Higher Thousand-grain Weight Research Articles

Genome-wide identification of phytosulfokine (PSK) peptide family reveals TaPSK1 gene involved in grain development of wheat (Triticum aestivum L.)

BackgroundPhytosulfokine (PSK) functions as a plant peptide growth factor that plays important and diverse roles in plant development and stress responses. Nevertheless, PSKs have not been systematically analyzed in wheat.ResultsA genome-wide comparative analysis of PSK genes in wheat was conducted and 15 TaPSKs were identified and divided into four subgroups in the wheat genome based on sequence similarity. The examination of motif compositions of TaPSK genes revealed the presence of the YIYTQ signature motif in the C-terminus of all TaPSK polypeptide precursors, with a highly conserved feature across different species. Exogenous application of TaPSK peptide promoted root growth in wheat. Quantitative RT-PCR analysis revealed that the TaPSKs exhibited preferential or tissue-specific expression patterns in wheat. In particular, three homologs of the TaPSK1 genes were specifically expressed in grains, with the strongest expression observed in the developing grains at 15 days after anthesis. Compared with wild type, transgenic rice lines overexpressing TaPSK1-A exhibited larger grain size and higher thousand-grain weight. The promoter region and genomic sequence of the wheat TaPSK1-A gene were cloned. Sequence polymorphism showed that five single-nucleotide polymorphisms (SNPs) were identified in the promoter region of TaPSK1-A. A Kompetitive Allele-Specific PCR (KASP) marker was developed for TaPSK1-A based on –806 bp SNP (C/T transition), and two haplotypes, TaPSK1-A-HapI and TaPSK1-A-HapII were detected in 260 wheat accessions collected from different regions. The expression of TaPSK1-A, promoter activity, and thousand-grain weight (TGW) in the TaPSK1-A-HapII haplotype were higher than those in the TaPSK1-A-HapI haplotype. Furthermore, yeast one-hybrid assays revealed the binding of TaNF-YB1 and TaERF39 to the promoter regions of the TaPSK1-A gene, and TaMADS29 could bind to the promoters of TaPSK1-B and TaPSK1-D genes.ConclusionsComparative genome-wide analysis of TaPSK peptide family revealed that the TaPSK1 gene is involved in wheat grain development, and the developed TaPSK1-A-KASP marker could be utilized for marker-assisted selection breeding of wheat.Graphical

A Novel Function of GW5 on Controlling the Early Growth Vigor and its Haplotype Effect on Shoot Dry Weight and Grain Size in Rice (Oryza sativa L.)

Strong early growth vigor is an essential target in both direct seeded rice breeding and high-yielding rice breeding for rice varieties with relatively short growth duration in the double-cropping region. Shoot dry weight (SDW) is one of the important traits associated with early growth vigor, and breeders have been working to improve this trait. Finding stable QTLs or functional genes for SDW is crucial for improving the early growth vigor by implementing molecular breeding in rice. Here, a genome-wide association analysis revealed that the QTL for SDW, qSDW-5, was stably detected in the three cultivation methods commonly used in production practice. Through gene-based haplotype analysis of the annotated genes within the putative region of qSDW-5, and validated by gene expression and knockout transgenic experiments, LOC_Os05g09520, which is identical to the reported GW5/GSE5 controlling grain width (GW) and thousand grain weight (TGW) was identified as the causal gene for qSDW-5. Five main haplotypes of LOC_Os05g09520 were identified in the diverse international rice collection used in this study and their effects on SDW, GW and TGW were analyzed. Phenotypic comparisons of the major haplotypes of LOC_Os05g09520 in the three subpopulations (indica, japonica and aus) revealed the same patterns of wider GW and higher TGW along with higher SDW. Furtherly, the haplotype analysis of 138 rice varieties/lines widely used in southern China showed that 97.8% of the cultivars/lines carry Hap2LOC_Os05g09520. These results not only provide a promising gene source for the molecular breeding of rice varieties with strong early growth vigor, but also elucidate the effect of the LOC_Os05g09520 haplotypes on SDW, GW, and TGW in rice. Importantly, this study provides direct genetic evidence that these three traits are significantly correlated, and suggests a breeding strategy for developing high-yielding and slender grain-shaped indica cultivars with strong early growth vigor.

Influence of natural and artificial selection on the yield differences among progeny derived from crossing between subspecies in cultivated rice

Extended natural and artificial selection have introduced pronounced differences throughout the genomes, morphological traits, and geographical distributions of the two predominant rice strains, Oryza sativa Xian/indica (XI) and O. sativa Geng/japonica (GJ). However, the unique roles of natural and artificial selection in strain differentiation remain uncharacterized. Therefore, we independently produced advanced inbreeding populations in typical GJ- and XI-cultivated areas, beginning from the F2 generation. We utilized pedigree and bulk-selective methods to characterize artificial and natural selection, respectively. Our examination uncovered preferences between artificial and natural selection. Artificial selection was favored by individuals with increased grain count per panicle but reduced panicle number, while natural selection favored traits like greater panicle number, reduced grains per panicle, and higher thousand-grain weight, especially in the XI cultivation areas. Notably, in the XI regions, natural selection produced a preference for wider grains, indicating that the elongated grains in XI rice may be caused by artificial rather than natural selection. Using de novo assembly of a high-quality parental genome, we observed an increased prevalence of the GJ genotype in GJ areas compared to XI areas, segregation distortion in the advanced inbreeding population could be attributed to loci regulating hybrid sterility. Notably, the semi-dwarf allele sd1-d, linked to the “Green Revolution”, was not favored in either selection paradigm in northern areas, possibly due to its disadvantages on grain yield per plant and NH4+ uptake rate. Our study offers novel perspectives into the contributions of natural and artificial selection to the divergence between XI and GJ subspecies.

Multi-omics analyses of the effect of carbon ion beam irradiation on cannabis fructus (Cannabis sativa L.) composition

Cannabis fructus (Cannabis sativa L.) belongs to the mulberry family, which is one of the earliest cultivated crops by humans and has industrial, agricultural, and medicinal values. In this study, the effect of carbon ion beam irradiation on the physiological and biochemical pathways of cannabis fructus was analyzed by multi-omics. Compared to the unirradiated cannabis fructus stem, 2,891 differentially-expressed genes, 1,145 differentially-expressed proteins, and 954 altered ubiquitination modification sites were identified in cannabis fructus irradiated by carbon ion beam irradiation (60 Gy dosage). Correlation analyses between proteomic, transcriptomic, and ubiquitin modification groups were carried out, and we found that the carbon fixation pathway, fatty acid synthesis pathway, JA synthesis, and glutathione metabolism pathway were enriched in relevant protein levels, and activates the MAPK signaling pathway regulated by gene expression. Further analysis found that the content of some proteins (LOX, AOC, and MDR) was correlated with their respective transcriptional levels, and the content of some proteins (APX, G-6-PD, GST, GAPHD, FBA, AOS, PR1, and ChiB) was correlated with their ubiquitination levels. In addition, M2 generation cannabis fructus seeds exhibited a larger volume, higher thousand-grain weight, and increased linolenic acid content compared to the control group, which was consistent with the trends of our multi-omics analyses. In summary, we demonstrated that 60 Gy carbon ion beam irradiation can enhance the carbon fixation pathway in cannabis fructus and enhance linolenic acid synthesis. Taken together, our multi-omics analysis of the response of cannabis fructus to carbon ion beam irradiation provides a theoretical foundation for readily producing cannabis fructus varieties with advantageous traits in the future.

Genetic architecture of quantitative trait loci (QTL) for FHB resistance and agronomic traits in a hard winter wheat population

Wheat resistance to Fusarium head blight (FHB) has often been associated with some undesirable agronomic traits. To study the relationship between wheat FHB resistance and agronomic traits, we constructed a linkage map of single nucleotide polymorphisms (SNPs) using an F6:8 population from G97252W × G97380A. The two hard winter wheat parents showed contrasts in FHB resistance, plant height (HT), heading date (HD), spike length (SL), spike compactness (SC), kernel number per spike (KNS), spikelet number per spike (SNS), thousand-grain weight (TGW) and grain size (length and width). Quantitative trait locus (QTL) mapping identified one major QTL (QFhb.hwwg-2DS) on chromosome arm 2DS for the percentage of symptomatic spikelets (PSS) in the spike, deoxynivalenol (DON) content and Fusarium damaged kernel (FDK). This QTL explained up to 71.8% of the phenotypic variation for the three FHB-related traits and overlapped with the major QTL for HT, HD, SL, KNS, SNS, TGW, and grain size. QTL on chromosome arms 2AL, 2DS, 3AL and 4BS were significant for the spike and grain traits measured. G97252W contributed FHB resistance and high SNS alleles at QFhb.hwwg-2DS, high KNS alleles at the QTL on 2AL and 2DS, and high TGW and grain size alleles at QTL on 3AL; whereas G97380A contributed high TGW and grain size alleles at the QTL on 2AL and 2DS, respectively, and the high KNS allele at the 4BS QTL. Combining QFhb.hwwg-2DS with positive alleles for spike and grain traits from other chromosomes may simultaneously improve FHB resistance and grain yield in new cultivars.

The Influence of Cropping Systems on Photosynthesis, Yield, and Grain Quality of Selected Winter Triticale Cultivars

Soil serves as the fundamental foundation for agricultural production; however, inappropriate utilization of soil in conventional cropping systems (CONV) coupled with agricultural practices focused on maximizing profit through the attainment of high-quality grain yield contributes to its degradation. An alternative to the CONV is the integrated cropping system (INTEG), which is based on sustainable plant cultivation by reducing the use of chemical pesticides and fertilizers. An interesting two-factor field experiment with winter triticale (×Triticosecale Wittm.) was conducted in 2019–2022 in south-eastern Poland. The experimental factors in this study included three winter triticale cultivars, namely Avokado, Medalion, Rotondo, and SU Liborius, and two cropping systems, CONV and INTEG. It was found that the use of the CONV brought out higher grain yield with higher protein and lower fat content. The cultivation of triticale grain under the CONV resulted in a higher thousand-grain weight (TGW), improved grain uniformity, and higher test weight values compared to INTEG. Additionally, the use of the CONV system resulted in improved values of physiological parameters (chlorophyll content and fluorescence and gas exchange), and that was caused by improved photosynthetic efficiency of triticale plants as a result of increased doses of mineral fertilizers, despite being cultivated in the INTEG; cv. SU Liborius achieved better values of yield parameters among the tested triticale cultivars grown in the CONV. Therefore, the selection of a suitable, efficient cultivar allows its growth under lower fertilization conditions and produces high grain yields of good quality. This knowledge can be valuable to farmers, as it would facilitate the selection of a triticale cultivar with the appropriate genetic profile for cultivation under specific agronomic conditions. Eventually, it would allow for the propagation of sustainable agricultural practices and contribute to enhancing soil biodiversity while maintaining the cost-effectiveness of agricultural production.

Precisely mapping a major QTL for grain weight on chromosome 5B of the founder parent Chuanmai42 in the wheat-growing region of southwestern China.

QTgw.saas-5B was validated as a major thousand-grain weight-related QTL in a founder parent used for wheat breeding and then precisely mapped to a 0.6 cM interval. Increasing the thousand-grain weight (TGW) is considered to be one of the most important ways to improve yield, which is a core objective among wheat breeders. Chuanmai42, which is a wheat cultivar with high TGW and a high and stable yield, is a parent of more than 30 new varieties grown in southwestern China. In this study, a Chuanmai42-derived recombinant inbred line (RIL) population was used to dissect the genetic basis of TGW. A major QTL (QTgw.saas-5B) mapped to the Xgwm213-Xgwm540 interval on chromosome 5B of Chuanmai42 explained up to 20% of the phenotypic variation. Using 71 recombinants with a recombination in the QTgw.saas-5B interval identified from a secondary RIL population comprising 1818 lines constructed by crossing the QTgw.saas-5B near-isogenic line with the recurrent parent Chuannong16, QTgw.saas-5B was delimited to a 0.6cM interval, corresponding to a 21.83Mb physical interval in the Chinese Spring genome. These findings provide the foundation for QTgw.saas-5B cloning and its use in molecular marker-assisted breeding.

Quantitative trait loci identification and breeding value estimation of grain weight-related traits based on a new wheat 50K single nucleotide polymorphism array-derived genetic map.

Mining novel and less utilized thousand grain weight (TGW) related genes are useful for improving wheat yield. In this study, a recombinant inbred line population from a cross between Zhongkemai 138 (ZKM138, high TGW) and Chuanmai 44 (CM44, low TGW) was used to construct a new Wheat 50K SNP array-derived genetic map that spanned 1,936.59 cM and contained 4, 139 markers. Based on this map, ninety-one quantitative trait loci (QTL) were detected for eight grain-related traits in six environments. Among 58 QTLs, whose superior alleles were contributed by ZKM138, QTgw.cib-6A was a noticeable major stable QTL and was also highlighted by bulked segregant analysis with RNA sequencing (BSR-Seq). It had a pyramiding effect on TGW enhancement but no significant trade-off effect on grain number per spike or tiller number, with two other QTLs (QTgw.cib-2A.2 and QTgw.cib-6D), possibly explaining the excellent grain performance of ZKM138. After comparison with known loci, QTgw.cib-6A was deduced to be a novel locus that differed from nearby TaGW2 and TaBT1. Seven simple sequence repeat (SSR) and thirty-nine kompetitive allele-specific PCR (KASP) markers were finally developed to narrow the candidate interval of QTgw.cib-6A to 4.1 Mb. Only six genes in this interval were regarded as the most likely candidate genes. QTgw.cib-6A was further validated in different genetic backgrounds and presented 88.6% transmissibility of the ZKM138-genotype and a 16.4% increase of TGW in ZKM138 derivatives. And the geographic pattern of this locus revealed that its superior allele is present in only 6.47% of 433 Chinese modern wheat varieties, indicating its potential contribution to further high-yield breeding.

Identification of Single Nucleotide Polymorphism in TaSBEIII and Development of KASP Marker Associated With Grain Weight in Wheat

Manipulation of genes involved in starch synthesis could significantly affect wheat grain weight and yield. The starch-branching enzyme (SBE) catalyzes the formation of branch points by cleaving the α-1,4 linkage in polyglucans and reattaching the chain via an α-1,6 linkage. Three types of SBE isoforms (SBEI, SBEII, and SBEIII) exist in higher plants, with the number of SBE isoforms being species-specific. In this study, the coding sequence of the wheat TaSBEIII gene was amplified. After the multiple sequence alignment of TaSBEIII genome from 20 accessions in a wheat diversity panel, one SNP was observed in TaSBEIII-A, which formed the allelic marker allele-T. Based on this SNP at 294 bp (C/T), a KASP molecular marker was developed to distinguish allelic variation among the wheat genotypes for thousand grain weight (TGW). The results were validated using 262 accessions of mini core collection (MCC) from China, 153 from Pakistan, 53 from CIMMYT, and 17 diploid and 18 tetraploid genotypes. Association analysis between TaSBEIII-A allelic variation and agronomic traits found that TaSBEIII-A was associated with TGW in mini core collection of China (MCC). The accessions possessing Allele-T had higher TGW than those possessing Allele-C; thus, Allele-T was a favorable allelic variation. By analyzing the frequency of the favorable allelic variation Allele-T in MCC, it increased from pre-1950 (25%) to the 1960s (45%) and increased continuously from 1960 to 1990 (80%). The results suggested that the KASP markers can be utilized in grain weight improvement, which ultimately improves wheat yield by marker-assisted selection in wheat breeding. The favorable allelic variation allele-T should be valuable in enhancing grain yield by improving the source and sink simultaneously. Furthermore, the newly developed KASP marker validated in different genetic backgrounds could be integrated into a breeding kit for screening high TGW wheat.

Pyramiding of High Grain Weight With Stripe Rust and Leaf Rust Resistance in Elite Indian Wheat Cultivar Using a Combination of Marker Assisted and Phenotypic Selection.

Wheat (Triticum aestivum L.) is an important cereal crop globally as well as in India and yield improvement programs encounter a strong impediment from ever-evolving rust pathogens. Hence, durable rust resistance is always a priority trait for wheat breeders globally. Grain weight, represented as thousand grain weight (TGW), is the most important yield-contributing trait in wheat. In the present study high TGW has been transferred into two elite Indian wheat cultivars PBW343 and PBW550 from a high TGW genotype, Rye selection 111, selected from local germplasm. In the background of PBW343 and PBW550, an increase in TGW upto 27.34 and 18% was observed, respectively (with respect to recipient parents), through conventional backcross breeding with phenotypic selections in 3 years replicated RBD trials. Resistance to leaf rust and stripe rust has been incorporated in the high TGW version of PBW550 through marker assisted pyramiding of stripe rust resistance gene Yr15 using marker Xuhw302, and a pair of linked leaf rust and stripe rust resistance genes Lr57-Yr40 using marker Ta5DS-2754099_kasp23. Improved versions of PBW550 with increased TGW ranging from 45.0 to 46.2 g (up to a 9% increase) and stacked genes for stripe and leaf rust resistance have been developed. This study serves as proof of utilizing conventional breeding and phenotypic selection combined with modern marker assisted selection in improvement of important wheat cultivars as a symbiont of conventional and moderan techniques.

Impact of high night temperature stress on different growth stages of wheat

Increasing temperature especially the High night temperature (HNT) is a major constriction for sustaining global food production under changing climate scenario. Recent climate change has resulted into events of abrupt night temperature rise either throughout the crop period or at specific growth stage. Historical temperature observations and model projections have predicted a more pronounced increase in night temperature [minimum temperature (Tmin.)] compared to that of day temperatures [maximum temperature (Tmax.)]. Hence, the present study was carried out to identify the most critical growth stage affected by HNT and their effects on physiological, growth and yield traits of wheat. A unique field-based movable temperature controlling system was designed to enhance the night temperature by 5 °C than ambient temperature. The three critical phenological stages of wheat i.e. at GS 15 (before tillering initiation), GS 45 (booting) and GS 75 (grain filling) of Zadok’s scale were separately subjected to HNT treatment. The results reveals that GS 75 (grain filling) was the most sensitive stage affected by HNT, which has recorded remarkable reduction in grain yield (GY), Harvest Index (HI), Thousand grain weight (TGW) and grain weight per spike (GWPS) compared to control treatment. Pearson correlation analysis showed that breeders can choose lines with higher TGW, GWPS and grain number per spike (GNPS) for obtaining better GY under HNT conditions and vegetative/booting stage HNT in field condition may not attribute for any significant yield loss.

TaCKX gene family, at large, is associated with thousand-grain weight and plant height in common wheat.

We used SMRT sequencing and explored the haplotypes of TaCKX genes, linked with thousand-grain weight and plant height, and developed the functionally validated markers, which can be used in the marker-assisted breeding program. Cytokinin oxidase/dehydrogenase (CKX) enzymes catalyze the permanent degradation of cytokinins. Identification of the TaCKX alleles associated with yield traits and the development of functional markers is the first step in using these alleles in marker-assisted breeding program. To identify the alleles, we sequenced the genome fragments, containing TaCKX genes from 48 wheat genotypes, by PacBio® sequencing. Six out of 22 TaCKX genes were found polymorphic, forming 14 distinct haplotypes. Functional markers were developed and validated for all the polymorphic TaCKX genes. Four specific haplotypes, i.e., TaCKX2A_2, TaCKX4A_2, TaCKX5A_3, and TaCKX9A_2, were found significantly associated with high thousand-grain weight (TGW) and short plant height (PH) in Chinese wheat micro-core collection (MCC) and GWAS open population (GWAS-OP), whereas TaCKX1B_2 in GWAS-OP and TaCKX11A_3 in MCC were significantly associated with high TGW and short PH. The mean values of TGW and PH for cumulative favorable haplotypes from chromosome 3A, i.e., TaCKX2A_2, TaCKX4A_2, and TaCKX5A_3, were significantly higher as compared to the cumulative unfavored haplotypes, and the change was additive in manner. Frequency distribution analysis revealed that since the 1960s, the frequency of the favorable haplotypes and TGW has gradually increased in Chinese wheat cultivars. Expression profiling in the seed tissue excised at 2, 4, 6, and 8days after anthesis depicted that the favorable haplotypes are significantly less expressive as compared to the unfavored haplotypes. We conclude that the functional markers developed in this study can be used to select the favorable haplotypes of TaCKX genes in wheat marker-assisted breeding programs.

Characterization, identification and evaluation of a novel wheat-Agropyron cristatum (L.) Gaertn. disomic addition line II-30-5

Agropyron cristatum (L.) Gaertn. (A. cristatum), which is an important wild relative of wheat (Triticum aestivum L.), exhibits multiple agronomic traits that are valuable for the genetic improvement of wheat. II-30-5 is derived from the BC3F1 progeny between wheat cv. Fukuhokomugi (Fukuho) and A. cristatum (2n = 4x = 28, PPPP). In this study, II-30-5 was proven to be a novel wheat-A. cristatum disomic 6P addition line by cytogenetics and A. cristatum expressed sequence tag-sequence-tagged site (EST-STS) markers. II-30-5 possessed several elite agronomic traits, such as good plant type, erect leaf position, short flag leaf, high thousand-grain weight, and high resistance to powdery mildew and leaf rust. Chromosomal rearrangements led to multiple addition lines in the same group of homologous group. Evaluation of the agronomic traits of different 6P addition lines showed that different 6P addition lines contain specific traits. In the same homologous group, the production of multiple addition lines increases the types of genetic variation for wheat improvement.

Diversity and sub-functionalization of TaGW8 homoeologs hold potential for genetic yield improvement in wheat

Grain Weight 8 (GW8) in rice is a SQUAMOSA Promoter-Binding Protein-Like (SPL) family transcription factor with multiple biological functions. In this study, three GW8 homoeologs were cloned from homoeologous group 7 chromosomes of wheat. Subcellular localization and trans-activation activity assays suggested that TaGW8 is a transcriptional activator. TaGW8 genes were preferentially expressed in young spikes and developing grains. Ectopic expressions of TaGW8 in Arabidopsis and rice resulted in enhanced vegetative growth, earlier flowering and larger seeds. TaGW8-7A was the most highly variable of the three homoeologs with four haplotypes (Hap-1/2/3/4). TaGW8-7B had two haplotypes (Hap-L/H). TaGW8-7A-Hap-2 was associated with high thousand-grain weight (TGW) and large kernel length and showed higher transcriptional levels and binding activity than the other haplotypes. The high frequency of TaGW8-7A-Hap-2 in Chinese wheat populations suggested that it had been extensively selected in breeding. This haplotype showed a high potential for exploitation in global wheat breeding because its frequency was low in exotic germplasm. TaGW8-7B-Hap-H produced abundant transcripts and was associated with early heading and maturity, less tiller number and high TGW. This haplotype might be suitable for multiple cropping systems due to short wheat season. In this study we identified sub-functionalization among TaGW8 homoeologs and obtained functional molecular markers that can be used in breeding for high grain yield.

DIFFERENTIAL RESPONSE OF WHEAT GENOTYPES TO HEAT STRESS DURING GRAIN FILLING

SUMMARYClimatic change affects global agriculture and is a severe threat to global food security due to variability of the three factors measuring climate change (CO2, temperature and precipitation) with temperature being the most crucial one. Wheat is severely affected by high temperatures with reproductive and grain-filling phases being most sensitive, impacting grain number, size and weight. Seed size and weight are the key agronomic traits subjected to artificial selection and involved in the domestication process since the origin of agriculture. Three genotypes Banks, EGA Gregory and Fang-60 with the latter known to be heat tolerant were grown under glass house conditions and subjected to heat stress for 3 days during early- (11–14 dpa – days post anthesis) and late- (27–30 dpa) grain filling stages in a mutually exclusive fashion. The impact of heat stress during early- and late- grain filling on the four major grain characteristics, thousand grain weight (TGW), grain length, grain width and grain thickness was assessed. The tolerant genotype Fang-60 exhibited significantly higher TGW during early-grain filling heat stress than the control possibly due to an ability to exploit the accelerated release of fertilizer under high temperature. Banks and EGA Gregory were moderately tolerant to susceptible to heat stress, respectively, at early- and late-grain filling with Fang-60 being tolerant to both early- and late- grain filling heat stress. This study confirms the availability of significant genetic variation in heat stress response in wheat that might be exploited to adapt wheat to higher growth temperatures.

Contributions of TaSUTs to grain weight in wheat under drought.

The homologous genes to OsSUT1-5 in wheat were identified and detailed analysed. TaSUT1 was the predominant sucrose transporter group and it illustrated the genotypic variations towards drought during grain filling. Sucrose transporters (SUT) play crucial roles in wheat stem water soluble carbohydrate (WSC) remobilization to grain. To determine the major functional SUT gene groups in shoot parts of wheat during grain development, drought tolerant varieties, Westonia and Kauz, were investigated in field drought experiments. Fourteen homologous genes to OsSUT1-5 were identified on five homeologous groups, namely TaSUT1_4A, TaSUT1_4B, TaSUT1_4D; TaSUT2_5A, TaSUT2_5B, TaSUT2_5D; TaSUT3_1A, TaSUT3_1D; TaSUT4_6A, TaSUT4_6B, TaSUT4_6D; TaSUT5_2A, TaSUT5_2B, and TaSUT5_2D, and their gene structures were analysed. Wheat plants above the ground were harvested from pre-anthesis to grain maturity and the stem, leaf sheath, rachis, lemma and developing grain were used for analysing TaSUT gene expression. Grain weight, thousand grain weight, kernel number per spike, biomass and stem WSC were characterized. The study showed that among the five TaSUT groups, TaSUT1 was the predominant sucrose transporting group in all organs sampled, and the expression was particularly high in the developing grain. In contrast to TaSUT1, the gene expression levels of TaSUT2, TaSUT3 and TaSUT4 were lower, except for TaSUT3 which showed preferential expression in the lemma before anthesis. The TaSUT5 gene group was very weakly expressed in all tissues. The upregulated gene expression of TaSUT1 Westonia type in stem and grain reveal a crucial role in stem WSC remobilization to grain under drought. The high TaSUT1 gene expression and the significant correlations with thousand grain weight (TGW) and kernel number per spike demonstrated the contribution in Kauz's high grain yield in an irrigated environment and high TGW in Westonia under drought stress. Further molecular level identification is required for gene marker development.

Gene editing and mutagenesis reveal inter-cultivar differences and additivity in the contribution of TaGW2\xa0homoeologues to grain size and weight in wheat

Key messageCRISPR-Cas9-based genome editing and EMS mutagenesis revealed inter-cultivar differences and additivity in the contribution of TaGW2 homoeologues to grain size and weight in wheat.The TaGW2 gene homoeologues have been reported to be negative regulators of grain size (GS) and thousand grain weight (TGW) in wheat. However, the contribution of each homoeologue to trait variation among different wheat cultivars is not well documented. We used the CRISPR-Cas9 system and TILLING to mutagenize each homoeologous gene copy in cultivars Bobwhite and Paragon, respectively. Plants carrying single-copy nonsense mutations in different genomes showed different levels of GS/TGW increase, with TGW increasing by an average of 5.5% (edited lines) and 5.3% (TILLING mutants). In any combination, the double homoeologue mutants showed higher phenotypic effects than the respective single-genome mutants. The double mutants had on average 12.1% (edited) and 10.5% (TILLING) higher TGW with respect to wild-type lines. The highest increase in GS and TGW was shown for triple mutants of both cultivars, with increases in 16.3% (edited) and 20.7% (TILLING) in TGW. The additive effects of the TaGW2 homoeologues were also demonstrated by the negative correlation between the functional gene copy number and GS/TGW in Bobwhite mutants and an F2 population. The highest single-genome increases in GS and TGW in Paragon and Bobwhite were obtained by mutations in the B and D genomes, respectively. These inter-cultivar differences in the phenotypic effects between the TaGW2 gene homoeologues coincide with inter-cultivar differences in the homoeologue expression levels. These results indicate that GS/TGW variation in wheat can be modulated by the dosage of homoeologous genes with inter-cultivar differences in the magnitude of the individual homoeologue effects.

Sequence Polymorphism and Cumulative Effect with 6 - SFT - A2 of Fructan Biosynthesis Gene 6 - SFT - D in Wheat

Gene 6-SFT encodes a key enzyme in fructan biosynthesis pathway in common wheat(Triticum aestivum L.). In this study, we analyzed the single nucleotide polymorphism(SNP) on 6-SFT-D locus in a diversity population of 23 hexaploid wheat(AABBDD) accessions and five wheat relative species(DD) by means of direct sequencing. Functional markers were developed according to the sequence polymorphism. The correlation between 6-SFT-D haplotypes and phenotypic traits and the cumulative effect of 6-SFT alleles were analyzed using a natural population consisting of 154 historical wheat accessions. Four SNPs were detected in 6-SFT-D sequences in the diversity population, forming three haplotypes. However, only two 6-SFT-D haplotypes were identified in the natural population. We developed a pair of allele-specific PCR markers based on a polymorphism(T/C) at the 2850 bp site. The results of haplotype–trait association analysis showed that 6-SFT-D was significantly associated with thousand-grain weight(TGW) and spike length under well-watered conditions. Hap I was a superior allele in improving TGW. Under drought stress and well-watered conditions, wheat materials carrying both superior allele of 6-SFT-D and 6-SFT-A2 had significantly higher TGW than other genotypes, suggesting that 6-SFT-D and 6-SFT-A2 have cumulative effect on TGW improvement.

TaTGW6-A1, an ortholog of rice TGW6, is associated with grain weight and yield in bread wheat

The TGW6 gene encodes a novel indole-3-acetic acid-glucose hydrolase and plays a significant role in improving thousand grain weight (TGW) and yield in rice. In the current study, a TGW6 ortholog from bread wheat chromosome 3AL was isolated and designated as TaTGW6-A1. Sequencing and alignment analysis showed that the cloned TaTGW6-A1 consists of a 987-bp intronless open reading frame. Three single-nucleotide polymorphisms (SNPs) were identified between cultivars with higher and lower TGW, forming two haplotypes, TaTGW6-A1a and TaTGW6-A1b. A derived cleaved amplified polymorphic sequence marker, TaTGW6-A1-CAPS, was developed to differentiate the two haplotypes. Linkage analysis using the wheat 90K iSelect SNP array mapped TaTGW6-A1-CAPS at 18 cM from BobWhite_c47304_56 in a RIL population derived from Zhou 8425B/Chinese Spring. A QTL for yield at the TaTGW6-A1 locus explained 17.4 % of the phenotypic variance in average yield over four environments. Association analysis on 242 Chinese and foreign cultivars indicated that TaTGW6-A1 was significantly associated with TGW. The cumulative frequency of the TaTGW6-A1a allele associated with higher TGW was approximately 80 %, indicating that the TaTGW6-A1a allele was positively selected in wheat breeding. In conclusion, TaTGW6-A1 in conjunction with its molecular marker may contribute as a valuable gene in improving yield potential in wheat.

Anatomy and Cytogenetic Identification of a Wheat-Psathyrostachys huashanica Keng Line with Early Maturation.

In previous studies, our research team successfully transferred the Ns genome from Psathyrostachys huashanica Keng into Triticum aestivum (common wheat cv. 7182) using embryo culture. In the present study, one of these lines, i.e., hybrid progeny 25-10-3, which matured about 10–14 days earlier than its wheat parent, was assessed using sequenced characterized amplified region (SCAR) analysis, EST-SSR and EST-STS molecular markers, and genomic in situ hybridization (GISH). We found that this was a stable wheat-P. huashanica disomic addition line (2n = 44 = 22 II) and the results demonstrated that it was a 6Ns disomic chromosome addition line, but it exhibited many different features compared with previously characterized lines, i.e., a longer awn, early maturation, and no twin spikelets. It was considered to be an early-maturing variety based on the early stage of inflorescence initiation in field experiments and binocular microscope observations over three consecutive years. This characteristic was distinct, especially from the single ridge stage and double ridge stage until the glume stage. In addition, it had a higher photosynthesis rate and economic values than common wheat cv. 7182, i.e., more spikelets per spike, more florets per spikelet, more kernels per spike, and a higher thousand-grain weight. These results suggest that this material may comprise a genetic pool of beneficial genes or chromosome segments, which are suitable for introgression to improve the quality of common wheat.