Rhizosphere Acidification as the Main Trait Characterizing the Differential In Vitro Tolerance to Iron Chlorosis in Interspecific Pyrus Hybrids

Abstract

Physiological responses of different interspecific Pyrus hybrids and an open pollinated Pyrus communis ‘Williams’ (Pcw) grown under in vitro culture conditions simulating lime induced chlorosis were studied. The hybrids were derived from crosses between the ‘Pyriam’ pear rootstock and four Pyrus species of the Mediterranean region, namely P. amygdaliformis Vill. (Pa), P. amygdaliformis persica Bornme. (Pap), P. communis cordata (Desv.) Hook. (Pcc), and P. elaeagrifolia Pall (Pe), all known for their higher field tolerance to iron-chlorosis than P. communis. Twenty hybrids and one open pollinated Pcw were micropropagated, and plantlets were in vitro characterized for their physiological responses to iron-deficiency conditions. Rooted plantlets were transferred to a culture medium with 2 µM Fe3+ DTPA and 10 or 20 mM NaHCO3. These physiological responses were scored at 1, 3, 7, and 28 days from the start of the in vitro assay. Leaf total chlorophyll content, the capacity of roots to acidify the medium, reduced iron, and exudates of phenolic acids and organic acids were analyzed in each media and time sample. Leaf chlorophyll levels for the clones derived from Pcc were the highest, especially under the highest bicarbonate concentration, followed by those derived from Pap and Pa. The higher chlorophyll content of Pcc clones were related with their higher capacity to acidify the media but not with their iron reduction capacity at the root level. On the other hand, hybrid clones derived from Pe showed a higher Fe3+ reduction ability than clones from all the other species during the whole assay but only when the bicarbonate concentration was lower. The exudation of phenolic acids by the roots was higher in Pcw than in the other species, and this response might explain why the total chlorophyll levels in Pcw clones are similar to those of Pe and Pa ones. These results with Pyrus spp. bring more evidence in support of the idea that iron reduction capacity at the root level is not directly related with a higher tolerance to iron deficiency caused by the high pH of calcareous soils. Instead, the ability to acidify the rhizosphere is the trait of choice for the selection of the pear hybrid clones better adapted to lime induced chlorosis. In addition, the in vitro assay to select the Pyrus clones for tolerance to iron chlorosis could be shortened to one week of culture in 10 mM NaHCO3, measuring the leaf chlorophyll level, acidification of the culture medium, and exudation of phenolic acids as the physiological responses to predict tolerance to lime-induced chlorosis.

GreenTray®, a TIS Bioreactor for Plant Micropropagation and Abiotic or Biotic Stress Bioassays

“…The differential capacities of the Prunus rootstocks Garnem ® and Rootpac ® 20 to grow in alkaline conditions and Ca 2+ depletion, reported herein, were in accordance with what is known from agronomic trials in calcareous soils [56,57]. Therefore, GreenTray ® could be useful for many nutritional studies, including the characterization and selection of plant materials [36].…”

“…Five Prunus hybrid rootstocks, namely Rootpac ® 20, Rootpac ® R (obtained by Agromillora) [33], Intensia ® (obtained by IRTA), Garnem ® (obtained by SIA-DGA), and GF667 (obtained by INRA); Adara, a plum rootstock (obtained by EEAD-CSIC); two Pyrus hybrid rootstocks, namely OHF87 TM (obtained by OSU) and Py170 , which is a hybrid between "OH11" [34] and Pyrus amygdaliformis, in the last phase of selection of the IRTA breeding program, oriented to have tolerance to iron chlorosis and reduced vigor [35,36]; three apple varieties, namely Carrandona (M0676), Florina (M0216), and Rozona (M0163) (collection of SERIDA) [37]; the Cannabis sativa variety "Ferimon" (obtained by HEMPit); and propagules of Kalanchoe daigremontiana were used for the present work (Table 1). Culture in the GreenTray ® was conducted in liquid Murashige and Skoog (MS) medium, supplemented with 30 g/L sucrose, 5 µM 6-Benzylaminopurine (BAP), and the pH adjusted to 4.7 before autoclaving [38].…”

GreenTray^®, a TIS Bioreactor for Plant Micropropagation and Abiotic or Biotic Stress Bioassays

“…The differential capacities of the Prunus rootstocks Garnem ® and Rootpac ® 20 to grow under alkaline and Ca2+ depletion, reported herein, were in accordance with what is known from agronomic trials in calcareous soils [57,58]. Therefore, GreenTray ® could be useful for many nutritional studies, including characterization and selection of plant materials [36].…”

“…Five Prunus hybrid rootstocks, namely Rootpac ® 20, Rootpac ® R (obtained by Agromillora) [34], Intensia ® (obtained by IRTA), Garnem ® (obtained by SIA-DGA), and GF667 (obtained by INRA); Adara, a plum rootstock (obtained by EEAD-CSIC); two Pyrus hybrid rootstocks, namely OHF87 TM (obtained by OSU) and Py170 , which is an hybrid between "OH11" [1] and Pyrus amygdaliformis, in the last phase of selection in IRTA breeding program, oriented to have tolerance to iron chlorosis and reduced vigor [35,36]; three apple varieties, namely Carrandona (M0676), Florina (M0216), and Rozona (M0163) (collection of SERIDA) [37]; the Cannabis sativa variety "Ferimon" (obtained by HEMPit), and propagules of Kalanchoe daigremontiana, were used for the present work.…”

The activation of iron deficiency responses of grapevine rootstocks is dependent to the availability of the nitrogen forms

“…Additionally, in various plant species exposed to Fe deficiency, increasing biosynthesis of organic acids in root tips has been shown as a strategy to mobilize Fe and increase nutrient acquisition by plants [ 10 , 84 , 85 ]. Indeed, also the values obtained in this study varied between rootstocks, while 3309 C considerably increased the accumulation of all organic acids in root tips with Fe deficiency, an interaction for the factors rootstock and iron availability as determined by Tow-Way ANOVA was only obtained for malic and citric acid for 3309 C (malic acid: F(1, 23) = 6.158, p = 0.022; citric acid: F(1, 23) = 9.766, p = 0.005,), while no interaction was determined for Fercal.…”