Francisco Javier Romera

Iron (Fe) is an essential micronutrient for crop productivity, but its low availability in alkaline and calcareous soils limits the growth of rice (Oryza sativa L.), which employs a combined strategy for its acquisition based on the release of phytosiderophores (PS) and the use of specific transporters. In this study, the effect of the rhizospheric bacterium Pseudomonas simiae WCS417 and the halotolerant yeast Debaryomyces hansenii CBS767 as inducers of responses to Fe deficiency in rice grown under hydroponic conditions was evaluated. Plants were inoculated in nutrient solutions with and without Fe, and PS production and the expression of genes associated with biosynthesis and transport were determined by qRT-PCR. The results showed that both microorganisms significantly increased PS production compared to controls, especially under Fe-deficient conditions, although P. simiae also exerted an effect under Fe sufficiency. Furthermore, induction of key genes (OsNAAT, OsIRO2, OsTOM1, OsYSL15, and OsIRT1), as well as genes related to the ethylene pathway (OsEIN2, OsACS2, and OsACO3), was observed, pointing to a regulatory role for this hormone in the response. In conclusion, P. simiae and D. hansenii act as inducers of Fe acquisition mechanisms in rice, offering a sustainable biotechnological approach to improve iron nutrition in limiting environments.

Verticillium wilt, caused by the soilborne fungus Verticillium dahliae , and iron (Fe) deficiency are major stresses impacting valuable crops in the Mediterranean basin. The plant response to these stresses consists of overlapping pathways that, coupled with the emerging use of endophytic fungi as biological control agents (BCAs) and plant biostimulants, enables the study of their potential to mitigate co‐occurring biotic stresses and nutritional deficiencies such as Fe deficiency. To this end, Arabidopsis thaliana plants of the “Columbia” ecotype grown in calcareous soil were inoculated with the non‐pathogenic strain Fusarium oxysporum FO12 and/or with the defoliating pathotype of V. dahliae strain V180. Plant growth, plant mineral uptake, and disease progression, along with the molecular quantification of FO12 and V180 fungal biomass and the relative expression of ISR‐, SAR‐, and Fe deficiency‐related genes in shoots, were evaluated. The results revealed that plants inoculated with the FO12 strain increased growth, leaf chlorophyll concentration (SPAD index), and phosphorus (P; 126%) and Fe (9%) contents, partially due to ethylene (ET) and Iron Man peptides (IMA). In addition, FO12 reduced the severity of symptoms by more than 85%. This reduction in disease severity may be attributed to a direct competitive effect on V180 progression and to the induction of several defence‐associated genes related to ET and SA synthesis and signaling. This bidirectional crosstalk between Fe deficiency responses and plant defences triggered by FO12 mainly through ET can improve plant resilience while reducing reliance on agrochemicals, offering a sustainable multifaceted tool for crop protection and nutrition management.

Calcareous soils, characterized by high pH and calcium carbonate content, often limit the availability of essential nutrients for crops such as rice (Oryza sativa L.), reducing yield and nutritional quality. In this study, we evaluated the effect of the halotolerant yeast Debaryomyces hansenii on the growth, nutrient uptake, and phosphorus acquisition mechanisms of rice plants cultivated in calcareous soil under controlled greenhouse conditions. Plants inoculated with D. hansenii, particularly via root immersion, exhibited significantly higher SPAD chlorophyll index, plant height, and grain yield compared to controls. A modest increase (~4%) in dry matter content was also observed under sterilized soil conditions. Foliar concentrations of Fe, Zn, and Mn significantly increased in plants inoculated with D. hansenii via root immersion in non-sterilized calcareous soil, indicating improved micronutrient acquisition under these specific conditions. Although leaf phosphorus levels were not significantly increased, D. hansenii stimulated acid phosphatase activity, as visually observed through BCIP staining, and upregulated genes involved in phosphorus acquisition under both P-sufficient and P-deficient conditions. At the molecular level, D. hansenii upregulated the expression of acid phosphatase genes (OsPAP3, OsPAP9) and a phosphate transporter gene (OsPTH1;6), confirming its influence on P-related physiological responses. These findings demonstrate that D. hansenii functions as a plant growth-promoting yeast (PGPY) and may serve as a promising biofertilizer for improving rice productivity and nutrient efficiency in calcareous soils, contributing to sustainable agricultural practices in calcareous soils and other nutrient-limiting environments.

Calcareous soils, characterized by high pH and calcium carbonate content, often limit the availability of essential nutrients for crops such as rice (Oryza sativa L.), reducing yield and nutritional quality. In this study, we evaluated the effect of the halotolerant yeast Debaryomyces hansenii (Dh) on the growth, nutrient uptake, and phosphorus acquisition mechanisms of rice plants cultivated in calcareous soil under controlled greenhouse conditions. Plants inoculated with Dh, particularly via root immersion, exhibited significantly higher SPAD chlorophyll index, plant height, dry matter content, and grain yield compared to controls. Foliar concentrations of Cu, Fe, Zn, and Mn also increased under certain conditions, suggesting improved micronutrient acquisition. Although phosphorus levels in leaves were not significantly enhanced, Dh stimulated acid phosphatase activity in both P-sufficient and P-deficient treatments. At the molecular level, Dh upregulated the expression of acid phosphatase genes (OsPAP3, OsPAP9) and a phosphate transporter gene (OsPTH1;6), confirming its influence on P-related physiological responses. These findings demonstrate that D. hansenii functions as a plant growth-promoting yeast (PGPY) and may serve as a promising biofertilizer for improving rice productivity and nutrient efficiency in calcareous soils, contributing to sustainable agricultural practices in challenging edaphic environments.

Iron (Fe) deficiency is among the most important agronomical concerns under alkaline conditions. Bicarbonate is considered an important factor causing Fe deficiency in dicot plants, mainly on calcareous soils. Current production systems are based on the use of high-yielding varieties and the application of large quantities of agrochemicals, which can cause major environmental problems. The use of beneficial rhizosphere microorganisms is considered a relevant sustainable alternative to synthetic fertilizers. The main purpose of this work has been to analyze the impact of the inoculation of tomato (Solanum lycopersicum L.) seedlings with the WCS417 strain of Pseudomonas simiae, in the presence or absence of bicarbonate, on plant growth and other physiological parameters. To conduct this research, three different inoculation methods were implemented: root immersion, foliar application, and substrate inoculation by irrigation. The results obtained show the ability of the P. simiae WCS417 strain to induce medium acidification in the presence of bicarbonate to increase the SPAD index and to improve the growth and development of the tomato plants in calcareous conditions provoked by the presence of bicarbonate, which indicates that this bacteria strain could have a great potential as an Fe biofertilizer.

The European “Green Deal” policies are shifting toward more sustainable and environmentally conscious agricultural practices, reducing the use of chemical fertilizer and pesticides. This implies exploring alternative strategies. One promising alternative to improve plant nutrition and reinforce plant defenses is the use of beneficial microorganisms in the rhizosphere, such as “Plant-growth-promoting rhizobacteria and fungi”. Despite the great abundance of iron (Fe) in the Earth’s crust, its poor solubility in calcareous soil makes Fe deficiency a major agricultural issue worldwide. Among plant promoting microorganisms, the yeast Debaryomyces hansenii has been very recently incorporated, for its ability to induce morphological and physiological key responses to Fe deficiency in plants, under hydroponic culture conditions. The present work takes it a step further and explores the potential of D. hansenii to improve plant nutrition and stimulate growth in cucumber plants grown in calcareous soil, where ferric chlorosis is common. Additionally, the study examines D. hansenii’s ability to induce systemic resistance (ISR) through a comparative relative expression study by qRT-PCR of ethylene (ET) biosynthesis (ACO1), or ET signaling (EIN2 and EIN3), and salicylic acid (SA) biosynthesis (PAL)-related genes. The results mark a significant milestone since D. hansenii not only enhances nutrient uptake and stimulates plant growth and flower development but could also amplify induced systemic resistance (ISR). Although there is still much work ahead, these findings make D. hansenii a promising candidate to be used for sustainable and environmentally friendly integrated crop management.

Rice (Oryza sativa L.) is a very important cereal worldwide, since it is the staple food for more than half of the world’s population. Iron (Fe) deficiency is among the most important agronomical concerns in calcareous soils where rice plants may suffer from this deficiency. Current production systems are based on the use of high-yielding varieties and the application of large quantities of agrochemicals, which can cause major environmental problems. The use of beneficial rhizosphere microorganisms is considered a relevant sustainable alternative to synthetic fertilizers. The main goal of this study was to determine the ability of the nonpathogenic strain Fusarium oxysporum FO12 to induce Fe-deficiency responses in rice plants and its effects on plant growth and Fe chlorosis. Experiments were carried out under hydroponic system conditions. Our results show that the root inoculation of rice plants with FO12 promotes the production of phytosiderophores and plant growth while reducing Fe chlorosis symptoms after several days of cultivation. Moreover, Fe-related genes are upregulated by FO12 at certain times in inoculated plants regardless of Fe conditions. This microorganism also colonizes root cortical tissues. In conclusion, FO12 enhances Fe-deficiency responses in rice plants, achieves growth promotion, and reduces Fe chlorosis symptoms.

Iron (Fe) is abundant in soils but with a poor availability for plants, especially in calcareous soils. To favor its acquisition, plants develop morphological and physiological responses, mainly in their roots, known as Fe deficiency responses. In dicot plants, the regulation of these responses is not totally known, but some hormones and signaling molecules, such as auxin, ethylene, glutathione (GSH), nitric oxide (NO) and S-nitrosoglutathione (GSNO), have been involved in their activation. Most of these substances, including auxin, ethylene, GSH and NO, increase their production in Fe-deficient roots while GSNO, derived from GSH and NO, decreases its content. This paradoxical result could be explained with the increased expression and activity in Fe-deficient roots of the GSNO reductase (GSNOR) enzyme, which decomposes GSNO to oxidized glutathione (GSSG) and NH₃. The fact that NO content increases while GSNO decreases in Fe-deficient roots suggests that NO and GSNO do not play the same role in the regulation of Fe deficiency responses. This review is an update of the results supporting a role for NO, GSNO and GSNOR in the regulation of Fe deficiency responses. The possible roles of NO and GSNO are discussed by taking into account their mode of action through post-translational modifications, such as S-nitrosylation, and through their interactions with the hormones auxin and ethylene, directly related to the activation of morphological and physiological responses to Fe deficiency in dicot plants.

Editorial: Role of shoot-derived signals in root responses to environmental changes

When plants suffer from Fe deficiency, they develop morphological and physiological responses, mainly in their roots, aimed to facilitate Fe mobilization and uptake. Once Fe has been acquired in sufficient quantity, the responses need to be switched off to avoid Fe toxicity and to conserve energy. Several hormones and signaling molecules, such as ethylene, auxin and nitric oxide, have been involved in the activation of Fe deficiency responses in Strategy I plants. These hormones and signaling molecules have almost no effect when applied to plants grown under Fe-sufficient conditions, which suggests the existence of a repressive signal related to the internal Fe content. The nature of this repressive signal is not known yet many experimental results suggest that is not related to the whole root Fe content but to some kind of Fe compound moving from leaves to roots through the phloem. After that, this signal has been named LOng-Distance Iron Signal (LODIS). Very recently, a novel family of small peptides, “IRON MAN” (IMA), has been identified as key components of the induction of Fe deficiency responses. However, the relationship between LODIS and IMA peptides is not known. The main objective of this work has been to clarify the relationship between both signals. For this, we have used Arabidopsis wild type (WT) Columbia and two of its mutants, opt3 and frd3, affected, either directly or indirectly, in the transport of Fe (LODIS) through the phloem. Both mutants present constitutive activation of Fe acquisition genes when grown in a Fe-sufficient medium despite the high accumulation of Fe in their roots. Arabidopsis WT Columbia plants and both mutants were treated with foliar application of Fe, and later on the expression of IMA and Fe acquisition genes was analyzed. The results obtained suggest that LODIS may act upstream of IMA peptides in the regulation of Fe deficiency responses in roots. The possible regulation of IMA peptides by ethylene has also been studied. Results obtained with ethylene precursors and inhibitors, and occurrence of ethylene-responsive cis-acting elements in the promoters of IMA genes, suggest that IMA peptides could also be regulated by ethylene.