The GmHMGR4 and GmHMGR6 cDNA sequences were cloned into the expression vector pC3300s directed by the CaMV 35S promoter, respectively. Briefly, we used primers with SacI and BamHI sites (underlined GmHMGR4, 5′-CTCTCGAG CTTTCGCGAGCTCCCCATTTCCCTTCCAATCT-3′ and 5′-CTGCAGGTCGACTCTAGAGGATCC CCCCACCATCATCAATACCA-3′; GmHMGR6, 5′-CTCTCGAG CTTTCGCGAGCTCAAACAAGGGTTGCACGCTCT-3′ and 5′-CTGCAGGTCGACTCTAGAGGATCC ACCCCCTC CCACCATCAAT-3′) to amplify the cDNA of GmHMGR4 and GmHMGR6. The above obtained PCR products were enzyme-digested and cloned into the vector pC3300s (named pC3300S-35S:GmHMGR4 and pC3300S-35S:GmHMGR6). The pC3300S-35S:GmHMGR4 and pC3300S-35S:GmHMGR6 were transformed into A. thaliana (Col-0, provided by Wuhan Towin Biotechnology Company Limited, China) using Agrobacterium GV3101. T0 transformants A. thaliana seeds were selected by MS containing glufosinate ammonium (100 mg∙mL−1). Then the corresponding insertions of transgenes were verified by PCR using the bar gene primer (Supplementary Fig. S5 ). After vernalizing at 4 ℃ for 24 h, the transgenic lines of A. thaliana grew in MS medium at 21℃ under 16-h-light/8-h-dark conditions. T3 transgenic homozygous lines were utilized to detect isoprenoid’s content and observe root growth. The chlorophyll content was analyzed by using fresh rosette leaves of 30-day-old A. thaliana. Other plants in this experiment were freeze-dried to analyze the content of squalene, sterols and tocopherols. Additionally, seeds were planted in square plates with MS and 1% agar for root growth tests, and the main root’s average length was measured daily.
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