生菜低温胁迫转录因子LsICE1的克隆、特征分析及其转化水稻

doi:10.3864/j.issn.0578-1752.2011.21.002

Abstract

Abstract: 【Objective】 Cloning, sequence analysis and transformation into rice with cold stress transcription factor LsICE1 from lettuce were conducted to elucidate the effects of overexpression of LsICE1 on low temperature stress tolerant capacities in rice.【Method】The highly conserved cDNA region of LsICE1 gene from Lactuca sativa L. was isolated by reverse transcription-PCR (RT-PCR) with a pair of degenerate primers, 3′-end and 5′-end were amplified by single oligonucleotide nested PCR (SON-PCR) and then the three cDNA segments were combined and constituted a full-length cDNA sequence of LsICE1 gene. Bioinformatics methods were used to analyze the obtained cDNA sequences and the deduced amino acid sequences. The gene expression levels at 4℃ were analyzed by semi-quantitative RT-PCR (s-qRT-PCR). With constructed plant expressing vector, LsICE1 gene was successfully transferred into rice by Agrobacterium-mediated transgenic technique. The regulation effects of LsICE1 gene with high expression levels on low temperature stress tolerant capacities were evaluated by the comparisons of survival rate and physiological parameters among the transgenic lines and control (CK) after low temperature treatment. 【Result】 Sequence analysis showed that the combined cDNA (designated as LsICE1, GenBank Accession No. HQ848932) fragment length was 1 622 bp containing a full coding region of 1 497 bp encoding 498 amino acid residues. The qRT-PCR result suggested that LsICE1 was differential expression gene under low temperature stress. The homology tree demonstrated that LsICE1 was at the same evolutionary branch with VvICE1. PCR and RT-PCR detection confirmed that LsICE1 gene had been integrated into rice genome. After low temperature stress treatment, compared with control, the transgenic lines with high expression levels of the LsICE1 gene had obvious higher survival rates and proline content. In the meantime, the accumulation rate of relative conductivity and malondialdehyde (MDA) content were much lower. 【Conclusion】 The cold stress transcription factor LsICE1 was firstly cloned from leaf of Lactuca sativa L. with cold tolerance. Overexpression of LsICE1 gene improved low temperature stress tolerant capacities in transgenic rice lines.

Key words: Lactuca sativa L., rice, LsICE1 gene, gene cloning, SON-PCR, sequence analysis

XIANG Dian-Jun, YIN Kui-De, MAN Li-Li, XU Zheng-Jin. Cloning and Characteristic Analysis of Cold Stress Transcription Factor LsICE1 from Lettuce and Transformation into Rice[J].Scientia Agricultura Sinica, 2011, 44(21): 4340-4349.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2011.21.002

https://www.chinaagrisci.com/EN/Y2011/V44/I21/4340

References

[1]Guy C L, Niemi K J, Brambl R. Altered gene expression during cold acclimation of spinach. Proceedings of the National Academy of Sciences of the USA, 1985, 82: 3673-3677.

[2]Wang Z B, Feng L R, Wang J J, Wang Z Y. Vitis amuerensis CBF3 gene isolation, sequence analysis and expression. Agricultural Sciences in China, 2010, 9(8): 1127-1132.

[3]陈儒钢, 朱文超, 巩振辉, 李大伟, 尹延旭, 逯明辉. 辣椒水通道蛋白基因CaAQP的克隆与序列分析. 中国农业科学, 2010, 43(20): 4323-4329.

Chen R F, Zhu W C, Gong Z H, Li D W, Yin Y X, Dai M H. Cloning and sequence analysis of the aquaporins gene CaAQP in pepper. Scientia Agricultura Sinica, 2010, 43(20): 4323-4329. (in Chinese)

[4]Navarro M, Marque G, Ayax C, Keller G, Borges J P, Marque C, Teulieres C. Complementary regulation of four eucalyptus CBF genes under various cold conditions. Journal of Experimental Botany, 2009, 60(9): 2713-2724.

[5]Wang Y, Hua J. A moderate decrease in temperature induces COR15a expression through the CBF signaling cascade and enhances freezing tolerance. The Plant Journal, 2009, 60: 340-349.

[6]Knox A K, Dhillon T, Cheng H, Tondelli A, Pecchioni N, Stockinger E J. CBF gene copy number variation at Frost Resistance-2 is associated with levels of freezing tolerance in temperate-climate cereals. Theoretical and Applied Genetics, 2010, 121: 21-35.

[7]Campoli C, Matus-Cádiz M A, Pozniak C J, Cattivelli L, Fowler D B. Comparative expression of CBF genes in the Triticeae under different acclimation induction temperatures. Molecular Genetics and Genomics, 2009, 282: 141-152.

[8]Zhao L F, Hu Y B, Chong K, Wang T. ARAG1, an ABA-responsive DREB gene, plays a role in seed germination and drought tolerance of rice. Annals of Botany, 2010, 105: 401-409.

[9]Yang Y F, Wu J, Zhu K, Liu L Q, Chen F D, Yu D Y. Identification and characterization of two chrysanthemum (Dendronthema× moriforlium) DREB genes, belonging to the AP2/EREBP family. Molecular Biology Reports, 2009, 36: 71-81.

[10]Xiao H G, Siddiqua M, Braybrook S, Nassuth A, Braybrook S. Three grape CBF/DREB1 genes respond to low temperature, drought and abscisic acid. Plant, Cell and Environment, 2006, 29: 1410-1421.

[11]Chinnusamy V, Ohta M, Kanrar S, Lee B H, Hong X H, Agarwal M, Zhu J K. ICE1: a regulator of cold-induced transcriptome and freezing tolerance in Arabidopsis. Genes and Development, 2003, 17: 1043-1054.

[12]Gilmour S J, Zarka D G, Stockinger E J, Salazar M P, Houghton J M, Thomashow M F. Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression. The Plant Journal, 1998, 16: 433-442.

[13]Badawi M, Reddy Y V, Agharbaoui Z, Tominaga Y, Danyluk J, Sarhan F, Houde M. Structure and functional analysis of wheat ICE (Inducer of CBF Expression) genes. Plant Cell Physiology, 2008, 49(8): 1237-1249.

[14]Liu L Y, Duan L S, Zhang J C, Zhang Z C, Mi G Q, Ren H Z. Cucumber (Cucumis sativus L.) over-expressing cold-induced transcriptome regulator ICE1 exhibits changed morphological characters and enhances chilling tolerance. Scientia Horticulturae, 2010, 124: 29-33.

[15]轩春雷, 肖向文, 李晓波, 祝建波. 盐芥ICE1转录因子的克隆及生物信息学分析. 生物技术通报, 2010(11): 108-114.

Xuan C L, Xiao X W, Li X B, Zhu J B. Cloning and bioinformatics analysis of a transcription factor ICE1 from Thellungiella halophila. Biotechnology Bulletin, 2010(11): 108-114. (in Chinese)

[16]林元震, 张志毅, 刘纯鑫, 郭海, 朱保庆, 陈晓阳. 甜杨抗冻转录因子ICE1基因的in silico克隆及其分析. 分子植物育种, 2007, 5(3): 424-430.

Lin Y Z, Zhang Z Y, Liu C X, Guo H, Zhu B Q, Chen X Y. in silico cloning and analyzing of PslCEl from Populus suaveolens, a freezing-resistant transcription factor. Molecular Plant Breeding, 2007, 5(3): 424-430. (in Chinese)

[17]郑银英, 崔百明, 常明进, 彭明. 转拟南芥 ICE1基因增强烟草抗寒性的研究. 西北植物学报, 2009, 29(1): 75-79.

Zheng Y Y, Cui B M, Chang M J, Peng M. Expression of AtICE1 gene in transgenic tobacco plants. Acta Botanica Boreali-Occidentalia Sinica, 2009, 29(1): 75-79. (in Chinese)

[18]张玉, 蒋欣梅, 于欣宏. ICE1基因表达载体的构建及对番茄的转化. 中国蔬菜, 2010(18): 27-33.

Zhang Y, Jiang X M, Yu X H. Construction of ICE1 gene expression vector and transformation into tomato (Lycopersicon esculentum Mill.). China Vegetables, 2010(18): 27-33. (in Chinese)

[19]Huang J Q, Sun Z H. Agrobacterium-mediated transfer of Arabidopsis ICEl gene into lemon (Citrus Limon L.). Agricultural Sciences in China, 2005, 4(9): 714-720.

[20]Miura K, Jin J B, Lee J, Hasegawa P M. SIZ1-mediated sumoylation of ICE1 controls CBF3/DREB1A expression and freezing tolerance in Arabidopsis. The Plant Cell, 2007, 19: 1403-1414.

[21]Hiei Y, Ohta S, Komar T, Kumashiro T. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. The Plant Journal, 1994, 6(2): 271-282.

[22]高明波, 柳展基, 于翠梅, 曹萍, 刘世强. 盐胁迫水稻愈伤组织的生理生化指标检测. 沈阳农业大学学报, 1998, 30(4): 404-407.

Gao M B, Liu Z Z, Yu C M, Cao P, Liu S Q. Determination of physiological and biochemical indexes of calli in selection of rice mutants resistant to sodium chloride. Journal of Shenyang Agricultural University, 1998, 30(4): 404-407. (in Chinese)

[23]王闵霞，马欣荣, 王天山, 谭红. 染色体步行PCR技术. 应用与环境生物学报, 2006, 12(3): 427-430.

Wang M X, Ma X R, Wang T X, Tan H. PCR techniques for chromosome walking. Chinese Journal of Applied and Environmental Biology, 2006, 12(3): 427-430. (in Chinese)

[24]Antal Z, Rascle C, Fevre M, Bruel C. Single oligonucleotide nested PCR: a rapid method for the isolation of genes and their flanking regions from expressed sequence tags. Current Genetics, 2004, 46: 240-246.

[25]邓洪渊, 王闵霞, 孙雪文, 马欣荣, 曹婷婷, 谭红. 一种改进的SON-PCR基因扩增方法. 应用与环境生物学报, 2006, 12(6): 857-860.

Deng H Y, Wang M X, Sun X W, Ma X R, Cao T T, Tan H. An improved SON-PCR method for gene amplification. Chinese Journal of Applied and Environmental Biology, 2006, 12(6): 857-860. (in Chinese)

[26]Lee B H, Henderson D A, Zhu J K. The Arabidopsis cold-responsive transcriptome and its regulation by ICE1. The Plant Cell, 2005, 17:3155-3175.

[27]Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. Two transcription factors, DREB1 and DREB2, with and EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought and low temperature-responsive gene expression, respectively, in Arabidopsis. The Plant Cell, 1998: 1391-1406.

[28]Gilmour S J, Sebolt A M, Salazar M P, Everard J D, Thomashow M F. Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple biochemical changes associated with cold acclimation. Plant Physiology, 2000, 124: 1854-1865.

Related Articles 15

[1]	XIAO DeShun, XU ChunMei, WANG DanYing, ZHANG XiuFu, CHEN Song, CHU Guang, LIU YuanHui. Effects of Rhizosphere Oxygen Environment on Phosphorus Uptake of Rice Seedlings and Its Physiological Mechanisms in Hydroponic Condition [J]. Scientia Agricultura Sinica, 2023, 56(2): 236-248.
[2]	ZHANG XiaoLi, TAO Wei, GAO GuoQing, CHEN Lei, GUO Hui, ZHANG Hua, TANG MaoYan, LIANG TianFeng. Effects of Direct Seeding Cultivation Method on Growth Stage, Lodging Resistance and Yield Benefit of Double-Cropping Early Rice [J]. Scientia Agricultura Sinica, 2023, 56(2): 249-263.
[3]	GU LiDan,LIU Yang,LI FangXiang,CHENG WeiNing. Cloning of Small Heat Shock Protein Gene Hsp21.9 in Sitodiplosis mosellana and Its Expression Characteristics During Diapause and Under Temperature Stresses [J]. Scientia Agricultura Sinica, 2023, 56(1): 79-89.
[4]	ZHANG Wei,YAN LingLing,FU ZhiQiang,XU Ying,GUO HuiJuan,ZHOU MengYao,LONG Pan. Effects of Sowing Date on Yield of Double Cropping Rice and Utilization Efficiency of Light and Heat Energy in Hunan Province [J]. Scientia Agricultura Sinica, 2023, 56(1): 31-45.
[5]	FENG XiangQian,YIN Min,WANG MengJia,MA HengYu,CHU Guang,LIU YuanHui,XU ChunMei,ZHANG XiuFu,ZHANG YunBo,WANG DanYing,CHEN Song. Effects of Meteorological Factors on Quality of Late Japonica Rice During Late Season Grain Filling Stage Under ‘Early Indica and Late Japonica’ Cultivation Pattern in Southern China [J]. Scientia Agricultura Sinica, 2023, 56(1): 46-63.
[6]	SANG ShiFei,CAO MengYu,WANG YaNan,WANG JunYi,SUN XiaoHan,ZHANG WenLing,JI ShengDong. Research Progress of Nitrogen Efficiency Related Genes in Rice [J]. Scientia Agricultura Sinica, 2022, 55(8): 1479-1491.
[7]	GUI RunFei,WANG ZaiMan,PAN ShengGang,ZHANG MingHua,TANG XiangRu,MO ZhaoWen. Effects of Nitrogen-Reducing Side Deep Application of Liquid Fertilizer at Tillering Stage on Yield and Nitrogen Utilization of Fragrant Rice [J]. Scientia Agricultura Sinica, 2022, 55(8): 1529-1545.
[8]	LIAO Ping,MENG Yi,WENG WenAn,HUANG Shan,ZENG YongJun,ZHANG HongCheng. Effects of Hybrid Rice on Grain Yield and Nitrogen Use Efficiency: A Meta-Analysis [J]. Scientia Agricultura Sinica, 2022, 55(8): 1546-1556.
[9]	HAN XiaoTong,YANG BaoJun,LI SuXuan,LIAO FuBing,LIU ShuHua,TANG Jian,YAO Qing. Intelligent Forecasting Method of Rice Sheath Blight Based on Images [J]. Scientia Agricultura Sinica, 2022, 55(8): 1557-1567.
[10]	GAO JiaRui,FANG ShengZhi,ZHANG YuLing,AN Jing,YU Na,ZOU HongTao. Characteristics of Organic Nitrogen Mineralization in Paddy Soil with Different Reclamation Years in Black Soil of Northeast China [J]. Scientia Agricultura Sinica, 2022, 55(8): 1579-1588.
[11]	ZHU DaWei,ZHANG LinPing,CHEN MingXue,FANG ChangYun,YU YongHong,ZHENG XiaoLong,SHAO YaFang. Characteristics of High-Quality Rice Varieties and Taste Sensory Evaluation Values in China [J]. Scientia Agricultura Sinica, 2022, 55(7): 1271-1283.
[12]	ZHAO Ling, ZHANG Yong, WEI XiaoDong, LIANG WenHua, ZHAO ChunFang, ZHOU LiHui, YAO Shu, WANG CaiLin, ZHANG YaDong. Mapping of QTLs for Chlorophyll Content in Flag Leaves of Rice on High-Density Bin Map [J]. Scientia Agricultura Sinica, 2022, 55(5): 825-836.
[13]	JIANG JingJing,ZHOU TianYang,WEI ChenHua,WU JiaNing,ZHANG Hao,LIU LiJun,WANG ZhiQin,GU JunFei,YANG JianChang. Effects of Crop Management Practices on Grain Quality of Superior and Inferior Spikelets of Super Rice [J]. Scientia Agricultura Sinica, 2022, 55(5): 874-889.
[14]	ZHANG YaLing, GAO Qing, ZHAO Yuhan, LIU Rui, FU Zhongju, LI Xue, SUN Yujia, JIN XueHui. Evaluation of Rice Blast Resistance and Genetic Structure Analysis of Rice Germplasm in Heilongjiang Province [J]. Scientia Agricultura Sinica, 2022, 55(4): 625-640.
[15]	WANG YaLiang,ZHU DeFeng,CHEN RuoXia,FANG WenYing,WANG JingQing,XIANG Jing,CHEN HuiZhe,ZHANG YuPing,CHEN JiangHua. Beneficial Effects of Precision Drill Sowing with Low Seeding Rates in Machine Transplanting for Hybrid Rice to Improve Population Uniformity and Yield [J]. Scientia Agricultura Sinica, 2022, 55(4): 666-679.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Cloning and Characteristic Analysis of Cold Stress Transcription Factor LsICE1 from Lettuce and Transformation into Rice

PDF

Cited