Effect of phosphorus limiting on phytase activity, proton efflux and oxygen consumption by nodulatedroots of common bean (Phaseolus vulgaris)

This work intended to measure the nodulated-roots oxygen consumption, proton efflux and phytase activity in 2 lines of common bean (Phaseolus vulgaris) (115, 147) at 2 levels of P supply. Rooted seedlings were inoculated with Rhizobium tropici CIAT 899 in hydroaeroponic cultivation under glasshouse. Phosphorus was supplied as KH2PO4 at 15 and 250 ìmol pl-1 week-1 (15P and 250P, respectively). Our results showed that plant growth nodulation and symbiotic nitrogen fixation were significantly affected by P limiting (15P) for the both lines, but this adverse effect was more pronounced in 147 than in 115. For the both lines, the phytase activity, higher in roots than in nodules, was significantly increased by P limiting, but 115 maintained higher values as compared to 147 line. Incotyledons, the phytase activity was higher in 115 than in 147. Phosphorus shortage increased the cumulated proton release only in 115, whereas it was lowered for 147. In this line, the proton release was linked to symbiotic nitrogen fixation. Under 15P, the proton efflux per unit of nodulated-root biomass was 25% greater for 115 than 147, suggesting that under P limitation, proton efflux may constitute an efficient way to increase P uptake in the tolerant line (115). 15P increased significantlynodulated-root O2 consumption per g nodule DW and nodule conductance, but to a higher extent in 147. As a whole, bean plants at P-deficient conditions increased the activity of phytases and proton efflux, thus maintaining the oxygen diffusion in nodules. This may represent an adaptive mechanism for N2- fixing legumes to respond to P deficiency, by increasing the utilisation and the uptake of phosphorus for symbiotic nitrogen fixation

Identification of a Phytase Gene in Barley (Hordeum vulgare L.)

Background: Endogenous phytase plays a crucial role in phytate degradation and is thus closely related to nutrient efficiency in barley products. The understanding of genetic information of phytase in barley can provide a useful tool for breeding new barley varieties with high phytase activity. Methodology/Principal Findings: Quantitative trait loci (QTL) analysis for phytase activity was conducted using a doubled haploid population. Phytase protein was purified and identified by the LC-ESI MS/MS Shotgun method. Purple acid phosphatase (PAP) gene was sequenced and the position was compared with the QTL controlling phytase activity. A major QTL for phytase activity was mapped to chromosome 5 H in barley. The gene controlling phytase activity in the region was named as mqPhy. The gene HvPAP a was mapped to the same position as mqPhy, supporting the colinearity between HvPAP a and mqPhy. Conclusions/Significance: It is the first report on QTLs for phytase activity and the results showed that HvPAP a, which shares a same position with the QTL, is a major phytase gene in barley grains

Root Proliferation, Proton Efflux, and Acid Phosphatase Activity in Common Bean (Phaseolus vulgaris) Under Phosphorus Shortage

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699International audienceThe impact of phosphorus (P) availability on root proliferation, proton efflux, and acid phosphatase activities in roots and leaves was investigated in two lines of common bean (Phaseolus vulgaris): BAT 477 and CocoT. Phosphorus was supplied as KH(2)PO(4) at 0 and 60 A mu mol per plant (0P and 60P, respectively). Under P shortage, the plant growth was more restricted in CocoT than in BAT 477, shoots being more affected than roots. The root area increased significantly at 0P in both lines. Up to 1 week following P shortage, the proton efflux increased in both lines despite a higher extent in BAT 477 as compared to CocoT. Root acid phosphatase activity was significantly higher under P limitation in the both lines, this trend being more pronounced in BAT 477 than in CocoT. This was also true for the leaf acid phosphatase. Regardless of the bean line, higher values were recorded for the old leaves as compared to the young ones for this parameter. Interestingly, a significant correlation between Pi content in old leaves and their acid phosphatase activity was found in P-lacking (0P) plants of the both bean lines, suggesting that acid phosphatase may contribute to increase the phosphorus use efficiency in bean through the P remobilization from the old leaves. As a whole, our results highlight the significance of the root H(+) extrusion and the acid phosphatase activity rather than the root proliferation in the relative tolerance of BAT 477 to severe P deficiency

Do nodule phosphatase and phytase link with the phosphorus use efficiency for n-2-dependent growth in Phaseolus vulgaris?

International audienceGrain legumes can contribute to cropping systems through their ability to fix atmospheric N2 in their root-nodules. However, the symbiotic nitrogen fixation process requiadditional phosphorus. Thus, the production of grain legumes, particularly common bean, is limited by P-deficiency in many soils, mostly in tropical and Mediterranean areas. Recombinant inbred lines (RILs — F8) from a cross of common bean parents BAT477 with DOR364 were selected for improved phosphorus-use efficiency and N2-dependent growth in glasshouse hydro-aeroponic culture and tested in farmers’ field. The present work with Phaseolus vulgarisas a model grain legume shows a significant difference in the overall phosphatase activity in nodules, including a relatively high level of phytase activity, between two recombinant inbred lines that contrast in their efficiency in utilization of P for symbiotic nitrogen fixation

Metabolic changes of iron uptake in N2_fixing common bean nodules during iron deficiency

Iron is an important nutrient in N 2-fixing legume nodules. The demand for this micronutrient increases during the symbiosis establishment, where the metal is utilized for the synthesis of various iron-containing proteins in both the plant and the bacteroid. Unfortunately, in spite of its importance, iron is poorly available to plant uptake since its solubility is very low when in its oxidized form Fe(III). In the present study, the effect of iron deficiency on the activity of some proteins involved in Strategy I response, such as Fe-chelate reductase (FC-R), H +-ATPase, and phosphoenolpyruvate carboxylase (PEPC) and the protein level of iron regulated transporter (IRT1) and H +-ATPase proteins has been investigated in both roots and nodules of a tolerant (Flamingo) and a susceptible (Coco blanc) cultivar of common bean plants. The main results of this study show that the symbiotic tolerance of Flamingo can be ascribed to a greater increase in the FC-R and H +-ATPase activities in both roots and nodules, leading to a more efficient Fe supply to nodulating tissues. The strong increase in PEPC activity and organic acid content, in the Flamingo root nodules, suggests that under iron deficiency nodules can modify their metabolism in order to sustain those activities necessary to acquire Fe directly from the soil solution

A novel method based on combination of semi-in vitro and in vivo conditions in Agrobacterium rhizogenes-mediated hairy root transformation of Glycine species

Despite numerous advantages of the many tissue culture-independent hairy root transformation protocols, the process is often compromised in the initial in vitro culture stage where inability to maintain high humidity and the delivery of nourishing culture medium decrease cellular morphogenesis and organ formation efficiency. Ultimately, this influences the effective transfer of produced plantlets during transfer from in vitro to in vivo conditions, where low survival rates occur during the acclimation period. We have developed an intermediate protocol for Agrobacterium rhizogenes transformation in Glycine species by combining a two-step in vitro and in vivo process that greatly enhances the efficiency of hairy root formation and which simplifies the maintenance of the transformed roots. In this protocol, cotyledonary nodes of Glycine max and Glycine canescens seedlings were infected by A. rhizogenes K599 carrying a reporter gene construct constitutively expressing green fluorescent protein (GFP). Glass containers containing sand and nutrient solution were employed to provide a moist clean microenvironment for the generation of hairy roots from inoculated seedlings. Transgenic roots were then noninvasively identified from nontransgenic roots based on the detection of GFP. Main roots and nontransgenic roots were removed leaving transgenic hairy roots to support seedling development, all within 1 mo of beginning the experiment. Overall, this protocol increased the transformation efficiency by more than twofold over traditional methods. Approximately 88% and 100% of infected plants developed hairy roots from G. max and G. canescens, respectively. On average, each infected plant produced 10.9 transformed hairy roots in G. max and 13–20 in G. canescens. Introduction of this simple protocol is a significant advance that eliminates the long and genotype-dependent tissue culture procedure while taking advantage of its optimum in vitro qualities to enhance the micropropagation rate. This research will support the increasing use of transient transgenic hairy roots for the study of plant root biology and symbiotic interactions with Rhizobium spp.Manijeh Mohammadi-Dehcheshmeh, Esmaeil Ebrahimie, Stephen D. Tyerman, Brent N. Kaise