欢迎访问作物学报,今天是

作物学报 ›› 2014, Vol. 40 ›› Issue (10): 1748-1755.doi: 10.3724/SP.J.1006.2014.01748

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝型油菜LPAT4基因的克隆与表达

肖旦望1,刘聪1,胡学芳1,陈社员2,官春云2,熊兴华1,2,*   

  1. 1湖南农业大学 / 作物基因工程湖南省重点实验室, 湖南长沙 410128; 2 湖南农业大学 / 国家油料改良中心湖南分中心, 湖南长沙 410128
  • 收稿日期:2014-03-10 修回日期:2014-07-06 出版日期:2014-10-12 网络出版日期:2014-07-25
  • 通讯作者: 熊兴华, E-mail: ndxiongene@yahoo.com, Tel: 13508487613
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2012AA101107-3)和湖南农业大学作物学开发基金资助项目(ZWKF201303)资助。

Cloning and Expression Analysis of LPAT4 Gene from Brassica napus

XIAO Dan-Wang1,LIU Cong1,HU Xue-Fang1,CHEN She-Yuan2,GUAN Chun-Yun2,XIONG Xing-Hua1,2,*   

  1. 1 Crop Gene Engineering Key Laboratory of Hunan Province, Hunan Agricultural University, Changsha 410128, China; 2 Hunan Branch of National Oilseed Crops Improvement Centre, Hunan Agricultural University, Changsha 410128, China
  • Received:2014-03-10 Revised:2014-07-06 Published:2014-10-12 Published online:2014-07-25
  • Contact: 熊兴华, E-mail: ndxiongene@yahoo.com, Tel: 13508487613

摘要:

植物溶血磷脂酸酰基转移酶(lysophospholipid acid actyltransferase, LPAT)是三酰甘油生物合成过程中的一个关键酶, 在脂质的合成、种子的发育以及生物膜的流动性等方面有重要作用。本研究采用同源克隆的方法, 获得LPAT4基因的2条全长CDS序列, 长度分别为1143 bp1140 bp。生物信息学分析表明, 它们均具有LPLAT_LCLAT1样结构域, 同属于LPLAT超基因家族, 并分别被命名为BnLPAT4-1BnLPAT4-2。时空表达分析表明, 它们均为组成型表达基因, 其中BnPAT4-1在叶中的表达量最高, BnPAT4-2在胚中的表达量最高。逆境分析表明, BnLPAT4-1BnLPAT4-2NaClPEG-4000、水渍、6BAABA的胁迫下呈现出不同的表达模式。极差分析显示, ABABnLPAT4-1的表达影响较大, BnLPAT4-2的表达却对PEG4000更敏感。为进一步研究油菜BnLPAT4基因功能奠定了基础。

关键词: LPAT4, 甘蓝型油菜, 时空表达, 非生物逆境, 表达分析

Abstract:

Lysophospholipid acid actyltransferase (LPAT) is a pivotal enzyme of triacylglycerol biosynthesis, which plays a key role in lipid synthesis, development of plant seeds and bio-membrane fluidity. Through the technology of homology-based cloning, two copies of LPAT4 full-length CDS sequences (1143 bp and 1140 bp) were cloned in this study, designated as BnLPAT4-1and BnLPAT4-2, respectively. Bioinformatics analysis revealed that they shared the LPLAT_LCLAT1 like domain and belonged to the LPLAT superfamily. Temporal and spatial expression results showed that BnLPAT4-1 and BnLPAT4-2 were constitutive expression genes. Among them, the highest expression of BnLPAT4-1 was in leaf, while that ofBnLPAT4-2 was in embryo. Stresses analysis indicated that BnLPAT4-1and BnLPAT4-2 presented different expression patterns under the treatments of NaCl, PEG4000, waterlogging, 6BA and ABA. Pole difference analysis displayed that ABA had a great effect on the expression of BnLPAT4-1, while BnLPAT4-2 was more sensitive to PEG-4000.The results provided a base for the research of regulation and function of BnLPAT4 in Brassica napus.

Key words: LPAT4, Brassica Napus, Temporal and spatial expression, Abiotic stress, Expression analysis

[1]Yu W L, Ansari W, Schoepp N G, Hannon M J, Mayfield S P, Burkart M D. Modifications of the metabolic pathways of lipid and triacylglycerol production in microalgae. Microb Cell Fact, 2011, 10: 1–11



[2]Li Y H, Basil S, Fred B, Mats X. A, Vincent A, Philip D. B, Sébastien B, David B, Allan D, Timothy P. D, Rochus B. F, Ian A. G, Kenta K, AmélieA. K, Tony L, Jonathan E. M, Martine M, Isabel M, Ikuo N, Owen R, Lacey S, Katherine M. S, Hajime W, Ruth W, Xu C C, Rémi Z, John O. Acyl-lipidmetabolism. Am Soc Plant Biol, 2013, 11: 1–70



[3]Baud S, Dubreucq B, Miquel M, Rochat C, Lepiniec L. Storage reserve accumulation in Arabidopsis: metabolic and developmental control of seed filling. Am Soc Plant Biol, 2008, 6: 1–24



[4]Kim H U, Li Y, Huang A H. Ubiquitous and endoplasmic reticulum-located lysophosphatidyl acyltransferase, LPAT2, is essential for female but not male gametophyte development in Arabidopsis. Plant Cell, 2005, 17: 1073–1089



[5]Lopez-Villalobos A, Dodds P F, Hornung R. Changes in fatty acid composition during development of tissues of coconut (Cocos nucifera L.) embryos in the intact nut and in vitro. J Exp Bot, 2001, 52: 933–942



[6]Knutzon D S, Hayes T R, Wyrick A, Xiong H, Maelor D H, Voelker T A. Lysophosphatidic acid acyltransferase from coconut endosperm mediates the insertion of laurate at the sn-2 position of triacylglycerols in lauric rapeseed oil and can increase total laurate levels. Plant Physiol, 1999, 120: 739–746



[7]Knutzon D S, Lardizabal K D, Nelsen J S, Bleibaum J L, Davies H M, Metz J G. Cloning of a coconut endosperm cDNA encoding a 1-acyl-sn-glycerol-3-phosphate acyltransferase that accepts medium-chain-length substrates. Plant Physiol, 1995, 109: 999–1006



[8]Bourgis F, Kader J C, Barret P, Renard M, Robinson D, Robinson C, Delseny M, Roscoe T J. A plastidial lysophosphatidic acid acyltransferase from oilseed rape. Plant Physiol, 1999, 120: 913–922



[9]Bernerth R, Frentzen M. Utilization of erucoyl-CoA by acyltransferases from developing seeds of Brassica napus (L.) involved in triacylglycerol biosynthesis. Plant Sci, 1990, 67: 21–28



[10]Taylor D C, Barton D L, Giblin E M, Mackenzie S L, Van Den Berg C, Mcvetty P. Microsomal lyso-phosphatidic acid acyltransferase from a Brassica oleracea cultivar incorporates erucic acid into the sn-2 position of seed triacylglycerols. Plant Physiol, 1995, 109: 409–420



[11]Lassner M W, Levering C K, Davies H M, Knutzon D S. Lysophosphatidic acid acyltransferase from meadowfoam mediates insertion of erucic acid at the sn-2 position of triacylglycerol in transgenic rapeseed oil. Plant Physiol, 1995, 109: 1389–1394



[12]Cao Y Z, Oo K C, Huang A H. Lysophosphatidate acyltransferase in the microsomes from maturing seeds of meadowfoam (Limnanthes alba). Plant Physiol, 1990, 94: 1199–1206



[13]Oo K C, Huang A H. Lysophosphatidate acyltransferase activities in the microsomes from palm endosperm, maize scutellum, and rapeseed cotyledon of maturing seeds. Plant Physiol, 1989, 91: 1288–1295



[14]Brown A P, Coleman J, Tommey A M, Watson M D, Slabas A R. Isolation and characterisation of a maize cDNA that complements a 1-acyl sn-glycerol-3-phosphate acyltransferase mutant of Escherichia coli and encodes a  protein which has similarities to other acyltransferases. Plant Mol Biol, 1994, 26: 211–223



[15]Ichihara K, Asahi T, Fujii S. 1-Acyl-sn-glycerol-3-phosphate acyltransferase in maturing safflower seeds and its contribution to the non-random fatty acid distribution of triacylglycerol. Eur J Biochem, 1987, 167: 339–347



[16]Griffiths G, Stobart A K, Stymne S. The acylation of sn-glycerol 3-phosphate and the metabolism of phosphatidate in microsomal preparations from the developing cotyledons of safflower (Carthamus tinctorius L.) seed. Biochem J, 1985, 230: 379–388



[17]Arroyo-Caro J M, Chileh T, Kazachkov M, Zou J, Alonso D L, Garcia-Maroto F. The multigene family of lysophosphatidate acyltransferase (LPAT)-related enzymes in Ricinuscommunis. Cloning and molecular characterization of two LPAT genes that are expressed in castor seeds. Plant Sci, 2013, 199/200: 29–40



[18]Chen S L, Huang J Q, Lei Y, Zhang Y T, Ren X P, Chen Y N, Jiang H F, Yan L Y, Li Y R, Liao B S. Identification and characterization of a gene encoding a putative lysophosphatidylacyltransferase from Arachis hypogaea. J Biosci, 2012, 37: 1029–1039



[19]Kim H U, Huang A H. Plastid lysophosphatidyl acyltransferase is essential for embryo development in Arabidopsis. Plant Physiol, 2004, 134: 1206–1216



[20]Ananda K. Ghosh N C S R. At4g24160, a Soluble acyl-coenzyme a-dependent lysophosphatidic acid acyltransferase. Plant Physiol, 2009, 151: 869–881



[21]Maisonneuve S, Bessoule J J, Lessire R, Delseny M, Roscoe T J. Expression of rapeseed microsomal lysophosphatidic acid acyltransferase isozymes  enhances seed oil content in Arabidopsis. Plant Physiol, 2010, 152: 670–684



[22]Rao S S, Hildebrand D. Changes in oil content of transgenic soybeans expressing the yeast SLC1 gene. Lipids, 2009, 44: 945–951



[23]戚维聪. 油菜发育种子中油脂积累与Kennedy途径酶活性的关系研究. 南京农业大学硕士学位论文, 江苏南京, 2008



Qi W C. Studies on correlations of developing seed lipid accumulation with Kennedy pathway enzyme activities in Brassica napus. MS Thesis of Nanjing Agriculture University, Nanjing, China, 2008 (in Chinese with English abstract)



[24]陈四龙, 黄家权, 雷永, 任小平, 文奇根, 陈玉宁, 姜慧芳, 晏立英, 廖伯寿. 花生溶血磷脂酸酰基转移酶基因的克隆与表达分析. 作物学报, 2012, 38: 245–255



Chen S L, Huang J Q, Lei Y, Ren X P, Wen Q G, Chen Y N, Jiang H F, Yan L Y, Liao B S. Cloning and expression analysis of lysophosphatidic acid acyltransferase (LPAT) encoding gene in peanut. Acta Agron Sin, 2012, 38: 245–255 (in Chinese with English abstract)



[25]Kim H U, Vijayan P, Carlsson A S, Barkan L, Browse J. A mutation in the LPAT1 gene suppresses the sensitivity of fab1 plants to low temperature. Plant Physiol, 2010, 153: 1135–1143



[26]Chen S L, Huang J Q, Lei Y, Zhang Y T, Ren X P, Chen Y N, Jiang H F, Yan L Y, Li Y R, Liao B S. Identification and characterization of a gene encoding a putative lysophosphatidyl acyltransferase from Arachis hypogaea. J Biosci, 2012, 37: 1029–1039



[27]Gong Q, Li P, Ma S, Indu R S, Bohnert H J. Salinity stress adaptation competence in the extremophile Thellungiella halophila in comparison with its relative Arabidopsis thaliana. Plant J, 2005, 44: 826–839



[28]Howell E C, Kearsey M J, Jones G H, King G J, Armstrong S J. A and C genome distinction and chromosome identification in Brassica napus by sequential fluorescence in situ hybridization and genomic in situ hybridization. Genetics, 2008, 180: 1849–1857

[1] 左同鸿, 张贺翠, 曾静, 朱利泉. 甘蓝自交不亲和相关基因BoPUB3L的克隆与表达分析[J]. 作物学报, 2026, 52(6): 1698-1710.
[2] 胡赵, 钱润, 谢丰璞, 应素平. 水稻SPX基因家族鉴定及响应磷处理的表达分析[J]. 作物学报, 2026, 52(6): 1902-1912.
[3] 田立涛, 丁宁, 王树林, 齐恩芳, 张荣, 王蕊蕊, 马利雯, 李建武, 杨江伟. 马铃薯Argonaute基因家族鉴定及晚疫病诱导表达分析[J]. 作物学报, 2026, 52(4): 1116-1126.
[4] 杨宗桃, 杨婷, 王禹童, 艾静, 李燕烨, 刘家勇, 邓军, 赵勇, 张跃彬. 甘蔗CLC基因家族鉴定与表达分析[J]. 作物学报, 2026, 52(3): 722-734.
[5] 孟成, 王哲. 玉米ZmPFK基因家族全基因组鉴定及响应胁迫表达分析[J]. 作物学报, 2026, 52(3): 764-779.
[6] 张力岚, 杨军, 王让剑. 基于WGCNA发掘茶树糖苷类香气前体含量性状相关的候选基因[J]. 作物学报, 2026, 52(2): 494-513.
[7] 马毅娜, 吴晓明玉, 李藕琪, 王圆, 陈丽, 张盈川, 赵伦, 文静, 傅廷栋, 沈金雄. Bna-miR1040-EIF3A模块调控油菜开花时间的功能研究[J]. 作物学报, 2026, 52(2): 349-362.
[8] 董丽华, 董成艳, 李正楠, 余静, 叶靓, 刘芳, 谭静. 玉米禾谷镰孢穗腐病抗性候选基因的筛选与鉴定[J]. 作物学报, 2026, 52(1): 131-147.
[9] 王彬, 蒙姜宇, 邱浩良, 贺亚军, 钱伟. 甘蓝型油菜BnaDUF579基因家族的鉴定与表达模式分析[J]. 作物学报, 2025, 51(8): 2100-2110.
[10] 沈傲, 刘敏, 倪迪安, 刘炜. 谷子m6A甲基转移酶基因SiMTA1的启动子序列特征和基因表达模式分析[J]. 作物学报, 2025, 51(7): 1969-1978.
[11] 郭腾达, 崔梦杰, 陈琳杰, 韩锁义, 郭敬坤, 吴晨迪, 付留洋, 黄冰艳, 董文召, 张新友. 花生磷脂酰肌醇转运蛋白基因AhSFH的克隆及其响应黄曲霉菌侵染的表达特征分析[J]. 作物学报, 2025, 51(6): 1489-1500.
[12] 夏琦, 郭滢, 王坤美, 王思忆, 巨建业, 彭雅雯, 刘忠松, 夏石头. 甘蓝型油菜种子和种皮中水杨酸含量与原花色素积累的关系研究[J]. 作物学报, 2025, 51(5): 1189-1197.
[13] 潘炬忠, 韦萍, 朱德平, 邵胜雪, 陈珊珊, 韦雅倩, 高维维. 水稻转录因子OsERF104的克隆和功能研究[J]. 作物学报, 2025, 51(4): 900-913.
[14] 王晓琳, 刘忠松, 康雷, 杨柳. 甘蓝型油菜角果长度和每角粒数基因定位以及角果皮转录组动态分析[J]. 作物学报, 2025, 51(4): 888-899.
[15] 张琴, 戴成, 马朝芝. 生长素响应报告基因转化甘蓝型油菜及各组织GUS动态信号分析[J]. 作物学报, 2025, 51(3): 667-675.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!