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[1]田磊,常斐斐,秦世尚,等.播娘蒿耐寒基因DsKIN1的克隆、序列分析及冷诱导表达分析[J].应用与环境生物学报,2017,23(03):467-473.[doi:2015.05019]
　TIAN Lei,CHANG Feifei,QIN Shishang,et al.Cloning and expression analysis of a cold-inducible gene (DsKIN1) from Descurainia sophia[J].Chinese Journal of Applied & Environmental Biology,2017,23(03):467-473.[doi:2015.05019]
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播娘蒿耐寒基因DsKIN1的克隆、序列分析及冷诱导表达分析()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
23卷
期数:
2017年03期
页码:
467-473
栏目:
研究论文
出版日期:
2017-06-25

文章信息/Info

Title:
Cloning and expression analysis of a cold-inducible gene (DsKIN1) from Descurainia sophia
作者:
田磊; 常斐斐; 秦世尚; 唐滋一; 陈麒名; 马晓瑞; 徐辉; 乔代蓉; 曹毅
四川大学生命科学学院，微生物与代谢工程四川省重点实验室 成都 610065
Author(s):
TIAN Lei;  CHANG Feifei;  QIN Shishang;  TANG Ziyi;  CHEN Qiming;  MA Xiaorui;  XU Hui;  QIAO Dairong;  & CAO Yi
Microbiology and Metabolic Engineering Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu 610065, China
关键词:
播娘蒿; DsKIN1; 基因克隆; 低温胁迫; Realtime-PCR
Keywords:
Descurainia sophia;  DsKIN1;  gene cloning;  cold stress;  realtime-PCR
分类号:
Q78 : Q945.78
DOI:
2015.05019
摘要:
播娘蒿是极端耐寒的冷诱导植物，研究其低温胁迫下的冷响应基因对于揭示其抗寒性具有重要意义. 根据拟南芥AtKIN1和AtKIN2同源比对，设计同源引物进行播娘蒿KIN基因克隆，通过RACE技术克隆得到DsKIN1基因，利用Vector NTI11.5软件对其进行序列分析，采用Realtime-PCR测定DsCBF1、DsCBF2、DsCOR和DsKIN1基因在不同处理方式[整株低温处理、同一植株局部低温处理和局部未受低温处理（两部分植株）]下各时间段（0.2 h、0.5 h、1 h、2 h、6 h、12 h、24 h）的表达，采用String数据库预测分析与DsKIN1相互作用的蛋白. 结果显示，DsKIN1基因序列全长为462 bp，开放阅读框ORF长度为198 bp，编码66个氨基酸，分子量（Mr）为6.61 × 103，理论等电点为9.10，亲水性好. 播娘蒿DsCBF1和DsCBF2在0.2 h开始表达，在2 h达到最大，随后下降. 在冷诱导0.2 h后，DsCBF调控的耐寒基因DsCOR和DsKIN1基因开始表达，均呈现上调的趋势. 整株、局部冷诱导植株与同一植株上未受低温胁迫部位的耐寒基因DsCOR和DsKIN1表达模式类似. 此外发现与DsKIN1相互作用的蛋白大部分为低温诱导蛋白、盐和渗透压胁迫响应的信号蛋白、与渗透调节有关的家族蛋白、RING-H2锌指蛋白2B以及未知蛋白等. 本研究说明播娘蒿局部冷诱导时，冷信号可进行传递，使未受冷诱导部位获得耐寒性能. （图7 表4 参27）
Abstract:
Descurainia sophia is a species of extremely hardy and cold-inducible plants. Research on cold-responsive genes of D. sophia under cold stress can help to elucidate the mechanisms underlying its cold resistance. Through homologous alignment of Arabidopsis thaliana AtKIN1 and AtKIN2, DsKIN1 was cloned based on homologous primers. The expression of DsKIN1 was examined by RT-PCR using different low-temperature processing methods (whole plant and partial plant treatment) and different periods (0.2 h, 0.5 h, 1 h, 2 h, 6 h, 12 h, and 24 h). Predicting and analyzing the proteins interacting with DsKIN1 involved the String database. Using RACE, a 462-bp DNA fragment was obtained as the full-length sequence of DsKIN1 containing a 198-bp open reading frame (ORF) encoding 66 amino acid residues. The sequence analysis in Vector NTI 11.5 showed that DsKIN1 has a molecular mass of 6.61 × 103, the theoretical isoelectric point of 9.10, and good hydrophilicity. The expression of DsCBF1 and DsCBF2 started at 0.2 h, reached the maximum at 2 h, and then dropped. The expression of cold-resistance genes DsCOR and DsKIN1 regulated by DsCBF started after cold induction for 0.2 h. This expression pattern showed an upregulation trend. In the whole-plant treatment group (various sites), partial-plant treatment group, and the group without treatment, expression patterns of DsCOR and DsKIN1 were similar. The proteins interacting with DsKIN1 were mostly low-temperature-inducible proteins, salt stress- and osmotic-stress-responsive signaling proteins, a family of proteins related to osmotic adjustment, RING-H2 type zinc-finger protein 2B, and some unknown proteins. This study indicates that a cold signal can be transmitted to give other sites (without cold induction) the capacity for cold resistance.

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更新日期/Last Update: 2017-06-25