Oliveira, V. A. et al. 98
Vol. 4, N.2: pp. 98-106, May, 2013 ISSN: 2179-4804
Journal of Biotechnology and Biodiversity
Use of seed treatment with fungicide in control of Colletotrichum truncatum and physiological quality of soybean seeds Glycine max
Vinícius Almeida Oliveira¹,*, Lorenzo Paradiso Martins², Rogério Cavalcante Gonçalves², Luíz Paulo Figueredo Benício³, Daniella Lima da Costa4, Juliane Ludwig 5
ABSTRACT
The fungus are the main microorganisms present in seeds, is the main cause of deterioration and loss in production. The anthracnose caused by Colletotrichum truncatum associated with soybean seeds as has main vehicle for introduction into the planting areas can be detected in all stages of crop development, from the cotyledons to the end of the cycle, being present in the stems, veins, leaflets and pods. Thus aimed to evaluate the influence of using different products fungicides as seed treatment, where the seeds were inoculated with the pathogenic fungus and treated with the chemicals they take Carbedazim + Fludioxonil + metalaxyl-M and carboxin + thiram. For each fungicide product was two treatments done using the doses recommended by the manufacturer and 75% of dose. We evaluated health, germination and promote plant (Plant growth, fresh weight and dry weight of root and shoot). This work concludes that the use of fungicide controls significantly seeds infected with Colletotrichum truncatum and presents a significant improvement as the development of structures seedling. Key-words: Germination, plant growth.
Uso de fungicida via tratamento de sementes no controle de Colletotrichum truncatum e na qualidade fisiológica de sementes de soja Glycine Max
RESUMO
Os fungos são os principais microrganismos presentes nas sementes, constituindo a principal causa da deterioração e perdas na produção. A antracnose causada pelo Colletotrichum truncatum associada à soja, tem as sementes como principal veículo de introdução em áreas de plantio podendo ser detectada em todas as fases de desenvolvimento da cultura, desde os cotilédones até o final do ciclo, estando presente nas hastes, nervuras, folíolos e vagens. Assim,
objetivou-se avaliar a influência do uso de diferentes produtos fungicidas via tratamento de sementes, onde as sementes foram inoculadas com o fungo patogênico e tratadas com os produtos Carbedazim + Tiram, Fludioxonil + Metalaxyl-M e Carboxina + Tiram. Para cada produto fungicida foi feito dois tratamentos, utilizando a dose indicada pelo fabricante e 75% da dose. Foram realizadas avaliações de sanidade, germinação, vigor e de crescimento de plantas. O presente trabalho conclui que o uso do fungicida controla significativamente sementes infectadas com Colletotrichum truncatum e apresenta melhora significativa quanto ao desenvolvimento das estruturas das plântulas.
Palavras-chaves: Geminação, crescimento de planta.
*Autor para correspondência.
¹,*Doutorando em Agronomia; Universidade Federal de Goiás; CP 13; 74001-970 Goiânia - GO – Brasil; mutamba8@hotmail.com
²Eng° Agrônomo Mestre em Agroenergia pela Universidade Federal do Tocantins.
³Doutorando em Solos e Nutrição de plantas Pela Universidade Federal de Viçosa; 36570-000; Viçosa – MG – Brasil.
4Instituto Federal de Educação, Ciência e Tecnologia do Tocantins - IFTO;77500-000; Porto Nacional – TO – Brasil.
5Departamento de Fitopatologia da UNIDERP; 79003-010; Campo Grande – MS – Brasil.
J. Biotec. Biodivers. v. 4, N.2: pp. 98-106, May. 2013
https://doi.org/10.20873/jbb.uft.cemaf.v4n2.oliveira
Oliveira, V. A. et al. 99
INTRODUCTION
It is estimated that the average rate of growth of
agricultural area in Brazil is 4.7% per annum over the next decade, one of the world's largest (IICA, 2007).
Dades of the Conab (2010) link the Brazil as one of the largest producers of the world, and the
production in the country, in last harvest, was approximately 67,86 million of tones harvested in area of 23,24 million of hectares.
But the economic exploitation of the potential yield (more of the 4000 kg/ha) is hardly achieved, due, among other factors, the onset of disease.
Barros (2008) reports that in the last years some disease of soybean have emerged as prominent as the incidence and the damage they have caused, among them anthracnose ( Colletotrichum dematium var. truncate). This disease affects all stages of crop development, may cause death of seedlings, necrotic spots on the petioles and
leaves, stems and pods (Galli et al., 2007), being
present in all the regions of soybean cultiv ation, and one major problems in soybean areas in the Cerrados.
The seeds are the main vehicle of transmission of
this pathogen, one time that fungus can affect the germination and seedling vigor, resulting in lower
emergence and the productivity (Patrício, et al., 1991; Sinclair, 1991). Is through of the seeds that
microorganism is introduced in new area, survive over the years and spread the plant population as
outbreaks of diseases.
The treatment of the seeds is mainly used with the order to allow germination of the infected seeds,
controlling pathogens transmitted by seeds and protect them of soil fungi (Henning, et al., 1994).
In addition to give protection the seeds, the chemical treatment this provides additional
assurance to the establishment of the crop at
reduced costs, less than 0,5% of cost in installation of the crop (Henning, et al., 2005).
Some studies have been performed with the
evaluate the efficiency of the treatment of seeds with fungicide in control Colletotrichum
truncatum, and being reported significant reductions of pathogen in seeds (Goulart, 2001), and this benefit is reflected increased plant productivity (Goulart, 1998). However, to make such assessments is imperative that there of pathogens in seeds, this fact being provided to
accomplish the artificial inoculation of the seeds with the pathogen focus of the study. The artificial inoculations of the soybean seeds with
Colletotrichum truncatum can be useful in
verifying the location of the pathogen seeds in determinations of its effect on the quality
physiological these and in preparation of samples with know effects of the pathogen (Galli et al.,
2005).
The analysis of growth is based, fundamentally, in fact to about 90%, in average, of dry matter
accumulation in plants throughout is development results of the photosynthetic; allowing to evaluate
the growth finish of plant with whole and the contribution of different organs in totally
development. Despite the complexity involved in
the growth of plants species, the analysis of growth is means a very accurate to avalue the development and measure the contribution of
different physiological processes about the plant behavior (Benincasa, 2003) and various works that
associate the reduction of accumulation of mass in the reduction of accumulation of mass in the presence of pathogen in seeds (Salustiano et al., 2005; Poletto et al., 2006).
This work has the objective to evaluate the effects
about the germination, vigor and growth of plants of the soybean derived of seeds infected artificiall y
with the fungus Colletotrichum truncatum and
treated with different fungicides rates via treatment of seeds.
MATERIALS AND METHODS
The work was conduced during the period of
March to June of 2010, in unit lang of university Anhaguera-Uniderp, the city of Campo Grande –
MS. The work was two steps, the first in laboratory and the second in vegetation`s house.
For the treatment of the seeds, these were weighed
and packed in transparent plastic bags and were soon added fungicides. After the addition of fungicides, the bags were manually agitated for
homogenization of the product and adherence to seeds. The amount of fungicide applied was
calculated from the recommendation for 100 kg of seeds, where it was made of three simple rule for determining the amount of fungicide to be used. It used seeds of soybean of the cultivar CD219RR. The fungicides used are described in the Table 1.
J. Biotec. Biodivers. v. 4, N.2: pp. 98-106, May. 201 3
Oliveira, V. A. et al. 100
Table 1. Products and doses used in treatment of the seeds soybean.
Fungicides
Dose for 100 Kg of Seeds i.a.* (g) p.c.** (ml)
A Carbendazin + Thiram 30 + 70 200
B Fludioxonil + Metalaxyl-M 35 + 10 100
C Carboxin + Thiram 50 + 50 250
*i.a. – ingredient active; **pc – produce commercial.
Isolated from a fungus Colletotrichum truncatum , there was a subculture to Petri dishes co ntaining BDA and placed in a chamber type BOD with 25°
for 15 days to formation of a fungal colony for subsequent seeds inoculation.
For seeds inoculation, this were dipped in a
solution of sodium hypochlorite 1% for three minutes for a disinfection prior. After the
disinfection the seeds was dried with filter paper,
then were mixed with fungi from the colony established from the isolated of the Colletotrichum truncatum and maintained in contact with the
fungus for 48 hours. After the inoculation the seed was separate for treatment and packed in bag of
paper.
The seeds treated and uninoculated was subjected
to a test of health, that proceeded as follows: The seeds was previously disinfested in solution of sodium hypochlorite the 1%, then distributed in
box GERBOX with two sheets of filter paper moistened. After this, the seeds was incubated the
a temperature of 22°C, under the scheme of light, with photoperiod corresponding the 12 hours of light and 12 hours in dark. After seven days of incubated, was made the evaluation of the microorganisms present in seeds, with the astereospic microscope and specialized
bibliography (Barnett & Hunter, 1999). After the identification was calculated the percentage of seeds contaminated by each gender in each treatment fungal. For the test of health were used four repetitions of 100 seeds for each treatment. The treatment were T1 – seeds untreated; T2 – Carbendazin + Thiram; T3 – Fludioxonil +
Metalaxyl – Mand T4 – Carboxin +Thiram.
At the analysis of laboratory was measured the germination and the index of emergency.
For the test of germination were used eight treatments with four repetitions of 50 seeds and the treatments: T0 – (Seeds untreated); T1 –
(Seeds inoculated untreated); T2 – (Fungicide A 75% of dose recommended); T3 – (Fungicide A
100% of dose recommended); T4 – (Fungicide B 75% of dose recommended); T5 – (Fungicide B 100% of dose recommended); T6 – (Fungicide C
75% of dose recommended) and T7 – (Fungicide
C 100% of dose recommended). The seeds were placed for germinate in germination paper
“germitest” previously moistened in water on volume corresponding the 2,5 times the weight of
dry paper. After the seeds were taken the germinator with temperature constant of 25°C.
The number of seeds germinated were evaluate daily for seven days, and considered germinated
the seeds that obtained radical protusion greater than 2mm. End of seventh day was accomplished
the calculation of percentage of seeds germinateds. For the index of speeds of emergence, used the methodology of coefficient speeds described for
Maguire (1962).
In test conducted vegetation`s home were evaluate
the following variables: index of plant growth, root length, shoot length and mass dry of shoot
and root.
The test was conducted in pots filled with a oxisol red dystrophic (Embrapa, 1999). The soil of vessels was corrected with limestone dolomitic, as the recommendations of analysis chemical of soil. After the correction of soil, each vessel received
20 mg.dm³ of N; 200mg.dm³ of P and 100mg.dm³ of K and 10% of volume of land manure of corral. In each vessel were planted three seeds, ten days after the plantations was accomplished the thinning leaving only plant for vessel. The experiment was conducted to the 25° day after the plantation.
The index of growth of the plants was evaluate
after the plants have reached 50% of germination. The plants were measures with help of a mare the stem of the plant to the apical meristem. The
plants were measure to the 25 days after the plantation (DAP).
For the length of the root, length of shoot, mass dry of soot and mass dry of root. To 25 DAP the
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Oliveira, V. A. et al. 101
plants were separate in root and shoot, by of a court accomplished with the scissors in collar height. After, with a ruler, was measurement made
of booths the parts for be determinate the length of shoot and the root.
For the variable fresh mass and mass dry of root
and of shoot, each one of the parts was placed in a bag’s paper, separately, and the same were taken
the greenhouse with air circulation, where
remained for 96 hours with temperature of 55°C. after, were weighed on an analytical balance for
determination of weigh in grams.
With exception of test of health, for the other tests,
the type of statistical desing used was fully randomized with eight treatment and four
repetitions. The data were submitted the analysis
of variance and the average submitted to test of Tukey the 5% of probability used the program SASM – Agri (Canteri et. al., 2001)
RESULTS AND DISCUSSION
In test of health of seeds there is that, even seeds
be of good origin and certifiel, still so, observed the presence of fungi pathogenic associated the same, and that may come to cause damage to
production. From the analysis performed, observed
the incidence of fungi Fusarium sp., Penicilium sp., Aspergillus sp., Phomopsis sp.,
Macrophomina sp. Mucor sp., Cladosporium sp., and bacteria in seeds untreated (Figure 1).
35
30
25
20
Fusarium Penicillium Aspergillus Phomopsis Macrophomina Mucor Cladosporium Bacteriose
15
10
5
0
T1 T2 T3 T4
Treatment
Figure 1 - Incidence of pathogens in soybean seeds treated with differents fungicides.
Among the fungicides evaluated, that showed
higher efficiency was the Carbendazin + Thiram (T2), reaching one control of 100%.
On the other hand, the treatment used Fludioxonil
+ Metalaxyl – M(T3) presented high performance, however was not effective in control of Phomopsis sp., one time that fungi is reported as one of more
important in soybean seeds (Henning & França
Neto, 1980). Already the treatment using the Carboxin + Thiran (T4), showed the less efficient,
one time that presented only control of Fusarium, Penicillium and Aspergillus (Figure 1).
On that refers to germination of seeds, observed
that the pathogen, decreased the germination as well index of velocity of germination seeds, one
time that seeds uninoculated for pathogen showed
germination significantly superior to inoculated untreated (Table 2). These results reinforce the obtained for Goulart (1997), that reports to be this
fungi able of develop quickly at the terms in that
test of germination is accomplished can affect the germination os seeds.
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Oliveira, V. A. et al. 102
Table 2. Germination in percentage and índex of velocity germination (IVG) of seeds inoculated with Colletotrichum truncatum and treated with 75 and 100% of dose different fungicide.
Treatment Germination IVG
T0 Simple seeds without treatment 97 a 21,17 b
T1 Seeds inoculated without treatment 57 b 13,75 c
T2 Carbendazin + Thiram (75% of dose) 96 a 24,00 ab
T3 Carbendazin + Thiram (100% of dose) 99 a 26,25 a
T4 Fludioxonil + Metalaxyl-M(75% of dose) 97 a 23,25 ab
T5 Fludioxonil + Metalaxyl-M(100% of dose) 96 a 24,25 ab
T6 Carboxin + Thiram (75% of dose) 95 a 23,22 ab
T7 Carboxin + Thiram (100% of dose) 93 a 21,75 b
C.V. 4,00% 5,95%
*Means followed of same letter in column not differ each other for testo f Tukey the 5% of probability.
Even with respect the germination and the index of
velocity germination, was possible observe also that all treatment increased those variables in
relative to witness inoculated without any treatment (Table 2), corroborating with the data
obtained for Yuyama & Henning (1997) where the
authors claim that treatment of seeds is used mainly with the intended to improve the germination of seeds infected. Important to
mention that, the treatment using 100% of dose
Carbendazin +Thiran (T3) possible increases significant in index of velocity germination
reaching the 24% in relative to averages of statistics analysis and tha witness that was not inoculated with Colletotrichum truncatum ,
demonstrating that product improves the
performance physiological of seeds (Pereira et. al., 2009) .
On that refers to índex of growth plants, the values mean obteined portray the efficiency of all
fungicide in control of main damage associated
with the incidence of Colletotrichum truncatum in seeds, that is decrease of emergency seeds (Table 3). This sense, works already demonstrated that
use of fungicide applied in seed improved the
quality physiological of seeds soybean (Lopes & Barros, 1997; Yuyama & Henning, 1997), corn
(Von Pinho et al., 1997), cotton (Goulart, 2002) and rice (Teixeira et al., 1997) .
Table 3. Index of velocity growth of plants (IVCP) originated in seeds inoculated with Colletotrichum truncatum and treated with 75 and 100% of dose of different fungicides.
Treatment IVCP
T0 Simple seeds without treatment 4,74 ab
T1 Seeds inoculated without treatment 1,29 b
T2 Carbendazin + Thiram (75% of dose) 5,40 a
T3 Carbendazin + Thiram (100% of dose) 5,96 a
T4 Fludioxonil + Metalaxyl-M(75% of dose) 6,54 a
T5 Fludioxonil + Metalaxyl-M(100% of dose) 6,07 a
T6 Carboxin + Thiram (75% of dose) 6,16 a
T7 Carboxin + Thiram (100% of dose) 6,42 a
C.V. 29,67%
*Means followed of same letter in column not differ each other for test of Tukey the 5% of probability.
To evaluate the final height of plants and the root length, again is observed the effect negative of
pathogen associate to seeds, and that seeds treated
showed more of 100% increment in both variables (Table 4). This event may associated to bigger protection of plants to some stress occurred among
the seeding and evaluation of test, on time that
condition may be aggravated case the treatment of seeds with fungicides not accomplished because to
fact of seeds infected/contaminated with pathogens
are more subject losses (Machado, 2000), leading reduction of growth plants, as observed in this work.
J. Biotec. Biodivers. v. 4, N.2: pp. 98-106, May. 201 3
Oliveira, V. A. et al. 103
Table 4. Length of shoot and the roots plants originated of seeds inoculated with Colletotrichum truncatum and treated with 75 and 100% of dose of different fungicides.
Treatment Shoot Length (cm) Root
T0 Simple seeds without treatment 7,25 ab 19,30 a
T1 Seeds inoculated without treatment 2,25 b 11,00 b
T2 Carbendazin + Thiram (75% of dose) 9,00 a 20,25 a
T3 Carbendazin + Thiram (100% of dose) 8,25 a 22,00 a
T4 Fludioxonil + Metalaxyl-M(75% of dose) 9,00 a 19,62 a
T5 Fludioxonil + Metalaxyl-M(100% of dose) 8,62 a 21,75 a
T6 Carboxin + Thiram (75% of dose) 9,37 a 20,25 a
T7 Carboxin + Thiram (100% of dose) 9,37 a 19,00 a’
C.V. 31,22% 27,32 %
*Means followed of same letter in column not differ each other for test of Tukey the 5% of probability.
On that refers the fresh mass of roots, the
treatment that said in relation to witness that not received treatment, was utilized 100% of dose of fungicide, however, this not varied other
treatment, except of treatment using 100% of dose Carboxin + Thiram (T7). To evaluate the fresh
mass of shoot, any of fungicides showed
significantly different each other, however, the treatment using 100% of dose Carboxin + Thiram showed same to treatment whose seeds were
inoculated, but untreated (Table 5).
Table 5. Fresh mass of shoot and plants roots originated of seeds inoculated with Colletotrichum truncatum and treated with 75 and 100% of dose of different fungicides.
Treatment
Shoot
Freshmass (g) Root
T0 Simple seeds without treatment 1,96 a 1,22 ab
T1 Seeds inoculated without treatment 0,29 b 0,08 c
T2 Carbendazin + Thiram (75% of dose) 2,66 a 1,26 ab
T3 Carbendazin + Thiram (100% of dose) 2,74 a 2,23 a
T4 Fludioxonil + Metalaxyl-M(75% of dose) 1,91 a 1,21 ab
T5 Fludioxonil + Metalaxyl-M(100% of dose) 2,57 a 1,64 ab
T6 Carboxin + Thiram (75% of dose) 2,25 a 1,63 ab
T7 Carboxin + Thiram (100% of dose) 1,70 ab 0,97 bc
C.V. 32,71% 37,30%
*Means followed of same letter in column not differ each other for testo f Tukey the 5% of probability.
About to accumulation of dry mass for root, observed the superiority provided for treatment using 100% of dose of Carbendazin + Thiram (T3), showing statistically same to treatment using 100% of fungicide Fludioxonil + Metalasyl- M (T5). Already, concerning the accumulation of dry
mass of shoot, any of treatment differed significantly each other, however, the treatment in
which was used 100% of dose of Carboxin + Thiram (T7) showed same to treatment whose seeds were inoculated, but untreated (Table 6). Such results may be credited to fact of fungicide have provided the control of fungi that deteriorate the seedling, as observed for Oliveira et al. (1993)
in corn and Teixeira (1997) in rice.
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Oliveira, V. A. et al. 104
Table 6. Dry mass of shoot and plants roots originated of seed inoculated with Colletotrichum t runcatum and treated with 75 and 100% of dose of different fungicides.
Treatment Shoot Drymass (g) Root
T0 Simple seeds without treatment 0,25 a 0,09 bc
T1 Seeds inoculated without treatment 0,04 b 0,01 c
T2 Carbendazin + Thiram (75% of dose) 0,35 a 0,10 b
T3 Carbendazin + Thiram (100% of dose) 0,36 a 0,21 a
T4 Fludioxonil + Metalaxyl-M(75% of dose) 0,26 a 0,11 b
T5 Fludioxonil + Metalaxyl-M(100% of dose) 0,34 a 0,12 ab
T6 Carboxin + Thiram (75% of dose) 0,29 a 0,10b
T7 Carboxin + Thiram (100% of dose) 0,24 ab 0,07 bc
C.V. 31,52 38,47
*Means followed of same letter in column not differ each other for test of Tukey the 5% of probability.
Leading in account the variables that determine the
potential germinal and physiological of seeds, observed that both agree with Nunes (2006), and the author observed there is effect of fungicide
about the germination of seeds, beyond the production of plants more stretching of root, at the
same time in which found more growth of shoot, so, fungicide could act as bioactivators and induce the production of crop regulators increasing, the activity enzymatic and allowing the expression of roots and plant.
SASM - Agri: Sistema para análise e separação de médias em experimentos agrícolas pelos métodos Scott - Knott, Tukey e Duncan. Revista Brasileira de Agrocomputação, v. 1, n. 2, p. 18 - 24, 2001.
CONAB- Companhia Nacional de Abastecimento. Acompanhamento de safra brasileira: grãos, quarto levantamento, janeiro 2010 / Companhia Nacional de Abastecimento – Brasília, 2010.
EMBRAPA - Empresa Brasileira De Pesquisa
CONCLUSIONS
Among the fungicides evaluated, Cabendazin +
Agropecuária. Sistema Brasileiro Classificação de Solos. P. 412, 1999.
de
Thiram was more efficient showing better performance on control of pathogen.
The treatment of seeds soybean with fungicides
generally, provided significant increase on potential germination and physiological of seeds improving the germination of seeds, increasing the
plant growth and its vigor.
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Recebido: 08/10/2012 Received: 11/08/2012
Aprovado: 02/02/2013 Approved: 02/02/2013
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