Heat and drought are among the major obstacles confronting crop production under climate change. The present study was conducted to evaluate 50 diverse wheat genotypes for cell membrane stability (CMS) and chlorophyll content at seedling and anthesis stages under heat and drought stress conditions, to understand the effect of the two abiotic factors and to find promising genotypes for future breeding. Experiments were conducted in the glasshouse (seedling stage) and the field (anthesis stage). Analysis of variance showed significant variation (P = 0.05) for all of the traits at seedling and anthesis stages. High levels of broad-sense heritability and genetic advance at 5% selection intensity indicated the presence of a high genetic component of variation and potential for genetic improvement through selection among the existing genetic variation. CMS showed a significant positive correlation with 1000-grain weight (TGW) under heat and drought conditions at both seedling and anthesis stages. Chlorophyll a/b ratio at seedling stage exhibited a significant negative correlation (r = –0.39, P < 0.05) with TGW under heat stress. Total chlorophyll content was significantly (r = 0.42, P < 0.05) correlated with TGW under heat stress at anthesis. Genotypes ETAD248 and ETAD7 showed the highest CMS and TGW values, whereas their chlorophyll a/b values were lowest, at both seedling and anthesis stages under heat and drought stress conditions. Higher CMS and total chlorophyll content, and lower chlorophyll a/b, were found to be useful indicators to identify genotypes with high TGW under heat and drought stress conditions. This study indicated the possibility of using seedling resistance as an indicator for later stage response in breeding for heat and drought resistance. The resistant genotypes identified can be used as potential germplasm in breeding programs.

Key message We developed breeder-friendly high-throughput and cost-effective KASP marker for marker-assisted selection for grain yield related traits in wheat. Abstract Plant-specific protein kinase, SnRK2s, is a major family of signaling genes associated with metabolic regulations, nutrient utilization and response to external stimuli. In the present study, three copies of TaSnRK2.9 were isolated from chromosomes 5A, 5B and 5D of wheat (Triticum aestivum L.). The coding regions of TaSnRK2.9-5A, TaSnRK2.9-5B and promoter region of TaSnRK2.9-5D were investigated for sequence polymorphism. Single nucleotide polymorphisms (SNPs) were identified for TaSnRK2.9-5A, while no polymorphism was identified in TaSnRK2.9-5B and TaSnRK2.9-5D. The nucleotide sequence of TaSnRK2.9-5A consisted of 2180 bp having eight introns and nine exons. Three SNPs were identified at 308 nt, 698 nt and 1700 nt. For high-throughput genotyping, two kompetitive allele-specific PCR (KASP) markers were developed. Four haplotypes Hap-5A-1, Hap-5A-2, Hap-5A-3 and Hap-5A-4 were detected in wheat populations collected from China, Europe and Pakistan. Association analysis was performed with mixed linear model in TASSEL (v 5.0). The results indicated that Hap-5A-1/2 of TaSnRK2.9-5A were significantly associated with high thousand kernel weight, while Hap-5A-4 with high grains per spike. Overexpressing transgenic rice also showed higher grains per spike which is in accordance with association analysis results. Geographic distribution and allelic frequency indicted that the favored haplotypes were positively selected in Chinese (Hap-5A-1/2), Pakistani (Hap-5A-1), east European (Hap-5A-1) and west European (Hap-5A-4) wheat breeding. The results suggest that the developed KASP markers can be utilized in yield improvement by marker-assisted selection in wheat breeding.

Reversible protein phosphorylation orchestrated by protein kinases and phosphatases is a major regulatory event in plants and animals. The SnRK2 subfamily consists of plant-specific protein kinases in the Ser/Thr protein kinase superfamily. Early observations indicated that SnRK2s are mainly involved in response to abiotic stress. Recent evidence shows that SnRK2s are multifarious players in a variety of biological processes. Here, we summarize the considerable knowledge of SnRK2s, including evolution, classification, biological functions and regulatory mechanisms at the epigenetic, post-transcriptional and post-translation levels.

Interrogations of local germplasm and landraces can offer a foundation and genetic basis for drought tolerance in wheat. Potential of drought tolerance in a panel of 30 wheat genotypes including varieties, local landraces, and wild crosses were explored under drought stress (DS) and well-watered (WW) conditions. Considerable variation for an osmotic adjustment (OA) and yield components, coupled with genotype and environment interaction was observed, which indicates the differential potential of wheat genotypes under both conditions. Reduction in yield per plant (YP), thousand kernel weight (TKW), and induction of OA was detected. Correlation analysis revealed a strong positive association of YP with directly contributing yield components under both environments, indicating the impotence of these traits as a selection-criteria for the screening of drought-tolerant genotypes for drylands worldwide. Subsequently, the association of OA with TKW which contributes directly to YP, indicates that wheat attains OA to extract more water from the soil under low water-potential. Genotypes including WC-4, WC-8 and LLR-29 showed more TKW under both conditions, among them; LLR-29 also has maximum OA and batter yield comparatively. Result provides insight into the role of OA in plant yield sustainability under DS. In this study, we figure out the concept of OA and its incredible role in sustainable plant yield in wheat.

Cereal crops have starch in their endosperm, which has provided calories to humans and livestock since the dawn of civilization to the present day. Starch is one of the important biological factors which is contributing to the yield of cereal crops. Starch is synthesized by different enzymes, but starch structure and amount are mainly determined by the activities of starch synthase enzymes (SS) with the involvement of starch branching enzymes (SBEs) and debranching enzymes (DBEs). Six classes of SSs are found in Arabidopsis and are designated as soluble SSI-V, and non-soluble granule bound starch synthase (GBSS). Soluble SSs are important for starch yield considering their role in starch biosynthesis in cereal crops, and the activities of these enzymes determine the structure of starch and the physical properties of starch granules. One of the unique characteristics of starch structure is elongated glucan chains within amylopectin, which is by SSs through interactions with other starch biosynthetic enzymes (SBEs and DBEs). Additionally, soluble SSs also have conserved domains with phosphorylation sites that may be involved in regulating starch metabolism and formation of heteromeric SS complexes. This review presents an overview of soluble SSs in cereal crops and includes their functional and structural characteristics in relation to starch synthesis.

High-throughput genotyping for functional markers offers an excellent opportunity to effectively practice marker-assisted selection (MAS) while breeding cultivars. We developed kompetitive allele-specific PCR (KASP) assays for genes conferring drought tolerance in common wheat (Triticum aestivum L.). In total, 11 KASP assays developed in this study and five already reported assays were used for their application in wheat breeding. We investigated alleles at 16 loci associated with drought tolerance among 153 Pakistani hexaploid wheat cultivars released during 1953–2016; 28 diploid wheat accessions (16 for AA and 12 for BB) and 19 tetraploid wheat (AABB) were used to study the evolutionary history of the studied genes. Superior allelic variations of the studied genes were significantly associated with higher grain yield. Favored haplotypes of TaSnRK2.3-1A, TaSnRK2.3-1B, TaSnRK2.9-5A, TaSAP-7B, and TaLTPs-1A predominated in Pakistani wheat germplasm indicating unconscious pyramiding and selection pressure on favorable haplotypes during selection breeding. TaSnRK2.8-5A, TaDreb-B1, 1-feh w3, TaPPH-7A, TaMOC-7A, and TaPARG-2A had moderate to low frequencies of favorable haplotype among Pakistani wheat germplasm pointing toward introgression of favorable haplotypes by deploying functional markers in marker-assisted breeding. The KASP assays were compared with gel-based markers for reliability and phenotypically validated among 62 Pakistani wheat cultivars. Association analyses showed that the favorable allelic variations were significantly associated with grain yield-contributing traits. The developed molecular marker toolkit of the genes can be instrumental for the wheat breeding in Pakistan.