Unravelling consensus genomic regions associated with quality traits in wheat using meta-analysis of quantitative trait loci.

MAIN CONCLUSION: Meta-analysis in wheat for three major quality traits identified 110 meta-QTL (MQTL) with reduced confidence interval (CI). Five GWAS validated MQTL (viz., 1A.1, 1B.2, 3B.4, 5B.2, and 6B.2), each involving more than 20 initial QTL and reduced CI (95%) (< 2 cM), were selected for quality breeding programmes. Functional characterization including candidate gene mining and expression analysis discovered 44 high confidence candidate genes associated with quality traits. A meta-analysis of quantitative trait loci (QTL) associated with dough rheology properties, nutritional traits, and processing quality traits was conducted in wheat. For this purpose, as many as 2458 QTL were collected from 50 interval mapping studies published during 2013-2020. Of the total QTL, 1126 QTL were projected onto the consensus map saturated with 249,603 markers which led to the identification of 110 meta-QTL (MQTL). These MQTL exhibited an 18.84-fold reduction in the average CI compared to the average CI of the initial QTL (ranging from 14.87 to 95.55 cM with an average of 40.35 cM). Of the 110, 108 MQTL were physically anchored to the wheat reference genome, including 51 MQTL verified with marker-trait associations (MTAs) reported from earlier genome-wide association studies. Candidate gene (CG) mining allowed the identification of 2533 unique gene models from the MQTL regions. In-silico expression analysis discovered 439 differentially expressed gene models with > 2 transcripts per million expressions in grains and related tissues, which also included 44 high-confidence CGs involved in the various cellular and biochemical processes related to quality traits. Nine functionally characterized wheat genes associated with grain protein content, high-molecular-weight glutenin, and starch synthase enzymes were also found to be co-localized with some of the MQTL. Synteny analysis between wheat and rice MQTL regions identified 23 wheat MQTL syntenic to 16 rice MQTL associated with quality traits. Furthermore, 64 wheat orthologues of 30 known rice genes were detected in 44 MQTL regions. Markers flanking the MQTL identified in the present study can be used for marker-assisted breeding and as fixed effects in the genomic selection models for improving the prediction accuracy during quality breeding. Wheat orthologues of rice genes and other CGs available from MQTLs can be promising targets for further functional validation and to better understand the molecular mechanism underlying the quality traits in wheat.

Strategies for accelerating genetic gains in crop plants: special focus on speed breeding.

Feeding 10 billion people sustainably by 2050 in the era of slow genetic progress has spurred urgent calls to bring more crops per unit time. Over the last century, crop physiologists and breeders have been trying to alter plant biology to investigate and intervene in developmental processes under controlled chambers. Accelerating the breeding cycle via "speed breeding" was the outcome of these experiments. Speed breeding accelerates the genetic gain via phenome and genome-assisted trait introgression, re-domestication, and plant variety registration. Furthermore, early varietal release through speed breeding offers incremental benefits over conventional methods. However, a lack of resources and species-specific protocols encumber the technological implementation, which can be alleviated by reallocating funds to establish speed breeding units. This review discusses the limitations of conventional breeding methods and various alternative strategies to accelerate the breeding process. It also discusses the intervention at various developmental stages to reduce the generation time and global impacts of speed breeding protocols developed so far. Low-cost, field-based speed breeding protocol developed by Punjab Agricultural University, Ludhiana, Punjab, India to harvest at least three generations of wheat in a year without demanding the expensive greenhouses or growth chambers is also discussed.

Application of potassium nitrate and salicylic acid improves grain yield and related traits by delaying leaf senescence in Gpc-B1 carrying advanced wheat genotypes.

Grain protein content (GPC) is an important quality trait that effectively modulates end-use quality and nutritional characteristics of wheat flour-based food products. The Gpc-B1 gene is responsible for the higher protein content in wheat grain. In addition to higher GPC, the Gpc-B1 is also generally associated with reduced grain filling period which eventually causes the yield penalty in wheat. The main aim of the present study was to evaluate the effect of foliar application of potassium nitrate (PN) and salicylic acid (SA) on the physiological characteristics of a set of twelve genotypes, including nine isogenic wheat lines carrying the Gpc-B1 gene and three elite wheat varieties with no Gpc-B1 gene, grown at wheat experimental area of the Department of Plant Breeding and Genetics, PAU, Punjab, India. The PN application significantly increased the number of grains per spike (GPS) by 6.42 grains, number of days to maturity (DTM) by 1.03 days, 1000-grain weight (TGW) by 1.97 g and yield per plot (YPP) by 0.2 kg/plot. As a result of PN spray, the flag leaf chlorophyll content was significantly enhanced by 2.35 CCI at anthesis stage and by 1.96 CCI at 10 days after anthesis in all the tested genotypes. Furthermore, the PN application also significantly increased the flag leaf nitrogen content by an average of 0.52% at booting stage and by 0.35% at both anthesis and 10 days after anthesis in all the evaluated genotypes. In addition, the yellow peduncle colour at 30 days after anthesis was also increased by 19.08% while the straw nitrogen content was improved by 0.17% in all the genotypes. The preliminary experiment conducted using SA demonstrated a significant increase in DTM and other yield component traits. The DTM increased by an average of 2.31 days, GPS enhanced by approximately 3.17 grains, TGW improved by 1.13g, and YPP increased by 0.21 kg/plot. The foliar application of PN and SA had no significant effect on GPC itself. The findings of the present study suggests that applications of PN and SA can effectively mitigate the yield penalty associated with Gpc-B1 gene by extending grain filling period in the wheat.

A Prospective Review on Selectable Marker-Free Genome Engineered Rice: Past, Present and Future Scientific Realm.

As a staple food crop, rice has gained mainstream attention in genome engineering for its genetic improvement. Genome engineering technologies such as transgenic and genome editing have enabled the significant improvement of target traits in relation to various biotic and abiotic aspects as well as nutrition, for which genetic diversity is lacking. In comparison to conventional breeding, genome engineering techniques are more precise and less time-consuming. However, one of the major issues with biotech rice commercialization is the utilization of selectable marker genes (SMGs) in the vector construct, which when incorporated into the genome are considered to pose risks to human health, the environment, and biodiversity, and thus become a matter of regulation. Various conventional strategies (co-transformation, transposon, recombinase systems, and MAT-vector) have been used in rice to avoid or remove the SMG from the developed events. However, the major limitations of these methods are; time-consuming, leftover cryptic sequences in the genome, and there is variable frequency. In contrast to these methods, CRISPR/Cas9-based marker excision, marker-free targeted gene insertion, programmed self-elimination, and RNP-based delivery enable us to generate marker-free engineered rice plants precisely and in less time. Although the CRISPR/Cas9-based SMG-free approaches are in their early stages, further research and their utilization in rice could help to break the regulatory barrier in its commercialization. In the current review, we have discussed the limitations of traditional methods followed by advanced techniques. We have also proposed a hypothesis, "DNA-free marker-less transformation" to overcome the regulatory barriers posed by SMGs.

Consensus genomic regions associated with multiple abiotic stress tolerance in wheat and implications for wheat breeding.

In wheat, a meta-analysis was performed using previously identified QTLs associated with drought stress (DS), heat stress (HS), salinity stress (SS), water-logging stress (WS), pre-harvest sprouting (PHS), and aluminium stress (AS) which predicted a total of 134 meta-QTLs (MQTLs) that involved at least 28 consistent and stable MQTLs conferring tolerance to five or all six abiotic stresses under study. Seventy-six MQTLs out of the 132 physically anchored MQTLs were also verified with genome-wide association studies. Around 43% of MQTLs had genetic and physical confidence intervals of less than 1 cM and 5 Mb, respectively. Consequently, 539 genes were identified in some selected MQTLs providing tolerance to 5 or all 6 abiotic stresses. Comparative analysis of genes underlying MQTLs with four RNA-seq based transcriptomic datasets unravelled a total of 189 differentially expressed genes which also included at least 11 most promising candidate genes common among different datasets. The promoter analysis showed that the promoters of these genes include many stress responsiveness cis-regulatory elements, such as ARE, MBS, TC-rich repeats, As-1 element, STRE, LTR, WRE3, and WUN-motif among others. Further, some MQTLs also overlapped with as many as 34 known abiotic stress tolerance genes. In addition, numerous ortho-MQTLs among the wheat, maize, and rice genomes were discovered. These findings could help with fine mapping and gene cloning, as well as marker-assisted breeding for multiple abiotic stress tolerances in wheat.

Ascertaining yield and grain protein content stability in wheat genotypes having the Gpc-B1 gene using univariate, multivariate, and correlation analysis.

The high performance and stability of wheat genotypes for yield, grain protein content (GPC), and other desirable traits are critical for varietal development and food and nutritional security. Likewise, the genotype by environment (G × E) interaction (GEI) should be thoroughly investigated and favorably utilized whenever genotype selection decisions are made. The present study was planned with the following two major objectives: 1) determination of GEI for some advanced wheat genotypes across four locations (Ludhiana, Ballowal, Patiala, and Bathinda) of Punjab, India; and 2) selection of the best genotypes with high GPC and yield in various environments. Different univariate [Eberhart and Ruessll's models; Perkins and Jinks' models; Wrike's Ecovalence; and Francis and Kannenberg's models], multivariate (AMMI and GGE biplot), and correlation analyses were used to interpret the data from the multi-environmental trial (MET). Consequently, both the univariate and multivariate analyses provided almost similar results regarding the top-performing and stable genotypes. The analysis of variance revealed that variation due to environment, genotype, and GEI was highly significant at the 0.01 and 0.001 levels of significance for all studied traits. The days to flowering, plant height, spikelets per spike, grain per spike, days to maturity, and 1000-grain weight were specifically affected by the environment, whereas yield was mainly affected by the environment and GEI. Genotypes, on the other hand, had a greater impact on the GPC than environmental conditions. As a result, a multi-environmental investigation was necessary to identify the GEI for wheat genotype selection because the GEI was very significant for all of the evaluated traits. Yield, 1000-grain weight, spikelet per spike, and days to maturity were observed to have positive correlations, implying the feasibility of their simultaneous selection for yield enhancement. However, GPC was observed to have a negative correlation with yield. Patiala was found to be the most discriminating environment for both yield and GPC and also the most effective representative environment for GPC, whereas Ludhiana was found to be the most effective representative environment for yield. Eventually, two NILs (BWL7508, and BWL7511) were selected as the top across all environments for both yield and GPC.