HMW-GSs 1Dx3+1Dy12 contribute to a suitable wheat gluten strength that confers superior Chinese steamed bread quality.

The aim of this study was to clarify the effects of the high-molecular-weight glutenin subunits (HMW-GSs) 1Dx3+1Dy12 (3+12) and 1Dx4+1Dy12 (4+12) at the Glu-D1 locus on gluten and Chinese steamed bread (CSB) quality. The grain protein content and composition, gluten content and gluten index, farinograph properties, and CSB quality were investigated using four wheat near-isogenic lines (NILs) carrying HMW-GSs 1Dx2+1Dy12 (2+12), 3+12, 4+12 and 1Dx5+1Dy10 (5+10), respectively. The unextractable polymeric protein (UPP) and glutenin macropolymer (GMP) content, gluten index, dough development time, stability time, and farinograph quality number of four NILs all ranked as 5+10 > 3+12 > 2+12/4+12, such as the gluten index ranked as 5+10(44.88%) > 3+12(40.07%) > 2+12(37.46%)/4+12(35.85%); however, their contributions to the quality of CSB were ranked as 3+12 > 5+10 > 2+12/4+12, such as the specific volume ranked as 3+12(2.64 mL/g) > 5+10(2.49 mL/g) > 2+12(2.36 mL/g)/4+12(2.35 mL/g), which indicated that a suitable gluten strength (3+12) was crucial to making high-quality CSB. In addition, subunits 4+12 had a similar quality performance to low-quality subunits 2+12. All these findings suggested that, except for the acknowledged high-quality subunits 5+10, the introduction of 3+12 at the Glu-D1 locus is an efficient way for quality improvement of gluten as well as CSB.

The wheat WRKY transcription factor TaWRKY1-2D confers drought resistance in transgenic Arabidopsis and wheat (Triticum aestivum L.).

The WRKY transcription factor family has been associated with a variety of plant biological processes, such as biotic and abiotic stress responses. In this study, 13 wheat TaWRKY DEGs in transcriptome data before and after drought stress, namely TaWRKY1 to TaWRKY8, including various copies, were identified and classified as Group I, II, or III. TaWRKY1-2D overexpression enhanced drought tolerance in transgenic Arabidopsis. Moreover, the AtRD29A, AtP5CS1, AtPOD1, AtCAT1, and AtSOD (Cu/Zn) genes, which are related to the stress response and antioxidant system, were significantly upregulated in TaWRKY1-2D transgenic Arabidopsis under drought stress. TaWRKY1-2 silencing in wheat increases the MDA content, reduces the contents of proline and chlorophyll and the activities of antioxidant enzymes, and inhibits the expression levels of antioxidant (TaPOD, TaCAT, and TaSOD (Fe))- and stress-related genes (TaP5CS) under drought stress. Yeast two-hybrid screening revealed TaDHN3 as an interaction partner of TaWRKY1-2D; their interaction was further confirmed using yeast two-hybrid and bimolecular fluorescence complementation. Furthermore, TaWRKY1-2D may play essential roles in wheat drought tolerance through posttranslational regulation of TaDHN3. Overall, these findings contribute to our knowledge of the WRKY family in wheat and identify TaWRKY1-2D as a promising candidate gene for improving wheat breeding to generate drought-tolerant wheat.

Genome-wide identification of foxtail millet's TRX family and a functional analysis of SiNRX1 in response to drought and salt stresses in transgenic Arabidopsis.

Thioredoxins (TRXs) are small-molecule proteins with redox activity that play very important roles in the growth, development, and stress resistance of plants. Foxtail millet (Setaria italica) gradually became a model crop for stress resistance research because of its advantages such as its resistance to sterility and its small genome. To date, the thioredoxin (TRX) family has been identified in Arabidopsis thaliana, rice and wheat. However, studies of the TRX family in foxtail millet have not been reported, and the biological function of this family remains unclear. In this study, 35 SiTRX genes were identified in the whole genome of foxtail millet through bioinformatic analysis. According to phylogenetic analysis, 35 SiTRXs can be divided into 13 types. The chromosome distribution, gene structure, cis-elements and conserved protein motifs of 35 SiTRXs were characterized. Three nucleoredoxin (NRX) members were further identified by a structural analysis of TRX family members. The expression patterns of foxtail millet's SiNRX members under abiotic stresses showed that they have different stress-response patterns. In addition, subcellular localization revealed that SiNRXs were localized to the nucleus, cytoplasm and membrane. Further studies demonstrated that the overexpression of SiNRX1 enhanced Arabidopsis' tolerance to drought and salt stresses, resulting in a higher survival rate and better growth performance. Moreover, the expression levels of several known stress-related genes were generally higher in overexpressed lines than in the wild-type. Thus, this study provides a general picture of the TRX family in foxtail millet and lay a foundation for further research on the mechanism of the action of TRX proteins on abiotic stresses.

Identification of TaBADH-A1 allele for improving drought resistance and salt tolerance in wheat (Triticum aestivum L.).

Drought and salt stress can strongly affect the growth and development of wheat. Wheat adapts to drought and salt stress through osmotic regulation. Betaine aldehyde dehydrogenase (BADH) is a key enzyme in the synthesis of betaine, an osmotic regulator. We cloned a region of the TaBADH-A1 promoter and genomic DNA that included the introns and exons, from four Chinese wheat cultivars. Following the analysis of TaBADH-A1 genomic DNA and promoter sequence polymorphisms of 4 cloned and 15 cultivars from the database, 7 haplotypes of TaBADH-A1 gene were identified. We divided the 7 haplotypes with a 254 bp insertion or deletion (indel) into two main alleles, BADH-A1a and BADH-A1b. Meanwhile, a molecular marker was developed based on the 254 bp indel of the third intron of TaBADH-A1 gene. Expression levels of BADH-A1b were found to be significantly higher than those of BADH-A1a under drought and salt stress conditions. Betaine accumulation was significantly higher in wheat containing BADH-A1b compared to BADH-A1a under drought and salt stress. We also identified that the average relative germination and survival rates of wheat with the BADH-A1b allele were significantly higher than wheat with the BADH-A1a allele. The results reveal that wheat containing BADH-A1b has stronger drought and salt tolerance than wheat with BADH-A1a. Meanwhile, the geographic distribution and frequency of the TaBADH-A1 locus alleles indicate that BADH-A1a has been preferred in Chinese wheat breeding programs, while BADH-A1b, associated with favorable stress tolerance, has been neglected. The results of this study provide evidence for an excellent candidate allele for marker-assisted selection of new wheat cultivars with increased salt tolerance and drought resistance.

Transcriptomic Identification of Wheat AP2/ERF Transcription Factors and Functional Characterization of TaERF-6-3A in Response to Drought and Salinity Stresses.

AP2/ERF (APETALA2/ethylene responsive factor) is a family of plant-specific transcription factors whose members are widely involved in many biological processes, such as growth, development, and biotic and abiotic stress responses. Here, 20 AP2/ERF genes were identified based on wheat RNA-seq data before and after drought stress, and classified as AP2, ERF, DREB, and RAV. The analysis of gene structure revealed that about 85% of AP2/ERF family members had lost introns, which are presumed to have been lost during the formation and evolution of the wheat genome. The expression of 20 AP2/ERF family genes could be verified by qRT-PCR, which further supported the validity of the RNA-seq data. Subsequently, subcellular localization and transcriptional activity experiments showed that the ERF proteins were mainly located in the nucleus and were self-activating, which further supports their functions as transcription factors. Furthermore, we isolated a novel ERF gene induced by drought, salt, and cold stresses and named it TaERF-6-3A. TaERF-6-3A overexpression increased sensitivity to drought and salt stresses in Arabidopsis, which was supported by physiological and biochemical indices. Moreover, the expression of stress- and antioxidant-related genes was downregulated in TaERF-6-3A-overexpressing plants. Overall, these results contribute to the further understanding of the TaERF-6-3A gene function in wheat.

Identification of the yield traits related haplotype combinations of transcription factor genes TaHDZ34 in common wheat.

A predominant objective in wheat breeding is improving yield-related traits. The homeodomain-leucine zipper (HD-Zip) transcription factor plays a significant role in plant growth and development. In this study, we cloned all homeologs of TaHDZ34, which is a member of the HD-Zip class IV transcription factor family in wheat (Triticum aestivum L.). Sequence polymorphism analysis showed that TaHDZ-A34, TaHDZ-B34, and TaHDZ-D34 formed five, six, and six haplotypes, respectively, and the genes were divided into two main haplotype groups. We also developed functional molecular markers. The TaHDZ34 genes were divided into eight main haplotype combinations. Association analysis and distinct population validation preliminarily indicated that TaHDZ34 genes modulate grain number per spike, effective spikelet number per spike, thousand kernel weight, and flag leaf area per plant in wheat. Hap-ABD was the most effective haplotype combination of TaHDZ34. Subcellular localization showed that TaHDZ-A34 was localized to the nucleus. The interacting proteins of TaHDZ-A34 were involved in protein synthesis/degradation, energy production and transportation, and photosynthesis. Geographic distribution and frequencies of TaHDZ34 haplotype combinations suggested that Hap-Abd and Hap-AbD were preferentially selected in Chinese wheat breeding programs. The high-yield-related haplotype combination Hap-ABD provided beneficial genetic resources for the marker-assisted selection of new wheat cultivars. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01298-5.

A Wheat TaTOE1-B1 Transcript TaTOE1-B1-3 Can Delay the Flowering Time of Transgenic Arabidopsis.

Flowering time is one of the most important agronomic traits in wheat production. A proper flowering time might contribute to the reduction or avoidance of biotic and abiotic stresses, adjust plant architecture, and affect the yield and quality of grain. In this study, TaTOE1-B1 in wheat produced three transcripts (TaTOE1-B1-1, TaTOE1-B1-2, and TaTOE1-B1-3) by alternative splicing. Compared to the longest transcript, TaTOE1-B1-1, TaTOE1-B1-3 has a deletion in the sixth exon (1219-1264 bp). Under long-day conditions, the heterologous overexpression of the TaTOE1-B1-3 gene delayed flowering, prolonged the vegetative growth time, and enlarged the vegetative body of Arabidopsis, but that of TaTOE1-B1-1 did not. As typical AP2 family members, TaTOE1-B1-1 and TaTOE1-B1-3 are mainly located in the nucleus and have transcriptional activation activities; the transcriptional activation region of TaTOE1-B1-3 is located in the C-terminal. In TaTOE1-B1-3 overexpression lines, the expression of flowering-related AtFT and AtSOC1 genes is significantly downregulated. In addition, this study confirms the protein-protein interaction between TaTOE1-B1-3 and TaPIFI, which may play an important role in flowering inhibition. These results provide a theoretical basis for the precise regulation of wheat flowering time.