RNA silencing construct
Transcription of the RNA silencing construct occurs from the Linum usitatissimum Linin gene promoter. The transcription of the Carthamus tinctorius palmitoyl-acyl carrier protein thioesterase (CtFATB) and delta(12)-fatty acid desaturase (CtFAD2.2) fragments occur first in the sense and then in the antisense orientation, separated by a spacer of Flaveria trinervia pyruvate orthophosphate dikinase intron (sense orientation) and Ricinus communis catalase 1 inton (in antisense orientation). Transcription terminates at the Agrobacterium tumefaciens octopine synthase gene terminator. The resulting transcript will form a hairpin structure (hairpin RNA - hpRNA) through the complemetary base pairing of sense and antisense sections of CtFATB and CtFAD2.2 and a loop of PDK int-1 and Cat-1. This structure can elicit RNA interference-mediated gene silencing (see below).

Seletion marker
The transcription of the Streptomyces sp. hygromycin B phosphotransferase gene occurs from the Cauliflower mosaic virus 35S promoter and terminates at the A. tumefaciens nopaline synthase gene terminator. Hygromycin B was used as a selectible marker for successful transformants during the development of the plant line.

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Note:
i) Only a portion of the full sequence was cloned into the gene silencing construct for both CtFATB and CtFAD2.2.
ii) Primer walking and southern blot analysis confirmed the presence of a single T-DNA insert into the safflower genome. The insertion of T-DNA generated a 69-basepair deletion within the genomic region. The entire Left Border T-DNA sequence (160 basepairs) was inserted into the genome, but only 41 basepairs of the 162 basepairs Right Border T-DNA sequence was inserted. Additionally, 91 basepairs of the replication of origin RiA4 from the binary vector pORE-CBIb was also inserted.