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Aims The ability to suppress soil nitrification through the release of nitrification inhibitors from plant roots is termed ‘biological nitrification inhibition’ (BNI). Here, we aimed at the quantification and characterization of the BNI function in sorghum that includes inhibitor production, their chemical identity, functionality and factors regulating their release. Methods Sorghum was grown in solution culture and root exudate was collected using aerated NH4Cl solutions. A bioluminescence assay using recombinant Nitrosomonas europaea was employed to determine the BNI activity. Activity-guided chromatographic fractionation was used to isolate biological nitrification inhibitors (BNIs). The chemical structure was analyzed using NMR and mass spectrometry; pH-stat systems were deployed to analyze the role of rhizosphere pH on BNIs release. Results Sorghum roots released two categories of BNIs: hydrophilic- and hydrophobic-BNIs. The release rates for hydrophilic- and hydrophobic- BNIs ranged from 10 to 25 ATU g−1 root dwt. d−1. Addition of hydrophilic BNIs (10 ATU g−1 soil) significantly inhibited soil nitrification (40 % inhibition) during a 30-d incubation test. Two BNI compounds isolated are: sakuranetin (ED80 0.6 μM; isolated from hydrophilic-BNIs fraction) and sorgoleone (ED80 13.0 μM; isolated from hydrophobic-BNIs fraction), which inhibited Nitrosomonas by blocking AMO and HAO enzymatic pathways. The BNIs release required the presence of NH4+ in the root environment and the stimulatory effect of NH4+ lasted 24 h. Unlike the hydrophobic-BNIs, the release of hydrophilic-BNIs declined at a rhizosphere pH >5.0; nearly 80 % of hydrophilic-BNI release was suppressed at pH ≥7.0. The released hydrophilic-BNIs were functionally stable within a pH range of 5.0 to 9.0. Sakuranetin showed a stronger inhibitory activity (ED50 0.2 μM) than methyl 3-(4-hydroxyphenyl) propionate (MHPP) (ED50 100 μM) (isolated from hydrophilic-BNIs fraction) in the in vitro culture-bioassay, but the activity was non-functional and ineffective in the soil-assay. Conclusions There is an urgent need to identify sorghum genetic stocks with high potential to release functional-BNIs for suppressing nitrification and to improve nitrogen use efficiency in sorghum-based production systems.

It was previously shown that pearl millet genotypes carrying a terminal drought tolerance quantitative trait locus (QTL) had a lower transpiration rate (Tr; g cm−2 d−1) under well-watered conditions than sensitive lines. Here experiments were carried out to test whether this relates to leaf abscisic acid (ABA) and Tr concentration at high vapour pressure deficit (VPD), and whether that leads to transpiration efficiency (TE) differences. These traits were measured in tolerant/sensitive pearl millet genotypes, including near-isogenic lines introgressed with a terminal drought tolerance QTL (NIL-QTLs). Most genotypic differences were found under well-watered conditions. ABA levels under well-watered conditions were higher in tolerant genotypes, including NIL-QTLs, than in sensitive genotypes, and ABA did not increase under water stress. Well-watered Tr was lower in tolerant than in sensitive genotypes at all VPD levels. Except for one line, Tr slowed down in tolerant lines above a breakpoint at 1.40–1.90 kPa, with the slope decreasing >50%, whereas sensitive lines showed no change in that Tr response across the whole VPD range. It is concluded that two water-saving (avoidance) mechanisms may operate under well-watered conditions in tolerant pearl millet: (i) a low Tr even at low VPD conditions, which may relate to leaf ABA; and (ii) a sensitivity to higher VPD that further restricts Tr, which suggests the involvement of hydraulic signals. Both traits, which did not lead to TE differences, could contribute to absolute water saving seen in part due to dry weight increase differences. This water saved would become critical for grain filling and deserves consideration in the breeding of terminal drought-tolerant lines.

Agriculture is the single largest geo-engineering initiative that humans have initiated on planet Earth, largely through the introduction of unprecedented amounts of reactive nitrogen (N) into ecosystems. A major portion of this reactive N applied as fertilizer leaks into the environment in massive amounts, with cascading negative effects on ecosystem health and function. Natural ecosystems utilize many of the multiple pathways in the N cycle to regulate N flow. In contrast, the massive amounts of N currently applied to agricultural systems cycle primarily through the nitrification pathway, a single inefficient route that channels much of this reactive N into the environment. This is largely due to the rapid nitrifying soil environment of present-day agricultural systems...

Nitrification and denitrification are the two most important processes that contribute to greenhouse gas emission and inefficient use of nitrogen. Suppressing soil nitrification through the release of nitrification inhibitors from roots is a plant function, and termed “Biological Nitrification Inhibition (BNI)”. We report here the role and contribution of sorgoleone release to sorghum-BNI function.

Pearl millet is the main component of traditional farming systems and a staple grain in the diet of sub-Saharan Africa and India. To facilitate breeding work in this crop, a genetic map consisting of single nucleotide polymorphism (SNP) markers was constructed using an F2 population of 93 progenies, from a wild × cultivated pearl millet cross. We used a modified genotyping-by-sequencing (GBS) protocol involving two restriction enzymes (PstI–MspI) and PCR amplification with primers including three selective bases to generate 3,321 SNPs. Of these, 2,809 high-quality SNPs exhibited a minor allele frequency ≥0.3. In total, 314 non-redundant haplotypes and 85 F2 individuals were used to construct a genetic map spanning a total distance of 640 cM. These SNPs were evenly distributed over seven linkage groups ranging considerably in size (62–123 cM). The average density for this map was 0.51 SNP/cM, and the average interval between SNP markers was 2.1 (±0.6) cM. Finally, to establish bridges between the linkage groups of this and previous maps, 19 SSR markers were examined for polymorphism between the parents of this population. We could only tentatively suggest a correspondence between four of our linkage groups and those of previous maps. Overall, GBS enabled us to quickly produce a genetic map with a density and uniformity of markers greater than previously published maps. The availability of such a map will be useful for the identification of genomic regions associated with Striga resistance and other important agronomic traits.

Reduced leaf senescence (stay-green) has been demonstrated to improve tolerance of post-flowering moisture stress in grain sorghum. A number of quantitative trait loci (QTLs) associated with stay-green have been identified in sorghum, to facilitate transfer of this trait into adapted genetic backgrounds. This study reports initial evaluations, in both well watered and post-flowering stress environments, following partial introgression (BC 2 F 3 /BC 1 F 4 generations) of four stable stay-green QTLs ( StgB , Stg1 , Stg3 and Stg4 ) from donor parent B35 to senescent variety R 16. The majority of the introgression lines had higher leaf chlorophyll levels at flowering (a distinctive trait of the donor parent) and a greater percentage green leaf area during the latter part of grain filling, than did R 16, indicating that the stay-green QTLs were expressed phenotypically in the R 16 background. None of the QTL introgression lines achieved the same level of stay-green as B35, however. Maintenance of a greater relative green leaf area during the latter half of grain filling was related to a greater relative grain yield in two of three post-flowering moisture deficit environments in which the materials were evaluated ( r 2  = 0.34 in 2004–2005 and r 2  = 0.76 in 2005–2006), as was a direct measure of leaf chlorophyll in one of the post-flowering stress environments in which this was measured ( r 2  = 0.42, P  < 0.05). Thus the study provided useful evidence that the marker-assisted backcross transfer of stay-green QTLs from B35 into an adapted, but senescent background has the potential to enhance tolerance of post-flowering drought stress in sorghum.

Pearl millet is an important component of food security in the semi-arid tropics and is assuming greater importance in the context of changing climate and increasing demand for highly nutritious food and feed. Molecular tools have been developed and applied for pearl millet on a limited scale. However, the existing tool kit needs to be strengthened further for its routine use in applied breeding programs. Here, we report enrichment of the pearl millet molecular linkage map by exploiting low-cost and high-throughput Diversity Arrays Technology (DArT) markers. Genomic representation from 95 diverse genotypes was used to develop a DArT array with circa 7,000 clones following PstI/BanII complexity reduction. This array was used to genotype a set of 24 diverse pearl millet inbreds and 574 polymorphic DArT markers were identified. The genetic relationships among the inbred lines as revealed by DArT genotyping were in complete agreement with the available pedigree data. Further, a mapping population of 140 F7 Recombinant Inbred Lines (RILs) from cross H 77/833-2 9 PRLT 2/89-33 was genotyped and an improved linkage map was constructed by integrating DArT and SSR marker data. This map contains 321 loci (258 DArTs and 63 SSRs) and spans 1148 cM with an average adjacent-marker interval length of 3.7 cM. The length of individual linkage groups (LGs) ranged from 78 cM (LG 3) to 370 cM (LG 2). This better-saturated map provides improved genome coverage and will be useful for genetic analyses of important quantitative traits. This DArT platform will also permit cost-effective background selection in marker-assisted backcrossing programs as well as facilitate comparative genomics and genome organization studies once DNA sequences of polymorphic DArT clones are available.

The sequencing and detailed comparative functional analysis of genomes of a number of select botanical models open new doors into comparative genomics among the angiosperms, with potential benefits for improvement of many orphan crops that feed large populations. In this study, a set of simple sequence repeat (SSR) markers was developed by mining the expressed sequence tag (EST) database of sorghum. Among the SSR-containing sequences, only those sharing considerable homology with rice genomic sequences across the lengths of the 12 rice chromosomes were selected. Thus, 600 SSR-containing sorghum EST sequences (50 homologous sequences on each of the 12 rice chromosomes) were selected, with the intention of providing coverage for corresponding homologous regions of the sorghum genome. Primer pairs were designed and polymorphism detection ability was assessed using parental pairs of two existing sorghum mapping populations. About 28% of these new markers detected polymorphism in this 4-entry panel. A subset of 55 polymorphic EST-derived SSR markers were mapped onto the existing skeleton map of a recombinant inbred population derived from cross N13 × E 36-1, which is segregating for Striga resistance and the stay-green component of terminal drought tolerance. These new EST-derived SSR markers mapped across all 10 sorghum linkage groups, mostly to regions expected based on prior knowledge of rice–sorghum synteny. The ESTs from which these markers were derived were then mapped in silico onto the aligned sorghum genome sequence, and 88% of the best hits corresponded to linkage-based positions. This study demonstrates the utility of comparative genomic information in targeted development of markers to fill gaps in linkage maps of related crop species for which sufficient genomic tools are not available

The stay-green trait is a reported component of tolerance to terminal drought stress in sorghum. To map quantitative trait loci (QTLs) for stay-green, two sorghum recombinant inbred populations (RIPs) of 226 F(3:5) lines each were developed from crosses (1) IS9830 x E36-1 and (2) N13 x E36-1. The common parental line, E36-1 of Ethiopian origin, was the stay-green trait source. The genetic map of RIP 1 had a total length of 1,291 cM, with 128 markers (AFLPs, RFLPs, SSRs and RAPDs) distributed over ten linkage groups. The map of RIP 2 spanned 1,438 cM and contained 146 markers in 12 linkage groups. The two RIPs were evaluated during post-rainy seasons at Patancheru, India, in 1999/2000 (RIP 2) and 2000/2001 (RIP 1). The measures of stay-green mapped were the green leaf area percentages at 15, 30 and 45 days after flowering (% GL15, % GL30 and % GL45, respectively). Estimated repeatabilities for % GL15, % GL30 and % GL45 amounted to 0.89, 0.81 and 0.78 in RIP 1, and 0.91, 0.88 and 0.85 in RIP 2, respectively. The number of QTLs for the three traits detected by composite interval mapping ranged from 5 to 8, explaining 31% to 42% of the genetic variance. In both RIPs, both parent lines contributed stay-green alleles. Across the three measures of the stay-green trait, three QTLs on linkage groups A, E and G were common to both RIPs, with the stay-green alleles originating from E36-1. These QTLs were therefore consistent across the tested genetic backgrounds and years. After QTL validation across sites and verification of the general benefit of the stay-green trait for grain yield performance and stability in the target areas, the corresponding chromosomal regions could be candidates for marker-assisted transfer of stay-green into elite materials.

Pearl millet marker-assisted selection (MAS) programs targeting adaptation to variable postflowering moisture environments would benefit from quantitative trait loci (QTLs) that improve grain yield across the full range of postflowering moisture conditions, rather than just in drought-stressed environments. This research was undertaken to identify such QTLs from an extensive (12-environment) phenotyping data set that included both stressed and unstressed postflowering environments. Genetic materials were test crosses of 79 F2-derived F4 progenies from a mapping population based on a widely adapted maintainer line (ICMB 841) x a postflowering drought-tolerant maintainer (863B). Three QTLs (on linkage group [LG] 2, LG 3, and LG 4) were identified as primary candidates for MAS for improved grain yield across variable postflowering moisture environments. The QTLs on LG 2 and LG 3 (the most promising) explained a useful proportion (13-25%) of phenotypic variance for grain yield across environments. They also co-mapped with QTLs for harvest index across environments, and with QTLs for both grain number and individual grain mass under severe terminal stress. Neither had a significant QTL x environment interaction, indicating that their predicted effects should occur across a broad range of available moisture environments. We have estimated the benefits in grain yield and accompanying changes in yield components and partitioning indices that would be expected as a result of incorporating these QTLs into other genetic backgrounds by MAS.