Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico
ilustraciones, diagramas, figuras
- Autores:
-
Ortiz Álvarez, Antonio
- Tipo de recurso:
- Fecha de publicación:
- 2023
- Institución:
- Universidad Nacional de Colombia
- Repositorio:
- Universidad Nacional de Colombia
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unal.edu.co:unal/85483
- Palabra clave:
- 580 - Plantas
570 - Biología
630 - Agricultura y tecnologías relacionadas
630 - Agricultura y tecnologías relacionadas::633 - Cultivos de campo y de plantación
Relaciones agua-planta
Plantas -- Efectos del cadmio
Cadmio
Plant-water relationships
Plants, Effect of cadmium on
Cadmium
Déficit hídrico
Theobroma cacao
Análisis del suelo
Estrés de sequia
Water shortages
Soil analysis
Drought stress
Relación planta-suelo
Plant-soil relationships
Cacao
Déficit hídrico
Estrés hídrico
Genotipo
Tolerancia
Acumulación de Cd
Translocación de Cd
Cocoa
Drought stress
Genotype
Tolerance
Cd accumulation
Cd translocation
- Rights
- openAccess
- License
- Atribución-NoComercial 4.0 Internacional
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UNACIONAL2 |
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Universidad Nacional de Colombia |
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|
| dc.title.spa.fl_str_mv |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| dc.title.translated.eng.fl_str_mv |
Evaluation of physiological response and cadmium accumulation in cocoa plants (Theobroma cacao L.) under water deficit conditions |
| title |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| spellingShingle |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico 580 - Plantas 570 - Biología 630 - Agricultura y tecnologías relacionadas 630 - Agricultura y tecnologías relacionadas::633 - Cultivos de campo y de plantación Relaciones agua-planta Plantas -- Efectos del cadmio Cadmio Plant-water relationships Plants, Effect of cadmium on Cadmium Déficit hídrico Theobroma cacao Análisis del suelo Estrés de sequia Water shortages Soil analysis Drought stress Relación planta-suelo Plant-soil relationships Cacao Déficit hídrico Estrés hídrico Genotipo Tolerancia Acumulación de Cd Translocación de Cd Cocoa Drought stress Genotype Tolerance Cd accumulation Cd translocation |
| title_short |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| title_full |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| title_fullStr |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| title_full_unstemmed |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| title_sort |
Respuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídrico |
| dc.creator.fl_str_mv |
Ortiz Álvarez, Antonio |
| dc.contributor.advisor.none.fl_str_mv |
Castaño Marín, Ángela María Magnitskiy, Stanislav |
| dc.contributor.author.none.fl_str_mv |
Ortiz Álvarez, Antonio |
| dc.contributor.orcid.spa.fl_str_mv |
Ortiz Álvarez, Antonio [0000000223580277] |
| dc.subject.ddc.spa.fl_str_mv |
580 - Plantas 570 - Biología 630 - Agricultura y tecnologías relacionadas 630 - Agricultura y tecnologías relacionadas::633 - Cultivos de campo y de plantación |
| topic |
580 - Plantas 570 - Biología 630 - Agricultura y tecnologías relacionadas 630 - Agricultura y tecnologías relacionadas::633 - Cultivos de campo y de plantación Relaciones agua-planta Plantas -- Efectos del cadmio Cadmio Plant-water relationships Plants, Effect of cadmium on Cadmium Déficit hídrico Theobroma cacao Análisis del suelo Estrés de sequia Water shortages Soil analysis Drought stress Relación planta-suelo Plant-soil relationships Cacao Déficit hídrico Estrés hídrico Genotipo Tolerancia Acumulación de Cd Translocación de Cd Cocoa Drought stress Genotype Tolerance Cd accumulation Cd translocation |
| dc.subject.lcc.spa.fl_str_mv |
Relaciones agua-planta Plantas -- Efectos del cadmio Cadmio |
| dc.subject.lcc.eng.fl_str_mv |
Plant-water relationships Plants, Effect of cadmium on Cadmium |
| dc.subject.agrovoc.spa.fl_str_mv |
Déficit hídrico Theobroma cacao Análisis del suelo Estrés de sequia |
| dc.subject.agrovoc.eng.fl_str_mv |
Water shortages Soil analysis Drought stress |
| dc.subject.lemb.spa.fl_str_mv |
Relación planta-suelo |
| dc.subject.lemb.eng.fl_str_mv |
Plant-soil relationships |
| dc.subject.proposal.spa.fl_str_mv |
Cacao Déficit hídrico Estrés hídrico Genotipo Tolerancia Acumulación de Cd Translocación de Cd |
| dc.subject.proposal.eng.fl_str_mv |
Cocoa Drought stress Genotype Tolerance Cd accumulation Cd translocation |
| description |
ilustraciones, diagramas, figuras |
| publishDate |
2023 |
| dc.date.issued.none.fl_str_mv |
2023 |
| dc.date.accessioned.none.fl_str_mv |
2024-01-29T18:11:48Z |
| dc.date.available.none.fl_str_mv |
2024-01-29T18:11:48Z |
| dc.type.spa.fl_str_mv |
Trabajo de grado - Maestría |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/masterThesis |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/acceptedVersion |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/TM |
| status_str |
acceptedVersion |
| dc.identifier.uri.none.fl_str_mv |
https://repositorio.unal.edu.co/handle/unal/85483 |
| dc.identifier.instname.spa.fl_str_mv |
Universidad Nacional de Colombia |
| dc.identifier.reponame.spa.fl_str_mv |
Repositorio Institucional Universidad Nacional de Colombia |
| dc.identifier.repourl.spa.fl_str_mv |
https://repositorio.unal.edu.co/ |
| url |
https://repositorio.unal.edu.co/handle/unal/85483 https://repositorio.unal.edu.co/ |
| identifier_str_mv |
Universidad Nacional de Colombia Repositorio Institucional Universidad Nacional de Colombia |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.relation.references.spa.fl_str_mv |
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U., Farooq, N., Naveed, M., Kucerik, J., Brtnicky, M., & Mustafa, A. (2022). Cadmium Phytotoxicity, Tolerance, and Advanced Remediation Approaches in Agricultural Soils; A Comprehensive Review. In Frontiers in Plant Science (Vol. 13). Frontiers Media S.A. https://doi.org/10.3389/fpls.2022.773815 |
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Atribución-NoComercial 4.0 Internacionalhttp://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Castaño Marín, Ángela María9b22edbde935ebc76d2ffb7d1bd19465Magnitskiy, Stanislav6c80f64a607d52111546f87810302ba2Ortiz Álvarez, Antonio33d2459ac9d78a9be23d0a51dccf40f1Ortiz Álvarez, Antonio [0000000223580277]2024-01-29T18:11:48Z2024-01-29T18:11:48Z2023https://repositorio.unal.edu.co/handle/unal/85483Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.co/ilustraciones, diagramas, figurasEl riesgo de sequía debido a la variabilidad climática y la presencia de cadmio (Cd) en el suelo es común en zonas cacaoteras alrededor del mundo. Hasta el momento no se encuentran estudios publicados sobre la acumulación de Cd en condiciones de déficit hídrico. Esta investigación tuvo como objetivo evaluar la respuesta fisiológica y la acumulación de cadmio en plantas de cacao bajo déficit hídrico. El estudio se realizó en casa de mallas en el Centro de Investigación Nataima de AGROSAVIA, municipio El Espinal, Tolima. Se utilizaron materiales con potencial como portainjertos de cacao, dos progenies (A2 y 1233) de hermanos completos obtenidas mediante cruzamiento dirigido, y un genotipo ampliamente utilizado como patrón en Colombia (IMC 67). Se analizaron diversos parámetros, incluyendo aspectos hídricos, de crecimiento, fisiológicos, bioquímicos y la acumulación de Cd en plantas de 7 meses de edad. Estas plantas crecieron en un suelo contaminado con Cd, sin añadir fuentes externas del metal, con una concentración inicial de Cd soluble de 0,356 mg kg-1. Se sometieron a déficit hídrico mediante la suspensión de riego durante períodos consecutivos de 19 y 27 días (D19 y D27), y luego se rehidrataron para evaluar su potencial de recuperación al estrés hídrico. El estrés hídrico redujo el potencial hídrico de las hojas (Ψhoja) con valores entre -1,51 y -2,09 MPa, siendo la progenie 1233 la más tolerante. Sin embargo, la recuperación de variables de intercambio gaseoso, el potencial hídrico y los pigmentos fotosintéticos tras la rehidratación, sugiere que las tres progenies poseen la capacidad de tolerar los niveles de estrés evaluados. La acumulación de Cd varió entre progenies, no entre niveles de estrés. La concentración de Cd se vio influenciada por la reducción de la biomasa (A2 y 1233) y la tasa de transpiración (IMC 67) causada por el déficit hídrico. La combinación de estrés hídrico y Cd estuvo relacionada con el contenido de clorofilas, el estrés oxidativo y acumulación de prolina en las hojas. Las progenies A2 y 1233 acumularon más Cd en la planta que IMC 67, con mayor concentración en las hojas. El factor translocación (FT) mostró que los órganos aéreos de las tres progenies estaban enriquecidos con Cd (FT>4). El déficit hídrico incrementó la translocación de Cd desde las raíces en A2 e IMC 67, a pesar de que este último acumuló menos Cd, mientras que en 1233 no hubo cambios significativos. La progenie 1233 se destacó como el portainjerto más prometedor debido a su tolerancia al estrés hídrico y estabilidad en la acumulación de Cd. (Texto tomado de la fuente)The risk of drought due to climate variability and the presence of cadmium (Cd) in the soil is common in cocoa-producing areas worldwide. Currently, there are no published studies on Cd accumulation under water deficit conditions. This research aimed to evaluate the physiological response and cadmium accumulation in cocoa plants under water deficit. The study was conducted in a nursery at the Nataima Research Center of AGROSAVIA, in El Espinal municipality, Tolima. Two potential cocoa rootstock materials, progenies A2 and 1233 from controlled crosses, and a commercially used rootstock genotype in Colombia (IMC 67), were used. Various parameters, including water-related aspects, growth, physiological, biochemical, and Cd accumulation in 7-month-old plants, were analyzed. The plants were grown in Cd-contaminated soil without the addition of external sources of the metal, with an initial soluble Cd concentration of 0.356 mg kg-1. They were subjected to water deficit by suspending irrigation for consecutive periods of 19 and 27 days (D19 and D27) and then rehydrated to assess their potential for water stress recovery. Water deficit reduced leaf water potential (Ψleaf) with values between -1.51 and -2.09 MPa, with progeny 1233 being the most tolerant. However, the recovery of gas exchange variables, water potential, and photosynthetic pigments after rehydration suggests that all three genotypes have the capacity to tolerate the stress levels evaluated. Cd accumulation varied among progenies, not among stress levels. The Cd concentration was influenced by biomass reduction (A2 and 1233) and transpiration rate (IMC 67) caused by water deficit. The combination of water deficit and Cd was associated with chlorophyll content, oxidative stress, and proline accumulation in the leaves. Progenies A2 and 1233 accumulated more Cd in the plant than IMC 67, with higher concentration in the leaves. The translocation factor (TF) indicated that the above-ground organs of all three progenies were enriched with Cd (TF>4). Water deficit increased Cd translocation from the roots in A2 and IMC 67, despite the latter accumulating less Cd, while 1233 showed no significant changes. Progeny 1233 stood out as the most promising rootstock due to its water stress tolerance and stable Cd accumulation.MaestríaMagíster en Ciencias Agrarias1.2 Materiales y métodos 1.2.1 Localización del experimento El experimento se realizó bajo condiciones de casa de malla en el Centro de Investigación Nataima – AGROSAVIA, ubicado en una zona de bosque seco tropical a 4°11’31’’N y 74°57’41’’W, a una altitud de 374 msnm en el municipio de El Espinal (Tolima, Colombia). 1.2.2 Material vegetal El material vegetal utilizado en este experimento hace parte de un programa de mejoramiento genético de cacao dirigido por la Corporación Colombiana de investigación agropecuaria – AGROSAVIA. Se utilizaron tres progenies de cacao: A2, 1233 e IMC 67. Las progenies A2 y 1233 son dos familias de hermanos completos obtenidos de cruzamientos dirigidos, las cuales presentan diferencias en la tolerancia al déficit hídrico, siendo 1233 el material tolerante, según estudios previos de AGROSAVIA (AGROSAVIA, 2021). La progenie IMC 67 es un genotipo comercial utilizado ampliamente el Colombia como patrón de cacao, obtenida a partir de libre polinización. Los cruzamientos se llevaron a cabo en el Centro de Investigación Palmira - AGROSAVIA (Valle del Cauca, Colombia) en condiciones de campo. Las semillas obtenidas se germinaron y se mantuvieron en un medio inerte (arena de río) durante 60 días en el Centro de Investigación Nataima - AGROSAVIA. Posteriormente, se trasplantaron plantas de mes de edad, con entre 6 y 8 hojas verdaderas, en bolsas plásticas que contenían suelo, en donde se mantuvieron hasta el final del experimento. El suelo utilizado presentaba un contenido promedio de 0,356 mg kg-1 de Cd soluble, determinado siguiendo la metodología de Bravo et al. (2021) (Información anexa 1). 1.2.3 Establecimiento del experimento Bajo condiciones de casa de malla, se realizó el trasplante en bolsas de plástico negro (60 cm alto x 30 cm de diámetro) con 40 kg de suelo franco, pH promedio de 5,29 y carbono orgánico de 5,1%. El suelo fue previamente homogenizado en tamiz de 12 mm. Considerando los resultados del análisis de suelos (Información anexa 2), cada planta, a los 4 meses de edad, se fertilizó con 5,7 g de Calcinit B (15,5% N; 26,0% Ca; 0,1% B), 8,6 de DAP (18,0% N; 46% P), 5,7 de ManuKiesek (3% K; 24% Mg; S 18%) y 9 ml de Transfer ionic (complejo orgánico de ácidos carboxílicos, gluconatos y ácido ascórbico), con el fin de corregir relaciones iónicas y saturación de bases en el suelo, de con acuerdo a los requerimientos nutricionales del cacao (Jaraba et al., 2021a). Durante el período comprendido entre el trasplante y la aplicación de los tratamientos de estrés hídrico, todas las plantas se hidrataron a capacidad de campo utilizando agua del acueducto. El riego se ajustaba según la humedad del suelo, la cual era monitoreada mediante sensores de humedad volumétrica tipo CS616 (Campbell Scientific®, USA). Se aplicaban entre 250 y 300 cc de agua por planta, con una frecuencia promedio de cada dos días, dependiendo de las condiciones de humedad detectadas. Los tratamientos de déficit hídrico se establecieron cuando las plantas tenían 6 meses de edad (dos Santos et al., 2014) y se distribuyeron en un diseño de bloques al azar con tres repeticiones en un arreglo factorial, donde el primer factor fueron las progenies: A2, IMC 67 y 1233, y el segundo factor los dos estados hídricos: plantas con estrés por déficit hídrico (ES) y plantas bien regadas (BR). Para el tratamiento BR, las plantas se regaron de manera óptima durante el periodo de evaluación, con el fin de mantener el contenido volumétrico de agua (CVA) del suelo entre 38 y 40% durante el periodo de evaluación (Figura 1). El valor de la capacidad de campo del suelo se determinó mediante curva de retención de humedad del suelo del experimento. El CVA se monitoreó con sensores de humedad volumétrica tipo CS616 (Campbell Scientific®, USA) previamente calibrados en laboratorio usando la metodología descrita en el manual del fabricante. Los sensores se instalaron a 25 cm de profundidad en el suelo, donde se encuentran las raíces secundarias y absorbentes de las plantas de cacao, encargadas de tomar agua y nutrientes del suelo (Jaraba et al., 2021b). 1.2.4 Tratamientos de estrés hídrico En condiciones de campo, el potencial hídrico foliar (Ψhoja) antes del amanecer se mantuvo entre -0,2 y -0,7 MPa, según valores reportados para plantas de cacao en estado hídrico óptimo (Dos Santos et al., 2014). En el tratamiento ES se suspendió el riego por 19 (D19) y 27 días (D27) consecutivos hasta que Ψhoja alcanzó valores entre -1,4 ± -0,4 y -2,05 ± 0,25 MPa, respectivamente, con el fin de someter las plantas a dos niveles de estrés, moderado y severo (De Almeida et al., 2016; Dos Santos et al., 2014; Rada et al., 2005), momentos en los cuales el CVA presentó el 30 y el 25% (Figura 1). Al final la fase de estrés D19 y D27, todas las plantas se regaron hasta alcanzar valores de capacidad de campo con 38-40% al tercer día, logrando recuperarse de la fase de estrés (fase de rehidratación). Las evaluaciones de la recuperación se realizaron al tercer día después de la rehidratación (DRH). Figura 1. Comportamiento del CVA (%) del suelo bajo dos niveles de déficit hídrico y rehidratación del mismo. Círculo abierto: CVA (%) del suelo cuando las plantas tenían un Ψhoja entre -1 y -1,8 MPa; Círculo cerrado: CVA (%) del suelo cuando las plantas de cacao tenían un Ψhoja menor a -1,8 MPa. BR, bien regado; ES, estrés por déficit hídrico; DRH, rehidratación de plantas sometidas a ES durante tres días consecutivos. Se evaluaron todos los parámetros en tres momentos diferentes: el día 19 y 27 de estrés hídrico (D19 y D27) y al final de la fase de rehidratación (DRH). Sin embargo, la biomasa y el contenido de Cd en la planta solo se determinaron en D19 y D27. Para realizar estas mediciones, las muestras de tejido foliar se trituraron en nitrógeno líquido y luego se conservaron a -80 °C. Posteriormente, se utilizaron estas muestras para determinar las variables fisiológicas y bioquímicas de las plantas de cacao. 1.2.5 Condiciones ambientales durante el experimento Durante el transcurso del experimento, se realizaron mediciones de variables ambientales clave, como la temperatura del aire (°C), la humedad relativa del aire (% HR) y la radiación solar (W m-2). Para este propósito, se utilizaron dos sensores ATMOS 14 de Meter Groups (EE. UU.), adaptados a un Data Logger EM50 y colocados dentro de la casa de mallas. Estos sensores registraron la temperatura y la humedad relativa diariamente, con una frecuencia de muestreo de 30 minutos. Además, para medir la radiación solar, se utilizó la estación meteorológica Vantage Pro2 (Davis Instruments, USA), ubicada a una distancia de 200 metros de la casa de malla. Los datos recopilados durante todo el experimento se muestran en la Figura 2. Figura 2. Condiciones de temperatura promedio (Temp), temperatura máxima (Temp máx.), temperatura mínima (Temp min.), humedad relativa (HR) y radiación solar (Rad solar) diaria, a la que fueron expuestas las plantas de cacao durante el tiempo del experimento. 1.2.6 Potencial hídrico de la hoja Se determinó Ψhoja entre las 2:00 a.m. y 4:00 a.m. tomando la tercera hoja madura de arriba hacia abajo (De Almeida et al., 2016) de nueve plantas por tratamiento (n = 9) con ayuda de una bomba de presión Schölander (PMS Model 615, Fresno, CA, United States). 1.2.7 Intercambio de gases Los parámetros de tasa fotosintética (A), conductancia estomática (gs) y transpiración (E) y carbono interno (Ci) se registraron de 9:00 a.m. a 12 m en la cuarta hoja madura de arriba hacia abajo (Osorio Zambrano et al., 2021) en doce plantas por tratamiento (n = 12), usando un sistema portátil de fotosíntesis LI-6800XT (LI-COR Biosciences Inc. NE, United States) con una concentración ambiental de CO2 de 400 µmol m-2 s-1 (J. De Almeida et al., 2016) y una densidad de flujo de fotones fotosintéticos de 600 µmol m-2 s-1, de acuerdo con los umbrales de respuesta de las curvas de luz para las tres progenies, realizadas previamente al establecimiento de los tratamientos (datos no mostrados). El equipo LI-6800XT también determinó los valores de déficit de presión de vapor (DPV) durante el tiempo de medición del intercambio gaseoso. 1.2.8 Acumulación de biomasa Se tomaron seis plantas por tratamiento (n = 6) y se dividieron en hojas+peciolos, tallos y raíces, se llevaron a una estufa a 60°C durante tres días y se determinó el peso seco de cada órgano en una balanza analítica. La relación raíz/parte aérea de la planta (R_S) se determinó dividiendo peso seco de la raíz entre el peso seco de las hojas+tallo. 1.2.9 Pigmentos fotosintéticos Se determinó el contenido de carotenoides (Car), clorofila a (Chla), clorofila b (Chlb) y clorofila total (ChlT) en la cuarta hoja madura de nueve plantas por tratamiento (n = 9). Se siguió la metodología propuesta por Warren (2008) y Nguyen et al. (2020) con algunas modificaciones. Después de extraer las clorofilas de dos discos (1,5 cm de diámetro) de la cuarta hoja madura con metanol (100%) y centrifugar a 5000 xg por 5 min, se usó el sobrenadante para determinar pigmentos con un lector de microplacas (Epoch BioTek) a 470, 652 y 665 nm. Para el cálculo de clorofilas y carotenoides se usaron las fórmulas propuestas por (Warren, 2008). 1.2.10 Prolina El contenido de prolina se determinó en la cuarta hoja madura, de arriba hacia abajo, en la misma hoja donde se evaluó el intercambio gaseoso. Se determinó en nueve plantas por tratamiento (n = 9) con base en el método de detección de (Bates et al., 1973) modificado por (Ábrahám et al., 2010) y se adicionan las modificaciones de (Barrera et al., 2010). Para obtener el extracto vegetal se usó ácido 5-sulfosalicílico 3% (p/v). La mezcla duró en agitación horizontal (200 rpm) durante 1 hora y se centrifugó a 14000 rpm durante 5 min para obtener el sobrenadante de interés. Se generó un mix de reacción a una relación 1:2:2 con ácido 5-sulfosalicílico 3% (p/v), ácido acético glacial y ninhidrina ácida, el cual se mezcló con el sobrenadante extraído y se dejó en baño María a 90°C por 60 min. La reacción se detuvo en una cama de hielo y posteriormente se le adicionó tolueno, se agitó en vórtex y se midió absorbancia a 520 nm de la fase orgánica de la mezcla. La concentración final de prolina se determinó a partir de la curva patrón de L-prolina (Barrera et al., 2010). 1.2.11 Peróxido de hidrógeno (H2O2) El contenido de H2O2 se determinó en la cuarta hoja madura de nueve plantas por tratamiento (n = 9) siguiendo en la metodología desarrollada por (Alexieva et al., 2001) con algunas modificaciones. Se tomaron 0,15 g de tejido foliar previamente congelado en nitrógeno líquido, y se homogenizaron con 1,5 ml de ácido tricloroacético (TCA) al 0,1% (p/v) durante 10 segundos en un vórtex. Posteriormente, se centrifugo a 12000 x g durante 15 min. Se recolectó 0,25 ml del sobrenadante y se le adicionó a 0,25 ml de buffer fosfato (100 mM) y 1 ml de yoduro de potasio (1 M KI p/v en H2O). Inmediatamente se incubó durante 1 hora en condiciones de oscuridad. El contenido de H2O2 se determinó usando un espectrofotómetro a una longitud de onda de 390 nm. La curva estándar de H2O2 se hizo preparando varias concentraciones de H2O2 puro, iniciando con una solución stock de 1000 ppm. 1.2.12 Malondialdehído (MDA) El acumulación de MDA, como producto de la peroxidación lipídica, se estimó siguiendo el método de Heath & Packer (1968). Se tomaron muestras de 0,2 g, previamente congeladas en nitrógeno líquido, de la cuarta hoja madura de nueve plantas por tratamiento (n = 9), se homogenizaron con TCA al 5%, se centrifugó 13000 x g durante 20 min. Se extrajo 0,5 ml del sobrenadante y se mezcló con 1 ml de solución de ácido tiobarbitúrico (TBA) al 0,5% y TCA al 20%, se llevó a baño María durante 20 min y posteriormente se centrifugó a 13000 x g durante 5 min y se leyó la absorbancia del sobrenadante a 532, 600 y 440 nm. El equivalente de malondialdehido (MDA) se calculó con las fórmulas de Heath & Packer (1968). 1.2.13 Contenido de cadmio en hojas Se determinó el contenido de Cd en las hojas, considerando que las variables fisiológicas y bioquímicas también se evaluaron en este órgano de las plantas. Además, se ha demostrado que el Cd presente en las hojas es un indicador predictivo del contenido de Cd presente en los granos de cacao (Wade et al., 2022), de ahí la importancia de determinar la acumulación de Cd en las hojas. Para determinar el contenido de Cd en las hojas de cacao, se recolectaron todas las hojas de la planta, utilizando tres plantas por progenie (n = 3) en los días 19 y 27 de estrés. Posteriormente, las muestras se sometieron a un proceso de secado en un horno a 70°C durante una semana, hasta alcanzar un peso constante. El contenido total de Cd en hojas se obtuvo utilizando la técnica de espectroscopia de emisión óptica con plasma acoplado inductivamente (ICP-OES) (Lanza et al., 2016). Los resultados del contenido de Cd en las hojas se expresaron tanto en términos de concentración en mg Cd kg de hojas, como en términos de acumulación en mg de Cd por órgano (hojas). 1.2.14 Análisis estadístico Se determinaron los supuestos de normalidad y homogeneidad de varianza de las variables evaluadas y se realizó un ANOVA de dos vías (progenie x tratamiento). Se hizo la comparación de medias para evaluar diferencias significativas a través de la prueba HSD de Tukey. Además, se realizó un análisis de correlaciones de Pearson con los variables para determinar posibles relaciones entre ellas (Figura anexa 2). Los análisis se llevaron a cabo usando el software R (R Studio Versión 4.0.1). Materiales y métodos Se empleó el mismo material vegetal descrito en el Capítulo 1 para evaluar la acumulación de cadmio en las plantas de las progenies evaluadas. A continuación, se describe la metodología para determinar las variables relacionadas con el Cd en el suelo y la planta. Contenido de cadmio en el suelo El suelo utilizado para el crecimiento de las plantas en el experimento fue extraído de un lote ubicado en el Centro de Investigación Tibaitatá - AGROSAVIA, el cual se dedica a la producción agropecuaria. Este suelo fue seleccionado específicamente por su contenido inicial de Cd pseudototal (4,85 mg kg-1) y por sus propiedades fisicoquímicas para el desarrollo de las plantas de cacao (Información anexa). Dado el uso del suelo durante las últimas décadas, se considera que el Cd tiene un origen antropogénico, debido a la actividad agropecuaria, específicamente al uso de fertilizantes fosfóricos que enriquecen el suelo con este metal. Con el fin de garantizar uniformidad, se homogeneizó una cantidad total de 10,08 toneladas de suelo, la cual se utilizó para llenar bolsas de plástico negro con una capacidad de 40 kg cada una, de 65 cm de alto por 28 cm de diámetro. Estas bolsas fueron utilizadas posteriormente para trasplantar las plántulas de cacao que tenían entre seis y ocho hojas verdaderas. El contenido de Cd en el suelo se determinó siguiendo la metodología descrita por (Bravo et al., 2021). La concentración de Cd pseudototal se determinó mediante espectrometría de plasma acoplado inductivamente con emisión óptica (ICP-OES) (Thermo Scientific ICAP 6000, Waltham, MA, EE. UU.). El término “pseudototal” se debe a que se usó el método de digestión pseudototal convencional (Lorentzen & Kingston, 1996), recomendado por la Agencia de Protección Ambiental (EPA 3050B), el cual solo extrae y digiere los metales pesados retenidos en las fracciones más lábiles del suelo, como la materia orgánica o los carbonatos (Bravo et al., 2021). La determinación de la concentración de Cd soluble en suelo, se realizó con curvas de calibración medidas con ICP-OES (Bravo et al., 2021). Se utilizó un material de referencia de cadmio puro (Merck-SRM solución estándar trazable de Cd (NO3)2 en HNO3 0,5 mol L-1-1000 mg kg-1 Cd Certipur. Referencia 1.19777.0500) para establecer las curvas de calibración estándar. Se prepararon curvas de calibración de bajo, medio y alto rango, con 10 puntos cada una, donde el límite de detección cuantitativa fue de 0,040 mg kg-1 de Cd2+, y se recuperó un 99.3% de Cd en las muestras de suelo. Según los resultados de esta determinación, se puede afirmar que las plantas de todos los tratamientos, incluyendo las plantas bien regadas, crecieron en suelo con un contenido promedio de 0,356 mg kg-1 de Cd soluble, valor considerado alto según Dutta et al. (2020). Contenido de Cd en la planta Para determinar el contenido de Cd en la planta, se separaron los órganos en hojas con peciolo, tallo y raíces de tres plantas por progenie (n = 3) en los días 19 y 27 de estrés. Posteriormente, las muestras se sometieron a un proceso de secado en un horno a 70°C durante una semana, hasta alcanzar un peso constante. El contenido de Cd en raíces, tallo y hojas, se obtuvo utilizando la técnica de espectroscopia de emisión óptica con plasma acoplado inductivamente (ICP-OES) (Lanza et al., 2016). Los resultados del contenido de Cd en la planta se expresaron tanto en términos de concentración en mg Cd por kg del órgano. Para determinar la acumulación de Cd por órgano de la planta, se multiplicó la concentración de Cd por la masa seca de cada órgano, expresado en mg de Cd por órgano. La acumulación Cd total en la planta se obtuvo sumando la acumulación de Cd en raíces, tallo y hojas. Factor de translocación de Cd (FT) El índice factor de translocación (FT) del Cd se determinó con la ecuación de Mattina et al. (2003) (ver Ecuación 1), utilizada por De Almeida et al. (2022) en plantas de cacao con 8 meses de edad. FT=(C parte aérea)/(C raíces) (Ecuación 1) Donde: C = concentración de Cd en mg kg-1 en la parte aérea (tallo + hojas) y las raíces de la planta. El objetivo de determinar el índice FT fue evaluar la capacidad de las progenies evaluadas para retener Cd en las raíces o translocarlo hacia los órganos de parte aérea. Este aspecto es relevante ya que los programas de mejoramiento genético de cacao buscan portainjertos con baja absorción y acumulación de Cd (Savvas et al., 2010). Análisis estadístico El efecto del estado hídrico de la planta en la acumulación de cadmio en las hojas por parte de las progenies de cacao se evaluó mediante un ANOVA de dos vías (progenie x tratamiento de estrés), permitiendo identificar diferencias significativas entre los tratamientos a través de la prueba HSD de Tukey. Además, se incluyó el contenido de cadmio en las hojas en el análisis de correlación de Pearson junto con los demás parámetros fisiológicos, con el objetivo de determinar la relación existente entre la acumulación de cadmio y dichas variables (Figura anexa 2).Fisiología de cultivosxxii, 99 páginasapplication/pdfspaUniversidad Nacional de ColombiaBogotá - Ciencias Agrarias - Maestría en Ciencias AgrariasFacultad de Ciencias AgrariasBogotá, ColombiaUniversidad Nacional de Colombia - Sede Bogotá580 - Plantas570 - Biología630 - Agricultura y tecnologías relacionadas630 - Agricultura y tecnologías relacionadas::633 - Cultivos de campo y de plantaciónRelaciones agua-plantaPlantas -- Efectos del cadmioCadmioPlant-water relationshipsPlants, Effect of cadmium onCadmiumDéficit hídricoTheobroma cacaoAnálisis del sueloEstrés de sequiaWater shortagesSoil analysisDrought stressRelación planta-sueloPlant-soil relationshipsCacaoDéficit hídricoEstrés hídricoGenotipoToleranciaAcumulación de CdTranslocación de CdCocoaDrought stressGenotypeToleranceCd accumulationCd translocationRespuesta fisiológica y acumulación de cadmio en cacao (Theobroma cacao L.) bajo déficit hídricoEvaluation of physiological response and cadmium accumulation in cocoa plants (Theobroma cacao L.) under water deficit conditionsTrabajo de grado - Maestríainfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/acceptedVersionTexthttp://purl.org/redcol/resource_type/TMColombiaAbbas, T., Rizwan, M., Ali, S., Adrees, M., Mahmood, A., Zia-ur-Rehman, M., Ibrahim, M., Arshad, M., & Qayyum, M. 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Frontiers Media S.A. https://doi.org/10.3389/fpls.2022.773815Corporación Colombiana de Investigación Agropecuaria AGROSAVIAEstudiantesInvestigadoresMaestrosPúblico generalLICENSElicense.txtlicense.txttext/plain; charset=utf-85879https://repositorio.unal.edu.co/bitstream/unal/85483/1/license.txteb34b1cf90b7e1103fc9dfd26be24b4aMD51ORIGINAL1094579566.2023.pdf1094579566.2023.pdfTesis de Maestría en Ciencias Agrariasapplication/pdf2208139https://repositorio.unal.edu.co/bitstream/unal/85483/2/1094579566.2023.pdf97675df90aee66fb37031695a79db161MD52THUMBNAIL1094579566.2023.pdf.jpg1094579566.2023.pdf.jpgGenerated Thumbnailimage/jpeg4584https://repositorio.unal.edu.co/bitstream/unal/85483/3/1094579566.2023.pdf.jpgcc2928b3a1023379a145fc9c9756f1dbMD53unal/85483oai:repositorio.unal.edu.co:unal/854832024-08-22 23:10:13.298Repositorio Institucional Universidad Nacional de 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