Biofortification of rice with zinc: assessment of the relative bioavailability of zinc in a Caco-2 cell model and suckling rat pups

Staple foods, such as rice, can now be enriched in micronutrients through conventional breeding (i.e., biofortification) to enhance dietary intake of vulnerable populations. The objectives of this study were (1) to establish a rapid, high capacity Caco-2 cell model to determine the relative bioavailability of zinc (Zn) from samples of staple food breeding lines for potential use as a guideline for selection/breeding and (2) to determine the relative bioavailability of Zn from conventional rice varieties and one Zn-biofortified type. Polished or undermilled, parboiled rice samples were digested in vitro with pepsin and pH adjustment, and by pancreatic enzymes. Zn uptake from digested samples was measured in Caco-2 cells in culture. A previously validated rat pup model was also used to assess Zn absorption in vivo, using gastric intubation and (65)Zn labeling. Pups were killed after 6 h, and radioactivity in tissues and in small intestine perfusate and cecum-colon contents was used to measure Zn bioavailability. A biofortified rice variety contained substantially more Zn than conventional varieties, with no change in phytate content. Absorbed Zn (μg/g rice) was significantly higher from the new variety in both the in vitro Caco-2 cell model (2.1-fold) and the rat pup model (2.0-fold). Results from the two models were highly correlated, particularly for the polished samples. Biofortification of rice with Zn results in significantly increased Zn uptake in both models. Since results from the Caco-2 cell model correlated well with those from rat pups, this cell model is likely to predict results in human populations and can be used for screening purposes.     

Assessment of iron bioavailability in whole wheat bread by addition of phytase-producing bifidobacteria

In this study, the influence of phytase-producing Bifidobacterium strains during the breadmaking process (direct or indirect) on final bread Fe dialyzability and ferritin formation in Caco-2 cell as a measure of cell Fe uptake was assessed. The addition of bifidobacteria significantly reduced the InsP(6) + InsP(5) concentrations compared to control samples. Fe-dialyzable contents for samples with bifidobacteria were increased 2.3-5.6-fold, and dialyzability was improved by 2.6-8.6% compared to controls. However, this was not reflected in an increase of Fe uptake by Caco-2 cells as was predicted by the phytate/Fe molar ratios. The results demonstrated the usefulness of phytase-producing bifidobacteria to reduce phytate during the breadmaking process and to increase Fe accessibility, although the effects appeared to be still insufficient to improve Fe bioavailability in Caco-2 cells. Further refinement of the use of phytase-producing bifidobacterial strains and/or breadmaking technological processes is deserved for improving Fe uptake.     

Genetic modification of low phytic acid 1-1 maize to enhance iron content and bioavailability

High phytate content in staple food crops is a major barrier to successful iron biofortification. We have exploited the low phytic acid 1-1 (lpa1-1) mutant of maize to generate transgenic plants with up-to 70 μg/g seed iron through the endosperm-specific overexpression of soybean ferritin, resulting in more than 2-fold improvement in iron bioavailability. The levels of bioavailable seed iron achieved in this study greatly exceed any achieved thus far and closely approach values estimated to have a nutritional impact on target populations. Gene expression studies reveal a large induction of the YS1 transporter in leaves and severe repression of an iron acquisition gene DMAS1 in roots, suggesting significant alterations in the iron homeostatic mechanisms in transgenic lpa1-1. Furthermore, preliminary tests show that the high-iron lpa1-1 seeds have higher germination rates and seedling vigor when compared to those of the nontransgenic seeds, which may help improve their value to plant breeders.     

Fermentation and lactic acid addition enhance iron bioavailability of maize

Maize is one of the most important cereal crops for human consumption, yet it is of concern due to its low iron bioavailability. The objective of this study was to determine the effects of processing on iron bioavailability in common maize products and elucidate better processing techniques for enhancing iron bioavailability. Maize products were processed to represent different processing techniques: heating (porridge), fermentation (ogi), nixtamalization (tortillas), and decortication (arepas). Iron and phytate contents were evaluated. Iron bioavailability was assessed using the Caco-2 cell model. Phytate content of maize products was significantly reduced by decortication (25.6%, p = 0.003) and nixtamalization (15%, p = 0.03), and iron content was reduced by decortication (29.1%, p = 0.002). The relative bioavailability (RBA, compared to 100% bioavailability of porridge with FeSO4) of ogi was significantly higher than that of other products when fortified with FeSO4 (p < 0.001) or reduced iron (p < 0.001). Addition of lactic acid (6 mg/g of maize) significantly increased iron solubility and increased bioavailability by about 2-fold (p < 0.01), especially in tortillas. The consumer panel results showed that lactic acid addition does not significantly affect the organoleptic characteristics of tortillas and arepas (p = 0.166 and 0.831, respectively). The results suggest that fermentation, or the addition of small amounts of lactic acid to unfermented maize products, may significantly improve iron bioavailability. Lactic acid addition may be more feasible than the addition of highly bioavailable but expensive fortificants. This approach may be a novel means to increase the iron bioavailability of maize products to reduce the incidence of iron deficiency anemia.     

Screening of iron bioavailability patterns in eight bean (Phaseolus vulgaris L.) genotypes using the Caco-2 cell in vitro model

The common bean ( Phaseolus vulgaris) is an important staple plant food in the diets of people of Latin America, East Africa,and other regions of the developing world. It is also a major source of dietary iron. The primary goal of this research was to use an in vitro digestion/Caco-2 model to study iron bioavailability in eight genotypes (three Mesoamerican and five Andean) that represent the diversity of grain types in this crop. Complementing this goal, we measured the distribution of both iron and phytate in different bean grain tissues (cotyledon, seed coats, and embryos). Seed coats were confirmed to be the exclusive tissue containing polyphenols. The removal of the seed coat and associated polyphenols improved Caco-2 iron bioavailability, and significant differences were observed between genotypes. The addition of ascorbate enhanced iron bioavailability and exposed additional differences in Fe availability among the genotypes. These results indicate that iron accumulation and in vitro iron bioavailability vary among bean genotypes and that polyphenols had greater inhibitory effects on Caco-2 iron bioavailability as compared to phytate.     

Zinc bioavailability response curvature in wheat grains under incremental zinc applications

Zinc application is generally recommended to enrich wheat grains with Zn; however, its influence on Zn bioavailability to humans has not received appreciable attention from scientists. In this pot experiment, seven Zn rates (from 0 to 18 mg kg^?1 soil) were applied to two wheat cultivars (Shafaq-2006 and Auqab-2000). Application of Zn significantly increased grain yield, grain Zn concentration and estimated Zn bioavailability, and significantly decreased grain phytate concentration and [phytate]:[Zn] ratio in wheat grains. The response of grain yield to Zn application was quadratic, whereas maximum grain yield was estimated to be achieved at 10.8 mg Zn kg^?1 soil for Shafaq-2006 and 7.4 mg Zn kg^?1 soil for Auqab-2000. These estimated Zn rates were suitable for increasing grain Zn concentration and Zn bioavailability (>2.9 mg Zn in 300 g grains) to optimum levels required for better human nutrition. Conclusively, Zn fertilization for Zn biofortification may be practiced on the bases of response curve studies aimed at maximizing grain yield and optimum Zn bioavailability. Moreover, additive Zn application progressively reduced the grain Fe concentration and increased the grain [phytate]:[Fe] ratio. However, a medium Zn application rate increased grain Ca concentration and decreased the grain [phytate]:[Ca] ratio. Hence, rate of Zn application for mineral biofortification needs to be carefully selected.     

Phytic acid,iron and zinc content in wheat ploidy levels and amphiploids: the impact of genotype and planting seasons

Micronutrient deficiency is one of the most common and widespread nutritional issues. Among the factors mitigating the bioavailability of Zn (zinc) and Fe (iron), phytic acid plays a key role; therefore, in order to scrutinize genetic alterations ?related to micronutrient and phytate contents, we examined the concentrations of zinc, iron, and phytic acid, as well as its mole ratio to ?zinc in various wheat species grown in two planting seasons. The concentrations of phytic acid and its mole ratio to zinc were 0.61?1.55 g kg^?1 dry weight and 1.88?4.17 for autumn, and 0.97?2.02 g kg^?1 dry weight and 2.10?4.05 for spring planting. There was a significant discrepancy among wheat species; tritipyrum had the highest concentration of iron, phytic acid and its mole ratio to zinc, and T. monococcum and T. aestivum recorded reasonable zinc bioavailability. Correlation studies between grain phytic acid concentrations and other measured traits revealed various relationships, denoting an irrefutable impact of planting season and wheat ploidy levels on modification of wheat genotypes. The characters contributing more positively with principal component (PC) 1 were Zn and Fe under spring planting and Fe under autumn planting. Spike number per square meter, biological yield and grain yield in spring cultivation, and grain zinc concentration in autumn cultivation were positively correlated to principal component (PC) 2. Given that the concentration of Fe and Zn in all the studied genotypes is relatively high and due to the existence of other desirable agronomic traits, this study believes that it could possibly enhance the applicability of some of these genotypes for breeding purposes.     

Comparison of iron bioavailability from 15 rice genotypes: studies using an in vitro digestion/caco-2 cell culture model

An in vitro digestion/Caco-2 model was used to compare iron bioavailability from 15 selected Fe-dense and normal genotypes of unpolished rice from the International Rice Research Institute. Iron uptake was determined using Caco-2 cell ferritin formation in response to exposure to a digest of the cooked rice. Iron bioavailabilities from all rice genotypes were ranked as a percent relative to a control variety (Nishiki). Iron concentration in the rice samples ranged from 14 to 39 microg/g. No correlation was observed between Fe uptake and grain-Fe concentration. Furthermore, phytic acid levels were not correlated with Fe bioavailability. Genotypes with low Fe bioavailability (Tong Lan Mo Mi, Zuchein, Heibao, and Xua Bue Nuo) were noticeably more brown to purple in color. The results suggest that certain unknown compounds related to rice grain color may be a major factor limiting Fe bioavailability from unpolished rice.     

Evaluation and utilization of <Emphasis Type="Italic">Aegilops</Emphasis> and wild <Emphasis Type="Italic">Triticum</Emphasis> species for enhancing iron and zinc content in wheat

Grains of 80 accessions of nine species of wild Triticum and Aegilops along with 15 semi-dwarf cultivars of bread and durum wheat grown over 2 years at Indian Institute of Technology, Roorkee, were analyzed for grain iron and zinc content. The bread and durum cultivars had very low content and little variability for both of these micronutrients. The related non-progenitor wild species with S, U and M genomes showed up to 3–4 folds higher iron and zinc content in their grains as compared to bread and durum wheat. For confirmation, two Ae. kotschyi Boiss. accessions were analyzed after ashing and were found to have more than 30% higher grain ash content than the wheat cultivars containing more than 75% higher iron and 60% higher zinc than that of wheat. There were highly significant differences for iron and zinc contents among various cultivars and wild relatives over both the years with very high broad sense heritability. There was a significantly high positive correlation between flag leaf iron and grain iron (r = 0.82) and flag leaf zinc and grain zinc (r = 0.92) content of the selected donors suggesting that the leaf analysis could be used for early selection for high iron and zinc content. ‘Chinese Spring’ (Ph ^I ) was used for inducing homoeologous chromosome pairing between Aegilops and wheat genomes and transferring these useful traits from the wild species to the elite wheat cultivars. A majority of the interspecific hybrids had higher leaf iron and zinc content than their wheat parents and equivalent or higher content than their Aegilops parents suggesting that the parental Aegilops donors possess a more efficient system for uptake and translocation of the micronutrients which could ultimately be utilized for wheat grain biofortification. Partially fertile to sterile BC₁ derivatives with variable chromosomes of Aegilops species had also higher leaf iron and zinc content confirming the possibility of transfer of required variability. Some of the fertile BC₁F₃ and BC₂F₂ derivatives had as high grain ash and grain ash iron and zinc content as that of the donor Aegilops parent. Further work on backcrossing, selfing, selection of fertile derivatives, leaf and grain analyses for iron and zinc for developing biofortified bread and durum wheat cultivars is in progress. Nidhi Rawat, Vijay K. Tiwari, and Neelam Singh have contributed equally to the work.     

Phytate, calcium, iron, and zinc contents and their molar ratios in foods commonly consumed in China

A total of 60 food samples commonly consumed in China were analyzed for phytate using the anion-exchange method and for calcium, iron, and zinc using atomic absorption spectrophotometry. The foods analyzed included those based on cereal grains and soybean. Phytate contents expressed on a wet weight basis ranged from 0 for foods made from starches to 1878 mg/100 g for dried stick-shaped soybean milk film. The calcium contents were between 2.08 mg/100 g for ground corn and 760.67 mg/100 g for diced fried soybean curd. The lowest values of iron and zinc were 0.04 mg/100 g for Panjin pearl rice cooked with discarding extra water and 0.08 mg/100 g for potato and bean starches, while the highest values of iron and zinc were observed in dried stick-shaped soybean milk film. Although many foods were relatively rich in calcium, zinc, and iron, many also contained a higher level of phytate. Of the 60 food samples, 34 foods had a phytate/calcium molar ratio >0.24, 53 foods had a phytate/iron molar ratio >1, 31 foods had a phytate/zinc molar ratio >15, and only 7 foods had a phytate x calcium/zinc >200. Phytate in foods impair the bioavailability of calcium, iron, and zinc, which to some extent depends upon food processing and cooking methods.     

Oxidation of polyphenols in phytate-reduced high-tannin cereals: effect on different phenolic groups and on in vitro accessible iron.

After reduction of phytate with phytase, water slurries of two high-tannin cereal flours were incubated with polyphenol oxidase (mushroom tyrosinase), and the effects on different phenolic groups and on in vitro accessible iron were studied. Enzyme incubation was also performed after cooking, soaking, and germination of the cereals. Phytase incubation significantly decreased the phytate content, and incubation with polyphenol oxidase had a reducing effect on the total phenol content, as well as on the amount of catechol and resorcinol groups. The in vitro accessible iron increased when the cereals were incubated with phytase and polyphenol oxidase, and the highest accessibility of iron was obtained when the germinated samples were incubated. The results from this study imply that oxidation of polyphenols in high-tannin cereals, after reduction of phytate, may be used to increase the bioavailability of iron in foods prepared from these cereals.     

缺铁胁迫对草莓幼苗光合特性及细胞器铁含量的影响

为了探讨缺铁胁迫对草莓（Fragaria ananassa Duch.）幼苗的光合特性及细胞器铁含量的影响,本研究选取4个草莓品种（红颜、 章姬、 甜查理、 童子一号）幼苗,采用溶液培养方法,设置Fe（Ⅱ）-EDTA浓度为0 mol/L、 110-4 mol/L两组处理,分别于处理后0、 4、 8、 12、 16 d对其叶绿素含量（SPAD）、 光合速率（Pn）、 叶绿体铁含量、 根系线粒体铁含量以及叶片铁含量、 根系铁含量、 生物量进行分析。结果表明,缺铁胁迫显著降低草莓幼苗叶绿素含量、 光合速率、 叶绿体铁含量、 叶片铁含量、 根系铁含量、 生物量,并且不同品种间差异达显著水平（P0.05）;缺铁胁迫对根系细胞线粒体铁含量影响较小。草莓的叶绿体铁含量与叶片铁含量、 叶片净光合速率和生物量呈极显著正相关（r=0.93**, r=0.87**, r= 0.72**）, 根系线粒体铁含量与叶片铁含量、 叶片净光合速率和生物量呈极显著正相关或显著正相关（r= 0.83**, r= 0.72**, r= 0.52*）。本试验条件下,供试草莓品种红颜受缺铁胁迫的影响大于其他3个品种。     

Effect of dietary ligands and food matrices on zinc uptake in Caco-2 cells: implications in assessing zinc bioavailability

The kinetics, depletion/repletion of zinc, and effects of dietary ligands/food matrices on (65)Zn uptake was studied in Caco-2 cells. The uptake of zinc showed a saturable and nonsaturable component, depending upon the media zinc concentrations. Intracellular depletion increased zinc uptake, whereas zinc loading did not. Phytic acid and histidine inhibited zinc uptake, while tannic acid, tartaric acid, arginine, and methionine increased zinc uptake. Tannic acid at a 1:50 molar ratio promoted zinc uptake from wheat- and rice-based food matrices. Further, Caco-2 cells responded similarly with zinc and iron uptake when fed Indian bread prepared from low- and high-extraction wheat flour, representing low and high phytate content. However, inclusion of tea extract or red grape juice as a source of polyphenols enhanced the uptake of zinc while decreasing that of iron. These results suggest that the Caco-2 cells predict the correct direction of response to dietary ligands even from complex foods.     

Bran Characteristics and Bread‐Baking Quality of Whole Grain Wheat Flour

Variations in physical and compositional bran characteristics among different sources and classes of wheat and their association with bread‐baking quality of whole grain wheat flour (WWF) were investigated with bran obtained from Quadrumat milling of 12 U.S. wheat varieties and Bühler milling of six Korean wheat varieties. Bran was characterized for composition including protein, fat, ash, dietary fiber, phenolics, and phytate. U.S. soft and club wheat brans were lower in insoluble dietary fiber (IDF) and phytate content (40.7–44.7% and 10.3–17.1 mg of phytate/g of bran, respectively) compared with U.S. hard wheat bran (46.0–51.3% and 16.5–22.2 mg of phytate/g of bran, respectively). Bran of various wheat varieties was blended with a hard red spring wheat flour at a ratio of 1:4 to prepare WWFs for determination of dough properties and bread‐baking quality. WWFs with U.S. hard wheat bran generally exhibited higher dough water absorption and longer dough mixing time, and they produced smaller loaf volume of bread than WWFs of U.S. soft and club wheat bran. WWFs of two U.S. hard wheat varieties (ID3735 and Scarlet) produced much smaller loaves of bread (<573 mL) than those of other U.S. hard wheat varieties (>625 mL). IDF content, phytate content, and water retention capacity of bran exhibited significant relationships with loaf volume of WWF bread, whereas no relationship was observed between protein content of bran and loaf volume of bread. It appears that U.S. soft and club wheat bran, probably owing to relatively low IDF and phytate contents, has smaller negative effects on mixing properties of WWF dough and loaf volume of bread than U.S. hard wheat bran.     

Improving iron bioavailability and nutritional value of maize (Zea mays L.) in sulfur-treated calcareous soil

Soil factors such as pH, calcium, carbonate, and bicarbonate precipitation products in calcareous soils reduce iron (Fe) availability to crops and limit grain Fe concentrations. In the present greenhouse study, we evaluated the potential of Fe fertilizer amendments combined with organic amendments, like biochar (BC) and poultry manure, in sulfur (S)-treated low pH calcareous soils (pH_S1) to assess Fe biofortification of maize. Elemental sulfur (S) was used both for lowering soil pH and Fe solubilization. Soil pH was successfully lowered down from 7.8 to 6.5 by S application at the rate of 2.5 g kg^?1 soil. Pot experiment results revealed that Fe fertilizer combined with BC and S (pH_S1) significantly increased root and shoot dry weight, grain weight, photosynthetic rate, transpiration rate, and stomatal conductance by 69%, 86%, 28%, 74%, 57%, and 33%, respectively, relative to the control. Similarly, combined application of Fe + BC in S-amended (pH_S1) soil increased starch (34%), protein (64%), and fat (1 fold) while antinutrient phytate and polyphenols were decreased up to 29% and 40%, respectively, over control. Regarding the maize nutrients profiles, application of Fe with BC gave the maximum increase of Fe and ferritin was increased 1.7 fold at pH_S1. The results of this study showed that Fe fertilization with BC at pH_S1 soil is beneficial for crop growth and Fe bioavailability.     

In vitro iron availability from iron-fortified whole-grain wheat flour

Iron deficiency is the most common nutritional disorder worldwide. Iron fortification of foods is considered to be the most cost-effective long-term approach to reduce iron deficiency. However, for fortified foods to be effective in reducing iron deficiency, the added iron must be sufficiently bioavailable. In this study, fortification of whole-grain wheat flour with different sources of iron was evaluated in vitro by measuring the amount of dialyzable iron after simulated gastrointestinal digestion of flour baked into chapatis and subsequent intestinal absorption of the released iron using Caco-2 cell layers. The dialyzability of iron from iron-fortified wheat flour was extremely low. Additions of 50 mg/kg iron to the flour in the form of ferrous sulfate, Ferrochel amino acid chelate, ferric amino acid chelate taste free (TF), Lipofer, ferrous lactate, ferrous fumarate, ferric pyrophosphate, carbonyl iron, or electrolytic iron did not significantly increase the amount of in vitro dialyzable iron after simulated gastrointestinal digestion. In contrast, fortification of flour with SunActive Fe or NaFeEDTA resulted in a significant increase in the amount of in vitro dialyzable iron. Relative to iron from ferrous sulfate, iron from SunActive Fe and NaFeEDTA appeared to be 2 and 7 times more available in the in vitro assay, respectively. Caco-2 cell iron absorption from digested chapatis fortified with NaFeEDTA, but not from those fortified with SunActive Fe, was significantly higher than from digested chapatis fortified with ferrous sulfate. On the basis of these results it appears that fortification with NaFeEDTA may result in whole-grain wheat flour that effectively improves the iron status.     

植物源铁生物有效性及评价方法研究进展

缺铁是个世界性的营养失衡问题,给人类健康和经济发展带来严重的负面影响。主要膳食中的铁缺乏或低生物有效性被认为是造成铁缺乏的主要原因。通过植物育种措施,尤其是提高植物源铁富集育种被认为是解决铁营养失衡最经济且有效的途径。然而,近年的研究表明,人体铁吸收与植物源有效铁量密切相关,而与铁积累总量没有相关性。快速、准确的评价植物源铁生物有效性对高有效性富铁作物育种意义重大。本文阐述了植物源铁富集和生物有效性的基因型差异及影响因素, 并分析了铁生物有效性评价方法的优缺点,为植物源铁生物有效性育种及评价提供参考。     

Organic acids influence iron uptake in the human epithelial cell line Caco-2

It has previously been suggested that organic acids enhance iron absorption. We have studied the effect of nine organic acids on the absorption of Fe(II) and Fe(III) in the human epithelial cell line Caco-2. The effect obtained was dose-dependent, and the greatest increase (43-fold) was observed for tartaric acid (4 mmol/L) on Fe(III) (10 micromol/L). Tartaric, malic, succinic, and fumaric acids enhanced Fe(II) and Fe(III) uptake. Citric and oxalic acid, on the other hand, inhibited Fe(II) uptake but enhanced Fe(III) uptake. Propionic and acetic acid increased the Fe(II) uptake, but had no effect on Fe(III) uptake. Our results show a correlation between absorption pattern and chemical structure; e.g. hydroxyl groups, in addition to carboxyls, were connected with a positive influence. The results may be important for elucidating factors affecting iron bioavailability in the small intestine and for the development of foods with improved iron bioavailability.     

Comparison of iron uptake from reduced iron powder and FeSO4 using the Caco-2 cell model: effects of ascorbic acid, phytic acid, and pH

The reduced iron powder has considerable potential for use as an iron fortificant because it does not change organoleptically during storage or food preparation for cereal flour, and its bioavailability is scarcely influenced by iron absorption inhibitors in foods. The objective of this article is to study the effects of ascorbic acid, phytic acid, and pH on iron uptake from reduced iron powder (43 microm) and FeSO 4, and to compare iron bioavailability of reduced iron powders among four selected granularity levels. The cell ferritin formation is used as a marker of iron uptake. Obviously, iron uptake of reduced iron powder is increased with decreasing of powder granularity and is much lower than FeSO 4 when the size is above 43 microm, but significantly higher at 40-60 nm. In the presence of ascorbic acid or phytic acid, Caco-2 cell iron absorption from reduced iron powder (43 microm) is significantly higher than that from FeSO 4. And iron uptake of Caco-2 cells is decreased with increasing of pH from 5.5 to 7.5. Moreover, the decrease trend is more obvious for reduced iron powder than for FeSO 4. Our results indicated that iron bioavailability of reduced iron powder by intestinal enterocytes is similar to that of iron salts, and reduced iron powder is more excellent than FeSO 4 as food fortificant, especially at ultramicroscopic granularity.     

Grain phosphorus and phytate contents of wheat genotypes released during last 6 decades and categorization of selected genotypes for phosphorus use efficiency

Grain phosphorus (P) and phytate contents are important for early growth and quality of wheat, respectively. We studied seed P and phytate contents of 57 wheat genotypes released during last 6 decades. The grain total P and phytate P concentrations ranged from 2.0 to 8.9 mg g^?1 and 1.6 to 4.7 mg g^?1, respectively. Grain P was higher in genotypes released in current or last decade than pre-green revolution period and it correlated well with grain phytate contents. Indicating that breeders worked only for increased grain P and did not consider the phytate contents. Twenty-five genotypes were selected based on differences in grain P and phytate contents to observe the P use efficiency under field conditions. Genotypes differed significantly for grain P and yield at both 0 and 60 kg ha^?1 P levels. In conclusion, phytate fraction in grains of recent wheat genotypes was more as compared to old genotypes and genotype Inqlab-91 and Chakwal-86 had maximum grain total P. Moreover, number of productive tillers, spikelet per spike, grain weight, harvest index and grain P concentration and uptake were found responsible for P tolerance under P deficiency.