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The main focus of this review is to discuss the current status of the use of GWAS for fodder quality and biofuel owing to its similarity of traits. Sorghum is a potential multipurpose crop, popularly cultivated for various uses as food, feed fodder, and biomass for ethanol. Production of a huge quantity of biomass and genetic variation for complex sugars are the main motivations not only to use sorghum as fodder for livestock nutritionists but also as a potential candidate for biofuel generation. Few studies have been reported on the knowledge transfer that can be used from the development of biofuel technologies to complement improved fodder quality and vice versa. With recent advances in genotyping technologies, GWAS became one of the primary tools used to identify the genes/genomic regions associated with the phenotype. These modern tools and technologies accelerate the genomic assisted breeding process to enhance the rate of genetic gains. Hence, this mini-review focuses on GWAS studies on genetic architecture and dissection of traits underpinning fodder quality and biofuel traits and their limited comparison with other related model crop species.

Background: Production of biofuels from lignocellulosic crop biomass is an alternative to reduce greenhouse gas emissions. The biofuel production involves collecting biomass, breaking down cell wall components followed by the conversion of sugars to ethanol. The lingo-cellulosic biomass comprises 40-50% cellulose, 20-30% hemicellulose, and 10-25% lignin. Sorghum is a widely adapted energy crop for biofuel production. Biomass with low lignin, high cellulose, and high hemicellulose contents are exploited to attain maximum biofuel production efficiency. Resistance to lodging, pest, disease, and abiotic stresses related to cell wall components is well documented, and quantitative trait loci were identified to understand these traits' genetic correlation. Selection for reduced lignin and increased cellulose content in stover can increase the ethanol yield. The Genome-Wide Association Studies (GWAS) is a complementary approach to evaluating the marker and phenotype associations among large diversity panels. Single nucleotide polymorphisms were scanned to identify loci associated with the traits of interest. In this study, the GWAS was performed on 245 sorghum minicore genotypes to analyze agronomic traits (days to 500lowering, fresh biomass yield, dry biomass yield) and cell wall components (cellulose, hemicellulose, and lignin). Further, in-silico validation of the candidate genes was performed in a global gene expression data from large-scale RNA sequencing studies in sorghum available in the NCBI GEO database was used. Objective: The objectives of this study are to evaluate native variations in biofuel related agronomic traits and stalk cell wall components and to identify significant SNPs or loci related to the cell wall components. Methods: In this article, an association mapping panel, comprising of 245 sorghum minicore germplasm accessions, was evaluated during two post rainy seasons of 2013 and 2014, and observations were recorded on the whole plot- for days to 50% flowering, fresh biomass yield (tha-1), and dry biomass yield (tha-1). The biomass of sun-dried plants from both seasons was collected separately, chopped, dried, and ground to powder. The cellulose, hemicellulose, and lignin contents were determined in the powdered. The content of each of these three components in sorghum was expressed in percent of dry matter. The data on agronomic traits and composition analysis was subjected to Analysis of Variance. For the current study, we remapped the raw GBS data with the sorghum assembly version v3.1. A total of 27,589 SNPs were obtained with a minor allele frequency (MAF)>1% and missing data Results: High broad-sense heritability was exhibited for all the traits in individual seasons along with significant genotype × environment interaction across seasons except lignin. Association mapping with a P-4 revealed genomic regions associated with the- (i) agronomic traits (days to 50% flowering, fresh and dry biomass), and (ii) biochemical traits (cellulose, hemicellulose, and lignin) associated with biofuels production, in individual seasons. Twelve significant SNPs for flowering time, 30 fresh biomass yields, and 24 for dry biomass yield, 25 for cellulose, 7 for hemicellulose, and 21 for lignin were identified. CIRCOS plot was constructed to identify and analyze similarities and differences while comparing the sorghum genome with different crops. For cellulose high similarity of >80% was observed for all sorghum gene sequences with the maize homologs. The overall similarity of sorghum homologs with foxtail millet was >65%, for Arabidopsis from 30.6% to 48.6%, and rice from 28.2% to 92.8%. SNPs for hemicellulose displayed maximum similarity to foxtail millet followed by maize. The sequence similarity of lignin SNPs in sorghum was highest with the maize genome followed by Arabidopsis. Both rice and foxtail millet showed >55% similarity to the sorghum genome. Conclusion: This study reports large variability for agronomic and biofuel traits in the sorghum minicore collection with high heritability. The genetic architecture of cell wall components using the GWAS approach was studied and candidate genes for each component were annotated. These results give a better understanding of the genetic basis of the sorghum cell wall composition. The association analysis identified regions of the genome that could be targeted to enhance the quality of biomass and yield along with the desired composition promoting breeding efficiency for enhanced biofuel yield.

Pearl millet is an important crop of the arid and semi-arid ecologies to sustain food and fodder production. The greater tolerance to drought stress attracts us to examine its cellular and molecular mechanisms via functional genomics approaches to augment the grain yield. Here, we studied the drought response of 48 inbreds representing four different maturity groups at the flowering stage. A set of 74 drought-responsive genes were separated into five major phylogenic groups belonging to eight functional groups, namely ABA signaling, hormone signaling, ion and osmotic homeostasis, TF-mediated regulation, molecular adaptation, signal transduction, physiological adaptation, detoxification, which were comprehensively studied. Among the conserved motifs of the drought-responsive genes, the protein kinases and MYB domain proteins were the most conserved ones. Comparative in-silico analysis of the drought genes across millet crops showed foxtail millet had most orthologs with pearl millet. Of 698 haplotypes identified across millet crops, MyC2 and Myb4 had maximum haplotypes. The protein–protein interaction network identified ABI2, P5CS, CDPK, DREB, MYB, and CYP707A3 as major hub genes. The expression assay showed the presence of common as well as unique drought-responsive genes across maturity groups. Drought tolerant genotypes in respective maturity groups were identified from the expression pattern of genes. Among several gene families, ABA signaling, TFs, and signaling proteins were the prospective contributors to drought tolerance across maturity groups. The functionally validated genes could be used as promising candidates in backcross breeding, genomic selection, and gene-editing schemes in pearl millet and other millet crops to increase the yield in drought-prone arid and semi-arid ecologies.

Pearl millet [Pennisetum glaucum (L.) R. Br.] is a staple food crop of arid and semi-arid regions of Asia and Africa. Forty-five pearl millet populations of Asian and African origin were assessed for genetic diversity using 29 simple sequence repeat (SSR) markers. The SSR-based clustering and structure analyses showed that Asian origin–Asian bred (As-As) and African origin–African bred (Af-Af) populations were distributed across seven clusters, indicating no strong relationship among populations with their geographical origin. Most of the African origin–Asian bred (Af-As) populations had a higher average number of alleles per locus than As-As or Af-Af populations, and the majority of them clustered separately from As-As or Af-Af populations, indicating that introgression of African origin breeding materials led to the development of new gene pools adapted to the Asian region. Fourteen populations representing seven clusters were crossed according to a diallel mating design to generate 91 population hybrids (seeds of direct and reciprocal crossesweremixed) and evaluated at three locations in 2016. All the 91 hybrids when partitioned into three groups based on genetic distance (GD) between parental combinations (low,moderate, and high), revealed no correlation between GD and panmictic midparent heterosis in any of the groups, indicating that grain yield heterosis cannot be predicted based on GD. Two population hybrids (GB 8735 × ICMP 87307 and Sudan I × Ugandi) exhibited high levels of yield heterosis over standard checks and can be further utilized using different breeding schemes to develop high-yielding pearl millet cultivars.

The genome sequences of 16 Streptomyces strains, showing potential for plant growth-promotion (PGP) activities in rice, sorghum, chickpea and pigeonpea, isolated from herbal vermicompost, have been decoded. The genome assemblies of the 16 Streptomyces strains ranged from 6.8 Mb to 8.31 Mb, with a GC content of 72 to 73%. The extent of sequence similarity (in terms of shared ortholog) in 16 Streptomyces strains showed 70 to 85% common genes to the closest publicly available Streptomyces genomes. It was possible to identify ~1,850 molecular functions across these 16 strains, of which close to 50% were conserved across the genomes of Streptomyces strains, whereas, ~10% were strain specific and the rest were present in various combinations. Genome assemblies of the 16 Streptomyces strains have also provided genes involved in key pathways related to PGP and biocontrol traits such as siderophores, auxin, hydrocyanic acid, chitinase and cellulase. Further, the genome assemblies provided better understanding of genetic similarity among target strains and with the publically available Streptomyces strains.

Iron deficiency chlorosis is an important abiotic stress affecting groundnut production worldwide in calcareous and alkaline soils with a pH of 7.5–8.5. To identify genomic regions controlling iron deficiency chlorosis resistance in groundnut, the recombinant inbred line population from the cross TAG 24 × ICGV 86031 was evaluated for associated traits like visual chlorosis rating and SPAD chlorophyll meter reading across three crop growth stages for two consecutive years. Thirty-two QTLs were identified for visual chlorosis rating (3.9%–31.8% phenotypic variance explained [PVE]) and SPAD chlorophyll meter reading [3.8%–11% PVE] across three stages over 2 years. This is the first report of identification of QTLs for iron deficiency chlorosis resistance- associated traits in groundnut. Three major QTLs (>10% PVE) were identified at severe stage, while majority of other QTLs were having small effects. Interestingly, two major QTLs for visual chlorosis rating at 60 days (2013) and 90 days (2014) were located at same position on LG AhXIII. The identified QTLs/markers after validation across diverse genetic material could be used in genomics-assisted breeding.

Indian dill seed (Anethum sowa Roxb. ex Fleming) is an important member of family Apiaceae. It is a significant seed spice having many medicinal properties. But, this aromatic herb is still orphan from breeding and crop improvement perspective as no serious attention has been given by breeders. To swift the breeding program, molecular markers play the main role. But due to the paucity of markers, breeding is slower in dill seed. Therefore, an attempt was made to develop the genomic SSR markers in dill seed through next-generation sequencing. A total of 2119.51 Mbp of raw data were generated on the Miseq NGS platform during survey sequencing. In 2,25,956 contigs created by Spades assembler, a total 48,951 repeat motifs were identified. A set of 20,294 primer pairs (dimer to hexamer repeats) were produced. Among detected repeat motifs, 48.89% was mononucleotides. Of 12 primers, 10 (83%) primers could be successfully amplified in dill seed and produced 11 amplicons. During cross-genera amplification of markers, 9 out of 10 primers could be successfully amplified in related genera. Developed markers can also be used to initiate the molecular breeding program, association mapping and to assess the evolutionary relationship among seed spices. Keywords: Anethum, Seed spices, SSR, Transferability

Livestock provides an additional source of income for marginal cropping farmers, but crop residues that are used as a main source of animal feed are characteristically low in digestibility and protein content. This reduces the potential livestock product yield and quality. The key trait, which influences the quality and the cost of animal feed, is digestibility. In this study, we demonstrate that sorghum breeding can be directed to achieve genetic gains for both fodder biomass and digestibility without any trade-offs. The genotypic variance has shown significant differences for biomass across years (13,035 in 2016 and 3,395 in 2017) while in vitro organic matter digestibility (IVOMD) showed significant genotypic variation in 2016 (0.253) under drought. A range of agronomic and fodder quality traits was found to vary significantly in the population within both the control and drought conditions and across both years of the study. There was significant genotypic variance (σg2) and genotypic × treatment variance (σgxt2) in dry matter production in a recombinant inbred line (RIL) population in both study years, while there was only significant σg2 and σgxt2 in IVOMD under the control conditions. There was no significant correlation identified between biomass and digestibility traits under the control conditions, but there was a positive correlation under drought. However, a negative relation was observed between digestibility and grain yield under the control conditions, while there was no significant correlation under drought population, which was genotyped using the genotyping-by-sequencing (GBS) technique, and 1,141 informative single nucleotide polymorphism (SNP) markers were identified. A linkage map was constructed, and a total of 294 quantitative trait loci (QTLs) were detected, with 534 epistatic interactions, across all of the traits under study. QTL for the agronomic traits fresh and dry weight, together with plant height, mapped on to the linkage group (LG) 7, while QTL for IVOMD mapped on to LG1, 2, and 8. A number of genes previously reported to play a role in nitrogen metabolism and cell wall-related functions were found to be associated with these QTL.

Groundnut rust caused by Puccinia arachidis Speg. is a major cause of yield and quality losses in groundnut (Arachis hypogaea L.) in the warm-humid tropics including Tanzania. Breeding and deployment of rust resistant cultivars with farmer-preferred attributes will bolster groundnut production and productivity. The objective of this study was to determine the combining ability effects and gene action controlling rust resistance in groundnut genotypes for breeding. Twelve selected and complementary parental lines were crossed in a diallel design, to develop F1 progenies, which were advanced to the F2 for individual plant selection. Thirty-three successful partial crosses and the 12 parents were field evaluated using a 5 × 9 alpha lattice designs with two replications over two seasons in Tanzania. The tested genotypes exhibited significant (P

Exploring the natural genetic variability and its exploitation for improved Nitrogen Use Efficiency (NUE) in sorghum is one of the primary goals in the modern crop improvement programs. The integrated strategies include high-throughput phenotyping, next generation sequencing (NGS)-based genotyping technologies, and a priori selected candidate gene studies that help understand the detailed physiological and molecular mechanisms underpinning this complex trait. A set of sixty diverse sorghum genotypes was evaluated for different vegetative, reproductive, and yield traits related to NUE in the field (under three N regimes) for two seasons. Significant variations for different yield and related traits under 0 and 50% N confirmed the availability of native genetic variability in sorghum under low N regimes. Sorghum genotypes with distinct genetic background had interestingly similar NUE associated traits. The Genotyping-By-Sequencing based SNPs (>89 K) were used to study the population structure, and phylogenetic groupings identified three distinct groups. The information of grain N and stalk N content of the individuals covered on the phylogenetic groups indicated randomness in the distribution for adaptation under variable N regimes. This study identified promising sorghum genotypes with consistent performance under varying environments, with buffer capacity for yield under low N conditions. We also report better performing genotypes for varied production use—grain, stover, and dual-purpose sorghum having differential adaptation response to NUE traits. Expression profiling of NUE associated genes in shoot and root tissues of contrasting lines (PVK801 and HDW703) grown in varying N conditions revealed interesting outcomes. Root tissues of contrasting lines exhibited differential expression profiles for transporter genes [ammonium transporter (SbAMT), nitrate transporters (SbNRT)]; primary assimilatory (glutamine synthetase (SbGS), glutamate synthase (SbGOGAT[NADH], SbGOGAT[Fd]), assimilatory genes [nitrite reductase (SbNiR[NADH]3)]; and amino acid biosynthesis associated gene [glutamate dehydrogenase (SbGDH)]. Identification and expression profiling of contrasting sorghum genotypes in varying N dosages will provide new information to understand the response of NUE genes toward adaptation to the differential N regimes in sorghum. High NUE genotypes identified from this study could be potential candidates for in-depth molecular analysis and contribute toward the development of N efficient sorghum cultivars.

Pearl millet (Pennisetum glaucumR. Br.) is an important staple and nutritious food crop in the semiarid and arid ecologies of South Asia (SA) and Sub-Saharan Africa (SSA). In view of climate change, depleting water resources, and widespread malnutrition, there is a need to accelerate the rate of genetic gains in pearl millet productivity. This review discusses past strategies and future approaches to accelerate genetic gains to meet future demand. Pearl millet breeding in India has historically evolved very comprehensively from open-pollinated varieties development to hybrid breeding. Availability of stable cytoplasmic male sterility system with adequate restorers and strategic use of genetic resources from India and SSA laid the strong foundation of hybrid breeding. Genetic and cytoplasmic diversification of hybrid parental lines, periodic replacement of hybrids, and breeding disease-resistant and stress-tolerant cultivars have been areas of very high priority. As a result, an annual yield increase of 4% has been realized in the last three decades. There is considerable scope to further accelerate the efforts on hybrid breeding for drought-prone areas in SA and SSA. Heterotic grouping of hybrid parental lines is essential to sustain long-term genetic gains. Time is now ripe for mainstreaming of the nutritional traits improvement in pearl millet breeding programs. New opportunities are emerging to improve the efficiency and precision of breeding. Development and application of high-throughput genomic tools, speed breeding, and precision phenotyping protocols need to be intensified to exploit a huge wealth of native genetic variation available in pearl millet to accelerate the genetic gains.

Sorghum (Sorghum bicolor L.) is a staple food crops in the arid and rainfed production ecologies. Sorghum plays a critical role in resilient farming and is projected as a smart crop to overcome the food and nutritional insecurity in the developing world. The development and characterisation of the sorghum pan-genome will provide insight into genome diversity and functionality, supporting sorghum improvement. We built a sorghum pan-genome using reference genomes as well as 354 genetically diverse sorghum accessions belonging to different races. We explored the structural and functional characteristics of the pan-genome and explain its utility in supporting genetic gain. The newly-developed pan-genome has a total of 35,719 genes, a core genome of 16,821 genes and an average of 32,795 genes in each cultivar. The variable genes are enriched with environment responsive genes and classify the sorghum accessions according to their race. We show that 53% of genes display presence-absence variation, and some of these variable genes are predicted to be functionally associated with drought adaptation traits. Using more than two million SNPs from the pan-genome, association analysis identified 398 SNPs significantly associated with important agronomic traits, of which, 92 were in genes. Drought gene expression analysis identified 1,788 genes that are functionally linked to different conditions, of which 79 were absent from the reference genome assembly. This study provides comprehensive genomic diversity resources in sorghum which can be used in genome assisted crop improvement.

Sorghum (Sorghum bicolorL.) is one of the most important food crops in the arid and rainfed production ecologies. It is a part of resilient farming and is projected as a smart crop to overcome the food and nutritional challenges in the developing world. The development and characterisation of the sorghum pan-genome will provide insight into genome diversity and functionality, supporting sorghum improvement. We built a sorghum pan-genome using reference genomes as well as 354 genetically diverse sorghum accessions belonging to different races. We explored the structural and functional characteristics of the pan-genome and explain its utility in supporting genetic gain. The newly-developed pan-genome has a total of 35,719 genes, a core genome of 16,821 genes and an average of 32,795 genes in each cultivar. The variable genes are enriched with environment responsive genes and classify the sorghum accessions according to their race. We show that 53% of genes display presence-absence variation, and some of these variable genes are predicted to be functionally associated with drought traits. Using more than two million SNPs from the pan-genome, association analysis identified 398 SNPs significantly associated with important agronomic traits, of which, 92 were in genes. Drought gene expression analysis identified 1,788 genes that are functionally linked to different conditions, of which 79 were absent from the reference genome assembly. This study provides comprehensive genomic diversity resources in sorghum which can be used in genome assisted crop improvement.

Sorghum is widely grown as a failsafe crop in semi-arid regions particularly in post rainy season. Though the effect of drought on crop performance is studied widely there are few studies illustrating the association of fodder quality and agronomic traits under drought. To study the interactions we evaluated a set of 24 cultivars under drought for three years in post rainy season. The effect of drought was evident in delayed flowering (by 2 days) and reduce plant height (by 0.98 cm) compared to control. The fodder digestibility traits were reduced (in vitro organic matter digestibility by 2.25 times) under drought. All the plant growth and yield parameters recorded higher heritability compared to fodder quality parameters (

Millets are the strategic food crops in arid and drought-prone ecologies. Millets, by virtue of nature, are very well-adapted to drought conditions and able to produce sustainable yield. Millets have important nutrients that can help prevent micro-nutrient malnutrition. As a result of the adverse effect of climate change and widespread malnutrition, millets have attained a strategic position to sustain food and nutritional security. Although millets can adapt well to the drought ecologies where other cereals fail completely, the yield level is very low under stress. There is a tremendous opportunity to increase the genetic potential of millet crops in dry lands when the genetics of the drought-tolerance mechanism is fully explained. MicroRNAs (miRNAs) are the class of small RNAs that control trait expression. They are part of the gene regulation but little studied in millets. In the present study, novel miRNAs and gene targets were identified from the genomic resources of pearl millet, sorghum, foxtail millet, finger millet, and proso millet through in silico approaches. A total of 1,002 miRNAs from 280 families regulating 23,158 targets were identified using different filtration criteria in five millet species. The unique as well as conserved structural features and functional characteristics of miRNA across millets were explained. About 84 miRNAs were conserved across millets in different species combinations, which explained the evolutionary relationship of the millets. Further, 215 miRNAs controlling 155 unique major drought-responsive genes, transcription factors, and protein families revealed the genetics of drought tolerance that are accumulated in the millet genomes. The miRNAs regulating the drought stress through specific targets or multiple targets showed through a network analysis. The identified genes regulated by miRNA genes could be useful in developing functional markers and used for yield improvement under drought in millets as well as in other crops.

Brown planthopper (BPH), one of the most important pests of the rice (Oryza sativa) crop, becomes catastrophic under severe infestations and causes up to 60% yield loss. The highly disastrous BPH biotype in the Indian sub-continent is Biotype 4, which also known as the South Asian Biotype. Though many resistance genes were mapped until now, the utility of the resistance genes in the breeding programs is limited due to the breakdown of resistance and emergence of new biotypes. Hence, to identify the resistance genes for this economically important pest, we have used a multi-parent advanced generation intercross (MAGIC) panel consisting of 391 lines developed from eight indica founder parents. The panel was phenotyped at the controlled conditions for two consecutive years. A set of 27,041 cured polymorphic single nucleotide polymorphism (SNPs) and across-year phenotypic data were used for the identification of marker&ndash, trait associations. Genome-wide association analysis was performed to find out consistent associations by employing four single and two multi-locus models. Sixty-one SNPs were consistently detected by all six models. A set of 190 significant marker-associations identified by fixed and random model circulating probability unification (FarmCPU) were considered for searching resistance candidate genes. The highest number of annotated genes were found in chromosome 6 followed by 5 and 1. Ninety-two annotated genes identified across chromosomes of which 13 genes are associated BPH resistance including NB-ARC (nucleotide binding in APAF-1, R gene products, and CED-4) domain-containing protein, NHL repeat-containing protein, LRR containing protein, and WRKY70. The significant SNPs and resistant lines identified from our study could be used for an accelerated breeding program to develop new BPH resistant cultivars.

Biofortification through plant breeding is a cost-effective and sustainable approach towards addressing micronutrient malnutrition prevailing across the globe. Screening cultivars for micronutrient content and identification of quantitative trait loci (QTLs)/genes and markers help in the development of biofortified varieties in chickpea (Cicer arietinum L.). With the aim of identifying the genomic regions controlling seed Fe and Zn concentrations, the F2:3 population derived from a cross between MNK-1 and Annigeri 1 was genotyped using genotyping by sequencing approach and evaluated for Fe and Zn concentration. An intraspecific genetic linkage map comprising 839 single nucleotide polymorphisms (SNPs) spanning a total distance of 1,088.04 cM with an average marker density of 1.30 cM was constructed. By integrating the linkage map data with the phenotypic data of the F2:3 population, a total of 11 QTLs were detected for seed Fe concentration on CaLG03, CaLG04, and CaLG05, with phenotypic variation explained ranging from 7.2% (CaqFe3.4) to 13.4% (CaqFe4.2). For seed Zn concentration, eight QTLs were identified on CaLG04, CaLG05, and CaLG08. The QTLs individually explained phenotypic variations ranging between 5.7% (CaqZn8.1) and 13.7% (CaqZn4.3). Three QTLs for seed Fe and Zn concentrations (CaqFe4.4, CaqFe4.5, and CaqZn4.1) were colocated in the "QTL-hotspot" region on CaLG04 that harbors several drought tolerance-related QTLs. We identified genes in the QTL regions that encode iron-sulfur metabolism and zinc-dependent alcohol dehydrogenase activity on CaLG03, iron ion binding oxidoreductase on CaLG04, and zinc-induced facilitator-like protein and ZIP zinc/iron transport family protein on CaLG05. These genomic regions and the associated markers can be used in marker-assisted selection to increase seed Fe and Zn concentrations in agronomically superior chickpea varieties.

Chickpea is a cheap source of protein and micronutrients to the small and marginal farmers living in south-Asia and sub-Saharan Africa. Due to changes in climatic conditions and cropping systems, the crop is being exposed to severe drought and heat stress during its reproductive period, which leads to significant yield losses and fluctuations in grain nutrient accumulation. The study was conducted with 140 diverse genotypes under non-stress, drought, and heat stress conditions to estimate their effects on grain nutrient (protein, Fe and Zn) contents and identify the marker-trait associations. Analysis of variance revealed highly significant differences among genotypes for nutrient content under respective planting conditions. The seed yield was negatively associated with the grain Fe (r = -0.37, -0.25, 0.11) and Zn (r = -0.49, -0.35, -0.72) under respective planting conditions. The PCA indicated that PC1 was positively associated with grain Fe in non-stress and heat, while Zn in all planting conditions, whereas PC2 was positively influenced by protein content followed by grain yield. Cluster analysis identified eight clusters, of which cluster VI showed maximum cluster means for Fe (73 mg kg-1) and Zn (48.1 mg kg-1). The Genome-wide association study revealed, a total of 181 MTAs for grain Fe, Zn, and protein content in all three growing conditions. A total of 4, 2, and 48 SNPs for grain Fe and Zn content, whereas, 66, 46, 15 SNPs for grain protein content have shown significant association under non-stress, drought, and heat stress, respectively. One SNP each on chr1 (S1_35622241; P≤3.47×10-6) and Chr4 (S4_44607232; P≤1.35×10-5) was co-associated under drought and non-stress conditions for protein and Fe, respectively. The identified robust MTAs will be validated and used in marker-assisted selection towards the rapid development of nutrient-rich varieties.

Seed protein content (SPC) is an important grain quality trait, which impacts the nutritional importance of pigeonpea seed in the diet of over a billion people globally. The present study was carried out to determine variation in SPC and its relationships with some agronomic traits among 23 parental lines of different types of pigeonpea mapping populations. The parental lines were evaluated under field conditions during 2014-2015 growing season. A randomised complete block design in two replications was used. Data were recorded on SPC, days to first flower (DTF), plant height at maturity (PltH), number of pods per plant (NPP), number of seeds per pod (NSP), hundred-seed weight (SW) and seed yield per plant (SY). There were significant differences among genotypes for all traits. Broad-sense heritability was 0.693 for SPC but ranged from 0.519 (NPP) to 0.999 (DTF) while genetic advance was 2.4% for SPC but ranged from 1.2 % (NSP) to 141.2 % (SY), and genetic gain ranged from 11.0 % (SPC) to 230.0 % (SY). Simple correlation showed that SPC is only significantly but negatively correlated with SW (r = -0.30, P < 0.05), while path analyses revealed that SPC is negatively associated SW and NPP but positively with DTF, PltH, NSP and SY. It is concluded that genetic variation for SPC and agronomic traits exist among pigeonpea genotypes studied. The variation is accompanied by both favourable and unfavourable relationships of SPC with the agronomic traits.

The present investigation was performed to assess genetic divergence and heterosis prediction in hybrid parents of pearl millet [Pennisetum glaucum (L.) R Br.] using simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers. Using 56 SSR loci, 412 alleles were detected in 147 lines with an average of 7.36 alleles per locus, and 75,007 SNP loci were detected in 117 lines. Both SSR- and SNP-based clustering and structure analysis partitioned all maintainer (B) and restorer (R) lines into two clear-cut separate groups, indicating the existence of two diverse gene pools, each representing the seed and restorer parents in pearl millet. Results of analysis of molecular variance and principal coordinate analysis also showed significant diversity between B and R lines. The correlation between parental genetic distances estimated based on SSRs and SNPs was high and significant (r = 0.58, p < .01). Similar clustering pattern of hybrid parents was observed with both marker systems, although the cost of genotyping was 41% less with SNPs than with SSRs, and the ratio of loci detected with SNPs was much higher (1:364 SSR/SNP), hence the use of SNPs is indicated over SSRs for germplasm characterization. A set of 136 hybrids (including all B × B, R × R, and B × R crosses) generated crossing 17 hybrid parents (nine B lines and eight R lines) in half diallel (without reciprocal) fashion, and evaluation at two locations revealed that the correlation between genetic distance and better parent heterosis for grain yield was moderate, positive, and significant (with SSR, 0.33, p < .01; with SNP, 0.35, p < .01), hence both SSRs and SNPs were found comparable in results for heterosis prediction.

Three bacteria, IC-59, IC-76A and IC-2002, isolated from the nodules of chickpea, were characterized for nodulation, nitrogen fixation, plant growth-promoting (PGP) and yield traits in five cultivars of chickpea such as BG256, RSG888, Subhra, K850 and ICCV2. All the bacteria produced cellulase, protease, β-1,3-glucanase, indole acetic acid, siderophore, hydro cyanic acid and 1-aminocyclopropane-1-carboxylate deaminase while none produced lipase and chitinase. The 16 S rDNA gene sequences of IC-59, IC-76A and IC-2002 were found to match closely with Rhizobium pusense, Paraburkholderia kururiensis and Stenotrophomonas maltophilia, respectively. The three bacteria nodulated all the cultivars of chickpea well, amplified nifH gene and fixed nitrogen. Under greenhouse conditions at 30 and 45 days after sowing, treatment of five cultivars of chickpea with bacterial cultures IC-59, IC-76A and IC-2002, enhanced the nodule number (up to 45%, 38% and 43%), nodule weight (up to 31%, 15% and 39%), shoot weight (11%, 16% and 14%) and root weight (37%, 48% and 62%), respectively, over the un-inoculated control. At crop maturity, IC-59, IC-76A and IC-2002 were found to enhance the shoot weight (16%, 40% and 26%), pod number (37%, 69% and 81%), pod weight (17%, 45% and 49%), seed number (21%, 31% and 39%) and seed weight (14%, 56% and 65%), respectively, over the un-inoculated control. Among the five cultivars, Subhra was found to enhance most of the PGP traits when treated with the three diazotrophic bacteria. It is concluded that the three diazotrophic bacteria could be potentially exploited for improving nodulation, nitrogen fixation, PGP and yields of chickpea.

Pearl millet is a non-model grain and fodder crop adapted to extremely hot and dry environments globally. In India, a great deal of public and private sectors’ investment has focused on developing pearl millet single cross hybrids based on the cytoplasmic-genetic male sterility (CMS) system, while in Africa most pearl millet production relies on open pollinated varieties. Pearl millet lines were phenotyped for both the inbred parents and hybrids stage. Many breeding efforts focus on phenotypic selection of inbred parents to generate improved parental lines and hybrids. This study evaluated two genotyping techniques and four genomic selection schemes in pearl millet. Despite the fact that 6× more sequencing data were generated per sample for RAD-seq than for tGBS, tGBS yielded more than 2× as many informative SNPs (defined as those having MAF > 0.05) than RAD-seq. A genomic prediction scheme utilizing only data from hybrids generated prediction accuracies (median) ranging from 0.73-0.74 (1000-grain weight), 0.87-0.89 (days to flowering time), 0.48-0.51 (grain yield) and 0.72-0.73 (plant height). For traits with little to no heterosis, hybrid only and hybrid/inbred prediction schemes performed almost equivalently. For traits with significant mid-parent heterosis, the direct inclusion of phenotypic data from inbred lines significantly (P < 0.05) reduced prediction accuracy when all lines were analyzed together. However, when inbreds and hybrid trait values were both scored relative to the mean trait values for the respective populations, the inclusion of inbred phenotypic datasets moderately improved genomic predictions of the hybrid genomic estimated breeding values. Here we show that modern approaches to genotyping by sequencing can enable genomic selection in pearl millet. While historical pearl millet breeding records include a wealth of phenotypic data from inbred lines, we demonstrate that the naive incorporation of this data into a hybrid breeding program can reduce prediction accuracy, while controlling for the effects of heterosis per se allowed inbred genotype and trait data to improve the accuracy of genomic estimated breeding values for pearl millet hybrids.

Post flowering stalk rot (PFSR) is one of the major biotic constraints to maize production in tropical and sub-tropical environments. It is a complex disease caused by multiple pathogens, among which Fusarium moniliforme and Macrophomina phaseolina are the major ones that cause severe yield losses in the Asian tropics. A set of maize inbred lines was evaluated at two locations for Fusarium stalk rot (FSR) and Macrophomina stalk rot (MSR). Based on line evaluation trials, resistant and susceptible lines were selected and crossed following a Diallel mating design IV to study the gene action for resistance to these stalk rots and the estimating the combining ability of inbred lines. A 9 × 9 diallel (Diallel-A) produced 36 hybrids for studying FSR resistance, and a 12 × 12 diallel (Diallel-B) produced 66 hybrids to analyse the resistance towards both FSR and MSR. These hybrids were evaluated at two locations for MSR and one location for FSR with artificial inoculation. The hybrids differed significantly for FSR (p

There is growing interest in the conservation and utilization of crop wild relatives (CWR) in international food security policy and research. Legumes play an important role in human health, sustainable food production, global food security, and the resilience of current agricultural systems. Pea belongs to the ancient set of cultivated plants of the Near East domestication center and remains an important crop today. Based on genome-wide analysis, P. fulvum was identified as a well-supported species, while the diversity of wild P. sativum subsp. elatius was structured into 5 partly geographically positioned clusters. We explored the spatial and environmental patterns of two progenitor species of domesticated pea in the Mediterranean Basin and in the Fertile Crescent in relation to the past and current climate. This study revealed that isolation by distance does not explain the genetic structure of P. sativum subsp. elatius in its westward expansion from its center of origin. The genetic diversity of wild pea may be driven by Miocene-Pliocene events, while the phylogenetic diversity centers may reflect Pleisto-Holocene climatic changes. These findings help set research and discussion priorities and provide geographical and ecological information for germplasm-collecting missions, as well as for the preservation of extant diversity in ex-situ collections.

The staple crops, maize, sorghum, bambara nut, groundnut, and sunflower common in semi-arid agro-pastoral farming systems of central Tanzania are prone to aflatoxin contamination. Consumption of such crop produce, contaminated with high levels of aflatoxin B1 (AFB1), affects growth and health. In this paper, aflatoxin contamination in freshly harvested and stored crop produce from central Tanzania was examined, including the efficacy of aflatoxin mitigation technologies on grain/kernal quality. A total of 312 farmers were recruited, trained on aflatoxin mitigation technologies, and allowed to deploy the technologies for 2 years. After 2 years, 188 of the 312 farmers were tracked to determine whether they had adopted and complied with the mitigation practices. Aflatoxigenic Aspergillus flavus and aflatoxin B1 contamination in freshly harvested and stored grains/kernels were assessed. A. flavus frequency and aflatoxin production by fungi were assayed by examining culture characteristics and thin-layer chromatography respectively. AFB1 was assayed by enzyme-linked immunosorbent assay. The average aflatoxin contamination in freshly harvested samples was 18.8 μg/kg, which is above the acceptable standard of 10 μg/kg. Contamination increased during storage to an average of 57.2 μg/kg, indicating a high exposure risk. Grains and oilseeds from maize, sorghum, and sunflower produced in aboveground reproductive structures had relatively low aflatoxin contamination compared to those produced in geocarpic structures of groundnut and bambara nut. Farmers who adopted recommended post-harvest management practices had considerably lower aflatoxin contamination in their stored kernels/grains. Furthermore, the effects of these factors were quantified by multivariate statistical analyses. Training and behavioral changes by farmers in their post-harvest practice minimize aflatoxin contamination and improve food safety. Moreover, if non-trained farmers receive mitigation training, aflatoxin concentration is predicted to decrease by 28.9 μg/kg on average.

Genetic diversity in the breeding materials is an essential component to improve the efficiency of any crop improvement programme. In present study, 150 seed and restorer parents were evaluated for grain yield and its component traits at two diverse agro-ecologies CCSHAU-Hisar and ICRISAT-Patancheru. Analysis of variance revealed significant differences among hybrid parents for all traits. Clustering of hybrid parents clearly partitioned almost all the B- and R-lines into separate groups indicating B- and R-lines to be genetically distant from each other. Further, seed (B-lines) and restorer (R-lines) parents were found distributed in four clusters each, and hybrid parents having same plant type (having similar set of specific traits) were found in same cluster. Significant numbers of hybrid parents having common parent in their pedigrees were found in same cluster. Cluster B-IV of B-lines and cluster R-III of R-lines had higher grain yield and above average performance for other grain linked traits. Grain yield was found to have highly positive significant correlation with all the traits under study for both B- and R-lines, except for effective tillers plant-1. Stover yield followed by panicle girth and panicle length had highest positive direct effect on grain yield.

Mungbean (Vigna radiata (L.) Wilczek) is an important pulse crop in India. A major constraint for improved productivity is the yield loss caused by mungbean yellow mosaic disease (MYMD). This disease is caused by several begomoviruses which are transmitted by the whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). The objective of this study was to identify the predominant begomoviruses infecting mungbean and the major cryptic species of B. tabaci associated with this crop in India. The indigenous B. tabaci cryptic species Asia II 1 was found dominant in Northern India, whereas Asia II 8 was found predominant in Southern India. Repeated samplings over consecutive years indicate a stable situation with, Mungbean yellow mosaic virus strains genetically most similar to a strain from urdbean (MYMV-Urdbean) predominant in North India, strains most similar to MYMV-Vigna predominant in South India, and Mungbean yellow mosaic India virus (MYMIV) strains predominant in Eastern India. In field studies, mungbean line NM 94 showed a high level of tolerance to the disease in the Eastern state of Odisha where MYMIV was predominant and in the Southern state of Andhra Pradesh where MYMV-Vigna was predominant, but only a moderate level of tolerance in the Southern state of Tamil Nadu. However, in Northern parts of India where there was high inoculum pressure of MYMV-Urdbean during the Kharif season, NM 94 developed severe yellow mosaic symptoms. The identification of high level of tolerance in mungbean lines such as ML 1628 and of resistance in black gram and rice bean provides hope for tackling the disease through resistance breeding.

Successful management and utilization of increasingly large genomic datasets is essential for breeding programs to increase genetic gain and accelerate cultivar development. To help with data management and storage, we developed a sorghum Practical Haplotype Graph (PHG) pangenome database that stores all identified haplotypes and variant information for a given set of individuals. We developed two PHGs in sorghum, one with 24 individuals and another with 398 individuals, that reflect the diversity across genic regions of the sorghum genome. 24 founders of the Chibas sorghum breeding program were sequenced at low coverage (0.01x) and processed through the PHG to identify genome-wide variants. The PHG called SNPs with only 5.9% error at 0.01x coverage - only 3% lower than its accuracy when calling SNPs from 8x coverage sequence. Additionally, 207 progeny from the Chibas genomic selection (GS) training population were sequenced and processed through the PHG. Missing genotypes in the progeny were imputed from the parental haplotypes available in the PHG and used for genomic prediction. Mean prediction accuracies with PHG SNP calls range from 0.57-0.73 for different traits, and are similar to prediction accuracies obtained with genotyping-by-sequencing (GBS) or markers from sequencing targeted amplicons (rhAmpSeq). This study provides a proof of concept for using a sorghum PHG to call and impute SNPs from low-coverage sequence data and also shows that the PHG can unify genotype calls from different sequencing platforms. By reducing the amount of input sequence needed, the PHG has the potential to decrease the cost of genotyping for genomic selection, making GS more feasible and facilitating larger breeding populations that can capture maximum recombination. Our results demonstrate that the PHG is a useful research and breeding tool that can maintain variant information from a diverse group of taxa, store sequence data in a condensed but readily accessible format, unify genotypes from different genotyping methods, and provide a cost-effective option for genomic selection for any species.

Genetic diversity of 130 forage-type hybrid parents of pearl millet was investigated based on multiple season data of morphological traits and two type of markers: SSRs (Simple sequence repeats) and GBS identified SNPs (Genotyping by sequencing-Single nucleotide polymorphism). Most of the seed and pollinator parents clustered into two clear-cut separate groups based on marker based genetic distance. Significant variations were found for forage related morphological traits at different cutting intervals (first and second cut) in hybrid parents. Across two cuts, crude protein (CP) varied from 11% to 15%, while in vitro organic matter digestibility (IVOMD) varied from 51% to 56%. Eighty hybrids evaluated in multi-location trial along with their parents for forage traits showed that significant heterosis can be realized for forage traits. A low but positive significant correlation found between SSR based genetic distance (GD between parents of hybrid) and heterosis for most of the forage traits indicated that SSR-based GD can be used for predicting heterosis for GFY, DFY and CP in pearl millet. An attempt was made to associate marker-based clusters with forage quality traits, to enable breeders select parents for crossing purposes in forage breeding programs.

This study was conducted to dissect the genetic basis and to explore the candidate genes underlying one of the important genomic regions on an SBI-10 long arm (L), governing the complex stay-green trait contributing to post-flowering drought-tolerance in sorghum. A fine-mapping population was developed from an introgression line cross&mdash, RSG04008-6 (stay-green) &times, J2614-11 (moderately senescent). The fine-mapping population with 1894 F2 was genotyped with eight SSRs and a set of 152 recombinants was identified, advanced to the F4 generation, field evaluated with three replications over 2 seasons, and genotyped with the GBS approach. A high-resolution linkage map was developed for SBI-10L using 260 genotyping by sequencing&mdash, Single Nucleotide Polymorphism (GBS&ndash, SNPs). Using the best linear unpredicted means (BLUPs) of the percent green leaf area (%GL) traits and the GBS-based SNPs, we identified seven quantitative trait loci (QTL) clusters and single gene, mostly involved in drought-tolerance, for each QTL cluster, viz., AP2/ERF transcription factor family (Sobic.010G202700), NBS-LRR protein (Sobic.010G205600), ankyrin-repeat protein (Sobic.010G205800), senescence-associated protein (Sobic.010G270300), WD40 (Sobic.010G205900), CPK1 adapter protein (Sobic.010G264400), LEA2 protein (Sobic.010G259200) and an expressed protein (Sobic.010G201100). The target genomic region was thus delimited from 15 Mb to 8 genes co-localized with QTL clusters, and validated using quantitative real-time (qRT)&ndash, PCR.

With high global demand, seeds and gum of cluster bean are an essential raw material for various industries. Worldwide the low yield and productivity of guar (Cyamopsis tetragonoloba) create a large gap between demand and supply of guar. Therefore, to upsurge guar production, there is a need to improve the guar at genetic level. The genetic improvement of cluster bean is slower due to insufficient genomic resources, co-dominant marker system especially simple sequence repeats (SSRs) and inadequate information on the variability of germplasm. For development of microsatellite markers in cluster bean, DNA survey sequencing was carried out using the Miseq NGS system. DNA sequencing generated 73,934 sequences which harboured microsatellite repeat sequences. Successfully, a total of 15,399 marker-pairs were designed. The amplicon size ranged between 101–385 base-pairs. To validate the primers and to analyse the diversity in released varieties and germplasm of cluster bean, successfully amplifiable 21 primers pairs were used. With a mean of 2.05 alleles per primers, 21 bands were detected. The polymorphism information content (PIC) for polymorphic markers fluctuated between 0.04 - 0.67 (mean = 0.3). With the average dissimilarity coefficient of 0.11, four clusters were noticed with NJ analysis. The grouping pattern of genotypes indicated a low genetic variability in the guar gene pool. The results of this study suggested that instead of re-circulation of parental lines, hybrid breeding using diverse parents followed by selection should be performed for genetic improvement of cluster bean. Keywords: Cluster bean, Diversity, DNA marker, NGS, SSR

Dry root rot (DRR) is an important emerging disease of mungbean caused by fungus Macrophomina phaseolina. The disease is seed and soil-borne and hence management is difficult. Exploitation of host resistance could be a good option to manage the disease. The objectives of this study were to characterize the isolates of M. phaseolina from three legume crops and to identify resistant sources against DRR of mungbean. Isolates of M. phaseolina from mungbean, urdbean, and vegetable soybean were identified using morphological characteristics and sequencing internal transcribed sequence (ITS) region of 18S rRNA. Isolates of M. phaseolina from urdbean and vegetable soybean were inoculated on susceptible check genotype of mungbean to assess their pathogenicity. Forty three mungbean genotypes were screened against M. phaseolina isolate of mungbean using the paper towel method to identify sources of resistance. Among these genotypes, 9 were resistant in repeated experiment, with disease score ranging from 1.9 to 3.0. Resistant genotypes showing lower disease score, namely IPM99-125, EC693368, and EC693369, were further screened using the sick pot method to confirm their resistance. Among these three genotypes, IPM99-125 showed consistently higher plant survival rate followed by EC693368 and EC693369 as compared to susceptible checks (VC3960-88, KPS1). In addition, these three genotypes were resistant when screened with two strains of M. phaseolina isolated from urdbean and vegetable soybean, with IPM99-125 having lower disease score. The mungbean genotype IPM99-125 could be useful in mungbean breeding programs to develop root rot-resistant varieties.

Chickpea (Cicer arietinum L.), a cool-season legume, is increasingly affected by heat-stress at reproductive stage due to changes in global climatic conditions and cropping systems. Identifying quantitative trait loci (QTLs) for heat tolerance may facilitate breeding for heat tolerant varieties. The present study was aimed at identifying QTLs associated with heat tolerance in chickpea using 292 F8-9 recombinant inbred lines (RILs) developed from the cross ICC 4567 (heat sensitive) &times, ICC 15614 (heat tolerant). Phenotyping of RILs was undertaken for two heat-stress (late sown) and one non-stress (normal sown) environments. A genetic map spanning 529.11 cM and comprising 271 genotyping by sequencing (GBS) based single nucleotide polymorphism (SNP) markers was constructed. Composite interval mapping (CIM) analysis revealed two consistent genomic regions harbouring four QTLs each on CaLG05 and CaLG06. Four major QTLs for number of filled pods per plot (FPod), total number of seeds per plot (TS), grain yield per plot (GY) and % pod setting (%PodSet), located in the CaLG05 genomic region, were found to have cumulative phenotypic variation of above 50%. Nineteen pairs of epistatic QTLs showed significant epistatic effect, and non-significant QTL &times, environment interaction effect, except for harvest index (HI) and biomass (BM). A total of 25 putative candidate genes for heat-stress were identified in the two major genomic regions. This is the first report on QTLs for heat-stress response in chickpea. The markers linked to the above mentioned four major QTLs can facilitate marker-assisted breeding for heat tolerance in chickpea.

Pearl millet is a climate resilient crop and one of the most widely grown millets worldwide. Heterotic hybrid development is one of the principal breeding objectives in pearl millet. In a maiden attempt to identify heterotic groups for grain yield, a total of 343 hybrid parental [maintainer (B-) and restorer (R-)] lines were genotyped with 88 polymorphic SSR markers. The SSRs generated a total of 532 alleles with a mean value of 6.05 alleles per locus, mean gene diversity of 0.55, and an average PIC of 0.50. Out of 532 alleles, 443 (83.27%) alleles were contributed by B-lines with a mean of 5.03 alleles per locus. R-lines contributed 476 alleles (89.47%) with a mean of 5.41, while 441 (82.89%) alleles were shared commonly between B- and R-lines. The gene diversity was higher among R-lines (0.55) compared to B-lines (0.49). The unweighted neighbor-joining tree based on simple matching dissimilarity matrix obtained from SSR data clearly differentiated B- lines into 10 sub-clusters (B1 through B10), and R- lines into 11 sub-clusters (R1 through R11). A total of 99 hybrids (generated by crossing representative 9 B- and 11 R- lines) along with checks were evaluated in the hybrid trial. The 20 parents were evaluated in the line trial. Both the trials were evaluated in three environments. Based on per se performance, high sca effects and standard heterosis, F1s generated from crosses between representatives of groups B10R5, B3R5, B3R6, B4UD, B5R11, B2R4, and B9R9 had high specific combining ability for grain yield compared to rest of the crosses. These groups may represent putative heterotic gene pools in pearl millet.

Yield under drought stress is a highly complex trait with large influence to even a minor fluctuation in the environmental conditions. Genomics-assisted breeding holds great promise for improving such complex traits more efficiently in less time, but requires markers associated with the trait of interest. In this context, a recombinant inbred line mapping population (TAG 24 × ICGV 86031) was used to identify markers associated with quantitative trait loci (QTLs) for yield and yield related traits at two important locations of West Africa under well watered and water stress conditions. Among the traits analyzed under WS condition, the harvest index (HI) and the haulm yield (HYLD) were positively correlated with the pod yield (PYLD) and showed intermediate broad sense heritability. QTL analysis using phenotyping and genotyping data resulted in identification of 52 QTLs. These QTLs had low phenotypic variance (

In cytoplasmic genetic male sterility-based (CGMS) hybrid seed production, instability of expression of male-sterility and fertility restoration across a wide range of environments are two of the major difficulties. Therefore, the present study was carried out to investigate the stability of male sterility of nine CGMS lines under three dates of sowing and the fertility restoration of 10 CGMS-based pigeonpea (Cajanus cajan (L.) Millsp.) hybrids at three different locations. Significant variability existed for pollen fertility among hybrids and sterility among cytoplasmic male sterile (CMS) lines. All the hybrids except ICPH 3494 and ICPH 3491 exhibited high (>80%) pollen fertility across locations. Hybrids ICPH 2671, ICPH 2740, and ICPH 3933 had 100% male-fertile plants across locations. All the CMS lines had completely male-sterile plants across sowing dates. The CMS lines BRG1 A, Hy3C A, BRG3 A, and TTB7 A exhibited 100% pollen sterility at different sowing dates. The pooled analysis revealed a signific...

Pearl millet is a climate resilient crop and one of the most widely grown millets worldwide. Heterotic hybrid development is one of the principal breeding objectives in pearl millet. In a maiden attempt to identify heterotic groups for grain yield, a total of 343 hybrid parental [maintainer (B-) and restorer (R-)] lines were genotyped with 88 polymorphic SSR markers. The SSRs generated a total of 532 alleles with a mean value of 6.05 alleles per locus, mean gene diversity of 0.55, and an average PIC of 0.50. Out of 532 alleles, 443 (83.27%) alleles were contributed by B-lines with a mean of 5.03 alleles per locus. R-lines contributed 476 alleles (89.47%) with a mean of 5.41, while 441 (82.89%) alleles were shared commonly between B- and R-lines. The gene diversity was higher among R-lines (0.55) compared to B-lines (0.49). The unweighted neighbor-joining tree based on simple matching dissimilarity matrix obtained from SSR data clearly differentiated B- lines into 10 sub-clusters (B1 through B10), and R- lines into 11 sub-clusters (R1 through R11). A total of 99 hybrids (generated by crossing representative 9 B- and 11 R- lines) along with checks were evaluated in the hybrid trial. The 20 parents were evaluated in the line trial. Both the trials were evaluated in three environments. Based on per se performance, high sca effects and standard heterosis, F1s generated from crosses between representatives of groups B10R5, B3R5, B3R6, B4UD, B5R11, B2R4, and B9R9 had high specific combining ability for grain yield compared to rest of the crosses. These groups may represent putative heterotic gene pools in pearl millet.

The low grain iron and zinc densities are well documented problems in food crops, affecting crop nutritional quality especially in cereals. Sorghum is a major source of energy and micronutrients for majority of population in Africa and central India. Understanding genetic variation, genotype × environment interaction and association between these traits is critical for development of improved cultivars with high iron and zinc. A total of 336 sorghum RILs (Recombinant Inbred Lines) were evaluated for grain iron and zinc concentration along with other agronomic traits for 2 years at three locations. The results showed that large variability exists in RIL population for both micronutrients (Iron = 10.8 to 76.4 mg kg−1 and Zinc = 10.2 to 58.7 mg kg−1, across environments) and agronomic traits. Genotype × environment interaction for both micronutrients (iron and zinc) was highly significant. GGE biplots comparison for grain iron and zinc showed greater variation across environments. The results also showed that G × E was substantial for grain iron and zinc, hence wider testing needed for taking care of G × E interaction to breed micronutrient rich sorghum lines. Iron and zinc concentration showed high significant positive correlation (across environment = 0.79; p < 0.01) indicating possibility of simultaneous effective selection for both the traits. The RIL population showed good variability and high heritabilities (>0.60, in individual environments) for Fe and Zn and other traits studied indicating its suitability to map QTL for iron and zinc.

article i nfo Advances in next-generation sequencing and genotyping technologies have enabled generation of large-scale genomic resources such as molecular markers, transcript reads and BAC-end sequences (BESs) in chickpea, pigeonpea and groundnut, three major legume crops of the semi-arid tropics. Comprehensive transcriptome assemblies and genome sequences have either been developed or underway in these crops. Based on these resources, dense genetic maps, QTL maps as well as physical maps for these legume species have also been developed. As a result, these crops have graduated from 'orphan' or 'less-studied' crops to 'genomic resources rich' crops. This article summarizes the above-mentioned advances in genomics and genomics-assisted breeding applications in the form of marker-assisted selection (MAS) for hybrid purity assessment in pigeonpea; marker-assisted backcrossing (MABC) for introgressing QTL region for drought-tolerance related traits, Fusarium wilt (FW) resistance and Ascochyta blight (AB) resistance in chickpea; late leaf spot (LLS), leaf rust and nematode resistance in groundnut. We critically present the case of use of other modern breeding approaches like marker-assisted recurrent selection (MARS) and genomic selection (GS) to utilize the full potential of genomics-assisted breeding for developing superior cultivars with enhanced tolerance to various environmental stresses. In addition, this article recommends the use of advanced-backcross (AB-backcross) breeding and development of specialized populations such as multi-parents advanced generation intercross (MAGIC) for creating new variations that will help in developing superior lines with broadened genetic base. In summary, we propose the use of integrated genomics and breeding approach in these legume crops to enhance crop productivity in marginal environments ensuring food security in developing countries.

The first sorghum [Sorghum bicolor (L.) Moench] hybrids based on West African Guinea-race-derived parents were created to enhance farmer’s food security and income through increased yields. To assess their performance, eight hybrids, six experimental pure-line cultivars, one pure-line check (Lata), and a highly adapted landrace cultivar (Tieble) were evaluated in 27 farmer-managed and two on-station yield trials in Mali, West Africa, from 2009 to 2011. The hybrids were confirmed to have photoperiod sensitivity similar to the well-adapted Guinea landrace check cultivar. Genotypic differences for on-farm grain yield were highly significant and genotype × environment crossover interactions were limited. The yield superiorities of individual hybrids, relative to the landrace check, ranged from 17 to 37% over the 27 on-farm trials. The three top yielding hybrids showed 30% yield advantages across productivity levels, with absolute yield advantages averaging 380 kg ha−1 under lower (1.0–1.5 t ha−1) and 660 kg ha−1 under higher (2.0–3.5 t ha−1) productivity conditions. A mean male-parent (better parent) heterosis of 26% was observed for the four hybrids having Lata as a male parent. As the hybrids studied here were obtained with a low intensity of selection using a limited number of parents, even greater yield superiorities may be attained with development of distinct parental pools and scaled-up hybrid breeding.

Transgenic research using the DREB group of transcription factors has received much attention in developing drought-tolerant and climate-ready varieties of crop plants. While many reports have demonstrated increased tolerance to water deficits under laboratory and greenhouse conditions, only a few have tested possible effects under field conditions, with limited success in most cases. Here, we present evidence of a transgenic solution for enhanced drought tolerance in peanut (Arachis hypogaea L.), which is an important grain legume and a valuable cash crop for smallholder and resource-poor farmers occupying the largest portion of the farming systems in low income food deficit countries. The presence, integration, expression and inheritance of the transgene in advancing generations of the transgenic peanut plants were assessed using PCR, Southern blot, inverse-PCR, RT-PCR and q-PCR techniques. Four trials were conducted in various water stress regimes under varying vapour pressure deficits (VPDs), and drought tolerance was studied using various component traits of drought. A substantial yield improvement of up to 24 % in drought trials under field conditions was achieved across a wide range of stress intensities and was related to higher harvest indices (HIs). All transgenic events had significantly higher seed filling under drought and displayed 20–30 % lower pod yield reduction than their untransformed counterpart under drought stress. Two transgenic events showed yield advantage under drought stress that consistently had higher pod and seed yield than the untransformed parent under drought stress across all trials, without displaying any yield penalty under irrigated conditions.

Fusarium wilt (FW) and Ascochyta blight (AB) are two important diseases of chickpea which cause 100 % yield losses under favorable conditions. With an objective to validate and/or to identify novel quantitative trait loci (QTLs) for resistance to race 1 of FW caused by Fusarium oxysporum f. sp. ciceris and AB caused by Ascochyta rabiei in chickpea, two new mapping populations (F2:3) namely ‘C 214’ (FW susceptible) × ‘WR 315’ (FW resistant) and ‘C 214’ (AB susceptible) × ‘ILC 3279’ (AB resistant) were developed. After screening 371 SSR markers on parental lines and genotyping the mapping populations with polymorphic markers, two new genetic maps comprising 57 (C 214 × WR 315) and 58 (C 214 × ILC 3279) loci were developed. Analysis of genotyping data together with phenotyping data collected on mapping population for resistance to FW in field conditions identified two novel QTLs which explained 10.4–18.8 % of phenotypic variation. Similarly, analysis of phenotyping data for resistance to seedling resistance and adult plant resistance for AB under controlled and field conditions together with genotyping data identified a total of 6 QTLs explaining up to 31.9 % of phenotypic variation. One major QTL, explaining 31.9 % phenotypic variation for AB resistance was identified in both field and controlled conditions and was also reported from different resistant lines in many earlier studies. This major QTL for AB resistance and two novel QTLs identified for FW resistance are the most promising QTLs for molecular breeding separately or pyramiding for resistance to FW and AB for chickpea improvement.

The Thar Desert, a very inhospitable place, accommodates only plant species that survive acute drought, unpredictable precipitation, and those can grow in the limited moisture of sandy soils. Capparis decidua is among one of the few plants able to grow well under these conditions. This species is highly exploited and has been naturally taken, as local people use it for various purposes like food, timber and fuel, although, no management or conservation efforts have been established. The present study was conducted in this arid area of Western Rajasthan (India) with the aim to obtain preliminary molecular information about this group of plants. We evaluated diversity among 46 samples of C. decidua using chemical parameters and random amplified polymorphic DNA (RAPD) markers. Fourteen chemical parameters and eight minerals (total 22 variables) of this species fruits were estimated. A total of 14 RAPD primers produced 235 band positions, of which 81.27% were polymorphic. Jaccard s similarity coefficients for RAPD primers ranged from 0.34 to 0.86 with a mean genetic similarity of 0.50. As per observed coefficient of variation, NDF (Neutral Detergent Fiber) content was found to be the most variable trait followed by starch and soluble carbohydrate. The Manhattan dissimilarity coefficient values for chemical parameters ranged between 0.02-0.31 with an average of 0.092. The present study revealed a very low correlation (0.01) between chemical parameters and RAPD-based matrices. The low correlation between chemical- and RAPD-based matrices indicated that the two methods were different and highly variable. The chemical-based diversity will assist in selection of nutritionally rich samples for medicinal purpose, while genetic diversity to face natural challenges and find sustainable ways to promote conservation for future use.El desierto de Thar, un lugar muy inhóspito, alberga sólo a las especies de plantas capaces de resistir a condiciones de sequía extrema, a las precipitaciones impredecibles, y a las plantas que pueden crecer en la humedad limitada de los suelos arenosos. Capparis decidua se encuentra entre una las pocas plantas capaces de crecer bien en estas condiciones. Esta especie es altamente explotada y se ha tomado de forma natural, así los habitantes locales las han usado para varios propósitos, como alimento, madera y combustible, aunque sin ningún programa de manejo o esfuerzo por conservación. El presente estudio se llevó a cabo en esta zona árida del oeste de Rajastán (India) con el objetivo de obtener información molecular preliminar sobre este grupo de plantas. Se evaluó la diversidad entre 46 muestras de C. decidua usando parámetros químicos y marcadores de ADN polimórfico amplificado al azar (RAPD por sus siglas en inglés). Catorce parámetros químicos y ocho minerales (22 variables en total) de los frutos de esta especie fueron estimados. Un total de 14 cebadores para RAPD produjeron 235 posiciones de bandas, de las cuales 81.27% fueron polimórficas. El coeficiente de similitud de Jaccard para los cebadores del RAPD varió entre 0.34 y 0.86 con un promedio de similitud genética de 0.50. De acuerdo con el coeficiente de variación observado, se encontró que el contenido de NDF fue el rasgo más variable, seguido por el almidón y los carbohidratos solubles. Los valores del coeficiente de disimilitud de Manhattan para los parámetros químicos osciló entre 0.02-0.31 con un promedio de 0.092. El presente estudio reveló una correlación muy baja (0.01) entre los parámetros químicos y las matrices basadas en RAPD. La baja correlación entre las matrices químicas y la basada en RAPD indicó que los dos métodos fueron diferentes y altamente variables. El estudio de la diversidad basada en su química ayudará en la selección de muestras nutricionalmente ricas para fines medicinales, mientras que la diversidad genética ayudará a enfrentar los desafíos naturales y encontrar formas sostenibles para promover la conservación de esta plana para uso futuro.

The low grain iron and zinc densities are well documented problems in food crops, affecting crop nutritional quality especially in cereals. Sorghum is a major source of energy and micronutrients for majority of population in Africa and central India. Understanding genetic variation, genotype × environment interaction and association between these traits is critical for development of improved cultivars with high iron and zinc. A total of 336 sorghum RILs (Recombinant Inbred Lines) were evaluated for grain iron and zinc concentration along with other agronomic traits for 2 years at three locations. The results showed that large variability exists in RIL population for both micronutrients (Iron = 10.8 to 76.4 mg kg−1 and Zinc = 10.2 to 58.7 mg kg−1, across environments) and agronomic traits. Genotype × environment interaction for both micronutrients (iron and zinc) was highly significant. GGE biplots comparison for grain iron and zinc showed greater variation across environments. The results also showed that G × E was substantial for grain iron and zinc, hence wider testing needed for taking care of G × E interaction to breed micronutrient rich sorghum lines. Iron and zinc concentration showed high significant positive correlation (across environment = 0.79; p < 0.01) indicating possibility of simultaneous effective selection for both the traits. The RIL population showed good variability and high heritabilities (>0.60, in individual environments) for Fe and Zn and other traits studied indicating its suitability to map QTL for iron and zinc.

Pigeonpea (Cajanus cajan L.) is an important food legume crop of rainfed agriculture. Owing to exposure of the crop to a number of biotic and abiotic stresses, the crop productivity has remained stagnant for almost last five decades at ca. 750 kg/ha. The availability of a cytoplasmic male sterility (CMS) system has facilitated the development and release of hybrids which are expected to enhance the productivity of pigeonpea. Recent advances in genomics and molecular breeding such as marker-assisted selection (MAS) offer the possibility to accelerate hybrid breeding. Molecular markers and genetic maps are pre-requisites for deploying MAS in breeding. However, in the case of pigeonpea, only one inter- and two intra-specific genetic maps are available so far. Here, four new intra-specific genetic maps comprising 59–140 simple sequence repeat (SSR) loci with map lengths ranging from 586.9 to 881.6 cM have been constructed. Using these four genetic maps together with two recently published intra-specific genetic maps, a consensus map was constructed, comprising of 339 SSR loci spanning a distance of 1,059 cM. Furthermore, quantitative trait loci (QTL) analysis for fertility restoration (Rf) conducted in three mapping populations identified four major QTLs explaining phenotypic variances up to 24 %. To the best of our knowledge, this is the first report on construction of a consensus genetic map in pigeonpea and on the identification of QTLs for fertility restoration. The developed consensus genetic map should serve as a reference for developing new genetic maps as well as correlating with the physical map in pigeonpea to be developed in near future. The availability of more informative markers in the bins harbouring QTLs for sterility mosaic disease (SMD) and Rf will facilitate the selection of the most suitable markers for genetic analysis and molecular breeding applications in pigeonpea. Electronic supplementary material The online version of this article (doi:10.1007/s00122-012-1916-5) contains supplementary material, which is available to authorized users.

Sweet sorghum is a multipurpose biofuel feedstock that offers grain for human consumption, fodder for livestock and ethanol for transportation purposes. The knowledge on sugar components at different phenological stages of crop growth and identification of appropriate stage of harvesting is critical for sweet sorghum commercialization and value chain sustenance. In this regard, sweet sorghum stalk yield, juice yield, Brix%, pH, sugars (sucrose, fructose and glucose) and their content were analyzed at three different phenological stages i.e. the dough stage, physiological maturity and post-physiological maturity. Variations in sugar content at different growth stages revealed that the sugar yield was high at physiological maturity, but highest at post-physiological maturity. Sucrose accounts for major fermentable sugar (about 70%) and it sharply increased by 146% from dough stage to post-physiological maturity. The variation in the monosaccharides content (glucose and fructose) is not statistically significant. This study points to the potential scope for widening the harvesting window of sweet sorghum, by cutting the stalks from physiological maturity stage and beyond up to 15 days (post-physiological maturity), thus helping the commercial distilleries by addressing a major impediment in sweet sorghum value chain. The entries SP 4495, SP 4511-3 and SPV 422 are suitable for harvesting in a wider window of time as the sugar levels are sustained at same level from physiological maturity to post-physiological maturity.

Axiom Cajanus SNP array revealed genetic architecture and temporal diversity in pigeonpea varieties. As one of the major outputs of next‐generation sequencing (NGS), a large number of genome‐wide single‐nucleotide polymorphisms (SNPs) have been developed in pigeonpea [Cajanus cajan (L.) Huth.]. However, SNPs require a genotyping platform or assay to be used in different evolutionary studies or in crop improvement programs. Therefore, we developed an Axiom Cajanus SNP array with 56K SNPs uniformly distributed across the genome and assessed its utility in a genetic diversity study. From the whole‐genome resequencing (WGRS) data on 104 pigeonpea lines, ∼2 million sequence variations (SNPs and insertion–deletions [InDels]) were identified, from which a subset of 56,512 unique and informative sequence variations were selected to develop the array. The Axiom Cajanus SNP array developed was used for genotyping 103 pigeonpea lines encompassing 63 cultivars released between 1960 and 2014 and 40 breeding, germplasm, and founder lines. Genotyping data thus generated on 103 pigeonpea lines provided 51,201 polymorphic SNPs and InDels. Genetic diversity analysis provided in‐depth insights into the genetic architecture and trends in temporal diversity in pigeonpea cultivars. Therefore, the continuous use of the high‐density Axiom Cajanus SNP array developed will accelerate high‐resolution trait mapping, marker‐assisted breeding, and genomic selection efforts in pigeonpea.

Ascochyta blight (AB, Ascochyta rabiei (Pass.) Lab.) is one of the most important foliar disease of chickpea (Cicer arietinum L.), globally. Chickpea is attacked by AB at any growth stage in cool and humid weather depending on the inoculum availability. However, the disease epidemics are most prominent during the flowering and podding growth stages. The main objective of this study was to determine the effect of growth stages of chickpea on the genetic resistance of AB and use this information in a resistance breeding program. Two susceptible and two moderately resistant chickpea cultivars were spray inoculated at seedling (GS1), post-seedling (GS2), vegetative (GS3), flowering (GS4) and podding (GS5) growth stages with A. rabiei conidial suspension under controlled environment conditions. Irrespective of crop cultivars the incubation period (IP) was shorter in GS1, GS4 and GS5 and was significantly extended in GS2 and GS3. Symptom development was delayed significantly in moderately resistant cultivars. The AB severity 10 days after inoculation ranged between 7 and 9 on susceptible cultivars and 3 and 5 on moderately resistant cultivars. Further the correlation coefficient of disease severity between GS1, GS4 and GS5 was highly significant (r = 0.95) indicating that, evaluation for resistance to AB can be done at GS 1 (seedling stage), and or GS4 (flowering stage) to GS5 (podding stage) growth stages of chickpea. This supports the evaluation for AB resistance using 10-day-old-seedlings in controlled environment at ICRISAT and adult plant field screening at hot-spot locations in Dhaulakuan and Ludhiana in India.

Knowledge of current genetic diversity and mating systems of crop wild relatives (CWR) in the Fertile Crescent is important in crop genetic improvement, because western agriculture began in the area after the cold-dry period known as Younger Dryas about 12,000 years ago and these species are also wild genepools of the world's most important food crops. Wild pea (Pisum sativum subsp. elatius) is an important source of genetic diversity for further pea crop improvement harbouring traits useful in climate change context. The genetic structure was assessed on 187 individuals of Pisum sativum subsp. elatius from fourteen populations collected in the northern part of the Fertile Crescent using 18,397 genome wide single nucleotide polymorphism DARTseq markers. AMOVA showed that 63% of the allelic variation was distributed between populations and 19% between individuals within populations. Four populations were found to contain admixed individuals. The observed heterozygosity ranged between 0.99 to 6.26% with estimated self-pollination rate between 47 to 90%. Genetic distances of wild pea populations were correlated with geographic but not environmental (climatic) distances and support a mixed mating system with predominant self-pollination. Niche modelling with future climatic projections showed a local decline in habitats suitable for wild pea, making a strong case for further collection and ex situ conservation.

Knowledge of current genetic diversity and mating systems of crop wild relatives (CWR) in the Fertile Crescent is important in crop genetic improvement, because western agriculture began in the area after the cold-dry period known as Younger Dryas about 12,000 years ago and these species are also wild genepools of the world's most important food crops. Wild pea (Pisum sativum subsp. elatius) is an important source of genetic diversity for further pea crop improvement harbouring traits useful in climate change context. The genetic structure was assessed on 187 individuals of Pisum sativum subsp. elatius from fourteen populations collected in the northern part of the Fertile Crescent using 18,397 genome wide single nucleotide polymorphism DARTseq markers. AMOVA showed that 63% of the allelic variation was distributed between populations and 19% between individuals within populations. Four populations were found to contain admixed individuals. The observed heterozygosity ranged between 0.99 to 6.26% with estimated self-pollination rate between 47 to 90%. Genetic distances of wild pea populations were correlated with geographic but not environmental (climatic) distances and support a mixed mating system with predominant self-pollination. Niche modelling with future climatic projections showed a local decline in habitats suitable for wild pea, making a strong case for further collection and ex situ conservation.