Seed-specific expression of TaYUC10 significantly increases auxin and protein content in wheat seeds.

KEY MESSAGE: Present study revealed that specific expression of TaYUC10.3 in wheat young seeds could increase the content of auxin, and protein. Auxin is a vital endogenous hormone in plants, which is involved in the regulation of various physiological and biochemical processes in plants. The flavin-containing monooxygenase encoded by the YUCCA gene is a rate-limiting enzyme in the tryptophan-dependent pathway of auxin synthesis. TaYUC10.3 was identified, cloned and found that it was abundantly expressed in wheat young seeds. In this study, a seed-specific expression vector of TaYUC10.3 was constructed with the promoter of 1Bx17 glutenin subunit gene and transformed wheat using the particle bombardment method. The quantitative RT-PCR showed that TaYUC10.3 was expressed in a large amount in young seeds of the transgenic lines. Plant hormone-targeted metabolomics showed that the auxin content of the transgenic lines was significantly increased compared with controls. The GC / MS non-targeted metabolite multiple statistical analyses showed that the variable importance in projection (VIP) of tryptophan reduced in the transgenic lines. Simultaneously, the VIP of indole acetic acid increased. The precursor amino acids for synthesizing some proteins and carbohydrates were upregulated in the transgenic lines. Subsequently, it was found that the protein content of the seeds of the transgenic TaYUC10.3 wheat was significantly higher than that of the control. The wet gluten content and sedimentation value of the transgenic TaYUC10.3 wheat were also high. This result indicated that TaYUC10.3 might participate in auxin synthesis and affects the protein content of wheat seeds.

Isolation and characterization of three TaYUC10genes from wheat.

YUCCA protein participates in a key rate-limiting step in the tryptophan-dependent pathway for auxin biosynthesis and is involved in numerous processes during plant development. In this study, the genomic and cDNA sequences of three TaYUC10 homoeologous genes were isolated. These sequences showed a very high conservation in coding region and the exon/intron structure, whereas their intron lengths were different. The cDNA and polypeptide chains of the three TaYUC10 genes were highly similar. These genes were most homologous to BdYUC10. Location analysis showed that TaYUC10.1 was present in chromosome 5BL. TaYUC10.3 was expressed in all parts of the wheat, but was predominant in the reproductive organs of mature wheat, such as flowering spikelets or fertilized embryos. In the fertilized embryos 28d post-anthesis, expression of TaYUC10.3 was clearly increased with the development of seeds. This indicates that TaYUC genes may play a vital role in seed development. TaYUC10.3 overexpressed in Arabidopsis had a typical phenotype, excessive auxin accumulation also seen in higher plants, and showed increased spacing of silique and downward curling of the blade margin. Sterility was observed in adult transgenic plants, becoming more severe in late development. The floral structures of sterile plants were not integrated. TaYUC10 may be required for numerous wheat growth processes, including flower and seed development.