Long-chain acyl-CoA synthetase (LACS), responsible for the conversion of free FAs into acyl-CoAs, is involved in multiple pathways of lipid metabolism. Although LACS genes in Arabidopsis have been well characterized, no detailed information concerning this family is available for wheat. In the present study, a systematic analysis was carried out for the wheat LACS family. As a result, 30 putative TaLACSs were identified. Expression analysis revealed that 22 Takacs were expressed in wheat anthers. Two orthologs of AtLACS1, TaLACS2 and TaLACS3, were repressed at the vacuolated stage in the cold-treated BS366 (a temperature-sensitive genic male-sterile line). Thus, TaLACS2 and TaLACS3 may function like AtLACS1 in wax biosynthesis in anthers, and the repression of both genes may be correlated with the male sterility of BS366. TaLACS5 is an ortholog of AtLACS5, which was expressed exclusively in anthers. TaLACS5 was repressed in the cold-treated BS366 at the tetrad, uninucleate, and vacuolated stages. The negative correlation between TaLACS5 and TaGAMYB-B, and the MYB domain found in the promoter sequence suggested that TaLACS5 may be negatively regulated by TaGAMYB-B to participate in wheat fertility. These findings will provide a valuable foundation for the understanding of the wheat LACS gene family in male fertility.

Sucrose transporters (SUTs) play vital roles in phloem sucrose unloading and transportation in wheat grains. However, the genomic information regarding the SUT gene family and their expression patterns in response to heat stress in grains of male-sterile wheat (Triticum aestivum L.) lines has not been systematically studied. In this study, a thorough examination of the wheat SUT gene family was conducted, focusing on their expression patterns in male-sterile lines under heat stress conditions in grain tissues. A total of 19 SUT genes were identified, with phylogenetic analysis indicating their classification into five distinct groups. Polyploidization was identified as a substantial factor in the expansion of SUT genes, with segmental duplication being the predominant mechanism driving the evolutionary expansion of the SUT gene family in wheat. Transcriptome data indicate that the expression levels of TaSUT1 and TaSUT2 were higher than other SUT genes in grains of male-sterile lines. The TaSUT1 expression showed a gradual decreasing trend, while TaSUT2 showed a reverse trend with the process of grain filling. After heat stress, the TaSUT1 expression in grains of male-sterile lines was first significantly increased and then significantly decreased with the filling stage extension, aligning with the observed trend of sucrose levels, indicating that heat stress may decrease the grain weight by reducing sucrose unloading and transportation process in grains. These results provide a systematic analysis of the SUT gene family and lay a theoretical foundation for us to study the grain filling of male-sterile lines in response to abiotic stress.

Thermo-sensitive cytoplasmic male sterility (TCMS) plays a crucial role in hybrid production and hybrid breeding; however, there are few studies on molecular mechanisms related to anther abortion in the wheat TCMS line. In this study, FA99, a new wheat thermo-sensitive cytoplasmic male sterility line, was investigated. Fertility conversion analysis showed that FA99 was mainly controlled by temperature, and the temperature-sensitive stage was pollen mother cell formation to a uninucleate stage. Further phenotypic identification and paraffin section showed that FA99 was characterized by indehiscent anthers and aborted pollen in a sterile environment and tapetum was degraded prematurely during the tetrad period, which was the critical abortion period of FA99. The contents of O²⁻, H₂O₂, MDA and POD were significantly changed in FA99 under a sterile environment by the determination of physiological indexes. Furthermore, through transcriptome analysis, 252 differentially expressed genes were identified, including 218 downregulated and 34 upregulated genes. Based on KOG function classification, GO enrichment and KEGG pathways analysis, it was evident that significant transcriptomic changes in FA99 under different fertility environments, and the major differences were “phenylalanine metabolism”, “phenylpropanoid biosynthesis”, “cutin, suberine and wax biosynthesis”, “phenylalanine, tyrosine and tryptophan biosynthesis” and “citrate cycle (TCA cycle)”. Finally, we proposed an intriguing transcriptome-mediated pollen abortion and male sterility network for FA99. These findings provided data on the molecular mechanism of fertility conversion in thermo-sensitive cytoplasmic male sterility wheat.

High-temperature (HT) stress frequently affects the early and middle stages of grain filling in hybrid seed production regions. Photo-thermo-sensitive male-sterile (PTMS) wheat lines, which play a critical role as female parents in hybrid seed production, face challenges under HT conditions. However, the mechanisms governing grain filling in PTMS lines under HT stress remain poorly understood. This study used the BS253 line to investigate the effects of HT on grain filling, primarily focusing on the transition from sucrose unloading to starch synthesis. The findings indicated that HT significantly reduced the grain starch content and weight by 7.65% and 36.35% at maturity, respectively. Further analysis revealed that the expression levels of TaSUT1 and TaSWEETs in grains initially increased after HT stress, paralleling the rise in sucrose content during the same period. The activities of ADP-glucose pyrophosphorylase, UDP-glucose pyrophosphorylase, granule-bound starch synthase, and soluble starch synthase were markedly decreased, indicating that impaired starch synthesis was a key factor limiting grain filling immediately after HT exposure. A total of 41 key regulatory genes involved in sucrose-to-starch metabolism were identified, with HT significantly reducing the expression of genes associated with pathways from sucrose unloading to starch synthesis during the middle and late stages post-HT. Based on the observed ultrastructural changes in the abdominal phloem and sucrose transporter expression levels under HT, we concluded that limited sucrose supply, degradation, and inhibition of starch synthesis collectively constrained grain filling during these stages. Additionally, 11 heat shock proteins and two catalase genes were identified and significantly upregulated during the initial phase post-HT, suggesting their potential role in enhancing sucrose supply at this critical time. More importantly, seven key genes involved in the sucrose-to-starch pathway were identified by weighted gene co-expression network analysis (WGCNA), which provides target genes for their functional research for starch synthase. These findings provide a comprehensive understanding of how HT limits grain filling, identify several genes involved in the sucrose-to-starch pathway, and offer a novel perspective for future research on HT-restricted grain filling across the entire process from sucrose unloading to starch synthesis in developing grains.

Maintaining stable male sterility is fundamental for ensuring the genetic purity and productivity of two-line hybrid wheat. However, unexpected heat events during the fertility-sensitive period can induce fertility restoration in photo-thermo-sensitive male sterile (PTMS) lines, posing a major threat to hybrid seed production. In this study, we identified two BS-type PTMS lines, BS166 and BS192, that consistently maintained sterility under heat stress in a seed-production environment, indicating strong heat-tolerant sterility. To uncover the molecular basis underlying this stability, we compared four BS-type PTMS lines exhibiting contrasting heat responses through field assessments, controlled heat treatments, transcriptome sequencing, and weighted gene co-expression network analysis (WGCNA). A total of 19,105 differentially expressed genes were identified, with the bisque4 module showing a significant correlation with seed setting rate. KEGG enrichment analysis revealed that starch and sucrose metabolism, cutin, suberin, and wax biosynthesis, fatty acid biosynthesis, and plant hormone signal transduction pathways were highly associated with heat-tolerant sterility. Core genes within these pathways displayed transcriptional stability in BS166 and BS192 but were strongly induced in heat-sensitive lines. In situ hybridization and RT-qPCR further confirmed tapetum-specific expression of TaBGLU32 and TaLACS1. Based on these findings, we propose a regulatory model explaining how PTMS lines maintain sterility stability under heat stress.