Potential of the physiological response of pea plants (Pisum sativum L. ) to iron deficiency (direct or lime- induced) / Potencial da resposta fisiológica das plantas de ervilha (Pisum sativum L. ) à deficiência de ferro (direta ou induzida pelo calcário)

Pea (Pisum sativum L.) is an important food crop in Tunisia, where calcareous soils represents the major limiting factor for agriculture production. In the present study a greenhouse experiment was conducted to assess the effects of direct and bicarbonate- induced iron deficiency on plant growth, chlorophyll fluorescence, photosynthesis, spad index and iron nutrition in two Tunisian pea genotypes (Pisum sativum L.). Plants were grown hydroponically and iron deficiency was induced for 3 weeks. Iron deficiency decreased all the above physiological parameters. The direct Fe deficiency is more drastic than bicarbonate- induced Fe deficiency. A close relationship between plant growth, photosynthesis and SPAD index was observed. Fe use efficiency for plant growth and Fe use efficiency for photosynthesis discriminates clearly the studied genotypes and seems to be the main reason of the tolerance of Kelvedon, as compared to Lincoln.

A ervilha (Pisum sativum L.) é uma cultura alimentar importante na Tunísia, onde os solos calcários representam o principal fator limitante para a produção agrícola. No presente estudo, foi conduzido um experimento em estufa para avaliar os efeitos da deficiência de ferro direta e induzida por bicarbonato sobre o crescimento de plantas, a fluorescência da clorofila, a fotossíntese, o índice SPAD e a nutrição de ferro em dois genótipos de ervilha da Tunísia (Pisum sativum L.). As plantas foram cultivadas hidroponicamente e a deficiência de ferro foi induzida durante 3 semanas. A deficiência de ferro diminuiu todos os parâmetros fisiológicos acima. A deficiência de Fe direta é mais drástica do que a deficiência de Fe induzida por bicarbonato. Observou-se uma estreita relação entre o crescimento das plantas, a fotossíntese e o índice SPAD. A eficiência de uso de Fe para o crescimento de plantas e a eficiência de uso de Fe para a fotossíntese discriminam claramente os genótipos estudados e parecem ser a razão principal da tolerância de Kelvedon, em comparação com a de Lincoln.

Differences in efficient metabolite management and nutrient metabolic regulation between wild and cultivated barley grown at high salinity.

Physiological and biochemical responses of Hordeum maritimum and H. vulgare to salt stress were studied over a 60-h period. Growth at increasing salinity levels (0, 100, 200 and 300 mM NaCl) was assessed in hydroponic culture. H. maritimum was shown to be a true halophyte via its typical behaviour at high salinity. Shoot growth of cultivated barley was gradually reduced with increasing salinity, whereas that of wild barley was enhanced at 100 and 200 mm NaCl then slightly reduced at 300 mM NaCl. The higher salt tolerance of H. maritimum as compared to H. vulgare was due to its higher capacity to maintain cell turgor under severe salinity. Furthermore, H. maritimum exhibited fine regulation of Na(+) transport from roots to shoots and, unlike H. vulgare, it accumulated less Na(+) in shoots than in roots. In addition, H. maritimum can accumulate more Na(+) than K(+) in both roots and shoots without the appearance of toxicity symptoms, indicating that Na(+) was well compartmentalized within cells and substituted K(+) in osmotic adjustment. The higher degree of salt tolerance of H. maritimum is further demonstrated by its economic strategy: at moderate salt treatment (100 mm NaCl), it used inorganic solutes (such as Na(+)) for osmotic adjustment and kept organic solutes and a large part of the K(+) for metabolic activities. Indeed, K(+) use efficiency in H. maritimum was about twofold that in H. vulgare; the former started to use organic solutes as osmotica only at high salinity (200 and 300 mm NaCl). These results suggest that the differences in salt tolerance between H. maritimum and H. vulgare are partly due to (i) differences in control of Na(+) transport from roots to shoots, and (ii) H. maritimum uses Na(+) as an osmoticum instead of K(+) and organic solutes. These factors are differently reflected in growth.

Iron deficiency tolerance traits in wild (Hordeum maritimum) and cultivated barley (Hordeum vulgare).

Phytosiderophores (PS) are Fe(III)-solubilizing compounds released by Poaceae roots under iron deficiency conditions. Several studies focused on the capacity of these plants to secrete PS as a center of their iron deficiency tolerance, and little information is available on other traits such as root/shoot biomass ratios, iron use efficiency, photosynthetic activity, and iron mobilization capacity that might also contribute to iron deficiency tolerance. In this study, we evaluated some traits other than PS release capacity that could be responsible for differences in iron deficiency tolerance in two barley species, Hordeum maritimum and Hordeum vulgare. Results showed that under iron starvation, biomass production was affected in both species, but H. maritimum kept higher root/shoot ratios due to the distribution efficiency of carbohydrates within the plant and the growth flexibility of its organs. Both species responded to iron starvation by an early release of PS, but they differed in their secretion capacity. In cultivated barley, the PS release rate was 1.5-2-fold higher than that of wild barley. This behavior was also concomitant with no modification in shoot iron concentration of the latter, which may lead to a low stimulation of its PS release as compared to the former. The amount of Fe(3+) mobilized by root exudates was determined at different pH values (between 5.6 and 8.6). Results showed a decrease in the mobilization capacity with the increasing pH, mainly in H. vulgare. At 8.6, it was reduced by 50% in H. vulgare and 30% in H. maritimum. These data suggest that differences in Poaceae tolerance to iron deficiency is attributed not only to PS secretion capacity, but also to carbohydrate distribution within the plant, Fe use efficiency, and root exudates capacity to mobilize Fe(III).

Interactive effects of salinity and iron deficiency in Medicago ciliaris.

In calcareous salt-affected soils, iron availability to plants is subjected to the effects of both sodium and bicarbonate ions. Our aim was to study interactive effects of salinity and iron deficiency on iron acquisition and root acidification induced by iron deficiency in Medicago ciliaris L., a species commonly found in saline ecosystems. Four treatments were used: C, control treatment, complete medium (CM) containing 30 microM Fe; S, salt treatment, CM with 75 mM NaCl; D, deficient treatment, CM containing only 1 microM Fe; DS, interactive treatment, CM containing 1 microM Fe with 75 mM NaCl. Our study showed that plant growth and chlorophyll content were much more affected by the interactive treatment than by iron deficiency or by the salt treatment, indicating an additive effect of these constraints in DS plants. These results could be partially explained by Na accumulation in shoots as well as a limitation of nutrient uptake such as Fe and K under salt stress, under iron deficiency, and especially under their combined effect. The study also showed that root acidification was deeply diminished when iron deficiency was associated with salinity. This probably explained the decrease of Fe uptake and suggested that root proton pump activity would be inhibited by salinity.

Effect of salt on physiological responses of barley to iron deficiency.

Iron chlorosis is very common on alkaline soils such as calcareous ones, since iron availability is limited by high pH. Under these conditions of iron deficiency, graminaceous plant species induce special mechanisms for iron acquisition, involving enhanced release of iron chelators called phytosiderophores. On the other hand, it is known that most of salt soils have alkaline pH. So, plants growing on this kind of soils are often subjected simultaneously to salinity and iron deficiency. This work aimed at (i) studying the physiological responses of barley (Hordeum vulgare L.) to iron deficiency, and (ii) evaluating the effect of salt on the iron nutrition and the phytosiderophore release. For this purpose, seedlings of Hordeum vulgare L. were cultivated under controlled conditions, either in a complete nutrient solution with or without NaCl, or in an iron free nutrient solution containing or not NaCl. The plant morphological aspect, chlorophyll content of young leaves, iron status, biomass production, and phytosiderophore release by roots were assessed. Plants subjected to Fe deficiency exhibited a severe chlorosis, accompanied by a significant biomass reduction. These plants developed more lateral roots than the control with a highly stimulated phytosiderophore release. However, the latter was greatly diminished when iron deficiency was associated to salinity. A depressive effect of salt on iron acquisition in plants subjected only to salt stress which was also observed and further confirmed by the important decrease of efficiency in iron acquisition. These results suggest that salinity may reduce capacity of plants to acquire iron from alkaline soils by inhibiting phytosiderophore release.

Genotypic variability within Tunisian grapevine varieties (Vitis vinifera L.) facing bicarbonate-induced iron deficiency.

Morpho-physiological responses to bicarbonate-induced Fe deficiency were investigated in five Vitis vinifera L. Tunisian varieties (Khamri, Blanc3, Arich Dressé, Beldi, and Balta4). One-month-old woody cuttings were cultivated for 85days on a free calcareous soil irrigated with tap water containing increasing bicarbonate levels (0, 4, 8, 12, and 16mM NaHCO(3)). After this screening, a second experiment compared root biochemical responses of two contrasting genotypes (tolerant-sensitive) dealing with bicarbonate-induced iron deprivation (20microM Fe+/-10mM HCO(3)(-)) for 75days. Using morpho-physiological criteria, grapevine tolerance to HCO(3)(-)-induced Fe shortage appeared to be genotype-dependent: Balta4 and Beldi varieties showed the highest leaf-chlorosis score (especially at the extreme HCO(3)(-) levels), in contrast to Khamri variety. Growth parameters (shoot height, total leaf area, leaf number, and biomass production) as well as juvenile leaf chlorophyll content were also differently affected depending on both genotype and bicarbonate dose. At 16mM HCO(3)(-), Khamri was the less sensitive variety, contrasting with Balta4. On the other hand, chlorophyll content correlated positively with HCl-extractible Fe content of the juvenile leaves, suggesting that the grapevine response to iron deficiency may partly depend on to the plant ability to adequately supply young leaves with this element. Root biochemical responses revealed a relatively higher root acidification capacity in Khamri (tolerant) under Fe-deficiency while no significant changes occurred in Balta4 (sensitive). In addition, Fe(III)-reductase and phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) activities were strongly stimulated by Fe-deficiency in Khamri, while remaining constant in Balta4. These findings suggest that biochemical parameters may constitute reliable criteria for the selection of tolerant grapevine genotypes to iron chlorosis.

Differential tolerance to iron deficiency of chickpea varieties and Fe resupply effects.

The consequences of direct iron deficiency and iron resupply were evaluated during development stages of two Tunisian chickpea varieties (INRAT88 and Chetoui) cultivated in continuously aerated solution with or without 20 muM Fe. The chlorosis score was estimated during culture. Growth parameters, chlorophyll concentration, acidification capacity and Fe concentration were measured every three days during the 21-day treatment. After three weeks of treatment, the chlorosis index was 3-fold higher in Chetoui than in INRAT88, and a considerable decrease of chlorophyll concentration was observed in Chetoui plants since the 6th day of -Fe deprivation. Iron deficiency significantly inhibited whole-plant biomass deposition in both varieties. However, the growth reduction appeared earlier, and was more pronounced in Chetoui than in INRAT88. The whole-plant Fe content decreased dramatically under deficient conditions, and we note an Fe enrichment in shoots at the expense of roots. The sensitivity of Chetoui as compared to INRAT88 was confirmed by the behaviour of resupplied (-Fe/+Fe) plants. In fact, the addition of iron to deficient plants had no significant effect in Chetoui, whereas it led to a total recovery in INRAT88. The capacity of INRAT88 to maintain plant growth and to preserve adequate chlorophyll synthesis under limited iron conditions is related to its better Fe-use efficiency, in addition to its capacity to rapidly recover from this stress.

Differences in responses to iron deficiency between two legumes: lentil (Lens culinaris) and chickpea (Cicer arietinum).

Two legumes, lentil and chickpea, were cultivated in nutrient solutions: Fe lacking or containing 30 microM Fe. After 12 days of Fe starvation, lentil showed a severe yellowing of young leaves, a large decrease in chlorophyll concentration, and a significant decline of plant biomass. Chickpea showed a better response than did lentil, primarily due to a stronger acidification capacity. In addition, no chlorosis symptoms were observed in chickpea until the end of treatment. There was no significant difference in potassium uptake between the two species, but an enrichment of the young leaves at the expense of the old ones was noted in chickpea, and at a lesser extent, in lentil, when they were exposed to Fe deficiency. Moreover, this constraint led to a significant decrease of iron content in the two legumes. However, chickpea displayed higher accumulation levels of HCl-extractible iron in young and old leaves than did lentil. This protection of young leaves against K(+) and Fe(2+) impoverishment confers to these organs the capacity to preserve their chlorophyll status and their photosynthetic integrity. Furthermore, the better performance of chickpea under conditions of low Fe availability could be partially related to its seed iron reserves, higher than those of lentil.

Physiological responses of Tunisian grapevine varieties to bicarbonate-induced iron deficiency.

Plants are frequently submitted to iron deficiency when growing on calcareous soils, which contain high concentrations of bicarbonate. The purpose of this study was to investigate the variability of physiological responses of Tunisian grapevine varieties to bicarbonate-induced iron chlorosis. Vine woodcuttings of seven autochthonous Tunisian varieties (Khamri, Mahdaoui, Blan3, Saouadi, Arich Dressé, Beldi and Balta4), two rootstocks (140Ru and S.O.4), and an introduced table variety (Cardinal) were cultivated on inert sand for 2 months using a complete nutrient solution (20 microM Fe) that was either well supplied or not supplied with 10 mM HCO3-. Young leaves of plants cultivated on bicarbonate-enriched medium showed characteristic symptoms of iron chlorosis, although the intensity of the symptoms depended on the variety and the rootstock. Chlorosis score confirmed these observations since the most sensitive varieties showed the highest values. This variability in tolerance to iron deficiency was also displayed when analysing the physiological parameters (shoot length, plant dry weight, and chlorophyll concentration) and the iron contents in the 4th leaf. Analysis of morphological and physiological parameters showed three behaviour groups. The first one corresponded to tolerant varieties (Khamri, Mahdaoui, and the root-stock: 140Ru), the second included moderately tolerant vines (Saouadi, Arich Dressé, Blanc3, and the rootstock: S.O.4) and the third represented the sensitive ones (Balta4, Beldi, and Cardinal).