茶树根系Actin基因克隆及表达分析

李远华; 陆建良; 范方媛; 石玉涛

doi:10.13305/j.cnki.jts.2015.04.005

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茶叶科学 ›› 2015, Vol. 35 ›› Issue (4) : 336-346. DOI: 10.13305/j.cnki.jts.2015.04.005

茶树根系Actin基因克隆及表达分析

李远华¹, 陆建良², 范方媛², 石玉涛¹

作者信息 +

Gene Cloning and Expression Analysis of Actin in Tea Plant Root

LI Yuanhua¹, LU Jianliang², FAN Fangyuan², SHI Yutao¹

Author information +

文章历史 +

摘要

采用SSH技术分析了VA菌根处理后福鼎大白茶根系基因差异表达情况,获得了差异序列,序列比对显示,在下调表达序列中可能包含了10种未知功能的基因;在上调表达序列中可能包含了5种可能的基因。采用RACE技术获得了Actin基因全长序列,Actin基因长1β606βbp (GenBank, 登录号KJ946252),具有1β131βbp开放阅读框（1^st~1β131^st）,编码377个氨基酸。分子生物信息学分析表明,Actin蛋白分子量约30.69βkD,等电点为5.27,定位于细胞核等亚细胞区位。研究还显示,Actin在不同品种中表达无显著差异,对非生物性胁迫响应也较弱。

Abstract

By using SSH, we analyzed differences in gene expression of root from Fuding white tea infected by VA mycorrhiza and obtained diversity sequences. The sequence alighment showed that the down-regulated expression sequence possibly contained 10 unknown genes and the up-regulated expression sequence possible contained 5 known genes. The Actin genic full-length sequence was obtained by using RACE. The length of Actin gene was 1β606βbp (GenBank Accession No. KJ946252), with 1β131βbp ORF (1^st-1β131^st), the sequence encoded 377 amino acid. Bioinformatics indicated that the Actin protein’s molecular weight was about 30.69 kD, IEP was 5.27, located in subcellular fraction area like cell nucleus. The study also showed Actin expressed no obvious difference in different cultivars and it responded weak to non-biological stress.

导出引用

李远华, 陆建良, 范方媛, 石玉涛. 茶树根系Actin基因克隆及表达分析[J]. 茶叶科学. 2015, 35(4): 336-346 https://doi.org/10.13305/j.cnki.jts.2015.04.005

LI Yuanhua, LU Jianliang, FAN Fangyuan, SHI Yutao. Gene Cloning and Expression Analysis of Actin in Tea Plant Root[J]. Journal of Tea Science. 2015, 35(4): 336-346 https://doi.org/10.13305/j.cnki.jts.2015.04.005

中图分类号： S571.1 Q52

参考文献

[1] 阎隆飞, 石德权. 高等植物中的收缩蛋白[J]. 生物化学与生物物理学报, 1963, 3: 490-495.
[2] Barrero R, Umeda M, Yamamura S, et al. Arabidopsis CAP regulates the actin cytoskeleton necessary for plant cell elongation and division[J]. The Plant Cell, 2002, 14(1): 149-163.
[3] Trelles-Sticken E, Adelfalk C, Loidl J, et al. Meiotic telomere clustering requires actin for its formation and cohesion for its resolution[J]. Cell Biology, 2005, 170(2): 213-223.
[4] Baluska F, Hlavacka A, Samaj J, et al. F-actin-dependentendocytosis of cell wall pectins in meristematic root cells Insights from brefeldin a induced compartments[J]. Plant Physiology, 2002, 130(9): 422-431.
[5] Franklin-Tong VE.Signaling and the modulation of pollen tube growth[J]. PlantCell, 1999, 11(4): 727-738.
[6] Maisch J, Nick P.Actin is involved in auxin-dependent patterning[J]. Plant Physiology, 2007, 143(4):1695-1704.
[7] Gestel KV, Kohler RH, Verbelen JP.Plant mitochondria move on Factin but their positioning in the cortical cytoplasm depends on both F-actin and microtubules[J]. Journal of Experimental Botany, 2002, 53(369): 659-667.
[8] Chaudhry F, Guerin C, Witsch M.Identification of Arabidopsis cyclase-associated protein 1 as the first nucleotide exchange factor for plant actin[J]. Molecular Biology of the Cell, 2007, 18(8): 3002-3014.
[9] Staiger CJ, Franklin-Tong VE.The actin cytoskeleton is a target of the self-incompatibility response in Papaver rhoeas[J]. Experimental Botany, 2003, 54(380): 103-113.
[10] Balasubramanian R, Karve A, Kandasamy M, et al. A role for F-actin in hexokinase-mediated glucose signaling[J]. Plant Physiology, 2007, 145(12): 1423-1434.
[11] Driss-Ecole D, Vassy J, Rembur J, et al. Immunolocalization of actin in root statocytes of Lensculinaris L[J]. Journal of Experimental Botany, 2000, 51(344): 521-528.
[12] Palmieri M, Kiss JZ.Disruption of the F-actin cytoskeleton lim its statolith movement in Arabidopsis hypocotyls[J]. Journal of Experimental Botany, 2005, 56(419): 2539-2550.
[13] Kandasamy M K, McKinney E C, Meagher R B. Functional nonequivalency of actin isovariants in Arabidopsis. Mol[J]. Biol Cell, 2002, 13: 251-261.
[14] 梁娜, 伍国强冯瑞军, 等. 甜菜Actin基因片段的克隆及序列分析[J]. 中国糖料, 2014, 4: 13-15.
[15] 伍国强, 席杰军, 包爱科, 等. 多浆旱生植物霸王Actin基因片段的克隆及序列分析[J]. 生物技术通报, 2008(2): 101-104.
[16] 马清, 周向睿, 伍国强, 等. 盐生植物碱蓬Actin基因片段的克隆及序列分析[J]. 生物技术, 2009, 19(1): 1-3.
[17] 吴永娜, 胡静, 王引权, 等. 当归肌动蛋白基因片段的克隆及序列分析[J]. 中草药, 2012, 42(12): 2485-2489.
[18] 田秀红, 高建明, 肖强, 等. 水花生Actin基因片段的克隆及序列分析[J]. 生物技术, 2006, 16(3): 13-14.
[19] 田云芳, 刘艺平, 李涵, 等. 蕙兰Actin基因的克隆及序列分析[J]. 河南农业科学, 2013, 42(6): 107-111.
[20] 袁秀云, 田云芳, 蒋素华, 等. 萼脊兰Actin 基因片段的克隆及序列分析[J]. 中国农学通报, 2012, 28(13): 243-248.
[21] 郭景康, 陈青云, 戢茜, 等. 拟南芥、水稻和杨树ACTIN 家族全基因组分析[J]. 上海大学学报, 2009, 15(4): 426-431.
[22] 陈鹏飞, 刘雪梅, 宋福南, 等. 白桦肌动蛋白(Actin)基因全长cDNA克隆与序列分析[J]. 植物研究, 2009, 29(3): 339-345.
[23] 陈永霞. 扁穗牛鞭草actin基因克隆及荧光定量PCR的建立[J]. 广东农业科学, 2012, 16: 158-161.
[24] 刘建新, 葛亚英, 田丹青, 等. 果子蔓凤梨Actin 基因的克隆与序列分析[J]. 分子植物育种, 2011, 9(1): 41-45.
[25] 张灵,段慧荣,李静,等. 旱生植物梭梭actin 基因片段的克隆及表达模式分析[J]. 基因组学与应用生物学, 2014, 33(3):598-603.
[26] 孔卫青, 杨金宏. 桑树肌动蛋白actin 基因全长序列的克隆与分析[J]. 广西植物, 2012, 32(3): 362-366.
[27] 马雄风, 喻春明, 唐守伟, 等. 苎麻Actin1 基因克隆及其在韧皮部纤维不同发育阶段的表达[J]. 作物学报, 2010, 36(1): 101-108.
[28] 吴菁华, 吴少华, 杨超. 建兰actin基因cDNA全长的克隆及其表达分析[J]. 生物技术通报, 2012, 12: 88-92.
[29] 江昌俊. 茶树育种学[M]. 北京: 中国农业出版社, 2005: 180-181.
[30] 李远华, 江昌俊, 杨顺利, 等. 茶树β-葡萄糖苷酶cDNA克隆和原核表达[J]. 农业生物技术学报, 2004, 12(6): 625-629.
[31] 李远华, 陆建良, 范方媛, 等. 茶树根系HGMR基因克隆及表达分析[J]. 茶叶科学, 2014, 34(6): 276-282.
[32] 李远华, 陆建良, 范方媛, 等. 茶树GAGP基因克隆及表达分析[J]. 茶叶科学, 2015, 35(1): 64-72.
[33] 李远华, 郑芳, 倪德江, 等. 茶树接种VA菌根的生理特性研究[J].茶叶科学,2011,31(6):504-512.
[34] Liang WH, Tang CR, Wu NH.Cloning and characterization of a new actin gene from Oryza sativa[J]. Prog Nat Sci, 2004, 14: 867-874.
[35] Kim H, Park M.Actin filaments play a critical role in vacular trafficking at the golgi complex in plant cells[J]. The Plant Cell, 2005, 17: 888-902.
[36] Meagher RB, Mckinney EC.Isovariant dynamics expand and buffer the responses of complex systems: the diverse plant actin gene family[J]. The Plant Cell, 1999, 11: 995-1005.
[37] Daz-camino C, Conde R. Actin expression is induced and three isoforms are differently expressed during germination in zeamays[J]. Journal of Experimental Botany, 2005, 56:557-565.
[38] Jiang YQ, Zhao WL.Expression and phylogenetic analysis of pea actin isoforms[J]. Acta Botanica Sinica, 2002, 44: 1456-1461.