Root-to-Shoot Hormonal Communication in Contrasting Rootstocks Suggests an Important Role for the Ethylene Precursor Aminocyclopropane-1-carboxylic Acid in Mediating Plant Growth under Low-Potassium Nutrition in Tomato

Selection and breeding of rootstocks that can tolerate low K supply may increase crop productivity in low fertility soils and reduce fertilizer application. However, the underlying physiological traits are still largely unknown. In this study, 16 contrasting recombinant inbred lines (RILs) derived from a cross between domestic and wild tomato species (Solanum lycopersicum × Solanum pimpinellifolium) have been used to analyse traits related to the rootstock-mediated induction of low (L, low shoot fresh weight) or high (H, high shoot fresh weight) vigor to a commercial F1 hybrid grown under control (6 mM, c) and low-K (1 mM, k). Based on hormonal and ionomic composition in the root xylem sap and the leaf nutritional status after long-term (7 weeks) exposure low-K supply, a model can be proposed to explain the rootstocks effects on shoot performance with the ethylene precursor aminocyclopropane-1-carboxylic acid (ACC) playing a pivotal negative role. The concentration of this hormone was higher in the low-vigor Lc and Lk rootstocks under both conditions, increased in the sensitive HcLk plants under low-K while it was reduced in the high-vigor Hk ones. Low ACC levels would promote the transport of K vs. Na in the vigorous Hk grafted plants. Along with K, Ca, and S, micronutrient uptake and transport were also activated in the tolerant Hk combinations under low-K. Additionally, an interconversion of trans-zeatin into trans-zeatin riboside would contribute to decrease ACC in the tolerant LcHk plants. The high vigor induced by the Hk plants can also be explained by an interaction of ACC with other hormones (cytokinins and salicylic, abscisic and jasmonic acids). Therefore, Hk rootstocks convert an elite tomato F1 cultivar into a (micro) nutrient-efficient phenotype, improving growth under reduced K fertilization.This research has received funding from the Spanish MINECO-FEDER (project AGL2014-59728-R) and from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no 289365 (project ROOTOPOWER).USD 2,116.5 APC fee funded by the EC FP7 Post-Grant Open Access PilotPeer reviewe

Improvement of yield and quality fruit in water stressed through selection of pepper rootstocks

[SPA] La reducción del agua disponible en el Sureste español está obligando a buscar alternativas que palien dichas disminuciones. En este trabajo se han estudiado los efectos de tres portainjertos comerciales (Atlante, Creonte y Terrano) sobre la respuesta agronómica y fisiológica de una variedad comercial de pimiento (cv Herminio) sometido a riego deficitario (50% de la capacidad de campo). Creonte produjo las plantas más productivas y eficientes en el uso del agua, registró la mayor actividad fotosintética y contenido de agua en las hojas, y el área foliar y la biomasa más estables bajo estrés hídrico. Atlante es un portainjertos vegetativo vigoroso y Terrano es más bien un portainjertos enano-reproductivo que produce plantas compactas eficientes sin efectos negativos en la calidad de la fruta. [ENG] The reduction of the water available in the southeastern Spanish is forcing to look for alternatives that alleviate these decreases. The effects of three commercial rootstocks (Atlante, Creonte and Terrano) on the agronomical and physiological responses of a commercial sweet pepper variety (cv Herminio) subject to deficit irrigation (50% of field capacity) have been studied. Although the three rootstocks increased total and marketable yield under control and deficit irrigation. Creonte produced the most productive and efficient plants in the use of water, recorded the highest photosynthetic activity and water content in the leaves, and the foliar area and biomass more stable under water stress. Atlante is a vigorous vegetative rootstock and Terrano is rather a dwarfing-reproductive rootstock that produces efficient compact plants without negative effects on fruit quality

Interaction between ABA and cytokinins in salinized tomato plants

Abscisic acid (ABA) and cytokinins (CKs) are phytohormones that mediate plant responses to abiotic stress and are mainly considered to be produced in roots. These two plant hormones have long been considered antagonists, but the role of this antagonism in mediating abiotic stress resistance remains unknown. In this study we have used reciprocal grafting of transgenic lines with altered hormone synthesis (ABA or CK synthesis) with the aim of assessing the importance of ABA and CK interaction in mediating plant salinity responses in tomato. Preliminary results have shown a negative effect in ABA and CK on tomato plant biomass under salinity. Results have also indicated that jasmonic acid (JA) might be involved in the negative impact on tomato growth caused by ABA and CK interaction under salt conditions.The authors thank Andrew J. Thompson, from Cranfield Univertity (UK), and Ann C. Smigocki from Molecular Plant Pathology Laboratory (EEUU) the NCED and IPT seeds sets respectively. This work was supported by CICYT AGL2011‐27996 and ROOTOPOWER (REF: 289365. FP7‐KBBE‐2011‐5 European Union)

ABA-overproduction response under salinity

[SPA] Con el fin de comprender la influencia de la fitohormona ácido abscísico (ABA) en la adaptación al riego salino, dos líneas transgénicas independientes de tomate (Solanum lycopersicum L.), sp12 y sp5, que sobreexpresan constitutivamente el gen NCED1 (codifica para la enzima que cataliza un paso limitante en la biosíntesis de ABA) y la variedad silvestre Ailsa Craig, se han estudiado en experimentos o bien i) como planta entera o ii) como portainjerto bajo condiciones control y de estrés salino. Aunque la expresión constitutiva de NCED disminuye el crecimiento bajo condiciones control, minimiza los efectos producidos por la sal (planta completa) y mejora significativamente el crecimiento cuando se usa como portainjerto. El análisis de la savia xilemática de raíz mostró que los fenotipos resultantes bajo las diferentes condiciones de cultivo eran difíciles de explicar en términos de sobreproducción de ABA. Para intentar explicar estos resultados se llevó a cabo un análisis de expresión de un conjunto de genes relacionados con hormonas y estrés mediante PCR cuantitativa, así como un estudio transcriptómico mediante microarrays en la raíz. Los resultados sugieren que la sobreexpresión de NCED parece alterar diversas rutas de señalización, derivando en una respuesta adaptativa al estrés que podría ayudar a explicar los fenotipos observados. [ENG] With the aim of better understanding the influence of the plan hormone abscisic acid (ABA) in adaptation to saline irrigation, two independent transgenic tomato (Solanum lycopersicum L.) lines, sp12 and sp5, overexpressing constitutively NCED1 (the enzyme that catalyzes a key rate-limiting step in ABA biosynthesis) and the wild type Ailsa Craig, have been studied in experiments either i) as whole plants or ii) as rootstocks under control and salinity conditions. While NCED overexpression penalizes growth under control conditions, it minimized the effect of salinity (whole plants) or significantly improved plant growth and yield when used as rootstocks. The analysis of the root xylem sap revealed that the phenotypes resulting under the different conditions were difficult to explain in terms of ABA overproduction. With the aim of explaining these results, the expression of a set of hormone and stress associated genes (analysed by real time PCR) as well as a transcriptomic analysis (by using one-color microarray) were performed in roots. The results suggest that NCED overexpression seems to alter several signalling pathways leading to stress adaptive responses that could help to explain the observed phenotypes.The authors thank Andrew J. Thompson from Cranfield University, the NCED seeds set. This work was supported by CICYT-FEDER (project AGL2011-27996) and European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no 289365(ROOTOPOWER project)

Root ABA signalling in salinized tomato

[SPA] Con el fin de comprender la influencia de la fitohormona ácido abscísico (ABA) en la adaptación al riego salino, dos líneas transgénicas independientes de tomate (Solanum lycopersicum L.), sp12 y sp5, que sobreexpresan constitutivamente el gen NCED1 (codifica para la enzima que cataliza un paso limitante en la biosíntesis de ABA) y la variedad silvestre Ailsa Craig, se han estudiado en experimentos o bien i) como planta entera o ii) como portainjerto bajo condiciones control y de estrés salino. Aunque la expresión constitutiva de NCED disminuye el crecimiento bajo condiciones control, minimiza los efectos producidos por la sal (planta completa) y mejora significativamente el crecimiento cuando se usa como portainjerto. El análisis de la savia xilemática de raíz mostró que los fenotipos resultantes bajo las diferentes condiciones de cultivo eran difíciles de explicar en términos de sobreproducción de ABA. Para intentar explicar estos resultados se llevó a cabo un análisis de expresión de un conjunto de genes relacionados con hormonas y estrés mediante PCR cuantitativa, así como un estudio transcriptómico mediante microarrays en la raíz. Los resultados sugieren que la sobreexpresión de NCED parece alterar diversas rutas de señalización, derivando en una respuesta adaptativa al estrés que podría ayudar a explicar los fenotipos observados. [ENG] With the aim of better understanding the influence of the plan hormone abscisic acid (ABA) in adaptation to saline irrigation, two independent transgenic tomato (Solanum lycopersicum L.) lines, sp12 and sp5, overexpressing constitutively NCED1 (the enzyme that catalyzes a key rate-limiting step in ABA biosynthesis) and the wild type Ailsa Craig, have been studied in experiments either i) as whole plants or ii) as rootstocks under control and salinity conditions. While NCED overexpression penalizes growth under control conditions, it minimized the effect of salinity (whole plants) or significantly improved plant growth and yield when used as rootstocks. The analysis of the root xylem sap revealed that the phenotypes resulting under the different conditions were difficult to explain in terms of ABA overproduction. With the aim of explaining these results, the expression of a set of hormone and stress associated genes (analysed by real time PCR) as well as a transcriptomic analysis (by using one-color microarray) were performed in roots. The results suggest that NCED overexpression seems to alter several signalling pathways leading to stress adaptive responses that could help to explain the observed phenotypes.The authors thank Andrew J. Thompson from Cranfield University, the NCED seeds set. This work was supported by CICYT-FEDER (project AGL2011-27996) and European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no 289365(ROOTOPOWER project).. -2010-5 European Union)

Efectos de la sobreproducción de ácido abscísico y citoquininas en la respuesta fisiológica y agronómica de tomate bajo estrés salino en invernadero

La salinidad limita la productividad de los cultivos y modifica el balance hormonal vegetal. El ácido abscísico (ABA) y las citoquininas (CKs) son fitohormonas que median la respuesta a estreses abióticos y se considera que son producidas principalmente en la raíz. Para evaluar el papel de estas dos hormonas en el crecimiento y productividad de los cultivos bajo salinidad, se realizaron injertos cuyo portainjerto eran unas variedades transgénicas de tomate que sobreproducían ácido abscísico (ABA) (líneas NCED) y citoquininas (líneas IPT), utilizando una variedad comercial de tomate cherry como injerto. Las plantas de tomate injertadas fueron cultivadas en un invernadero comercial durante 6 meses en condiciones de salinidad moderada. Las plantas cuyo portainjerto era una variedad transgénica eran más vigorosas y presentaban mayor producción que las que tenían como portainjerto una planta wild type o la propia parte aérea injertada. Los análisis hormonales no explican completamente estos datos, por lo que otros tipos de análisis (genéticos, enzimáticos) son necesarios para explicar completamente estos datos.Los autores de la presente comunicación desean agradecer a Andrew J. Thompson de la Universidad de Cranfield (Reino Unido) y a Ann C. Smigocki del Molecular Plant Pathology Laboratory (EEUU) por los sets de semillas NCED e IPT respectivamente. Este trabajo ha sido financiado por CICYT AGL2011‐27996 y ROOTOPOWER (REF: 289365. FP7‐KBBE‐2011‐5 European Union

ABA-overproduction response under salinity

[SPA] Con el fin de comprender la influencia de la fitohormona ácido abscísico (ABA) en la adaptación al riego salino, dos líneas transgénicas independientes de tomate (Solanum lycopersicum L.), sp12 y sp5, que sobreexpresan constitutivamente el gen NCED1 (codifica para la enzima que cataliza un paso limitante en la biosíntesis de ABA) y la variedad silvestre Ailsa Craig, se han estudiado en experimentos o bien i) como planta entera o ii) como portainjerto bajo condiciones control y de estrés salino. Aunque la expresión constitutiva de NCED disminuye el crecimiento bajo condiciones control, minimiza los efectos producidos por la sal (planta completa) y mejora significativamente el crecimiento cuando se usa como portainjerto. El análisis de la savia xilemática de raíz mostró que los fenotipos resultantes bajo las diferentes condiciones de cultivo eran difíciles de explicar en términos de sobreproducción de ABA. Para intentar explicar estos resultados se llevó a cabo un análisis de expresión de un conjunto de genes relacionados con hormonas y estrés mediante PCR cuantitativa, así como un estudio transcriptómico mediante microarrays en la raíz. Los resultados sugieren que la sobreexpresión de NCED parece alterar diversas rutas de señalización, derivando en una respuesta adaptativa al estrés que podría ayudar a explicar los fenotipos observados. [ENG] With the aim of better understanding the influence of the plan hormone abscisic acid (ABA) in adaptation to saline irrigation, two independent transgenic tomato (Solanum lycopersicum L.) lines, sp12 and sp5, overexpressing constitutively NCED1 (the enzyme that catalyzes a key rate-limiting step in ABA biosynthesis) and the wild type Ailsa Craig, have been studied in experiments either i) as whole plants or ii) as rootstocks under control and salinity conditions. While NCED overexpression penalizes growth under control conditions, it minimized the effect of salinity (whole plants) or significantly improved plant growth and yield when used as rootstocks. The analysis of the root xylem sap revealed that the phenotypes resulting under the different conditions were difficult to explain in terms of ABA overproduction. With the aim of explaining these results, the expression of a set of hormone and stress associated genes (analysed by real time PCR) as well as a transcriptomic analysis (by using one-color microarray) were performed in roots. The results suggest that NCED overexpression seems to alter several signalling pathways leading to stress adaptive responses that could help to explain the observed phenotypes.The authors thank Andrew J. Thompson from Cranfield University, the NCED seeds set. This work was supported by CICYT-FEDER (project AGL2011-27996) and European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no 289365(ROOTOPOWER project)

Impact of overexpression of 9-cis-epoxycarotenoid dioxygenase on growth and gene expression under salinity stress

To better understand abscisic acid (ABA)’s role in the salinity response of tomato (Solanum lycopersicum L.), two independent transgenic lines, sp5 and sp12, constitutively overexpressing the LeNCED1 gene (encoding 9-cis-epoxycarotenoid dioxygenase, a key enzyme in ABA biosynthesis) and the wild type (WT) cv. Ailsa Craig, were cultivated hydroponically with or without the addition of 100 mM NaCl. Independent of salinity, LeNCED1 overexpression (OE) increased ABA concentration in leaves and xylem sap, and salinity interacted with the LeNCED1 transgene to enhance ABA accumulation in xylem sap and roots. Under control conditions, LeNCED1 OE limited root and shoot biomass accumulation, which was correlated with decreased leaf gas exchange. In salinized plants, LeNCED1 OE reduced the percentage loss in shoot and root biomass accumulation, leading to a greater total root length than WT. Root qPCR analysis of the sp12 line under control conditions revealed upregulated genes related to ABA, jasmonic acid and ethylene synthesis and signalling, gibberellin and auxin homeostasis and osmoregulation processes. Under salinity, LeNCED1 OE prevented the induction of genes involved in ABA metabolism and GA and auxin deactivation that occurred in WT, but the induction of ABA signalling and stress-adaptive genes was maintained. Thus, complex changes in phytohormone and stress-related gene expression are associated with constitutive upregulation of a single ABA biosynthesis gene, alleviating salinity-dependent growth limitation

Overproduction of ABA in rootstocks alleviates salinity stress in tomato shoots

To determine whether root-supplied ABA alleviates saline stress, tomato (Solanum lycopersicum L. cv. Sugar Drop) was grafted onto two independent lines (NCED OE) overexpressing the SlNCED1 gene (9-cis-epoxycarotenoid dioxygenase) and wild type rootstocks. After 200 days of saline irrigation (EC = 3.5 dS m−1), plants with NCED OE rootstocks had 30% higher fruit yield, but decreased root biomass and lateral root development. Although NCED OE rootstocks upregulated ABA-signalling (AREB, ATHB12), ethylene-related (ACCs, ERFs), aquaporin (PIPs) and stress-related (TAS14, KIN, LEA) genes, downregulation of PYL ABA receptors and signalling components (WRKYs), ethylene synthesis (ACOs) and auxin-responsive factors occurred. Elevated SlNCED1 expression enhanced ABA levels in reproductive tissue while ABA catabolites accumulated in leaf and xylem sap suggesting homeostatic mechanisms. NCED OE also reduced xylem cytokinin transport to the shoot and stimulated foliar 2-isopentenyl adenine (iP) accumulation and phloem transport. Moreover, increased xylem GA3 levels in growing fruit trusses were associated with enhanced reproductive growth. Improved photosynthesis without changes in stomatal conductance was consistent with reduced stress sensitivity and hormone-mediated alteration of leaf growth and mesophyll structure. Combined with increases in leaf nutrients and flavonoids, systemic changes in hormone balance could explain enhanced vigour, reproductive growth and yield under saline stress