Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
DOI: https://doi.org/10.5902/1980509833322
ISSN 1980-5098
Artigos
Submissão: 27/06/2018
Aprovação: 21/01/2020 Publicação: 04/06/2020
Biological inputs in promoting the growth of Bauhinia forficata Link.
seedlings
Insumos biológicos na promoção do crescimento de mudas de Bauhinia forficata
Link.
Aline PeccattiI, Ana Paula Moreira RovedderII, Gerusa Pauli Kist SteffenIII, Joseila
MaldanerIV, Betina CamargoV, Luna Parode DalculVI, Luana Camila CapitaniI,
Rafaela Badinelli HummelVII, Frederico NeeuenschwanderI
Abstract
Crops of medicinal plants demand careful attention with agrochemicals in order to avoid changing
the composition of its active principles. Biological inputs are more recommended for this purpose. We
tested the effects of Trichoderma and vermicompost on Bauhinia forficata Link. seedlings, one of the most
exploited species for medicinal purposes. Two trials were installed in a completely randomized design with
40 replicates in a greenhouse in southern Brazil. We tested two Trichoderma asperelloides strains (T1 and
T2) and two Trichoderma harzianum strains (T13 and T33) inoculated on substrate and a control treatment
(substrate without inoculation). The vermicompost assay tested the proportions 0, 20, 40, 50, 60 and 80 %
vermicompost (T1, T2, T3, T4, T5 and T6 respectively). The variables of height, collar diameter, chlorophyll
content and leaf number were evaluated at 45, 90 and 135 days after seeding. We analyzed seedlings survival,
leaf area, shoot and root total dry biomass, and Dickson Quality Index at 135 days after seeding. Leaf area
was submitted to the Tukey test (α = 0.05). Other variables did not present normality and homogeneity
of variances and were compared by Kruskal-Wallis (α = 0.05). Biological inputs positively influenced the
initial growth of Bauhinia forficata. Height, collar diameter, leaf area and try biomass (total, shoot and root)
were higher in relation to control treatments for both trials. Among Trichoderma strains, T13 presented
the best results in relation to T33. Seedlings produced with larger percentages of vermicompost (50, 60 and
80%) presented statistically higher development for height and collar diameter 90 days after seeding when
compared to the control treatment. However, there was no clear pattern of differences between the other
doses of this entry, requiring further dosing tests. Trichoderma and vermicompost showed to be promising
for the production of Bauhinia forficata for medicinal purposes.
Keywords: Trichoderma; Vermicompost; Medicinal species; Pata-de-vaca; Fabaceae
I
II
III
IV
V
VI
VII
Engenheira(o) Florestal, Ma., Universidade Federal de Santa Maria, Av. Roraima, 1000, CEP 97105-900, Santa Maria (RS), Brasil. aline.peccatti@
gmail.com (ORCID: 0000-0001-9453-7658) / lumilacapitan@gmail.com (ORCID: 0000-0001-8751-5717) / frede.1990@hotmail.com (ORCID: 00000002-3520-7680)
Engenheira Florestal, Drª., Professora do Departamento de Ciências Florestais da Universidade Federal de Santa Maria, Av. Roraima, 1000, CEP
97105-900, Santa Maria (RS), Brasil. anarovedder@gmail.com (ORCID: 0000-0002-2914-5954)
Engenheira Agrônoma, Drª., Pesquisadora no Centro de Pesquisas em Florestas do Departamento de Diagnóstico e Pesquisa Agropecuária, BR 287,
Acesso VCR 830, Boca do Monte, Caixa Postal 346, CEP 97001-970, Santa Maria (RS), Brasil. gerusa-steffen@seapi.rs.gov.br (ORCID: 0000-00020464-567X)
Bióloga, Drª., Pesquisadora do Centro de Pesquisas em Florestas do Departamento de Diagnóstico e Pesquisa Agropecuária, BR 287, Acesso VCR
830, Boca do Monte, Caixa Postal 346, CEP 97001-970, Santa Maria (RS), Brasil. jomaldaner@gmail.com (ORCID: 0000-0002-3008-5047)
Engenheira Florestal, Universidade Federal de Santa Maria, Av. Roraima, 1000, CEP 97105-900, Santa Maria (RS), Brasil. betinacamargo93@gmail.
com (ORCID: 0000-0001-5526-1783)
Engenheira Florestal, Pesquisadora Autônoma. Rua Borges de Medeiros, 889, apto 101, CEP 97400-000, São Pedro do Sul (RS), Brasil. lunadalcul@
hotmail.com (ORCID: 0000-0002-1580-9967)
Engenheira Florestal, Drª., Pesquisadora Autônoma, Linha 21 Norte, CEP 98750-000, Ajuricaba (RS), Brasil. rafaela.hummel@gmail.com (ORCID:
0000-0002-3251-6031)
Esta obra está licenciada sob uma Creative Commons Attribution-NonCommercial 4.0 Unported License.
Peccatti, A.; Rovedder, A. P. M.; Steffen, G. P. K.; Maldaner, J.; Camargo, B.;
Dalcul, L. P.; Capitani, L. C.; Hummel, R. B.; Neeuenschwander, F.
368
Resumo
O cultivo de plantas medicinais exige cuidado na aplicação de agroquímicos a fim de evitar a alteração
da composição de seus princípios ativos. Os insumos biológicos são mais recomendáveis para esse
propósito. Foi testado o efeito de Trichoderma spp. e vermicomposto em mudas de Bauhinia forficata Link.,
uma das espécies florestais mais exploradas para fins medicinais, a partir de dois ensaios instalados em
delineamento inteiramente casualizado com 40 repetições, em casa de vegetação. Testamos duas estirpes
de Trichoderma asperelloides (T1 e T2) e duas estirpes de Trichoderma harzianum (T13 e T33) inoculadas
em substrato e um tratamento controle (substrato sem inoculação). O ensaio com vermicomposto testou
as proporções de 0, 20, 40, 50, 60 e 80 % de vermicomposto (T1, T2, T3, T4, T5 e T6 respectivamente). As
variáveis altura, diâmetro do coleto, teor de clorofila e número de folhas foram avaliadas aos 45, 90 e 135
dias após a semeadura. Analisamos também a sobrevivência, área foliar, biomassa seca total, da parte
aérea e raízes e Índice de Qualidade de Dickson das mudas aos 135 dias após a semeadura. A área foliar foi
submetida ao teste de Tukey (α = 0,05). Outras variáveis não apresentaram normalidade e homogeneidade
de variâncias e foram comparadas por Kruskal-Wallis (α = 0,05). Os insumos biológicos influenciaram
positivamente no crescimento inicial de Bauhinia forficata. A altura, o diâmetro do coleto, a área foliar e a
biomassa seca (total, aérea e radicular) foram superiores em relação aos tratamentos controle, para ambos
os ensaios. Dentre as estirpes de Trichoderma spp., T13 apresentou melhores resultados para o crescimento
das mudas em relação à T33. Mudas produzidas com proporções maiores de vermicomposto (50, 60 e 80
%) apresentaram desenvolvimento estatisticamente superior em altura e diâmetro do coleto, a partir dos
90 dias após semeadura, quando comparado ao tratamento controle. No entanto, não houve um padrão
claro de diferenças entre as outras doses testadas, exigindo mais testes de dosagem. Trichoderma spp. e
vermicomposto mostraram-se promissores para produção de Bauhinia forficata para fins medicinais.
Palavras-chave: Trichoderma; Vermicomposto; Espécies medicinais; Pata-de-vaca; Fabaceae
Introduction
Native forest species with medicinal potential are explored through predatory extractivism, consisting
a practice incompatible with the premises of sustainability and genetic conservation. Moreover, the product
supply is scarce and/or improper for human consumption. To reverse this scenario, multidisciplinary
research aimed at conservation and development of alternative agricultural production methods of these
species have been encouraged by the Brazilian government through its National Policy on Integrative and
Complementary Practices and the National Policy on Medicinal and Phytotherapeutic Plants (CARVALHO
et al., 2011).
In this context, agro-ecological practices and management allow for diversifying the economic
activities of small rural properties and promote natural resource conservation (ROVEDDER et al., 2016).
Trichoderma spp. is an important fungus in the development of ecological technologies for plant production,
mainly for medicinal and food purposes since they reduce the application of agrochemicals. In addition,
they promote plant growth through mechanisms that increase the nutrient uptake, root system expansion,
survival in adverse conditions and the induction of plant defense responses (CONTRERAS-CORNEJO et
al., 2016).
Vermicompost is another alternative that has been widely used as a biological input in seedling
production systems. Its application has proven effects due to the presence of humic acids and plant growth
regulating hormones which act to improve the soil physicochemical and biological conditions through the
availability of nutrients, increased aeration, water storage and cation exchange capacity (MAJI et al., 2017). In
addition to the reducing production costs, its use contributes for improving the environment and the quality
of life.
Among the forest species with medicinal potential, Bauhinia forficata Link. (Fabaceae) stands
out for being widely known in folk medicine, mainly being indicated in treatments against diabetes and
kidney problems (TROJAN-RODRIGUES et al., 2012). It is one of the most exploited species for medicinal
purposes, and therefore it is considered a priority for conservation in the southern region of Brazil (SANTOS;
SIMINSKI, 2011), reinforcing the need for technological demands that aid in its propagation.
In this sense, the objective of this study was to evaluate the potential of Trichoderma spp. and
vermicompost in the initial growth of Bauhinia forficata seedlings under nursery conditions, aiming at
developing techniques for producing native forest species which are compatible with medicinal use.
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Biological inputs in promoting the growth of Bauhinia forficata Link. ...
369
Material and methods
Study location
Two experiments were conducted in the greenhouse of the Centro de Pesquisa em Florestas
of the Departamento de Diagnóstico e Pesquisa Agropecuária (DDPA) of the Secretaria da Agricultura,
Pecuária e Desenvolvimento Rural do RS (SEAPDR/RS) located in the municipality of Santa Maria,
RS state, Brazil (29°41’25 “S and 53°48’42” W), in the period of September 2015 to February 2016.
Experimental procedures
Trichoderma spp. isolates
The experiments were installed in a completely randomized design with four treatments
consisting of inoculation of Trichoderma spp. in substrate ad a control treatment without fungal
isolate for comparison. Forty (40) repetitions were installed for each treatment, considering one
plant per tube as repetition and tested four Trichoderma spp. strains (T1, T2, T13 and T33). T1
and T2 strains corresponded to Trichoderma asperelloides species; T13 and T33 corresponded to
Trichoderma harzianum species. The Laboratory of Biochemical Phytopathology of the Biological
Institute from the University of São Paulo (USP) performed the species level identification of
fungal isolates.
Multiplication of the fungal isolates was performed according to Steffen and Maldaner
(2017), through inoculating rice grains colonized by pure cultures of microorganisms in a mixture
of Mecplant® commercial substrate and non-sterile sieved soil in a 1:1 (v/v) proportion. The soil is
classified as Acrisols with sand texture (SOIL SURVEY STAFF, 2014).
The center called ‘Centro de Pesquisa em Florestas’ provided the Trichoderma spp. strains.
T1 and T2 were isolated from native forest soil. T13 was isolated from the oat rhizosphere and
T33 was isolated from a soil sample of a soybean plantation. T1 and T2 were isolated of soil with
native forest while T13 was isolated from the oat rhizosphere and T33 was isolated from a soil
sample of a soybean plantation.
Proportions of vermicompost
The experiment was conducted in a completely randomized design with six treatments
and 40 replicates per treatment. We tested different proportions of vermicompost and nonsterile sifted soil from the Acrisols with sand texture with native vegetation cover, characterized
as: T1: Control treatment, without application of vermicompost (100% A horizon of Argisol soil);
T2: 20% vermicompost + 80% A horizon of Argisol; T3: 40% vermicompost + 60% A horizon of
Argisol; T4: 50% vermicompost + 50% A horizon of Argisol; T5: 60% vermicompost + 40% A
horizon of Argisol; and T6: 80% vermicompost + 20% A horizon of Argisol.
The Centro de Pesquisas em Florestas produced the vermicompost from transforming tanned
bovine manure by the Eisenia andrei Bouché earthworm species. After 100 days, the material
was sieved in a 2 mm mesh to separate the worms from the vermicompost and homogenize the
material. The Soil Analysis Laboratory of the Federal University of Santa Maria (UFSM) carried
out the chemical analysis of vermicompost and soil accorging to Embrapa (1997) (Table 1).
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Peccatti, A.; Rovedder, A. P. M.; Steffen, G. P. K.; Maldaner, J.; Camargo, B.;
Dalcul, L. P.; Capitani, L. C.; Hummel, R. B.; Neeuenschwander, F.
370
Table 1 – Chemical properties of soil and vermicompost used in the composition of
substrates for the production of Bauhinia forficata seedlings
Tabela 1 – Propriedades químicas do solo e vermicomposto utilizados na composição dos
substratos para.produção de mudas de Bauhinia forficata
pH
OM
Ca
Mg
-
m/v
Soi
4.5
1.8
3.4
1.3
Ver
6.4
16.7
12.3
Al
BS
P
Al
H+Al
CTC
0.7
8.7
5.7
13.5
0
2.8
27.8
K
Cu
Zn
B
cmol dm³
%
mg/dm³
Soi
13.6
35.7
11.5
134.7
1.0
1.7
0.1
Ver
0
90.9
500
800
3.7
35.5
0.2
Source: Authors (2019)
Abbreviations: Soi = soil; Ver = vermicompost.
General Procedures
The seeds of Bauhinia forficata were obtained from the seed bank of the Fepagro Seed
Analysis Laboratory, stored for 5 months in a dry cold chamber at 6° C (± 2). The lot was
constituted by seeds obtained from five matrices in the Central Depression of RS state, presenting
germination power of 59.5 % at the time of seeding.
The seeding was done in tubes with a volume of 180 cm³ and filled with the respective
substrates of each test. The overcoming dormancy was performed with immersion of the seeds
in sulfuric acid for 5 minutes and later washed in running water.
After seeding, the plastic grids containing the tubes remained inside the greenhouse.
Irrigation was performed daily throughout the experiment. Seedling thinning occurred
approximately 30 days after seeding, leaving one seedling per tube.
Evaluated variables
The following variables were evaluated at 45, 90 and 135 days after seeding (DAS): height
(H) with graded ruler (cm), collar diameter (CD) with digital pachymeter (mm), chlorophyll
content (CC) of fully expanded apical leaves with digital chlorophyllometer (chlorofiLOG® CFL
1030, Falker) and leaf number (LN) expanded completely.
The survival rate of seedlings (SVL) (%) was determined at 45 days after seedling. The leaf
area (LA) (cm³) through of the QUANT version 1.0.2 program (OLIVEIRA, 2007), shoot (SDB)
and root dry biomass (RDB), total dry biomass (TDB), SDB/RDB ratio and Dickson Quality Index
(DQI) (DICKSON; LEAF; HOSNER, 1960) were determined at 135 days after seeding. DQI were
estimated by the formula:
DQI = TDB(g) ∕ [(H(cm) ∕ CD(mm))+(SDB(g) ∕ RDB(g))]
In which: DQI = Dickson Quality Index; TDB = total dry biomass; H = height; CD = collar diameter; SDB =
shoot dry biomass; RDB = root dry biomass.
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Biological inputs in promoting the growth of Bauhinia forficata Link. ...
371
A destructive evaluation was carried out to obtain the LA, SDB, RDB and TDB, randomly
selecting eight individuals by treatment at 135 days. The individuals had separate shoot and root
parts, conditioned in paper bags and placed in a forced air ventilation oven at 60°C for 72 hours.
After drying, the material was weighed in a 0.001g precision scale. The results were expressed in
grams/plant.
Statistical analysis
The data were processed and analyzed with the Microsoft Excel (Action supplement) and
Assistat version 7.7 pt. The mean values of the height, collar diameter, leaf number, chlorophyll
content, survival, shoot dry biomass, root dry biomass, total dry biomass and Dickson Quality
Index were analyzed by the Kruskall-Wallis test at 5 % probability of error. The non-parametric
test was chosen because the data did not show normal distribution as verified by the Lilliefors and
Shapiro-Wilk tests and homogeneity of the variances by the Bartlett test. The leaf area attended
the normality and homogeneity assumptions of the variances, using ANOVA performed with the
Tukey test at a 5% significance level.
Results and discussion
Trichoderma spp.
Trichoderma asperelloides T1, Trichoderma asperelloides T2 and Trichoderma harzianum
T13 promoted significant effects on growth in height and collar diameter of Bauhinia forficata
seedlings (Table 2). It is believed that the promotion of plant growth is related to the combination
of one or more action mechanisms of Trichoderma spp., such as increase in nutrient absorption
and synthesis of hormones that stimulate growth. Effects of growth promotion on height (H) and
collar diameter (CD) by Trichoderma spp. were also observed in other tree species of the family
Fabaceae as Lysilona behanensis L., Caesalpinia violacea L. and Albizia procera (Roxb.) (GONZÁLEZ;
REINALDO; ORTIZ, 2015) while for Leucaena leococephala (Lam.) de Wit this effect was observed
only for the CD (DIAZ; GONZÁLES, 2018).
Some studies have demonstrated that an initial period is necessary for Trichoderma
spp. action mechanisms to effectively influence plant growth. For Jacaranda micrantha Cham.
(Bignoniaceae) seedlings this period was verified at 90 DAS (AMARAL et al., 2017) while for
Grevillea robusta A. Cunn. Ex. R. Br. (Proteaceae) seedlings was at 30 DAS (UMASHANKAR et al.,
2012). In this study, this moment occurred at 90 DAS when T1, T2 and T13 differed significantly
from the control. At 135 DAS, T13 differed statistically from T33 for the same variables (Table 2).
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Peccatti, A.; Rovedder, A. P. M.; Steffen, G. P. K.; Maldaner, J.; Camargo, B.;
Dalcul, L. P.; Capitani, L. C.; Hummel, R. B.; Neeuenschwander, F.
372
Table 2 – Height and collar diameter of Bauhinia forficata seedlings at 45, 90 and 135 days
after seeding and survival at 45 days after seeding with different Trichoderma spp. isolates
inoculated on the substrate
Tabela 2 – Altura e diâmetro do coleto de mudas de Bauhinia forficata aos 45, 90 e 135 dias após
semeadura e sobrevivência de mudas aos 45 dias após semeadura com diferentes isolados de
Trichoderma spp. inoculados no substrato
Days after seeding
Treatments*
45
H (cm)
90
SVL
(%)
135
CD (mm)
H (cm)
CD (mm)
H (cm)
CD (mm)
Control
5.5 a**
1.6 a
8.0 b
1.9 b
9.7 c
2.1 c
65.0 ab
T1
6.3 a
1.6 a
10.5 a
2.4 a
12.2 ab
2.5 ab
67.5 ab
T2
6.0 a
1.6 a
9.6 a
2.3 a
11.8 ab
2.5 ab
70.0 ab
T13
5.9 a
1.6 a
10.6 a
2.5 a
13.1 a
2.7 a
77.5 a
T33
6.2 a
1.5 a
9.5 ab
2.1 ab
11.1 bc
2.2 bc
50.0 b
CV (%)
22.2
12.3
23.7
22.1
19.1
21.8
17.1
Source: Authors (2019)
In which: H (height); CD (collar diameter); SVL (seeding and survival).
(*) Control: non-inoculation; T1: inoculation with Trichoderma asperelloides strain T1; T2: inoculation with
Trichoderma asperelloides strain T2; T13: inoculation with Trichoderma harzianum strain T13; T33: inoculation
with Trichoderma harzianum strain T33; CV = Coefficient of variation.
(**) Values followed by the same letter in the column did not differ significantly by the Kruskal-Wallis test
(α=0.05).
Stewart and Hill (2014) support the hypothesis that the Trichoderma spp. promotes plant
growth by releasing phytohormones and by phytohormone synthesis induction in the plant.
Rootstocks of Prunus cerasus x Prunus canescens (Rosaceae) cultivated in vitro showed root and
shoot growth due to the levels of indole acetic acid and gibberellic acid produced by plants and
not by the Trichoderma spp. However, Trichoderma harzianum induced hormone production in the
plants (SOFO; SCOPA; MANFRA, 2011).
Regarding the seedlings survival, the influence of Trichoderma spp. was not so distinct
(Table 2). The study carried by Sofo, Milella and Tataranni (2010) Trichoderma spp. increased the
survival percentage of Prunus spp. seedlings during the acclimation phase by 92%, whereas it was
approximately 80% for the control.
The leaf numbers per plant was not influenced by Trichoderma spp. (Table 3). Similar results
were observed for Jacaranda micrantha (AMARAL et al., 2017), Grevillea robusta (UMASHANKAR
et al., 2012) and Leucaena leococephala (DÍAZ; GONZÁLES, 2018) species. Variations in LN have
been reported as responses to stress factors through the activation of physiological plant defense
mechanisms (TAIZ et al., 2017). Despite this, there was an increase in the leaf area of T1, T2 and
T13 in relation to the control (Table 3). This result is promising since the leaves constitute the
main part of the plant used for medicinal purposes.
Regarding the chlorophyll content in the leaves, T13 stood out statistically in relation
to the control and T33, only in the 45 DAS. Although there were no differences in following
evaluations, this result suggests that there was a direct expression of this variable in the leaf area
increase in the plants with the T13 isolate at 135 days (Table 3), demonstrating the existence of
a close relation between these two variables. According to Taiz et al. (2017), this relation occurs
due to the greater or lesser amount of photoassimilates produced and accumulated by the plants.
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Table 3 – Leaf numbers and leaf chlorophyll content of Bauhinia forficata seedlings at 45, 90
and 135 days after seeding and leaf area obtained at 135 days after seeding with different
Trichoderma spp. isolates inoculated on the substrate
Tabela 3 – Número de folhas e teor de clorofila foliar em mudas de Bauhinia forficata aos 45,
90 e 135 dias após semeadura e área foliar obtida aos 135 dias após semeadura com diferentes
isolados de Trichoderma spp. inoculados no substrato
Days after seeding
Treatments*
45
LN
90
135
LA (cm²)
CC
LN
CC
LN
CC
Control
3 a**
24.5 b
5a
23.8 a
7a
33.1 a
70.44 c
T1
3a
26.1 ab
6a
25.4 a
7a
32.2 a
96.89 ab
T2
3a
26.7 ab
6a
24.9 a
7a
32.9 a
96.58 ab
T13
3a
30.4 a
6a
25.2 a
7a
34.1 a
117.31 a
T33
3a
22.5 b
6a
24.9 a
7a
31.5 a
72.17 bc
CV (%)
15.0
21.1
15.8
17.2
17.9
14.5
24.9
Source: Authors (2019)
In which: LN (leaf numbers); CC (chlorophyll content); LA (leaf area).
(*) Control: non-inoculation; T1: inoculation with Trichoderma asperelloides strain T1; T2: inoculation with
Trichoderma asperelloides strain T2; T13: inoculation with Trichoderma harzianum strain T13; T33: inoculation
with Trichoderma harzianum strain T33; CV = Coefficient of variation.
(**) Values followed by the same letter in the column did not differ significantly by the Kruskal-Wallis test
(α=0.05) for the variables LN and CC and the Tukey test (α=0.05) for the variable LA.
For the results of total dry biomass, shoot and root dry biomass, T13 was significantly
higher in relation to the control treatment and T33 for the three parameters (Table 4). A
similar effect was observed in Salix fragilis L. (Salicaceae) seedlings grown with Trichoderma
harzianum (ADAMS; DE-LEJI; LYNCH, 2007) and in the root dry biomass of Theobroma cacao
L. (Malvaceae) with the application of Trichoderma hematum (BAE; SICHER; KIM, 2009). For
Leucaena leucocephala, however, there was no increase in SDB and RDB of the plants with the
application of Trichoderma harzianum (DIAZ; GONZÁLES, 2018). This result suggests that the
ability of Trichoderma spp. in colonize the rhizospheric environment and promote increase in
biomass (STEWART; HILL, 2014) may vary between the different strains and plant species.
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Peccatti, A.; Rovedder, A. P. M.; Steffen, G. P. K.; Maldaner, J.; Camargo, B.;
Dalcul, L. P.; Capitani, L. C.; Hummel, R. B.; Neeuenschwander, F.
374
Table 4 – Total dry biomass, shoot dry biomass, root dry biomass, shoot dry biomass and
root dry biomass ratio and Dickson Quality Index of Bauhinia forficata seedlings with
different Trichoderma spp. isolates inoculated on the substrate after 135 days of seeding
Tabela 4 – Biomassa seca total, biomassa seca aérea, biomassa seca radicular, relação biomassa
seca aérea e biomassa seca radicular e Índice de Qualidade de Dickson de mudas de Bauhinia
forficata com diferentes isolados de Trichoderma spp. inoculados no substrato 135 dias após
semeadura
Treatments*
TDB (g)
SDB (g)
RDB (g)
SDB/RDB
DQI
Control
0.93 b**
0.34 b
0.59 b
0.60 a
0.20 ab
T1
1.03 ab
0.47 ab
0.56 b
0.94 a
0.18 ab
T2
1.13 ab
0.43 b
0.70 ab
0.71 a
0.20 ab
T13
1.63 a
0.66 a
0.96 a
0.71 a
0.29 a
T33
0.95 b
0.40 b
0.55 b
0.85 a
0.15 b
33.5
33.4
38.7
37.6
39.8
CV (%)
Source: Authors (2019)
In which: TDB (total dry biomass); SDB (shoot dry biomass); RDB (root dry biomass); SDB/RDB (shoot dry
biomass and root dry biomass ratio); DQI (Dickson Quality Index).
(*) Control: non-inoculation; T1: inoculation with Trichoderma asperelloides strain T1; T2: inoculation
with Trichoderma asperelloides strain T2; T13: inoculation with Trichoderma harzianum strain T13; T33:
inoculation with Trichoderma harzianum strain T33; CV = Coefficient of variation.
(**) Values followed by the same letter in the column did not differ significantly by the Kruskal-Wallis test
(α=0.05).
Trichoderma spp. did not present a significant effect on the DQI when compared to the
control (Table 4), which is contrary to the results found by Amaral et al. (2017) in Jacaranda micrantha
seedlings. However, T13 differed significantly from T33. The lack of a comparative methodology
to evaluate DQI makes it difficult to obtain conclusions about the quality of seedlings of native
species based only on this parameter. However, it is understandable that these values have not
been standardized against the genetic, physiological and adaptive diversity which regulate the
growth of these species.
Differences observed between T13 and T33 strains (Trichoderma harzianum) for most of
the studied variables (height, collar diameter, survival, leaf area, chlorophyll content at 45 days,
dry biomass and DQI) suggest that there is variability between strains of the same species as
Trichoderma spp. These variations may occur due to the ability of a strain expressing higher
levels of some action mechanism over the other (MARZANO; GALLO; ALTOMARE, 2013),
or because there are different adaptation degrees to biotic and abiotic factors between strains
(SARIAH et al., 2005).
Another hypothesis that may be associated and which should be better explored in later
studies is the influence of the origin site of the isolates, since T13 and T33 were isolated from
distinct surfaces (oat plant rhizosphere and soil from a soybean plantation, respectively). Li et al.
(2017) also point to the need for a more detailed description of the variables that characterize the
origin site of Trichoderma spp. strains in order to better explore such information.
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Vermicompost
Effects of the vermicompost on the height and collar diameter were more expressive from
90 DAS, possibly because the seedlings used their nutritive reserves during the initial period
(Table 5). The use of vermicompost in the substrate formulation presents favorable conditions
for tree species growth, since they are materials containing a great amount of nutrients and the
availability occurs gradually for a longer period of time (ANTUNES et al., 2016). In addition, it
is also believed that the improvement of the chemical characteristics of the substrate due to the
addition of the humic material strongly influenced these results (Table 1).
Table 5 – Height and collar diameter of Bauhinia forficata seedlings at 45, 90 and 135 days
after seeding and percentage of seedlings survival at 45 days with different proportions of
vermicompost in the substrate
Tabela 5 – Altura e diâmetro do coleto de mudas de Bauhinia forficata aos 45, 90 e 135 dias após
semeadura e percentual de sobrevivência das mudas aos 45 dias com diferentes proporções de
vermicomposto no substrato
Days after seeding
Treatments*
45
90
135
SVL (%)
H (cm)
CD (mm)
H (cm)
CD (mm)
H (cm)
CD (mm)
T1
5.9 b**
1.8 a
8.7 b
2.3 b
11.1 c
2.5 b
80.0 a
T2
6.4 ab
1.8 a
10.7 a
2.6 ab
12.6 bc
2.6 ab
82.5 ab
T3
5.9 b
1.7 a
11.2 a
2.6 a
13.5 ab
2.7 ab
75.0 ab
T4
6.4 ab
1.8 a
11.3 a
2.5 ab
13.1 ab
2.6 ab
82.5 a
T5
6.4 ab
1.8 a
11.7 a
2.6 a
14.3 ab
2.7 a
75.0 ab
T6
7.2 a
1.8 a
12.2 a
2.8 a
15.0 a
2.9 a
70.0 b
CV (%)
20.4
12.9
20.8
19.8
19.7
17.5
6.4
Source: Authors (2019)
In which: H (height); CD (collar diameter); SVL (seeding and survival).
(*) T1 = Control; T2 = 20%; T3 = 40%; T4 = 50%; T5 = 60% e T6 = 80% of vermicompost used in the substrate
composition; CV = Coefficient of Variation.
(**) Values followed by the same letter in the column did not differ significantly by the Kruskal-Wallis test
(α=0.05).
Other authors obtained similar results as in this study when formulating a substrate for
seedling production of different tree species with different vermicompost proportions, suggesting
different nutritional requirements among species (DANNER et al., 2007; STEFFEN et al., 2011).
In the study present, the results were observed with the highest proportions of vermicomposto
(60 and 80%).
The differences were not very evident in the survival analysis. Only T6 differed from T1
and T4 (Table 5). This result suggests that the higher volume of vermicompost may have influenced
the physical properties of the substrate. According to Singh et al. (2013), the vermicompost
increases the total porosity of the substrate, providing higher micropore content responsible
for water retention capacity, and lower macropore content responsible for water infiltration,
drainage capacity and aeration capacity in the substrate. The addition of 80% of vermicompost in
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Peccatti, A.; Rovedder, A. P. M.; Steffen, G. P. K.; Maldaner, J.; Camargo, B.;
Dalcul, L. P.; Capitani, L. C.; Hummel, R. B.; Neeuenschwander, F.
376
the substrate possibly reduced the macropore content, making it difficult to oxygenate the roots
and to establish the seedlings (STEFFEN et al., 2011).
For the leaf number, there was no effect of adding different proportions of vermicompost
in the three evaluated periods (Table 6).
Table 6 – Leaf numbers and leaf chlorophyll content of Bauhinia forficata seedlings at
45, 90 and 135 days and leaf area obtained at 135 days, with different proportions of
vermicompost in the substrate
Tabela 6 – Número de folhas e teor de clorofila foliar de mudas de Bauhinia forficata cultivadas
em substratos compostos por diferentes proporções de vermicomposto bovino e solo aos 45, 90
e 135 dias e área foliar obtida aos 135 dias após semeadura
Days after seeding
45
Treatments*
90
135
LA (cm²)
LN
CC
LN
CC
LN
CC
T1
3 a**
31.3 a
5a
32.9 a
6a
33.3 a
90.51 b*
T2
3a
28.1 ab
6a
28.2 b
7a
36.1 a
116.48 ab
T3
3a
27.6 ab
6a
28.2 b
7a
36.5 a
118.79 ab
T4
3a
27.8 ab
6a
26.3 b
6a
34.5 a
120.69 a
T5
3a
28.0 ab
6a
27.7 b
8a
35.5 a
123.89 a
T6
3a
29.9 ab
6a
26.5 b
7a
32.8 a
129.59 a
CV (%)
15.4
15.4
19.4
17.0
21.3
17.6
18.9
Source: Authors (2019)
In which: LN (leaf numbers); CC (chlorophyll content); LA (leaf area).
(*) T1 = Control; T2 = 20%; T3 = 40%; T4 = 50%; T5 = 60% e T6 = 80% of vermicompost used in the substrate
composition; CV = Coefficient of Variation.
(**) Values followed by the same letter in the column did not differ significantly by the Kruskal-Wallis test
(α=0.05) for the variables LN and CC, and by the Tukey test (α=0.05) for the variable LA.
For leaf area, the proportions of 50, 60 and 80% vermicompost (T4, T5 and T6 respectively)
had a positive influence, presenting higher mean values than the control treatment (Table 6). The
increase in LA provides a larger area for the production and accumulation of photoassimilates
and therefore a direct relation with the increase in the growth variables (TAIZ et al., 2017).
In relation to the chlorophyll content, there was a statistical difference only at 90 DAS. In
this period, T1 differed from the other treatments presenting higher content of photosynthetic
pigments, but this result did not persist at 135 DAS, during which time T1 presented lower average
leaf area (Table 6). We expected to find a direct relationship between leaf area and chlorophyll
content. However, the inverse results demonstrate the performance of other physiological
mechanisms, such as degradation by the light effect (TAIZ et al., 2017).
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Table 7 – Total dry biomass, shoot dry biomass, root dry biomass, shoot dry biomass and
root dry biomass ratio and Dickson Quality Index of Bauhinia forficata seedlings obtained at
135 days, with different proportions of vermicompost on the substrate
Tabela 7 – Biomassa seca total, biomassa seca aérea, biomassa seca radicular, relação biomassa
seca aérea e biomassa seca radicular e Índice de Qualidade de Dickson de mudas de Bauhinia
forficata obtidas aos 135 dias, com diferentes proporções de vermicomposto no substrato
Treatments*
TDB (g)
SDB (g)
RDB
SDB/RDB
DQI
T1
1.10 b**
0.41 b
0.70 b
0.58 a
0.22 a
T2
1.47 ab
0.51 ab
0.95 ab
0.54 a
0.28 a
T3
1.46 ab
0.50 ab
0.96 ab
0.56 a
0.29 a
T4
1.76 a
0.60 a
1.16 a
0.52 a
0.34 a
T5
1.67 ab
0.60 a
1.07 ab
0.56 a
0.31 a
T6
1.73 a
0.63 a
1.09 ab
0.58 a
0.31 a
23.6
24.5
26.3
37.5
27.9
CV (%)
Source: Authors (2019)
In which: TDB (total dry biomass); SDB (shoot dry biomass); RDB (root dry biomass); SDB/RDB (shoot dry
biomass and root dry biomass ratio); DQI (Dickson Quality Index).
(*) T1 = Control; T2 = 20%; T3 = 40%; T4 = 50%; T5 = 60% e T6 = 80% of vermicompost used in the substrate
composition; CV = Coefficient of Variation.
(**) Values followed by the same letter in the column did not differ significantly by the Kruskal-Wallis test
(α=0.05).
Concerning to the morphological parameters of total dry biomass (TDB), only the T4
and T6 treatments differed significantly from the control. However, T4, T5 and T6 presented
higher allocation of shoot dry biomass (SDB) in relation to the control, while for the root dry
biomass (RDB) only T4 was superior significantly to the control (TABLE 7). Similar effects
of increase of RDB after addition of vermicompost was verified in Acacia mearnsii (Fabaceae)
seedlings (ANTUNES et al., 2016). The shoot and root dry biomass are considered important
parameters of seedling quality and are related to the rusticity and survival of the seedlings in
the field, respectively (GOMES; PAIVA, 2011). In this sense, the survival results (Table 4) and
RDB obtained for T4 were shown to be related even in nursery conditions, indicating that the
seedlings of this treatment present greater chances of establishment in the field.
There were no differences between the treatments regarding the values of SDB/RDB and
DQI (Table 7).
Despite this, it can be said that the DQI obtained were satisfactory for all treatments,
ranging from 0.28 to 0.31 at 135 DAS. It should be noted, however, that there is still no DQI
pattern for native forest species, which is acceptable given the immense genetic variability
among populations, to different seedling production techniques and at the age at which the
seedlings were evaluated. For Eugenia involucrata DC. (Myrtaceae), for example, the DQI varied
between 0.37 and 0.44 (SOUZA et al., 2015) while for Enterolobium contortisiliquum (Vell.) Morong
(Fabaceae) this index varied between 0.49 and 0.66 respectively (ABREU et al., 2015).
Ci. Fl., Santa Maria, v. 30, n. 2, p. 367-379, abr./jun. 2020
Peccatti, A.; Rovedder, A. P. M.; Steffen, G. P. K.; Maldaner, J.; Camargo, B.;
Dalcul, L. P.; Capitani, L. C.; Hummel, R. B.; Neeuenschwander, F.
378
Conclusion
For the conditions of this study, it was concluded that Trichoderma spp. and vermicompost
biological inputs promoted the initial growth of Bauhinia forficata, indicating wide applicability
in seedling production. The variables that best express this effect for both inputs were height
and collar diameter from 90 days, leaf area and dry biomass (shoot and root) at 135 days.
The Trichoderma spp. strains tested promoted the growth of the seedlings and T13
presented better results than T33 for most of the analyzed variables; a fact that may be related to
the genetic variability, the mechanism actions and the origin site of the isolates.
The proportions of vermicompost used have few differences between them. However,
there is a tendency among treatments with higher proportions (50, 60 e 80 %) to present better
results in the studied variables.
Considering the importance of the Bauhinia forficata species, we suggest that new studies
be carried out contemplating the use of technologies for the production of seedlings with
ecological inputs.
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