拟南芥叶边缘锯齿状突变体的分离与鉴定

doi:10.3724/SP.J.1143.2012.11103

摘要/Abstract

摘要：

叶的极性建立直接决定叶的平展性发育，极性改变导致叶形态异常，影响植物体的各种正常生理活动。利用反向遗传学方法，从拟南芥基因激活标签突变体库中分离到一个叶片边缘锯齿状表型的突变体(命名为pCB1294)，该突变体同时表现出叶表皮腺毛形态发育异常。通过TailPCR方法成功定位突变基因为At5g41663，该基因编码miR319b基因。Real time PCR显示，pCB1294突变体植株中miR319b基因的表达量是野生型（col）植株的11倍多。所得结果为进一步研究miRNA调控叶极性的分子机制和进一步分析miR319b与叶形态发生的关系奠定了基础。

关键词: 拟南芥, 突变体, 叶锯齿, Tail-PCR, miRNA

Abstract:

Leaf polarity determines leaf flatness development directly, and abnormal polarity usually results in many abnormal leaves, which subsequently affects many physiological functions of plants. So the normal leaf development is important to plants. Here, an abnormal serration leaf margin mutant with abnormal leaf trichome development, named pCB1294, was isolated from an activation tagging Arabidopsis mutant pool through reverse genetics. By TailPCR, the mutant gene loci At5g41663 encoding miR319b was successfully identified. Real time PCR shows the relative expression level of miR319b gene in the pCB1294 mutant is eleven times of higher than that of the wild (col). Our study lay the foundation for further studying the genetic mechanism of leaf polarity and investigating the interaction between miR319b and leaf morphology.

Key words: Arabidopsis thaliana, Mutant, Leaf margin serration, Tail-PCR, miRNA

中图分类号:

Q 75

李婉莎, 王春涛, 胡向阳. 拟南芥叶边缘锯齿状突变体的分离与鉴定[J]. Plant Diversity, 2012, 34(01): 28-32.

LI Wan-Sha, WANG Chun-Tao, HU Xiang-Yang. Isolation and Identification of an Arabidopsis thaliana Mutant with Serration Leaf Margin[J]. Plant Diversity, 2012, 34(01): 28-32.

参考文献

〔参考文献〕
Baker CC, Sieber P, Wellmer F et al., 2005. The early extra petals1 mutant uncovers a role for microRNA miR164c in regulating petal number in Arabidopsis［J］. Current Biology, 15: 303—315
Guan YL (关艳龙), Li WS (李婉莎), Yin KD (殷奎德) et al., 2010. Constructing and phenotypic analyzing an activation tagging Arabidopsis mutant pool［J］. Acta Botanica Yunnanica (云南植物研究), 32(1): 53—59
Guo HS, Xie Q, Fei JF et al., 2005. MicroRNA directs mRNA cleavage of the transcription factor NAC1 to downregulate auxin signals for Arabidopsis lateral root development［J］. The Plant Cell, 17: 1376—1386
Huang H, 2003. Recent progresses from studies of leaf development［J］. Chinese Bulletin of Botany (植物学通报), 20 (4): 416—422
Hudson A, 1999. Axioms and axes in leaf formation［J］. Current Opinion in Plant Biology, 2: 56—60
Kidner CA, Martienssen RA, 2004. Spatially restricted microRNA directs leaf polarity through ARGONAUTE1 ［J］. Nature, 428: 81—84
Kim J, Jung JH, Reyes JL et al., 2005. microRNAdirected cleavage of ATHB15 mRNA regulates vascular development in Arabidopsis inflorescence stems［J］. The Plant Journal,42: 84—94
Liu YG, Mitsukawa N, Oosumi T et al., 1995. Efficient isolation and mapping of Arabidopsis thaliana TDNA insert junctions by thermal asymmetric interlaced PCR［J］. The Plant Journal, 8: 457—463
Liu DM (刘冬梅), Yang FX (杨凤玺), Yu DQ (余迪求), 2009. Overexpression of miR396 miRNAS caused flower stigma curved in Arabidopsis thaliana［J］. Acta Botanica Yunnanica (云南植物研究), 31 (4): 353—356
Nath U, Crawford BC, Carpenter R et al., 2003. Genetic control of surface curvature［J］. Science, 299: 1404—1407
Nikovics K, Blein T, Peaucelle A et al., 2006. The balance between the MIR164A and CUC2 genes controls leaf margin serration in Arabidopsis［J］. The Plant Cell, 18 (11): 2929—2945
Ori N, Cohen AR, Etzioni A et al., 2007. Regulation of LANCEOLATE by miR319 is required for compoundleaf development in tomato［J］. Nature Genetics, 39 (6): 787—791
Palatnik JF, Allen E, Wu X et al., 2003. Control of leaf morphogenesis by microRNAs ［J］. Nature, 425: 257—263
Palatnik JF, Wollmann H, Schommer C et al., 2007. Sequence and expression differences underlie functional specialization of Arabidopsis microRNAs miR159 and miR319［J］. Developmental Cell,13 (1): 115—125
Scanlon MJ, 2000. Developmental complexities of simple leaves［J］. Current Opinion in Plant Biology, 3: 31—36
Schwab R, Palatnik JF, Riester M et al., 2005. Specific effects of microRNAs on the plant transcriptome［J］. Developmental Cell, 8: 517—527
Smith LG, Hake S, 1992. The initiation and determination of leaves［J］. The Plant Cell, 4 (9): 1017—1027
Tsukaya H, 2005. Leaf shape: Genetic controls and environmental factors［J］. International Journal of Developmental Biology, 49: 547—555
VarkonyiGasic E, Wu R, Wood M et al., 2007. Protocol: a highly sensitive RTPCR method for detection and quantification of microRNAs［J］. Plant Methods, 3 (1): 12
Williams L, Grigg SP, Xie M et al., 2005. Regulation of Arabidopsis shoot apical meristem and lateral organ formation by microRNA miR166g and its AtHDZIP target genes［J］. Development, 132: 3657—3668
Yu L, Yu XH, Shen RJ et al., 2005. HYL1 gene maintains venation and polarity of leaves［J］. Planta, 221: 223—422

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