Identification and Expression Analysis of Foreign NAC086 Gene in Chinese Cabbage (Brassica campestris ssp. pekinensis)-Cabbage (B. oleracea var. capitata) Translocation Line
SHANG Jin-Meng2,*, WANG Ru-Ying1,*, XUAN Shu-Xin1,**, JIANG Dan2, FEI De-Qing1, WANG Yan-Hua1, FENG Da-Ling2, SHEN Shu-Xing1,**
1 College of Horticulture, Hebei Agricultural University/Key Laboratory for Vegetable Germplasm Enhancement and Utilization of Hebei/Collaborative Innovation Center of Vegetable Industry in Hebei, Baoding 071001, China; 2 College of Life Science, Hebei Agricultural University, Baoding 071001, China
Abstract：The NAC (NAM, ATAF, and CUC) transcription factors (TFs), which constitute one of the largest plant-specific transcription factor family, play an important role in resisting stress response, regulating plant growth and development, participating in organ model building, auxin transduction and leaf apoptosis, etc. In order to identify the foreign NAC086 gene of Chinese cabbage (Brassica campestris ssp. pekinensis)-cabbage (B. oleracea var. capitata) double haploid translocation line AT1-18, the cloning and expression of NAC086 genes were carried out from the translocation line AT1-18 and its parents by PCR, RT-PCR (reverse transcription-PCR) and qRT-PCR (real-time quantitative PCR) methods. The results showed that the gene Bol034440 was amplified with the specific primers in cabbage and translocation line, however, the gene Bra006392 was only obtained in Chinese cabbage. Sequence analysis showed that the identity was as high as 99% between the sequence of PCR product from the translocation line using Bol40-1 primers and CDS sequence of gene Bol034440. It was proved that Bra006392 gene of Chinese cabbage was replaced by Bol034440 gene of cabbage in the translocation line. RT-PCR analysis showed that there was almost no Bol034440 expression in the different tissues of Chinese cabbage, but the high relative expression was in the different tissues of the translocation line, followed by roots>leafs>buds>flowers>stems, with significant difference between roots and flowers, buds, stems. Bra006392 had almost no expression in the tissues (except roots) of the translocation line, but had the highest expression in the leaves of Chinese cabbage, which was significantly higher than that in other tissues. Bra008567 had high expression level in different tissues of Chinese cabbage and translocation lines, and the highest expression was in the root of Chinese cabbage, which was significantly higher than other tissues. Whereas, the highest expression was in the leaves of the translocation line, which was also significantly higher than other tissues except roots. There were significant differences between the two materials and from other tissues. qRT-PCR analysis showed that Bol034440 and Bra008567 responding to exogenous auxin treatment were negative and responding to the salt treatment was positive, the response time of both was 2 h after treatment, and Bol034440 was dominant in the response process. The results offered insight into the functional validation of NAC086 and studying the interaction and regulation of cabbage NAC086 gene in the genetic background of Chinese cabbage.
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