI PBS 1.6 a 2.9 a 27.4 a 0.7 c 14.3 c
3 NI PB 1.6 a 2.7 a 28.0 a 0.8 bc 15.0 c
4 Ll PVP 1.5 a 2.7 a 30.1 a 1.0 ab 33.1 a
5 Ll PBS 1.5 a 2.8 a 29.0 a 0.8 bc 25.9 ab
6 Ll PB 1.5 a 2.7 a 29.4 a 0.8 bc 26.1 ab
7 Pt PVP 1.6 a 2.5 a 31.7 a 1.2 a 31.4 a
8 Pt PBS 1.5 a 2.8 a 28.1 a 0.8 bc 24.6 ab
9 Pt PB 1.4 a 2.7 a 29.0 a 0.9 bc 23.9 abc
Dierent leers in the same column indicate statistical dierences (Tukey, p ≤ 0.05). T: treatment; I:
mycorrhizal inoculant; S: substrate; DQI: Dickson quality index; MC: mycorrhizal colonization; NI:
no inoculum; Ll: Laccaria laccata; Pt: Pisolithus tinctorius; PVP: peat-vermiculite-perlite; PBS: peat-
bark-sawdust; PB: peat-bark.
Table 3. Morphological characteristics in nursery-grown Pinus durangensis Mart. plants 13 months
after sowing.
T I S Survival
(%)
Height
(cm)
Diameter
(mm)
Dry biomass (g)
Aerial Root Total
All PVP 93.9 b 15.7 a 5.6 a 2.5 a 1.7 a 4.2 a
S All PBS 98.7 a 14.3 b 5.0 b 1.9 b 1.2 b 3.1 b
All PB 96.9 ab 13.5 b 5.0 b 1.9 b 1.3 b 3.2 b
I
NI All 95.8 a 15.1a 5.2 a 2.0 a 1.3 b 3.4 b
Ll All 96.5 a 14.1a 5.2 a 2.0 a 1.4 ab 3.5 ab
Pt All 96.6 a 14.3 a 5.3 a 2.2 a 1.5 a 3.7 a
1 NI PVP 94.3 ab 17.6 a 5.6 ab 2.5 ab 1.6 ab 4.1 ab
2 NI PBS 99.5 a 14.5 b 5.0 bc 1.8 d 1.2 c 3.0 d
3 NI PB 93.8 ab 13.3 b 5.0 bc 1.9 d 1.2 c 3.1 d
4 Ll PVP 92.7 b 14.8 ab 5.5 abc 2.4 abc 1.6 ab 4.0 abc
5 Ll PBS 98.4 ab 14.2 b 5.1 bc 1.9 cd 1.3 c 3.2 cd
6 Ll PB 98.4 ab 13.4 b 5.0 bc 2.0 cd 1.3 bc 3.3 cd
7 Pt PVP 94.8 ab 14.7 b 5.8 a 2.8 a 1.8 a 4.6 a
8 Pt PBS 98.4 ab 14.5 b 5.1 bc 2.0 cd 1.3 bc 3.3 cd
9 Pt PB 97.9 ab 13.7 b 5.1 bc 2.0 bcd 1.4 bc 3.4 bcd
Dierent leers in the same column indicate statistical dierences (Tukey, p ≤ 0.05); T: treatment; I:
mycorrhizal inoculum; S: substrate; NI: no inoculum; Ll: Laccaria laccata; Pt: Pisolithus tinctorius; PVP:
peat-vermiculite-perlite; PBS: peat-bark-sawdust; PB: peat-bark.
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 8
and perlite, in dierent proportions, with and without inoculation of L. laccata (1.5 g
per plant). These authors observed that plant growth due to mycorrhization was not
very evident, although the inoculated treatments had 44 to 64 % mycorrhization and
the uninoculated individuals had between 14 and 40 % mycorrhization. The same
happened with Baltasar-Martínez et al. (2007) when inoculating spores of Rhizopogon
roseolus Corda on Pinus ponderosa Dougl. ex C. Laws. plants subjected to three
fertilization levels, two inoculation times, and two nursery harvest dates, nding no
dierences in plant growth or mycorrhizal colonization, with values lower than 39 %
in all cases.
Substrate-mycorrhizal inoculum interaction
The interaction of substrate-mycorrhizal inoculum factors was signicant in the
variables evaluated (p ≤ 0.05), with the exception of the of the aerial part/root ratio,
robustness index, and lignication index. Survival only showed dierences between
treatments 2 (PBS-SI) (99.5 %) and 4 (PVP-Ll) (92.7 %) (Table 3). Salcido-Ruiz et al.
(2020) obtained similar results, with 80 to 96 % survival in Pinus engelmannii Carr.
plants grown in a substrate based on peat, vermiculite, and perlite (57:23:20) eight
months after being inoculated with ectomycorrhizal fungi (HECM). Plant survival in
the forest nursery at INIFAP’s Guadiana Valley Experimental Field was satisfactory (>
90 %) due to relatively controlled environmental conditions.
In the present study, dierences (p ≤ 0.05) were found in morphological variables.
Height ranged from 13.3 to 17.6 cm, with treatments 1 (PVP-SI) and 4 (PVP-Ll) in
the top statistical group (although there were no dierences between the two, nor
between the laer and the rest of the treatments) (p > 0.05). Diameter ranged from 5
to 5.8 mm, with the highest value in treatment 7 (PVP-Pt), with no dierences with
treatments 1 and 4; the rest of the treatments had average data from 5 to 5.1 mm
(Table 3). The height variable is important because it is associated with the number of
needles produced in the plants, which in turn is related to photosynthetic capacity and
transpiration area; meanwhile, the diameter of the collar is related to the robustness of
the plants (Ritchie et al., 2010).
The values of the morphological parameters evaluated comply with the ranges
recommended in NMX-AA-170-SCFI-2016 (DOF, 2016), which establishes a height
of 15 to 20 cm and neck diameter greater than 4 mm; both parameters are met in
treatment 1 (PVP-SI), while the second variable is met in all treatments evaluated.
The most prominent substrate (PVP) is the most widely used in Mexico; however,
its cost is higher than the other two evaluated (PBS and PB), which is why the laer
are beginning to be widely used in nurseries located in central and northern Mexico
(Aguilera-Rodríguez et al., 2016).
The production of dry biomass of the aerial part uctuated from 1.8 to 2.8 g, with the
formation of diverse statistical groups, where treatment 7 (PVP-Pt) stood out without
statistical dierences with treatments 1 (PVP-SI) and 4 (PVP-Ll) and values of 2.8, 2.5,
and 2.4 g, respectively; the other treatments produced from 1.8 to 2 g. For root dry
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Scientic article 9
biomass, the averages ranged from 1.2 to 1.8 g, with the same behavior as the aerial
biomass variable (treatments 7, 1, and 4); total biomass varied from 3 to 4.6 g, and the
best results were obtained in the same treatments as for aerial and root biomass (Table
3).
For the aerial/root ratio, the index ranged from 1.4 to 1.6 (Table 4), while the robustness
index ranged from 2.5 to 3.1. This implies a robust and balanced plant between root
biomass and the aerial part, which favors its quality (Ritchie et al., 2010). As for the
lignication index, the percentages ranged from 27.4 to 31.7 %, with no dierences (p
> 0.05) between treatments. For the Dickson quality index, values from 0.7 to 1.2 were
found, with dierences (p > 0.05) between treatments. Treatment 7 (PVP-Pt) showed
beer results, with no signicant dierences with treatments 4 (PVP-Ll) and 1 (PVP-
SI). In general, the PVP substrate stood out (Table 4).
When evaluating Pinus cooperi Blanco, produced on four substrates based on S1: peat
and bark (54:46); S2: raw sawdust, composted bark, and peat (50:20:30); S3: sawdust,
bark, and peat (50:20:30); and S4: sawdust, bark, and peat (50:25:25), González-Orozco
et al. (2018) reported satisfactory results in S1 in the variables height (15.1 cm), root
collar diameter (4 mm), and total biomass (2.8), although S2 and S3 produced plants
with acceptable morphological characteristics in height (16.4 and 16.2 cm), diameter
(3.8 and 3.7 mm), and total biomass (2.5 and 2.4 g). Castro-Garibay et al. (2018) found
similar results in plant quality due to the use of sawdust when producing Pinus greggii
plants on sawdust, bark, and peat-based substrates (60:20:20).
Mycorrhizal colonization (MC) (14.3 to 33.1 %) showed dierences (p ≤ 0.05), forming
several statistical groups where treatments 4 to 9 (inoculated with ectomycorrhizal
fungi, regardless of the substrate used, and MC from 23.9 to 33.1 %) stood out,
although in several cases without dierences with the rest of the treatments, which
had colonization percentages from 14.3 to 16.9 % (Table 4). Inoculation with spore
suspensions with P. tinctorius and L. laccata allowed mycorrhization in the root systems
of P. durangensis 13 months after sowing and eight months after inoculation. However,
the values were less than 34 %, considered medium to low, including the percentage
of undesirable mycorrhization observed in the control treatment (uninoculated). This
eect is dicult to avoid in experiments of this nature, since the inoculum can arrive
through the air, from the mycorrhizal treatments, or even from outside the experiment;
however, it was always signicantly lower compared to the inoculated treatments.
The mycorrhizal colonization resulting from this trial is lower than that recorded by
Salcido-Ruiz et al. (2020), who inoculated Pinus engelmannii plants with two mixtures
of HECM spores, one composed of Amanita rubescens (Pers. ex Fr.) Gray, Amanita sp.,
Lactarius indigo (Schwein.), Ramaria sp., and Boletus sp., and another by a mixture of
P. tinctorius and Scleroderma citrinum Pers; in addition, they added controlled release
fertilizer (3 and 6 g L-1 of substrate), obtaining from 15 to 71 % mycorrhization, with
an inverse trend in plant quality as a function of the fertilization dose applied. Proper
substrate selection is a key element because it favors the production of aerial and root
biomass compared to uninoculated plants (Rentería-Chávez et al., 2017).
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In general, mycorrhizal fungi are aerobic. Saif (1981) indicates that oxygen concentrations
between 12 and 16 % are ideal for both mycorrhization and the benecial expression
of this symbiosis in plant growth. In the present study, the lowest MC percentages
coincide with the substrates with the highest percentage of moisture retention (PBS
and PB) and the highest with the one with the highest aeration porosity (PVP).
This study’s mycorrhization percentages were modest, and the mycorrhizal component
had lile eect on the morphological growth of Pinus durangensis. This was similar to
Vicente-Arbona et al. (2019) on Pinus greggii with L. laccata and Baltasar-Martínez et al.
(2007) with Pinus ponderosa and Rhizopogon roseolus, who found slight dierences in
plant growth and mycorrhizal colonization due to mycorrhizal inoculum application.
Both studies agree that this issue should be further studied in nurseries and plantations.
At the laer site, positive eects of mycorrhizal inoculation on initial growth are more
likely to be found. This situation was corroborated by Barroetaveña et al. (2016), who
found that after ve years of inoculating P. ponderosa plants in nursery conditions with
Suillus luteus (L) Roussel, Rhizopogon roseolus, Hebeloma mesophaeum (Louis), and leaf
lier, the rst two species signicantly favored the growth of the plants under study
during the period after planting in the eld, despite the prevailing conditions of water
stress.
The factor with the greatest inuence on the results was the substrate, where the PVP
combination stood out, but with close values in the other two substrates (PBS and PB).
This was largely because the three substrates evaluated, in general, meet the criteria
for total porosity, aeration porosity, and moisture holding capacity recommended by
Landis (1990). In relation to mycorrhization, although it was not a determining factor
in the quality of the plants evaluated, the plant material inoculated with P. tinctorius
stood out.
Based on the results obtained, treatment 7 (PVP-Pt) stands out with the best values
in the variables evaluated, and above all, it complies with the recommendations of
NMX-AA-170-SCFI-2016 (DOF, 2016), which establishes the minimum morphological
parameters to consider that the plant produced in nursery conditions will have the
necessary aributes for adequate survival and initial growth after planting. In the case
of the other two substrates evaluated (PB and PBS in interaction with Pt and Ll), they
also comply with the minimum values suggested by the standard.
Substrate costs
Based on the price of each component used to prepare the substrates evaluated, the
cost per cubic meter was determined (Table 5), with the PVP combination being the
highest, followed by PB and PBS (Figure 1). Of the three substrates evaluated, the rst
was 28.4 % more expensive than PB and 34.6 % higher than PBS, which proved to be
the most economical. The dierence between the two most economical substrates (PB
and PBS) was 8.6 %, with PBS being the least expensive of all.
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 11
Madrid-Aispuro et al. (2020) evaluated four substrates based on composted pine
bark, raw pine sawdust, and peat, mixed in dierent proportions, and found that
the substrate based on raw sawdust, composted pine bark, and peat is an alternative
to produce quality forest plants at a lower production cost, as long as sawdust is
incorporated up to a maximum of 50 % in the mixture.
Aguilera-Rodríguez et al. (2016) compared in Pinus montezumae Lamb. the quality of
the plant produced and production costs due to the use of two substrates: composted
pine sawdust, composted pine bark, and vermiculite (70:15:15), and peat, perlite, and
vermiculite (60:20:20); in both substrates, plant quality was favorable. However, they
recommend the former because it is 45.8 % cheaper. González-Orozco et al. (2018)
compared costs on four substrates: S1: peat and composted pine bark (46:54); S2:
Table 5. Costs of the materials used in the substrate mixtures evaluated in the production of Pinus
durangensis in nursery.
Component Volume
presentation (L)
Cost per bulk
(USD)¶
Bulks per cubic
meter
Cost per cubic
meter ($USD)
Peat 160+45.26 6.25 282.9
Vermiculite 114 35.14 8.77 308.2
Agrolite 100 10.65 10 106.5
Bark 1000 69.22 1 69.2
Sawdust 1000 10.64 1 10.6
+Volume of the bulk once the peat has been decompacted. ¶Dollar costs as of January 31, 2023, based
on three quotes in Durango, Durango, Mexico ($18.78 MXN per $1 USD).
Figure 1. Cost of the evaluated substrates per 160 mL cavity.
Agrociencia 2024. DOI: hps://doi.org/10.47163/agrociencia.v58i4.2960
Scientic article 12
peat, composted bark, and raw pine sawdust (30:20:50); S3: peat, bark, and sawdust
(25:25:50); and S4: peat, bark, and sawdust (20:30:50). Although in the rst substrate
they obtained favorable results in plant quality, S2 and S3 also produced acceptable
growth, with lower production costs by 39.8 and 43.1 % in relation to S1.
CONCLUSIONS
The combination of the substrate composed of peat, vermiculite, and perlite (50:25:25)
and the inoculation with spores of Pisolithus tinctorius had superior values in the
morphological characteristics of Pinus durangensis plants produced in nursery
conditions. The substrate based on peat, vermiculite, and perlite (50:25:25) gave the best
results in plant growth; however, the addition to peat of alternative substrates, such
as composted bark and raw sawdust (50:25:25), and peat plus composted bark (50:50),
also generated good-quality plants, reducing costs by 28. 4 % when using the mixture
of peat and bark, and by 34.6 % with the combination of peat, bark, and sawdust.
Mycorrhization of Pinus durangensis with Pisolithus tinctorius produced under nursery
conditions generated beer quality plants, followed using Laccaria laccata.
ACKNOWLEDGMENTS
To the Universidad Juárez del Estado de Durango, for its support to the author of the
correspondence “in the exercise of the sabbatical year during the period August 2022-July 2023”.
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