Phenotypic Analysis and Transcriptomic Profiling

Based on the phenotypes of PH and NNMS, we selected extremely high materials R19 (the PH is 118.25 cm, the NNMS is 19.25) and R200 (122.4 cm, 22.6) and extremely small materials R120 (56.2 cm, 15) and R155 (56.4 cm, 15.8) and SN14 (81.0 cm, 18.1). There are significant phenotypic differences at the P = 0.05 level between extremely high line and extremely small lines on PH and NNMS (Supplementary Tables S10 and S11). Extraction of shoot tip tissue from the ternate compound-leaf stage for RNA extraction using TRIzol Reagent (Invitrogen, 15596-026, Carlsbad, CA, USA) was performed. Reverse transcription of extracted RNA into cDNA using the Tianhe Real-time quantitative PCR (RT-qPCR) kit was performed using SYBR qPCR Mix (Vazyme, Q711, Vazyme biotech, Nanjing, China) on the Light Cycler 480 System (Roche, Roche Diagnostics, Basel, Switzerland). The expression levels of candidate genes were calculated with GmUKN1 as an internal reference according to the following formula [65 (link)]:

The qRT-PCR primer sequences specific for candidate genes were designed using Primer Premier 5.0 (http://www.premierbiosoft.com/primerdesign/index.html).