Novel use of trinexapac-ethyl to study weed seed germination
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Huarte, Héctor Roberto ; Ruiz Carmona, Antonio Jesús ; Zapiola, María Luz Novel use of trinexapac-ethyl to study weed seed germination Artículo publicado en Weed Technology, 2014 Este documento está disponible en la Biblioteca Digital de la Universidad Católica Argentina, repositorio institucional desarrollado por la Biblioteca Central “San Benito Abad”. Su objetivo es difundir y preservar la producción intelectual de la Institución. La Biblioteca posee la autorización del autor para su divulgación en línea. Cómo citar el documento: Huarte, H. R., A. J. Ruiz Carmona y M. L. Zapiola. 2014. Novel use of trinexapac-ethyl to study weed seed germination [en línea]. Weed Technology. 28. DOI: 10.1614/WT-D-13-00178.1 Disponible en: http://bibliotecadigital.uca.edu.ar/repositorio/investigacion/novel-use-trinexapac-ethyl.pdf [Fecha de consulta: ….]
Weed Technology 2014 28:000–000
Novel Use of Trinexapac-Ethyl to Study Weed Seed Germination
Héctor Roberto Huarte, Antonio Jesús Ruiz Carmona, and Marı́a Luz Zapiola*
Understanding seed biology and onset of germination requirements is a key point for designing
effective weed management programs. Gibberellins (GAs) are known to play a role in onset of
germination of several species. Onset of germination often requires an increase in de novo GA
content or an increase in sensitivity to GAs. Reduced germination of seeds incubated in solutions
containing compounds that inhibit GA synthesis provide evidence that GAs are required to trigger
germination. Trinexapac-ethyl (TE), a GA synthesis inhibitor, is frequently used as a plant growth
regulator in crop production. However, to the best of our knowledge, TE has not been used to study
the requirement of GAs in onset of germination. Germination studies were conducted using seeds of
artichoke thistle and common teasel under a range of TE concentrations (0 to 500 lM TE), a
combined solution of 125 lM TE þ 100 lM GA3 for artichoke, and 250 lM TE þ 100 lM GA3 for
common teasel. Germination tests were conducted at 20/10 C (12-h thermoperiod) in darkness for
artichoke and at constant 15 C with 12 h of light for common teasel. Germination of artichoke in
125 lM TE was reduced to 47% when compared with the check (95%), but the combined TE þ
GA3 solution maintained germination at 84%. Germination of common teasel in 250 lM TE was
reduced to 10% when compared with the check (91%), and the combined TE þ GA3 solution
increased germination to 63%. These results demonstrate the utility of TE to study the role of GAs in
onset of germination. This novel use of TE is a valuable option to study germination requirements of
weed species, and therefore contributes to the design of weed management programs.
Nomenclature: Gibberellins; trinexapac-ethyl; prohexadione-calcium; artichoke thistle, Cynara
cardunculus L. CYCA; common teasel, Dipsacus fullonum L. DIFU2.
Key words: Dormancy, germination, plant growth regulators, seed biology, weed biology, weed
management.
El entender la biologı́a de las semillas y los requisitos para la germinación son puntos clave para el diseño de programas de
manejo de malezas efectivos. Se sabe que gibberellins (GAs) juegan un papel en el inicio de la germinación en varias
especies. El inicio de la germinación a menudo requiere de un aumento en el contenido de GA de novo o un aumento en la
sensibilidad a GAs. Germinación reducida de semillas incubadas en soluciones que contienen compuestos que inhiben la
sı́ntesis de GA brindan evidencia de que GAs son requeridas para promover la germinación. Trinexapac-ethyl (TE) es un
inhibidor de la sı́ntesis de GA que es frecuentemente usado como regulador del crecimiento vegetal en la producción de
cultivos. Sin embargo, hasta donde conocemos, TE no ha sido usado para el estudio del requisito de GAs en el inicio de la
germinación. Se realizaron estudios de germinación usando semillas de Cynara cardunculus y Dipsacus fullonum con una
variedad de concentraciones de TE (0 a 500 lM TE), una solución combinada de 125 lM TE þ 100 lM GA3 para C.
cardunculus, y 250 lM TE þ 100 lM GA3 para D. fullonum. Las pruebas de germinación se realizaron a 20/10 C (perı́odo
termal de 12-h) en oscuridad para C. cardunculus y a 15 C constantes con 12 h de luz para D. fullonum. La germinación de
C. cardunculus en 125 lM TE se redujo a 47% al compararse con el testigo (95%), pero la solución combinada de TE þ
GA3 mantuvo la germinación a 84%. La germinación de D. fullonum en 250 lM TE se redujo a 10% en comparación con
el testigo (91%), y la solución combinada de TE þ GA3 aumento la germinación a 63%. Este uso novedoso de TE es una
opción valiosa para el estudio de los requisitos de germinación de especies de malezas, y por lo tanto contribuye al diseño
de programas de manejo de malezas.
Designing a weed management program requires However, knowledge about seed dormancy and
knowledge of seed biology and onset of germina- germination requirements is limited or nonexistent
tion requirements of the weed species involved. for many weed species. Every tool that contributes
to the study of weed seed germination is a useful
DOI: 10.1614/WT-D-13-00178.1 tool for weed management.
* Professor, Graduate Student, and Professor, Department
of Crop Sciences, School of Agricultural Sciences, Universidad Dormancy can be defined as the lack of capacity
Cat ólica Argentina (UCA). Corresponding author’s E- of a seed to germinate in a specified period of time
mail::robertohuarte@uca.edu.ar. under any combination of normal physical envi-
Huarte et al.: Huarte et al.: Novel use of trinexapac-ethyl 0
//titan/production/w/wete/live_jobs/wete-28-03/wete-28-03-17/layouts/wete-28-03-17.3d 27 August 2014 8:16 pm Allen Press, Inc. Customer MS# WT-D-13-00178.1 Page 1ronmental factors that are otherwise favorable for its to studies of weed germination requirements when
germination (Baskin and Baskin 2007). A dynamic PGRs normally used are not commercially avail-
balance between gibberellins (GAs) and abscisic acid able. Having the ability to use the ai formulated as
is key in seed dormancy regulation and the a commercial product increases the chances of
completion of germination (Kucera et al. 2005). obtaining the compound and reduces the cost of
Onset of germination is associated with an increase the experiment.
in GA content and greater sensitivity to GA (Finch- Artichoke thistle and common teasel are two
Savage and Leubner-Metzger 2006). Reduced weeds that have invaded thousands of hectares of
germination of seeds incubated in solutions of Argentinean right-of-way and grasslands (Marzoca,
compounds that reduce GA synthesis provides 1976). Since 1963, artichoke thistle was declared a
evidence of de novo GA requirements for onset of national pest. In contrast, common teasel turned
germination (Arana et al. 2006; Huarte and into a highly problematic weed species just in
Benech-Arnold 2010). recent years, and still no official records certified
GAs are plant hormones synthesized in higher this fact. Artichoke thistle is a herbaceous perennial
plants that are essential for many processes of plant plant native to the Mediterranean basin that
growth and development, including breaking seed produces several thousands of wind-dispersed seeds
dormancy and seed germination (Yamaguchi each year (Potts et al. 2008; White and Holt
2008). GAs occur in different types of plants with 2005). On the other hand, common teasel is a
different levels of activity. In general, GA synthesis herbaceous biennial plant native to Europe and
involves three stages: ent-kaurene synthesis from North Africa that disperses more than 3,000seeds
geranylgeranyl diphosphate, GA 12 and GA 53 each year (Werner 1975). Local populations of
synthesis from ent-kaurene, and finally, the both species are known to have seed dormancy
conversion of these compounds into GA4 and (Huarte and Benech-Arnold 2005; La Greca
GA1 (Yamaguchi 2008). GA4 and GA1 are the 2010). Artichoke thistle seeds require fluctuating
main active GAs in higher plants (Sawada et al. temperatures for germination (Huarte 2006). On
2008). the other hand, common teasel seeds require either
Plant growth regulators (PGRs) like trinexapac- fluctuating temperatures or light for germination
ethyl (TE) and prohexadione-calcium (prohexa- (Huarte and Zapiola 2011). To design a weed
dione-Ca) are effective in reducing active GA management program, the possibility of reduced
synthesis (Rademacher 2000). These PGRs inter- germination and emergence processes using non-
fere with later steps of GA synthesis (Yamaguchi et chemical methods is important. To do these,
al. 2007). Both TE and prohexadione-Ca are knowledge about weed seed germination behavior
frequently used as growth regulators in grass seed is required.
and cereal crop production, for growth control in The objective was to evaluate the potential use of
turf grasses, and for reduction of vegetative growth TE to confirm the requirement of de novo GA
in fruit trees (Qian and Engelke 1999; Rademacher synthesis in onset of germination of artichoke thistle
1995; Rajala and Peltonen-Sainio 2001; Zapiola et and common teasel, and therefore identify an
al. 2006). Incubation of seeds in prohexadione-Ca option for use in germination and seed dormancy
and other PGRs with different sites of action like studies in countries where traditionally used
paclobutrazol has been frequently used to study the compounds are not available or are too expensive.
involvement of de novo GAs in onset of We hypothesized that TE would reduce germina-
germination (Arana et al. 2006; Yang et al. 1995; tion in cases where onset of germination depends on
Zehhar et al. 2002). Nevertheless, to the best of de novo GA synthesis.
our knowledge, TE has not been used to study
involvement of GAs in onset of germination.
Various PGRs are commercially available in several Materials and Methods
countries. In some countries, TE, but not Seeds. Heads of artichoke thistle were collected at
prohexadione-Ca, can be found as ai in commer- an infested roadside in Saldungaray (38811 0 S,
cial growth regulators. Therefore, the potential use 61846 0 W), province of Buenos Aires, Argentina, in
of TE in this type of study is a valuable alternative January 2009 and at an infested research plot at the
0 Weed Technology 28, July–September 2014
//titan/production/w/wete/live_jobs/wete-28-03/wete-28-03-17/layouts/wete-28-03-17.3d 27 August 2014 8:16 pm Allen Press, Inc. Customer MS# WT-D-13-00178.1 Page 2School of Agricultural Sciences, Universidad
Católica Argentina (UCA) (34835 0 S, 58827 0 W),
city of Buenos Aires, in January 2013. Common
teasel seeds were collected at an infested research
plot at the School of Agricultural Sciences, UCA in
January 2009, 2011, and 2013. Seeds were hand
threshed and kept in paper bags at 18C until use
for germination experiments.
Germination Experiments. The effect of TE on
seed germination was evaluated. TE treatment
levels evaluated for artichoke thistle were 0, 10, 25,
50, 100, 125, 250, and 500 lM TE using
Moddust 250 EC (Syngenta, Switzerland). An
additional treatment consisted of 125 lM TE þ
100 lM GA3 (Sigma Chemical Company, St.
Louis, MO). Three replicates of 25 seeds each per Figure 1. Germination of artichoke thistle seeds at various
treatment level were placed on blotting paper in 9- trinexepac-ethyl (TE) concentrations and a mixed solution of
cm petri dishes with 6 ml of each corresponding 125 lM TE þ 100 lM gibberellin3 (GA3). Vertical bars indicate
solution. Petri dishes were placed in germination the standard error. Final germination count was at 14 d.
Different letters at the top of each bar indicate significant
chambers at fluctuating temperatures (10/20 C, 12 differences according Tukey’s test (a ¼ 0.05).
h) in darkness. Germination was scored daily for
14 d. 250, and 500 lM TE treatments reduced germina-
TE treatment levels evaluated for common teasel tion of artichoke to 47, 33, and 25%, respectively,
were 0, 62.5, 125, 250, and 500 lM TE. An when compared with the control (95%). Incubation
additional treatment consisted of 250 lM TE þ 500
in a solution of 125 lM TE þ 100 lM GA3
lM GA3. Three replicates of 30 seeds each per
maintained germination at a level comparable with
treatment level were placed on blotting paper in 9-
cm petri dishes with 8 ml of each corresponding that of the control (84%).
solution. Petri dishes were placed in germination Common teasel germination was also affected by
chambers at constant temperature (15 C) and 12-h TE (Figure 2) (P , 0.05). The 125, 250, and 500
light. Germination was scored daily for 24 d. lM TE reduced germination to 44, 10, and 1%,
For both species, seeds with visible radicle respectively, when compared with the control
protrusion were considered germinated and were (91%). Incubation in a solution of 250 lM TE þ
removed. All germination experiments were repeat- 500 lM GA3 only increased germination to 63%,
ed. The data presented were averaged across two 31% below that of the control.
and three experimental runs for artichoke and The fact that incubation of seeds in TE reduced
common teasel, respectively, because there was no germination in both species confirms their require-
significant experiment-by-treatment interaction (P ment of de novo GA synthesis for onset of
¼ 0.86 and 0.67) for artichoke and common teasel, germination. However, greater TE concentrations
respectively. Germination was expressed as percent- were needed to reduce germination in common
age of total seeds germinated by the end of the teasel than in artichoke thistle, suggesting that the
experiment per petri dish. Data were analyzed by effective TE concentration to prevent germination is
ANOVA using Statistix 8.0 Analytical Software species specific. The effect of TE was suppressed by
(Tallahassee, FL). Tukey’s test at 5% level of coincubation with GA3 in both species, demon-
probability was used for mean comparisons. strating that the reduction in germination found
with TE was a result of de novo GA synthesis
inhibition and not of other causes. These results
Results and Discussion provide evidence of the efficacy of using TE to study
There was an effect of TE on germination of the requirement of GAs in onset of germination.
artichoke thistle (Figure 1) (P , 0.05). The 125, Consequently, TE is a valuable option to study
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Huarte et al.: Huarte et al.: Novel use of trinexapac-ethyl 0
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