Biofortification Traits Research Articles

Validation of quantitative trait loci for biofortification traits and variability research on agro-morphological, physiological, and quality traits in dicoccum wheat (Triticum dicoccum Schrank.)

Validation of quantitative trait loci for biofortification traits and variability research on agro-morphological, physiological, and quality traits in dicoccum wheat (Triticum dicoccum Schrank.)

GGE biplot analysis of biofortification traits in relation to grain yield in landraces of tetraploid wheat (Triticum turgidum ssp. dicoccum)

GGE biplot analysis of biofortification traits in relation to grain yield in landraces of tetraploid wheat (Triticum turgidum ssp. dicoccum)

Multi-Environment Analysis of Biofortification Traits in Pre-bred Bread Wheat (Triticum aestivum L.)

Multi-Environment Analysis of Biofortification Traits in Pre-bred Bread Wheat (Triticum aestivum L.)

Molecular dissection of connected rice populations revealed important genomic regions for agronomic and biofortification traits.

Breeding staple crops with increased micronutrient concentration is a sustainable approach to address micronutrient malnutrition. We carried out Multi-Cross QTL analysis and Inclusive Composite Interval Mapping for 11 agronomic, yield and biofortification traits using four connected RILs populations of rice. Overall, MC-156 QTLs were detected for agronomic (115) and biofortification (41) traits, which were higher in number but smaller in effects compared to single population analysis. The MC-QTL analysis was able to detect important QTLs viz: qZn5.2, qFe7.1, qGY10.1, qDF7.1, qPH1.1, qNT4.1, qPT4.1, qPL1.2, qTGW5.1, qGL3.1 , and qGW6.1 , which can be used in rice genomics assisted breeding. A major QTL (qZn5.2 ) for grain Zn concentration has been detected on chromosome 5 that accounted for 13% of R2. In all, 26 QTL clusters were identified on different chromosomes. qPH6.1 epistatically interacted with qZn5.1 and qGY6.2 . Most of QTLs were co-located with functionally related candidate genes indicating the accuracy of QTL mapping. The genomic region of qZn5.2 was co-located with putative genes such as OsZIP5, OsZIP9, and LOC_OS05G40490 that are involved in Zn uptake. These genes included polymorphic functional SNPs, and their promoter regions were enriched with cis-regulatory elements involved in plant growth and development, and biotic and abiotic stress tolerance. Major effect QTL identified for biofortification and agronomic traits can be utilized in breeding for Zn biofortified rice varieties.

Molecular Mapping of Biofortification Traits in Bread Wheat (Triticum aestivum L.) Using a High-Density SNP Based Linkage Map.

A set of 188 recombinant inbred lines (RILs) derived from a cross between a high-yielding Indian bread wheat cultivar HD2932 and a synthetic hexaploid wheat (SHW) Synthetic 46 derived from tetraploid Triticum turgidum (AA, BB 2n = 28) and diploid Triticum tauschii (DD, 2n = 14) was used to identify novel genomic regions associated in the expression of grain iron concentration (GFeC), grain zinc concentration (GZnC), grain protein content (GPC) and thousand kernel weight (TKW). The RIL population was genotyped using SNPs from 35K Axiom® Wheat Breeder's Array and 34 SSRs and phenotyped in two environments. A total of nine QTLs including five for GPC (QGpc.iari_1B, QGpc.iari_4A, QGpc.iari_4B, QGpc.iari_5D, and QGpc.iari_6B), two for GFeC (QGfec.iari_5B and QGfec.iari_6B), and one each for GZnC (QGznc.iari_7A) and TKW (QTkw.iari_4B) were identified. A total of two stable and co-localized QTLs (QGpc.iari_4B and QTkw.iari_4B) were identified on the 4B chromosome between the flanking region of Xgwm149-AX-94559916. In silico analysis revealed that the key putative candidate genes such as P-loop containing nucleoside triphosphatehydrolase, Nodulin-like protein, NAC domain, Purine permease, Zinc-binding ribosomal protein, Cytochrome P450, Protein phosphatase 2A, Zinc finger CCCH-type, and Kinesin motor domain were located within the identified QTL regions and these putative genes are involved in the regulation of iron homeostasis, zinc transportation, Fe, Zn, and protein remobilization to the developing grain, regulation of grain size and shape, and increased nitrogen use efficiency. The identified novel QTLs, particularly stable and co-localized QTLs are useful for subsequent use in marker-assisted selection (MAS).

Transition From Targeted Breeding to Mainstreaming of Biofortification Traits in Crop Improvement Programs.

Biofortification breeding for three important micronutrients for human health, namely, iron (Fe), zinc (Zn), and provitamin A (PVA), has gained momentum in recent years. HarvestPlus, along with its global consortium partners, enhances Fe, Zn, and PVA in staple crops. The strategic and applied research by HarvestPlus is driven by product-based impact pathway that integrates crop breeding, nutrition research, impact assessment, advocacy, and communication to implement country-specific crop delivery plans. Targeted breeding has resulted in 393 biofortified crop varieties by the end of 2020, which have been released or are in testing in 63 countries, potentially benefitting more than 48 million people. Nevertheless, to reach more than a billion people by 2030, future breeding lines that are being distributed by Consultative Group on International Agricultural Research (CGIAR) centers and submitted by National Agricultural Research System (NARS) to varietal release committees should be biofortified. It is envisaged that the mainstreaming of biofortification traits will be driven by high-throughput micronutrient phenotyping, genomic selection coupled with speed breeding for accelerating genetic gains. It is noteworthy that targeted breeding gradually leads to mainstreaming, as the latter capitalizes on the progress made in the former. Efficacy studies have revealed the nutritional significance of Fe, Zn, and PVA biofortified varieties over non-biofortified ones. Mainstreaming will ensure the integration of biofortified traits into competitive varieties and hybrids developed by private and public sectors. The mainstreaming strategy has just been initiated in select CGIAR centers, namely, International Maize and Wheat Improvement Center (CIMMYT), International Rice Research Institute (IRRI), International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), International Institute of Tropical Agriculture (IITA), and International Center for Tropical Agriculture (CIAT). This review will present the key successes of targeted breeding and its relevance to the mainstreaming approaches to achieve scaling of biofortification to billions sustainably.

Iron, Zinc and Phytic Acid Retention of Biofortified, Low Phytic Acid, and Conventional Bean Varieties When Preparing Common Household Recipes.

Biofortification is an effective method to improve the nutritional content of crops and nutritional intake. Breeding for higher micronutrient mineral content in beans is correlated with an increase in phytic acid, a main inhibitor of mineral absorption in humans. Low phytic acid (lpa) beans have a 90% lower phytic acid content compared to conventional beans. This is the first study to investigate mineral and total phytic acid retention after preparing common household recipes from conventional, biofortified and lpa beans. Mineral retention was determined for two conventional, three biofortified and two lpa bean genotypes. Treatments included soaking, boiling (boiled beans) and refrying (bean paste). The average true retention of iron after boiling was 77.2–91.3%; for zinc 41.2–84.0%; and for phytic acid 49.9–85.9%. Soaking led to a significant decrease in zinc and total phytic acid after boiling and refrying, whereas for iron no significant differences were found. lpa beans did not exhibit a consistent pattern of difference in iron and phytic acid retention compared to the other groups of beans. However, lpa beans had a significantly lower retention of zinc compared to conventional and biofortified varieties (p < 0.05). More research is needed to understand the underlying factors responsible for the differences in retention between the groups of beans, especially the low retention of zinc. Combining the lpa and biofortification traits could further improve the nutritional benefits of biofortified beans, by decreasing the phytic acid:iron and zinc ratio in beans.

Advances in Genomic Interventions for Wheat Biofortification: A Review

Wheat is an essential constituent of cereal-based diets, and one of the most significant sources of calories. However, modern wheat varieties are low in proteins and minerals. Biofortification is a method for increasing the availability of essential elements in the edible portions of crops through agronomic or genetic and genomic interventions. Wheat biofortification, as a research topic, has become increasingly prevalent. Recent accomplishments in genomic biofortification could potentially be helpful for the development of biofortified wheat grains, as a sustainable solution to the issue of “hidden hunger”. Genomic interventions mainly include quantitative trait loci (QTL) mapping, marker-assisted selection (MAS), and genomic selection (GS). Developments in the identification of QTL and in the understanding of the physiological and molecular bases of the QTLs controlling the biofortification traits in wheat have revealed new horizons for the improvement of modern wheat varieties. Markers linked with the QTLs of desirable traits can be identified through QTL mapping, which can be employed for MAS. Besides MAS, a powerful tool, GS, also has great potential for crop improvement. We have compiled information from QTL mapping studies on wheat, carried out for the identification of the QTLs associated with biofortification traits, and have discussed the present status of MAS and different prospects of GS for wheat biofortification. Accelerated mapping studies, as well as MAS and GS schemes, are expected to improve wheat breeding efficiency further.

Genome-Wide Association Mapping in a Rice MAGIC Plus Population Detects QTLs and Genes Useful for Biofortification.

The development of rice genotypes with micronutrient-dense grains and disease resistance is one of the major priorities in rice improvement programs. We conducted Genome-wide association studies (GWAS) using a Multi-parent Advanced Generation Inter-Cross (MAGIC) Plus population to identify QTLs and SNP markers that could potentially be integrated in biofortification and disease resistance breeding. We evaluated 144 MAGIC Plus lines for agronomic and biofortification traits over two locations for two seasons, while disease resistance was screened for one season in the screen house. X-ray fluorescence technology was used to measure grain Fe and Zn concentrations. Genotyping was carried out by genotype by sequencing and a total of 14,242 SNP markers were used in the association analysis. We used Mixed linear model (MLM) with kinship and detected 57 significant genomic regions with a -log10 (P-value) ≥ 3.0. The PH1.1 and Zn7.1 were consistently identified in all the four environments, ten QTLs qDF3.1, qDF6.2 qDF9.1 qPH5.1 qGL3.1, qGW3.1, qGW11.1, and qZn6.2 were detected in two environments, while two major loci qBLB11.1 and qBLB5.1 were identified for Bacterial Leaf Blight (BLB) resistance. The associated SNP markers were found to co-locate with known major genes and QTLs such as OsMADS50 for days to flowering, osGA20ox2 for plant height, and GS3 for grain length. Similarly, Xa4 and xa5 genes were identified for BLB resistance and Pi5(t), Pi28(t), and Pi30(t) genes were identified for Blast resistance. A number of metal homeostasis genes OsMTP6, OsNAS3, OsMT2D, OsVIT1, and OsNRAMP7 were co-located with QTLs for Fe and Zn. The marker-trait relationships from Bayesian network analysis showed consistency with the results of GWAS. A number of promising candidate genes reported in our study can be further validated. We identified several QTLs/genes pyramided lines with high grain Zn and acceptable yield potential, which are a good resource for further evaluation to release as varieties as well as for use in breeding programs.

Identification of genomic regions associated with agronomic and biofortification traits in DH populations of rice

Rice provides energy and nutrition to more than half of the world’s population. Breeding rice varieties with the increased levels of bioavailable micronutrients is one of the most sustainable approaches to tackle micronutrient malnutrition. So, high zinc and iron content in the grain are primary targets in rice biofortification breeding. In this study, we conducted QTL mapping using doubled haploid (DH) populations, PSBRc82 x Joryeongbyeo and PSBRc82 x IR69428, phenotyped for agronomic traits and micronutrients during two growing seasons and using genotypic information from analysis with the 6K SNP chip. A number of DH lines were identified as having high grain Zn and Fe content in polished rice. Importantly, we identified 20 QTLs for agronomic traits and 59 QTLs for a number of biofortification traits. Of the 79 QTLs, 12 were large-effect QTLs (>25% PVE), nine QTLs were consistent across seasons in either population, and one QTL was identified in both populations. Moreover, at least two QTLs were clustered in defined regions of chromosomes 1, 2, 3, 4, 5, 7 and 9. Eight epistatic interactions were detected for Cu, Mg, Na, and Zn in population 1. Furthermore, we identified several candidate genes near QTLs for grain Zn (OsNRAMP, OsNAS, OsZIP, OsYSL, OsFER, and OsZIFL family) and grain yield (OsSPL14 and OsSPL16). These new QTLs and candidate genes help to further elucidate the genetic basis for grain micronutrient concentration, and may prove useful for marker assisted breeding for this important trait.

English

Actinomycetes have been demonstrated for plant growth-promotion, antagonistic against plant pathogens and insect pests and biofortification traits in many agriculturally important crops. The present investigation was conducted to characterize probiotic properties of actinomycete(s) isolated from pearl millet flour and batter samples. A media selective and specific were used for isolation, actinomycetes isolation agar (AIA), and the most prominent actinomycete (found abundantly in the AIA plate) was isolated and maintained on AIA slants at 4°C for further investigation. The most prominent actinomycete was characterized for traits including Gram staining, morphology (such as color, margin, size, shape, elevation, form and surface), biochemistry (such as urease, catalase, oxidase, hydrogen sulphide, nitrogen reduction, gelatin liquefaction, starch hydrolysis and carbohydrate utilization), IMViC tests (such as indole, methyl red, Voges Proskauer and citrate utilization), probiotic potentials (such as acid [pH 2, 3], bile [0.5%], NaCl [6 and 9%], phenol tolerance [0.4%]), antibiotic tolerance (such as tetracycline, streptomycin, kanamycin, chloramphenicol, ciprofloxacin, ampicillin, penicillin, erythromycin and vancomycin) and antimicrobial activities against human pathogens (such as Escherichia coli, Staphylococcus aureus and Salmonella typhi). It was possible to isolate only one probiotic actinomycete based on the properties. The sequences of 16s rDNA gene of the actinomycete was matched with Streptomyces species in BLAST analysis. The sequences of the Streptomyces spp. were submitted to NCBI and accession number obtained. This study indicated that the selected Streptomyces spp. could be used to develop new probiotic foods. Key words: Probiotics, pearl millet, Streptomyces species, product development.

Genetic dissimilarity of common bean lines for agronomic and biofortification traits

The evaluation of the genetic dissimilarity of common bean lines obtained by the breeding program is significant to identify duplicate accessions, to select superior lines, and to plan new hybrid combinations. The objectives of this study were to evaluate the genetic dissimilarity of common bean genotypes for agronomic and mineral biofortification traits, select superior lines, and identify promising hybrid combinations for biofortification programs. For this purpose, 169 common bean genotypes (F6:8 inbred lines and parental cultivars) were evaluated in two experiments conducted in a simple lattice design in Santa Maria-RS, Brazil. The following agronomic traits were evaluated: insertion of the first pod, insertion of the last pod, cycle, mass of 100 grains, grain yield, and concentrations of potassium, phosphorus, zinc, and copper in grains. The genotypes assessed presented genetic variability for all traits, except for copper concentration in grains. The traits that showed the highest contribution to the genetic dissimilarity were the concentrations of phosphorus, potassium, and zinc in grains. In cluster analyses, six groups by principal components and 33 groups by the Tocher optimization method were identified. These methods partially agreed in the formation of different groups of common bean genotypes. Controlled crossings between the lines 17 and 162 with lines of groups XX, XXI, XXIV, and XXXIII are recommended to obtain recombinants with high grain yield and biofortified for phosphorus, potassium, and zinc.

Genetic diversity and selection of common bean lines based on technological quality and biofortification.

The development of common bean cultivars with high technological quality that are biofortified with minerals, is required to meet the demand for food with health benefits. The objectives of this study were to evaluate whether common bean genotypes differ in terms of technological and mineral biofortification traits, to study the correlations between these characters, to analyze the genetic dissimilarity of common bean genotypes, and to select superior lines for these traits. For this, 14 common bean genotypes were evaluated in experiments conducted in three growing seasons in the Rio Grande do Sul State, Brazil. A significant genotype x environment interaction was observed for technological quality (mass of 100 grains and cooking time) and biofortification traits (concentration of potassium, phosphorus, calcium, iron, zinc, and copper). Positive correlation estimates were obtained between phosphorus and potassium (r = 0.575), iron and zinc (r = 0.641), copper and iron (r = 0.729), and copper and phosphorus (r = 0.533). In the main component cluster analysis, four groups of genotypes were formed. The following lines are recommended for selection: LP 11-363 for fast-cooking, CNFC 11 948 for high iron concentration, and LEC 03-14 for high potassium, phosphorus, and calcium concentrations in grains. Common bean lines with high phosphorus and iron concentrations in grains can be indirectly selected based on higher potassium, copper, and zinc concentrations. Controlled crossings between LP 11-363 x CNFC 11 948 and LP 11-363 x LEC 03-14 are recommended to obtain segregating lines that are fast-cooking and biofortified with minerals.

Plant growth-promotion and biofortification of chickpea and pigeonpea through inoculation of biocontrol potential bacteria, isolated from organic soils.

Seven strains of bacteria [Pseudomonas plecoglossicida SRI-156, Brevibacterium antiquum SRI-158, Bacillus altitudinis SRI-178, Enterobacter ludwigii SRI-211, E. ludwigii SRI-229, Acinetobacter tandoii SRI-305 and Pseudomonas monteilii SRI-360; demonstrated previously for control of charcoal rot disease in sorghum and plant growth-promotion (PGP) in rice] were evaluated for their PGP and biofortification traits in chickpea and pigeonpea under field conditions. When treated on seed, the seven selected bacteria significantly enhanced the shoot height and root length of both chickpea and pigeonpea over the un-inoculated control. Under field conditions, in both chickpea and pigeonpea, the plots inoculated with test bacteria enhanced the nodule number, nodule weight, root and shoot weights, pod number, pod weight, leaf weight, leaf area and grain yield over the un-inoculated control plots. Among the seven bacteria, SRI-229 was found to significantly and consistently enhance all the studied PGP and yield traits including nodule number (24 and 36%), nodule weight (11 and 44%), shoot weight (22 and 20%), root weight (23 and 16%) and grain yield (19 and 26%) for both chickpea and pigeonpea, respectively. When the harvested grains were evaluated for their mineral contents, iron (up to 18 and 12%), zinc (up to 23 and 5%), copper (up to 19 and 8%), manganese (up to 2 and 39%) and calcium (up to 22 and 11%) contents in chickpea and pigeonpea, respectively, were found enhanced in test bacteria inoculated plots over the un-inoculated control plots. This study further confirms that the selected bacterial isolates not only have the potential for PGP in cereals and legumes but also have the potential for biofortification of mineral nutrients.

Genetic architecture of biofortification traits in soybean (Glycine max L. Merr.) revealed through association analysis and linkage mapping

Seed mineral elements are essential not only for seed germination and seedling morphological formation but also for human health. The genetic dissection of seed mineral accumulation is important for mineral biofortification in soybean. However, the molecular mechanisms controlling mineral element accumulation are genetically complex because a number of genetic loci are involved in the metabolic pathways of mineral accumulation in seeds. The objective of this study was to detect the genetic loci for mineral concentrations in soybean seeds, including calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn) and phosphorus (P). Quantitative trait loci (QTL) mapping for the corresponding traits was performed in 184 recombinant inbred lines (RILs) and 219 cultivated soybean accessions. The data for each year and the average across 2 years were used for identification and mapping of QTL controlling seed mineral concentrations. Linkage mapping in the RILs identified totally 35 significant QTL for the five seed mineral concentrations in three cases, some of which were co-localized. Association mapping in the 219 accessions detected 28 single-nucleotide polymorphisms associated with the seed mineral concentrations. Among these, BARC-018099-02516, which was associated with seed Zn concentration, was located close to qZn-11-2. There were 20 putative mineral-related genes in interesting regions of mineral QTL. Three QTL for seed weight were mapped in the RILs, two QTL for seed weight were co-localized with seed Ca, Zn and P concentration QTL which anchored to the same region on chromosome 11. These results will provide a profound understanding of the genetic basis for seed mineral accumulation in soybean seeds and the foundation for mineral biofortification through marker-assisted selection breeding.

Quantitative Trait Loci for Biofortification Traits in Maize Grain

Detecting genes that influence biofortification traits in cereal grain could help increase the concentrations of bioavailable mineral elements in crops to solve the global mineral malnutrition problem. The aims of this study were to detect the quantitative trait loci (QTLs) for phosphorus (P), iron (Fe), zinc (Zn), and magnesium (Mg) concentrations in maize grain in a mapping population, as well as QTLs for bioavailable Fe, Zn, and Mg, by precalculating their respective ratios with P. Elemental analysis of grain samples was done by coupled plasma-optical emission spectrometry in 294 F(4) lines of a biparental population taken from field trials of over 3 years. The population was mapped using sets of 121 polymorphic markers. QTL analysis revealed 32 significant QTLs detected for 7 traits, of which some were colocalized. The Additive-dominant model revealed highly significant additive effects, suggesting that biofortification traits in maize are generally controlled by numerous small-effect QTLs. Three QTLs for Fe/P, Zn/P, and Mg/P were colocalized on chromosome 3, coinciding with simple sequence repeats marker bnlg1456, which resides in close proximity to previously identified phytase genes (ZM phys1 and phys2). Thus, we recommend the ratios as bioavailability traits in biofortification research.