Quantitative Expression Analysis of TaELPs

The Quick RNA isolation Kit (Tiangen Biochemical Technology Co., Ltd., Beijing, China) was used to extract RNA according to the manufacturer’s instructions, and DNase I treatment was used to remove DNA contamination. Synthesis of the first strand of cDNA was carried out according to the instructions of the kit (Takara, Japan). To measure the expression of TaELPs, gene-specific primers were designed using WheatOmics 1.0 PrimersServer Database (http://202.194.139.32/PrimerServer/, accessed on 21 October 2021). qRT-PCR analysis was performed with specific primers (Table S10). qRT-PCR reactions were conducted using the following protocol: 95 °C for 2 min, followed by 40 cycles of 95 °C for 20 s and 60 °C for 20 s and 72 °C for 20 s. We verified the specificity of each primer’s amplicon via melting curve analysis, and the Elongation factor 1a (TaEF-1a) was used as an internal reference gene (GenBank accession no. Q03033) [85 (link)]. The threshold values (CT) were generated using the ABI PRISM 7500 system (Applied Biosystems, Foster City, CA, USA), and the transcription level of TaELPs was assessed using the comparative 2^−ΔΔCT method [86 (link)].