| Gene and promoter cloning and gene expression analysis of NCED3 from winter rapeseed (Brassica rapa L.) |
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| DOI：10.7606/j.issn.1000-7601.2020.05.10 |
| Key Words: NCED3 gene winter rapeseed cold resistance promoter cloning sequence analysis stress relative expression |
| Author Name | Affiliation | | WU Junyan | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | MA Li | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | FANG Yan | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | ZHANG Yahong | Tianshui Institute of Agricultural Sciences, Tianshui, Gansu 741001， China | | MA Xuecai | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | LIU Lijun | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | NIU Zaoxia | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | LI Xuecai | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China | | LI Aiguo | Institute of Dry Farming, Hebei Academy of Agriculture and Forestry Sciences, Hengshui, Hebei 053500， China | | SUN Wancang | College of Agronomy, Gansu Agricultural University， Rapeseed Engineering Research Center of Gansu Province, Lanzhou, Gansu 730070, China |
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| Abstract: |
| For studying the role of NCED3 gene in winter rapeseed (Brassica rapa L.), we cloned NCED3 from two different cold\|tolerant winter rapeseeds cultivars, aligned the sequences, and identified its expression in leaves and roots. Results showed that the open reading frame (ORF) of NCED3 had 1 794 bp encoding 597 amino acids. NCED3 from Longyou 6 had a calculated molecular mass of 65.74 kD and theoretical isoelectric point of 5.81 while it from Tianyou 2 was 65.78 kD and 5.94. The two proteins were both hydrophilic proteins with a hydrophobic peak. Compared with NCED3 from Brassica napus, some bases changed and resulted in the changes of amino acids. Results showed that promoters had such hormone\|responsive elements as ABRE, TGA\|element, CGTCA\|motif, and TGACG\|element. In addition, low\|temperature responsiveness element (LTR), defense and stress responsiveness element (TC\|rich repeats) were identified. The similarity of the two promoters was 99.38% and there was only one different cis acting element, circadian, was speculated to be related to circadian rhythm in them. The expression of NCED3 in leaves and roots after low temperature, PEG, and ABA was higher than that in the control group with an increase range of 0.07~6.48, and reached a peak at 8 h. Compared with the expression characteristics in roots, the expression in leaves significantly increased at 2 h after stress treatment, and the increase range in Tianyou 2 (1.54~6.00) was greater than that in Longyou 6 (0.04~1.95). |
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