velopment based on the SNPs are described in Appendix S1. The sequences of TaCYP78A5-2A of 43 αvβ5 Species landraces and 42 cultivars (Table S7) had been obtained from GVM database ( bigd.huge.ac.cn/gvm, GVM000082; Zhou et al., 2020), along with the molecular diversity p values and Tajima’s D have been computed working with DnaSP5.0 (Librado and Rozas, 2009).Total RNA isolation and cDNA synthesisWheat tissue/organ samples kept at 0 had been used for RNA isolation. Total RNA was extracted making use of the Steady Pure Plant RNA Extraction Kit (Accurate Biotechnology, Changsha, China) as outlined by the manufacturer’s protocol. The good quality of RNA samples was determined by agarose gel electrophoresis and RNA concentration was measured employing a Nanodrop 2000 spectrophotometer (ND2000; Thermo Scientific, Wilmington, NC). The cDNA was synthesized using the Evo M-MLVRT Premix (Correct Biotechnology, Changsha, China).Association evaluation involving TaCYP78A5 haplotypes and agronomic traits of wheat accessionsThe genotypic information depending on genetic variations of TaCYP78A52A and phenotypic information of agronomic traits from the 323 wheat accessions at 16 environmental websites (E1 16) more than three years were analysed together with the TASSEL5.1 software (Bradbury et al., 2007). The Information from the 16 environmental internet sites and agronomic trait measurement of wheat accessions are described in Appendix S1.cDNA cloning and sequence alignment of TaCYP78A5 in wheatTo clone wheat TaCYP78A5, the mRNA sequence of Arabidopsis KLU/CYP78A5 (AT1G13710) was used to blast against wheat genome reference sequences to obtain the putative TaCYP78A5. Distinct primers had been made depending on the putative sequences of TaCYP78A5, and the cDNA synthesized in the mixed RNA samples of roots, leaves, young panicles and grains of Xiaoyan 6 was used as the template for TaCYP78A5 amplification. The primers utilized are shown in Table S8. The amplified items have been constructed into Vps34 custom synthesis pMD19T vector (Takara, Japan) and more than 10 clones were randomly chosen for sequencing. The resulted sequences had been compared with IWGSC Ref v 1.0 to obtain fulllength cDNA of three homoeologs of TaCYP78A5 in wheat. DNAMAN8 (lynnon) was applied to examine these sequences. The tree plot of TaCYP78A5 and its homologues were investigated in Triticeae-Gene Tribe (http://wheat.cau.edu.c n/TGT/) (Chen et al., 2020).Determination of TaCYP78A5-2A promoter activityThe promoter activity was detected in accordance with the instruction of Dual-Luciferase Reporter Assay Method (Promega), plus the process is shown in Appendix S1.BSMV-mediated TaCYP78A5 gene silencing in wheatThe BSMV-VIGS strategy applied for knockdown the expression of TaCYP78A5 in developmental grains of wheat was performed as previously reported (Ma et al., 2012). The protocol of BSMVderived vector building and BSMV-mediated TaCYP78A5 gene silencing is described in Appendix S1.Building of TaCYP78A5 overexpression vectors and improvement of transgenic wheat linesThe protocol for constructing TaCYP78A5 overexpression vectors UBI::TaCYP78A5-2A-GFP/GUS and pINO::TaCYP78A5-GUS is described in Appendix S1. The constructed vectors have been, respectively, introduced into Agrobacterium tumefaciens strain EHA105. The embryogenic calli of the immature embryo 15 days immediately after fertilization (DAF) of wheat line JW1 had been made use of as recipient components. Wheat transformation was carried out by A. tumefaciens-mediated strategy as described previously (Ishida et al., 2015; Zhang et al., 2018). The positive transgenic plants have been identified by leaf daubing with
