Micronutrient malnutrition, especially deficiency of two mineral elements, iron [Fe] and zinc [Zn] in the developing world needs urgent attention. Pearl millet is one of the best crops with many nutritional properties and is accessible to the poor. We report findings of the first attempt to mine favorable alleles for grain iron and zinc content through association mapping in pearl millet. An association mapping panel of 130 diverse lines was evaluated at Delhi, Jodhpur and Dharwad, representing all the three pearl millet growing agro-climatic zones of India, during 2014 and 2015. Wide range of variation was observed for grain iron (32.3–111.9 ppm) and zinc (26.6–73.7 ppm) content. Genotyping with 114 representative polymorphic SSRs revealed 0.35 mean gene diversity. STRUCTURE analysis revealed presence of three sub-populations which was further supported by Neighbor-Joining method of clustering and principal coordinate analysis (PCoA). Marker-trait associations (MTAs) were analyzed with 267 markers (250 SSRs and 17 genic markers) in both general linear model (GLM) and mixed linear model (MLM), however, MTAs resulting from MLM were considered for more robustness of the associations. After appropriate Bonferroni correction, Xpsmp 2261 (13.34% R2-value), Xipes 0180 (R2-value of 11.40%) and Xipes 0096 (R2-value of 11.38%) were consistently associated with grain iron and zinc content for all the three locations. Favorable alleles and promising lines were identified for across and specific environments. PPMI 1102 had highest number (7) of favorable alleles, followed by four each for PPMFeZMP 199 and PPMI 708 for across the environment performance for both grain Fe and Zn content, while PPMI 1104 had alleles specific to Dharwad for grain Fe and Zn content. When compared with the reference genome Tift 23D2B1-P1-P5, Xpsmp 2261 amplicon was identified in intergenic region on pseudomolecule 5, while the other marker, Xipes 0810 was observed to be overlapping with aspartic proteinase (Asp) gene on pseudomolecule 3. Thus, this study can help in breeding new lines with enhanced micronutrient content using marker-assisted selection (MAS) in pearl millet leading to improved well-being especially for women and children.

Finger millet, an orphan crop, possesses immense potential in mitigating climate change and could offer threefold security in terms of food, fodder, and nutrition. It is mostly cultivated as a subsistence crop in the marginal areas of plains and hills. Considering the changes in climate inclusive of recurrent weather vagaries witnessed every year, it is crucial to select stable, high-yielding, area-specific, finger millet cultivars. Sixty finger millet varieties released across the country were evaluated over six consecutive rainy seasons from 2011 to 2016 at the Agricultural Research Station, Vizianagaram. The genotype × environment interaction (GEI) was found to be significant in the combined ANOVA. Furthermore, the Additive Main effects and Multiplicative Interaction (AMMI) analysis asserted that genotypes and the GEI effects accounted for approximately 89% of the total variation. Strong positive associations were observed in an estimated set of eleven stability parameters which were chosen to identify stable genotypes. Furthermore, Non-parametric and Parametric Simultaneous Selection indices (NP-SSI and P-SSI) were calculated utilizing AMMI-based stability parameter (ASTAB), modified AMMI stability value (MASV), and Modified AMMI Stability Index (MASI) to identify stable high yielders. Both methods had inherent difficulties in ranking genotypes for SSI. To overcome this, the initial culling [i.e., SSI with culling strategy (C-SSI)] of genotypes was introduced for stability. In the C-SSI method, the top ten genotypes were above-average yielders, while those with below-average yield were observed in NP-SSI and P-SSI methods. Similarly, the estimation of best linear unbiased prediction (BLUP)-based simultaneous selections, such as harmonic mean of genotypic values (HMGV), relative performance of genotypic values (RPGV), and harmonic mean of relative performance of genotypic values (HMRPGV), revealed that none of the top ten entries had below-average yield. The study has proven that C-SSI and BLUP-based methods were equally worthy in the selection of high-yielding genotypes with stable performance. However, the C-SSI approach could be the best method to ensure that genotypes with a considerable amount of stability are selected. The multi-year trial SSI revealed that entries Indaf-9, Sri Chaitanya, PR-202, and A-404; and VL324 and VL146 were ascertained to be the most stable high-yielding genotypes among medium-to-late and early maturity groups, respectively.

Micronutrient malnutrition, especially the paucity of iron (Fe) and zinc (Zn) is posing a big threat to the world affecting nearly 25% of worldwide population. Pearl millet is endowed with huge amount of variability for micronutrients especially for grain Fe and Zn content. Micronutrient enrichment in pearl millet is possible by identifying stable genotypes for high levels of micronutrients and utilising them in breeding programme. In this context, a set of 40 pearl millet genotypes along with one check, Dhanshakti (G30), were evaluated at three different agro climatic zones during the year 2014 for grain iron (Fe) and zinc (Zn) contents using Atomic Absorption Spectrometry. The genotypes contributed 58.3% and 52.8% of the total variation for grain Fe and Zn content, respectively. The magnitude of variation contributed by interaction component was also relatively high (39.7% and 32.5% for grain Fe and Zn). Both AMMI and GGE biplot analysis identified desirable genotypes; PPMI 708 (G40), PPMI 1102 (G25) and PPMI 683 (G39) for grain Fe content, whereas PPMI 708 (G40), PPMI 1116 (G24) and PPMI 683 (G39) for grain Zn content. The Pearson correlation coefficient for grain Fe and Zn content showed that both traits are highly associated (r = 0.8, p <0.01) and these traits did not associate significantly with grain yield. Hence, there is possibility for simultaneous improvement of both grain Fe and Zn content without compromising for grain yield.

Biofortification of lines of pearl millet (Pennisetum glaucum (L.) R.Br.) with increased iron (Fe) and zinc (Zn) will have great impact because pearl millet is an indispensable component of food and nutritional security of inhabitants of arid and semi-arid regions. The aim of the present study was to assess the stability of Fe and Zn content in recombinant inbred lines (RILs) developed for grain Fe and Zn content, and to use these lines in developing micronutrient-rich pearl millet hybrids. A mapping population consisting of 210 RILs along, with parents and checks, was assessed in three consecutive years (2014–16) under rainfed conditions at the same experimental location in an alpha design with two repetitions. Significant differences were observed in genotype, environment and genotype × environment interaction mean squares for all variables, particularly grain micronutrients. The first two principal components of an interaction principal component analysis cumulatively explained 100% of the total variation; respective contributions of the first and second components were 64.0% and 36.0% for Fe, and 58.1% and 41.9% for Zn. A positive and moderately high correlation (0.696**) between Fe and Zn contents suggests good prospects of simultaneous improvement for both micronutrients. Among the 210 RILs, RIL 69, RIL 186, RIL 191, RIL 149 and RIL 45 were found to be more stable with higher mean micronutrient content, additive main effects and multiplicative interaction stability value (ASV) and genotype selection index (GSI) under rainfed condition. These RILs are promising and can be tested further for their combining ability for yield as well as grain micronutrient content for developing superior biofortified, heterotic pearl millet hybrids.

Pearl millet [Pennisetum glaucum (L.) R. Br.] is grown under both arid and semi-arid conditions in India, where other cereals are hard to grow. Pearl millet cultivars, hybrids, and OPVs (open pollinated varieties) are tested and released by the All India Coordinated Research Project on Pearl Millet (AICRP-PM) across three zones (A1, A, and B) that are classified based on rainfall pattern. Except in locations with extreme weather conditions, hybrids dominate pearl millet growing areas, which can be attributed to hybrid vigor and the active role of the private sector. The importance of OPVs cannot be ruled out, owing to wider adaptation, lower input cost, and timely seed availability to subsidiary farmers cultivating this crop. This study was conducted to scrutinize the presently used test locations for evaluation of pearl millet OPVs across India, identify the best OPVs across locations, and determine the variation in grain Fe and Zn contents across locations in these regions. Six varieties were evaluated across 20 locations in A1 and A (pooled as A) and B zones along with three common checks and additional three zonal adapted checks in the respective zones during the 2019 rainy season. Recorded data on yield and quality traits were analyzed using genotype main effects and genotype × environment interaction biplot method. The genotype × environment (G × E) interaction was found to be highly significant for all the grain yield and agronomic traits and for both micronutrients (iron and zinc). However, genotypic effect (G) was four (productive tillers) to 49 (grain Fe content) times that of G × E interaction effect for various traits across zones that show the flexibility of OPVs. Ananthapuramu is the ideal test site for selecting pearl millet cultivars effectively for adaptation across India, while Ananthapuramu, Perumallapalle, and Gurugram can also be used as initial testing locations. OPVs MP 599 and MP 600 are identified as ideal genotypes, because they showed higher grain and fodder yields and stability compared with other cultivars. Iron and zinc concentration showed highly significant positive correlation (across environment = 0.83; p &lt; 0.01), indicating possibility of simultaneous effective selection for both traits. Three common checks were found to be significantly low yielders than the test entries or zonal checks in individual zones and across India, indicating the potential of genetic improvement through OPVs.

Pearl millet is a climate-resilient, nutritious crop with low input requirements that could provide economic returns in marginal agro-ecologies. In this study, we report quantitative trait loci (QTLs) for iron (Fe) and zinc (Zn) content from three distinct production environments. We generated a genetic linkage map using 210 F6 recombinant inbred line (RIL) population derived from the (PPMI 683 × PPMI 627) cross using genome-wide simple sequence repeats (SSRs). The molecular linkage map (seven linkage groups) of 151 loci was 3,273.1 cM length (Kosambi). The content of grain Fe in the RIL population ranged between 36 and 114 mg/Kg, and that of Zn from 20 to 106 mg/Kg across the 3 years (2014–2016) at over the three locations (Delhi, Dharwad, and Jodhpur). QTL analysis revealed a total of 22 QTLs for grain Fe and Zn, of which 14 were for Fe and eight were for Zn on three consecutive years at all locations. The observed phenotypic variance (R2) explained by different QTLs for grain Fe and Zn content ranged from 2.85 (QGFe.E3.2014–2016_Q3) to 19.66% (QGFe.E1.2014–2016_Q3) and from 2.93 (QGZn.E3.2014–2016_Q3) to 25. 95% (QGZn.E1.2014–2016_Q1), respectively. Two constitutive expressing QTLs for both Fe and Zn co-mapped in this population, one on LG 2 and second one on LG 3. Inside the QTLs candidate genes such as Ferritin gene, Al3+ Transporter, K+ Transporters, Zn2+ transporters and Mg2+ transporters were identified using bioinformatics approaches. The identified QTLs and candidate genes could be useful in pearl millet population improvement programs, seed, restorer parents, and marker-assisted selection programs.

Biofortification of pearl millet (Pennisetum glaucum (L.) R. Br.) with improved iron (Fe) and zinc (Zn) will have great impact as it is an indispensable component of nutritional security of inhabitants of arid and semi-arid regions. Ten genotypes along with checks were evaluated in RBD in six locations during kharif, 2016 under rainfed conditions. Significant differences were observed in genotype, environment and genotype × environment interaction mean squares for grain Fe and Zn contents, indicating differential nutrient accumulation by the genotypes. The first two principal components obtained in AMMI analysis were significant and cumulatively explained the total variation were 81.47 % for Fe and 73.97 % for Zn. A positive and moderately high correlation (r=0.6) between Fe and Zn contents suggests good prospects of simultaneous improvement for both micronutrients. Among the ten genotypes, PPMI 953 was found to be more stable with high mean Fe (90 ppm) and Zn (59 ppm) contents. On crossing with designated A lines of pearl millet, the line PPMI 953 found to be restorer for A1 system with complete fertility restoration of F1 panicle of the cross, ICMA(1) 863 x PPMI 953 under bagged condition and resulting F1 with 78-84% fertility measured by seed setting % under bag. The F2 individuals showed 9:7 fertility-sterility ratio (χ 2 value=0.002, P value=0.964). The promising line, PPMI 953 may be used as source for further genetic improvement with respect to grain micronutrient content or can be directly used as male parent in development of high iron pearl millet hybrids.

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