Nitrogen use efficiency in bread wheat: Genetic variation and prospects for improvement.

Nitrogen (N) is one of the primary macronutrients required for crop growth and yield. This nutrient is especially limiting wheat yields in the dry and low fertile agro-ecologies having low N in the root zone soil strata. Moreover, majority of farmers in India and South Asia are small to marginal with meagre capacity to invest in costly nitrogen fertilizers. Therefore, there is an immense need to identify lines that use nitrogen efficiently. A set of 50 diverse wheat genotypes consisting of indigenous germplasm lines (05), cultivars released for commercial cultivation (23) and selected elite lines from CIMMYT nurseries (22) were evaluated in an alpha-lattice design with two replications, a six-rowed plot of 2.5m length for 24 agro morphological, physiological and NUE related traits during two consecutive crop seasons in an N-depleted precision field under two different N levels of 50%-N50 (T1) and 100%-N100 (T2) of recommended N, i.e., 100 kg/ha. Analysis of variance revealed significant genetic variation among genotypes for all the traits studied. About 11.36% yield reduction was observed at reduced N levels. Significant correlations among NUE traits and yield component traits were observed which indicated pivotal role of N remobilization to the grain in enhancing yield levels. Among N-insensitive genotypes identified based on their yielding ability at low N levels, UASBW13356, UASBW13358, UASBW13354, UASBW13357 and KRL1-4 showed their inherent genotypic plasticity toward N application. The genotypes with more yield and high to moderate NUtE can be used as parents for the breeding of N efficient genotypes for marginal agro-ecologies. Low N tolerant genotypes identified from the current investigation may be further utilized in the identification of genomic regions responsible for NUE and its deployment in wheat breeding programs. The comprehensive data of 24 traits under different nitrogen levels for diverse genotypes from India and global sources (mainly CIMMYT) should be useful for supporting breeding for NUE and thus will be of great help for small and marginal farmers in India and South Asia.

First report of Fusarium poae Causing Fusarium head Blight of Wheat in India.

Wheat is one of major cereal crops with paramount importance that is cultivated across the globe. Fusarium head blight (FHB) is a catastrophic disease of wheat which has recently risen to prominence due to its direct impact on the quality and quantity of wheat on a global scale. During a field survey conducted in Rabi 2021-22, wheat spikes showing characteristic symptoms of head blight were observed in northern parts of the Karnataka, India, in the districts Bagalkote, Belagavi and Dharwad. The infected spikelets from the heads with symptoms of infection were washed well in distilled water and surface sterilized using 1% sodium hypochlorite solution. They were further washed using sterilized distilled water to remove the traces of sodium hypochlorite. These spikelets were then transferred to sterile potato dextrose agar (PDA) plates under aseptic conditions. The plates were incubated at a temperature of 27±1°C for ten days to obtain good fungal growth. The fungus produced white to orangish pink, dense mycelia and hyaline septate hyphae. Macroconidia were sickle-shaped measuring 35.7 to 52.6 x 3.2 to 5.1 µm, dorsoventrally curved with an elongated basal cell ending in a prominent long foot; the apical cell was also elongated, tapered, slightly curved. Spores had 3 to 4 cells and formed on monophialide. Microconidia and chlamydospores were present only in a few isolates. Fungal genomic DNA was extracted from all the established isolates using CTAB (Cetyl-trimethyl ammonium bromide) method (Murray and Thompson, 1980). The ITS region of r-DNA and translation elongation factor-1 alpha (TEF-1α) genes of the ten isolates were amplified using ITS1/ITS4 primer pair (White et al. 1990) and the species-specific EF1F/EF1R primer pair (O'Donnell et al., 1998) respectively, to detect and distinguish within Fusarium species. The results exhibited 95% similarity with Fusarium poae with GenBank Accession No. XMO44849482.1. which was previously reported as a causal agent of Fusarium head blight of wheat in Georgia, USA. The TEF1-α sequences were deposited in the GenBank and the accession numbers from OP716756 to OP716765 were obtained. For the pathogenicity test, spore suspension containing a load of 108 conidia ml-1 was prepared from ten days old Fusarium poae culture and sprayed on the healthy wheat heads of the susceptible variety UAS-304 during anthesis stage by using hand atomizer. Control plants were sprayed in a similar manner with sterile distilled water. In order to enhance disease development and increase the accuracy of the evaluation, humidity was maintained for 72 hours post-inoculation period by covering each spike with a plastic bag and misting at least once daily. The characteristic head blight symptoms were observed in the inoculated wheat spikes. No symptoms were noticed in the water-treated control. The plants initially showed bleaching in single spikelet after seven days of inoculation, which eventually spread to the entire spike. After fourteen days of inoculation, the spikes showed blighted appearance with pinkish or orangish mass of mycelia grown on the affected region. The pathogen was further reisolated from the infected plants and examined under the microscope. The similar morphological features as that of the originally inoculated pathogen was observed, hence fulfilling Koch's postulates. To our knowledge, this is the first report of Fusarium poae causing head blight of wheat in India. Keeping the current climate change scenario in view, the plausibility of this pathogen causing a major havoc in the near future must not be ignored, considering the fact that it has a wide host range and highly evolving nature. The ubiquity of the Fusarium head blight recently in northern parts of Karnataka urges the need to conduct further studies on the variability, distribution and management of the pathogen.

Genome-wide association study for grain yield and component traits in bread wheat (Triticum aestivum L.).

Genomic regions governing days to heading (DH), grain filling duration (GFD), grain number per spike (GNPS), grain weight per spike (GWPS), plant height (PH), and grain yield (GY) were investigated in a set of 280 diverse bread wheat genotypes. The genome-wide association studies (GWAS) panel was genotyped using a 35K Axiom Array and phenotyped in five environments. The GWAS analysis showed a total of 27 Bonferroni-corrected marker-trait associations (MTAs) on 15 chromosomes representing all three wheat subgenomes. The GFD showed the highest MTAs (8), followed by GWPS (7), GY (4), GNPS (3), PH (3), and DH (2). Furthermore, 20 MTAs were identified with more than 10% phenotypic variation. A total of five stable MTAs (AX-95024590, AX-94425015, AX-95210025 AX-94539354, and AX-94978133) were identified in more than one environment and associated with the expression of DH, GFD, GNPS, and GY. Similarly, two novel pleiotropic genomic regions with associated MTAs i.e. AX-94978133 (4D) and AX-94539354 (6A) harboring co-localized QTLs governing two or more traits were also identified. In silico analysis revealed that the SNPs were located on important putative candidate genes such as F-box-like domain superfamily, Lateral organ boundaries, LOB, Thioredoxin-like superfamily Glutathione S-transferase, RNA-binding domain superfamily, UDP-glycosyltransferase family, Serine/threonine-protein kinase, Expansin, Patatin, Exocyst complex component Exo70, DUF1618 domain, Protein kinase domain involved in the regulation of grain size, grain number, growth and development, grain filling duration, and abiotic stress tolerance. The identified novel MTAs will be validated to estimate their effects in different genetic backgrounds for subsequent use in marker-assisted selection (MAS).

Genetic dissection of grain iron and zinc, and thousand kernel weight in wheat (Triticum aestivum L.) using genome-wide association study.

Genetic biofortification is recognized as a cost-effective and sustainable strategy to reduce micronutrient malnutrition. Genomic regions governing grain iron concentration (GFeC), grain zinc concentration (GZnC), and thousand kernel weight (TKW) were investigated in a set of 280 diverse bread wheat genotypes. The genome-wide association (GWAS) panel was genotyped using 35 K Axiom Array and phenotyped in five environments. The GWAS analysis showed a total of 17 Bonferroni-corrected marker-trait associations (MTAs) in nine chromosomes representing all the three wheat subgenomes. The TKW showed the highest MTAs (7), followed by GZnC (5) and GFeC (5). Furthermore, 14 MTAs were identified with more than 10% phenotypic variation. One stable MTA i.e. AX-95025823 was identified for TKW in both E4 and E5 environments along with pooled data, which is located at 68.9 Mb on 6A chromosome. In silico analysis revealed that the SNPs were located on important putative candidate genes such as Multi antimicrobial extrusion protein, F-box domain, Late embryogenesis abundant protein, LEA-18, Leucine-rich repeat domain superfamily, and C3H4 type zinc finger protein, involved in iron translocation, iron and zinc homeostasis, and grain size modifications. The identified novel MTAs will be validated to estimate their effects in different genetic backgrounds for subsequent use in marker-assisted selection. The identified SNPs will be valuable in the rapid development of biofortified wheat varieties to ameliorate the malnutrition problems.

Whole Genome Sequencing and Comparative Genomics of Indian Isolates of Wheat Spot Blotch Pathogen Bipolaris sorokiniana Reveals Expansion of Pathogenicity Gene Clusters.

Spot blotch is a highly destructive disease in wheat caused by the fungal pathogen Bipolaris sorokiniana (teleomorph, Cochliobolus sativus). It is prevalent in warm and humid areas, including Africa, Asia, Latin America, and the USA. In the present study, twelve isolates of B. sorokiniana were collected from wheat fields in three different geographical locations in India. The pathogenicity of seven sporulating isolates was assessed on 'DDK 1025', a spot blotch-susceptible wheat variety under greenhouse conditions. The isolate 'D2' illustrated the highest virulence, followed by 'SI' and 'BS52'. These three isolates were sequenced using the Illumina HiSeq1000 platform. The estimated genome sizes of the isolates BS52, D2, and SI were 35.19 MB, 39.32 MB, and 32.76 MB, with GC contents of 48.48%, 50.43%, and 49.42%, respectively. The numbers of pathogenicity genes identified in BS52, D2, and SI isolates were 2015, 2476, and 2018, respectively. Notably, the isolate D2 exhibited a relatively larger genome with expanded arsenals of Biosynthetic Gene Clusters (BGCs), CAZymes, secretome, and pathogenicity genes, which could have contributed to its higher virulence among the tested isolates. This study provides the first comparative genome analysis of the Indian isolates of B. sorokiniana using whole genome sequencing.

Variations in straw fodder quality and grain-Straw relationships in a mapping population of 287 diverse spring wheat lines.

A wheat association mapping population consisting of 287 diverse spring wheat lines were evaluated for three years in one location (Varanasi) and out of these for one year across three locations (Karnal, Dharwad and Varanasi) in India. Straw fodder quality traits analyzed were nitrogen (N) content, neutral (NDF) and acid (ADF) detergent fiber, acid detergent lignin (ADL), ash (ASH), in vitro organic matter digestibility (IVOMD) and metabolizable energy (ME) content. Grain yield (GY) and straw yield (SY) were also recorded. Highly significant (P < 0.0001) differences among lines were observed for all traits except for ADF and ADL in the three years trials conducted at Varanasi. However, year and location had strong (P < 0.0001) effects on all traits. Compared to line-dependent variations in GY and SY variation in straw fodder quality traits were small. Proportionally greatest variations between lines were observed for straw N where lowest and highest N varied by about 30%. Difference for NDF and ADF between lines were at most 4% units and below 3% units for IVOMD. Grain yield and straw yield were positively correlated (P < 0.0001) with GY accounting for 26% of the variation in SY. Straw N, IVOMD and ME were weakly but significantly (P < 0.05) negatively associated with GY and SY. Straw NDF and ADF were significantly (P < 0.05) positively correlated with GY but the association was again weak. Straw NDF, ADF and ADL were also weakly but significantly positively correlated with SY. Genome-wide association studies (GWAS) were applied to detect significant marker- straw fodder quality trait associations. Five genomic regions contributed for six traits (ADF, ADL, ASH, IVOMD, ME and NDF). ADF and ADL mapped in the common QTL region on chromosome 2B. Similarly, for the IVOMD and ME QTLs on chromosome 5B were associated with SNP marker, wsnp_Ku_c35090_44349517. While some associations were detected for ADF, ALD, ASH, IVOMD, ME and NDF on chromosomes 1A, 2B, 3A, 5A and 5B, the phenotypic variation explained was low to medium by individual QTL. A likely contributing factor was the comparatively small difference in straw fodder quality traits among the lines. It is interesting to note that line dependent variations in GY and SY were about two-fold. In other words, strong genotypic variations of GY and SY do exist. The lack of any similar variations in straw fodder quality traits is intriguing and requires further research.

Correction to: Strategic crossing of biomass and harvest index-source and sink-achieves genetic gains in wheat.

[This corrects the article DOI: 10.1007/s10681-017-2040-z.].