[1]秦文斌,戴忠良,山溪,等.甘蓝冷胁迫相关基因BobHLH18克隆与表达分析[J].江苏农业学报,2019,(01):149-156.[doi:doi:10.3969/j.issn.1000-4440.2019.01.022]
　QIN Wen-bin,DAI Zhong-liang,SHAN Xi,et al.Molecular cloning and expression analysis of cold stress-related gene BobHLH18 in cabbage (Brassica oleracea var. capitata L.)[J].,2019,(01):149-156.[doi:doi:10.3969/j.issn.1000-4440.2019.01.022]
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甘蓝冷胁迫相关基因BobHLH18克隆与表达分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2019年01期
页码:
149-156
栏目:
园艺
出版日期:
2019-02-26

文章信息/Info

Title:
Molecular cloning and expression analysis of cold stress-related gene BobHLH18 in cabbage (Brassica oleracea var. capitata L.)
作者:
秦文斌1; 戴忠良1; 山溪1; 唐君2; 王神云2; 李建斌2
(1.江苏丘陵地区镇江农业科学研究所，江苏句容212400；2.江苏省高效园艺作物遗传改良重点实验室/江苏省农业科学院蔬菜研究所，江苏南京210014)
Author(s):
QIN Wen-bin1; DAI Zhong-liang1; SHAN Xi1; TANG Jun2; WANG Shen-yun2; LI Jian-bin2
(1.Zhenjiang Institute of Agricultural Sciences of the Ning-Zhen Hilly District, Jurong 212400, China;2.Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement/Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
甘蓝; BobHLH18; 转录因子; 冷胁迫
Keywords:
cabbage; BoHLH18; transcription factors; cold stress
分类号:
S635
DOI:
doi:10.3969/j.issn.1000-4440.2019.01.022
文献标志码:
A
摘要:
利用同源克隆方法在耐寒,迟抽薹甘蓝自交系Y923中克隆到一个甘蓝响应冷胁迫bHLH转录因子基因BobHLH18的DNA和cDNA全长，基因组搜索分析结果表明，该基因位于甘蓝1号染色体上，属于LF1亚基因组编码基因；序列分析结果表明，该基因含有4个外显子和3个内含子，编码338个氨基酸，蛋白质分子量为38 380，等电点为6.80，其编码蛋白质的N端含有一个bHLH结构域；亚细胞定位分析指出该基因编码蛋白质定位在细胞核上，表明该基因编码蛋白质为核定位蛋白质，与其为转录因子特征相符；序列比对结果显示，BobHLH18蛋白与白菜和拟南芥中的bHLH蛋白具有较高的同源性，相似度分别为90.5%和89.0%；聚类分析指出BobHLH18及其同源蛋白分别聚类成2个进化分支，且来自十字花科植物的bHLH18同源蛋白质都聚在同一进化分支上；qRT-PCR分析结果表明BobHLH18基因受冷胁迫诱导，能在叶片中被较高的诱导表达，表明该基因可能在甘蓝叶片应答冷胁迫过程中起重要作用。
Abstract:
In this study, a gene full-length sequence of the cold stress-related bHLH transcription factor BobHLH18, was isolated from the cabbage inbred line Y923 with cold and bolting resistance by homologous cloning methods. Genome search analysis results showed that this gene was located on C01 chromosome of cabbage, which belonged to the LF1 sub-genome. Sequence analysis results showed that this gene contained four exons and three introns, and its encoded protein contained 338 amino acids with the molecular weight of 38 380 and the isoelectric point of 6.80, its N terminal contained bHLH domain. Subcellular localization predication analysis results indicated that the protein encoded by BobHLH18 was located on the nucleus and had nuclear protein features, which was consistent with transcription factor characteristics. Homology analysis results showed that BobHLH18 protein had high homology with bHLH18 protein in Chinese cabbage and Arabidopsis, and the similarities were 90.5% and 89.0%, respectively. Cluster analysis results showed that BobHLH18 and its homologous proteins could be divided into two evolutionary branches, and the bHLH18 homologs from the cruciferous plants could be clustered into the same division. qRT-PCR analysis results indicated that BobHLH18 gene was induced by cold stress and had high expression levels at 12 h after cold treatment, suggesting that BobHLH18 gene may play an important role in the response of cabbage leaves to cold stress.

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备注/Memo

备注/Memo:
收稿日期：2018-03-16 基金项目：国家重点研发计划项目(2016YFD0101702)；江苏省重点研发计划项目（BE2017379）；江苏省农业科技自主创新基金项目［CX(15)1016］；宁波市科技计划项目(2015C110008) 作者简介：秦文斌(1971-)，男，江苏句容人，本科，副研究员，从事蔬菜栽培育种研究。(E-mial)qinwenbinbin@126.com 通讯作者：李建斌，(E-mial)jbli0518@163.com
更新日期/Last Update: 2019-02-27