Primers used to isolate/analyze GL1, GL2, GL3, TTG1 and TRY genes from B. villosa.

1Forward (F) primers were designed at the 5′ coding end and reverse (R) primers to the 3′ end of cDNA sequences from B. napus for GL1 and GL2 and from B. rapa for EGL3 and TTG1. Forward primers for TRY were designed 72 nucleotides before coding region and reverse primer at 3′ end of the cDNA from B. rapa. Underlined sequences indicate incorporated restriction enzyme sites; BamHI and PstI sites in the GL1, GL2, EGL3 and TRY forward and reverse primers, respectively; BamHI and KpnI sites in the TTG1 forward and reverse primers, respectively.2Primers for Q-PCR were designed to conserved regions based on alignments between the A. thaliana homologue and the homologues from four Brassica species. B nEF1F/BnEF1R are endogenous reference gene primers.1. 正向（F）引物设计于甘蓝型油菜（B. napus）GL1与GL2的互补DNA（cDNA）序列的编码区5′端，反向（R）引物设计于其3′端；针对芸薹（B. rapa）EGL3与TTG1的cDNA序列，亦采用相同策略设计引物。针对拟南芥（A. thaliana）同源基因TRY的正向引物，设计于芸薹cDNA编码区上游72个核苷酸位点处，反向引物设于其cDNA的3′端。下划线序列代表引入的限制性酶切位点（restriction enzyme sites）：GL1、GL2、EGL3及TRY的正向引物与反向引物分别引入BamHI与PstI酶切位点；TTG1的正向引物与反向引物分别引入BamHI与KpnI酶切位点。 2. 实时定量PCR（Q-PCR）引物设计于保守区域，该区域基于拟南芥（A. thaliana）同源序列与4种芸薹属物种的同源序列比对结果确定。BnEF1F/BnEF1R为内参基因引物。