Phytic acid is considered as one of the major antinutritional compounds in cereal and legume seeds. The development of lpa (low phytic acid) grains, resulting in increased mineral cation availability, is considered a major goal in the improvement of the nutritional quality of seed crops, especially those largely consumed in developing countries. From a mutagenized population of common bean we isolated a homozygous lpa mutant line (lpa-280-10) showing, compared to wild type, a 90% reduction of phytic acid, a 25% reduction of raffinosaccharides and a much higher amount of free or weakly bound iron cations in the seed. Genetic analysis showed that the lpa character is due to a recessive mutation that segregates in a monogenic, Mendelian fashion. Germination tests performed using varying ageing or stress conditions, clearly showed that the bean line lpa-280-10 has a better germination response than the wild type. These data, together with those obtained from 2 years of agronomic trials showing that the mutant seed yield is close to that of its parents and other evidence, indicate that the new lpa-280-10 mutation might be the first devoid of visible macroscopic negative effects in plants, pods and seeds.
Key words: inositol 1,4,5-tris-phosphate kinase, low phytic acid, myo-inositol-1,2,3,4,5,6-hexakisphosphate (InsP 6 ), myo-inositol-phosphate monophosphatase (IMP), myo-inositol-3-phosphate synthase (MIPS), multidrug resistance protein (MRP) type ATP-binding cassette (ABC) transporter, Phaseolus vulgaris (common bean). Summary• We previously identified the lpa1 (low phytic acid) 280-10 line that carries a mutation conferring a 90% reduction in phytic acid (InsP 6 ) content. In contrast to other lpa mutants, lpa1(280-10) does not display negative pleiotropic effects. In the present paper, we have identified the mutated gene and analysed its impact on the phytic acid pathway.• Here, we mapped the lpa1(280-10) mutation by bulk analysis on a segregating F 2 population, an then, by comparison with the soybean genome, we identified and sequenced a candidate gene. The InsP 6 pathway was analysed by gene expression and quantification of metabolites.• The mutated Pvmrp1(280-10) cosegregates with the lpa1(280-10) mutation, and the expression level of several genes of the InsP 6 pathway are reduced in the lpa1(280-10) mutant as well as the inositol and raffinosaccharide content. PvMrp2, a very similar paralogue of PvMrp1 was also mapped and sequenced.• The lpa1 mutation in beans is likely the result of a defective Mrp1 gene (orthologous to the lpa genes AtMRP5 and ZmMRP4), while its Mrp2 paralog is not able to complement the mutant phenotype in the seed. This mutation appears to down-regulate the InsP 6 pathway at the transcriptional level, as well as altering inositol-related metabolism and affecting ABA sensitivity.Abbreviations: ABC, ATP-binding cassette; MRP, multidrug resistance-associated protein; InsP 6, myo-inositol-1,2,3,4,5,6-hexakisphosphate; IMP, myo-inositolphosphate monophosphatase; MIPS, myo-inositol-3-phosphate synthase; MIK, myo-inositol kinase; IPK2, inositol 1,4,5-tris-phosphate kinase; ITPK, inositol 1,3,4-triphosphate 5 ⁄ 6-kinase; IPK1, inositol 1,3,4,5,6 pentakisphosphate 2-kinase.
Iron bioavailability from common beans is negatively influenced by phytic acid (PA) and polyphenols (PPs). Newly developed low-PA (lpa) beans with 90% less PA and variable PPs might improve iron bioavailability. The aim of this study was to evaluate the influence of lpa beans on iron bioavailability in women (n = 20). We compared iron absorption from 4 different beans using a paired, double meal, crossover design. Iron absorption was measured as erythrocyte incorporation of stable iron isotopes (Fe(57), Fe(58)) from 2 lpa bean lines, one high in PPs (means ± SDs; PA = 124 ± 10 mg/100 g; PPs = 462 ± 25 mg/100 g) and one low in PPs (PA = 70 ± 10 mg/100 g; PPs = 54 ± 2 mg/100 g). The other 2 beans used were their parents with a normal PA concentration, one high in PPs (PA = 1030 ± 30 mg/100 g; PPs = 676 ± 19 mg/100 g) and one low in PPs (PA = 1360 ± 10 mg/100 g; PPs = 58 ± 1 mg/100 g). Fractional iron absorption from the lpa bean high in PPs was 6.1% (95% CI: 2.6, 14.7), which was 60 and 130% higher compared with the parent high in PPs (P < 0.001) and low in PPs (P < 0.001), respectively. The total amount of iron absorbed per test meal from the lpa bean high in PPs (372 μg; 95% CI: 160, 890) was 60 and 163% higher compared with the parent high in PPs (P < 0.001) and low in PPs (P < 0.001), respectively. Fractional iron absorption from the lpa line low in PPs (4%; 95% CI: 1.8, 8.7) was 50% higher and the total amount of iron absorbed per test meal (261 μg; 95% CI: 120, 570) was 85% higher than iron from the parent low in PPs (P < 0.001). There was no difference between the lpa beans high or low in PPs or between the parents high or low in PPs. A 90% reduction in PA leads to an increase in bioavailable iron from beans, independent of the PP concentration. The lpa mutation could be a key tool for improving iron bioavailability from beans.
We introduced into Rhizobium leguminosarum bv. viciae LPR1105 a new pathway for the biosynthesis of the auxin, indole-3-acetic acid (IAA), under the control of a stationary phase-activated promoter active both in free-living bacteria and bacteroids. The newly introduced genes are the iaaM gene from Pseudomonas savastanoi and the tms2 gene from Agrobacterium tumefaciens. Free-living bacteria harbouring the promoter-iaaMtms2 construct release into the growth medium 14-fold more IAA than the wild-type parental strain. This IAA overproducing R. l. viciae, the RD20 strain, elicits the development of vetch root nodules containing up to 60-fold more IAA than nodules infected by the wild-type strain LPR1105. Vetch root nodules derived from RD20 are fewer in number per plant, heavier in terms of dry weight and show an enlarged and more active meristem. A significant increase in acetylene reduction activity was measured in nodules elicited in vetch by RD20.
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