Primescript rtase mix

Total RNA was extracted from the roots of I. lactea using the Trizol kit (TaKaRa, Beijing, China), then the first strand cDNA was synthesized from 1 μg of total RNA using a 5×Primescript RTase mix (Takara, Beijing, China). The open reading frame (ORF) of IlHMA2 (GenBank accession: KY696282) was amplified by the primers P1 (5′-ATGGAATTCTTCCAAGTACTA-3′) and P2 (5′-TTATTCGCTTCCACTCCCATG-3′). The PCR products were purified from agarose gels and cloned into pMD-19T cloning vector followed by sequencing (Sangon, Shanghai, China). Then, the ORF of IlHMA2 with P3 (5′-CATGGTAGATCTATGGAATTCTTCCAAGTACTA-3′) and P4 (5′-ATTCGAGCTGGTCACCTTATTCGCTTCCACTCCCATG-3′) was amplified and cloned into the Bgl II/BstE II site of the pCAMBIA3301 vector to construct the 35S:IlHMA2 plasmid using infusion cloning technology.

Total RNA was extracted from the roots of I. lactea using the Trizol kit (TaKaRa, Beijing, China), then the first strand cDNA was synthesized from 1 μg of total RNA using a 5×Primescript RTase mix (Takara, Beijing, China). The open reading frame (ORF) of IlHMA2 (GenBank accession: KY696282) was amplified by the primers P1 (5′-ATGGAATTCTTCCAAGTACTA-3′) and P2 (5′-TTATTCGCTTCCACTCCCATG-3′). The PCR products were purified from agarose gels and cloned into pMD-19T cloning vector followed by sequencing (Sangon, Shanghai, China). Then, the ORF of IlHMA2 with P3 (5′-CATGGTAGATCTATGGAATTCTTCCAAGTACTA-3′) and P4 (5′-ATTCGAGCTGGTCACCTTATTCGCTTCCACTCCCATG-3′) was amplified and cloned into the Bgl II/BstE II site of the pCAMBIA3301 vector to construct the 35S:IlHMA2 plasmid using infusion cloning technology.