利用TAILPCR技术从中国砂梨品种‘金花’、‘懋功’及白梨品种‘鸭梨’基因组DNA中分 别扩增出长度为854、1 448和1 137 bp的S13-、S12-、S21-RNase基因5′端上游序列，并提交 GenBank，序列号为HM047239、HM047240、HM047241。经PLACE和PlantCARE软件顺式调控元件预测发 现，这3个启动子均具有典型核心启动子TATA 盒和 CAAT 盒，且在上游均存在光应答调控元件(box 4，Gbox)、脱落酸应答元件ABRE及水杨酸应答元件TCAelement等，由此可知其可能受光照、脱 落酸、水杨酸等多种条件的共同调节。此外，与已知日本梨S2-、S3-、S4-、S5-RNase基因启动子序 列比对发现，SRNase启动子TATA盒上游存在一约200 bp的同源区域。以MEG 4.0构建系统发育树表 明，梨属和苹果属SRNase等位基因多态性可能在亚科的分化之前就已形成。
Abstract
Using TAILPCR, 5′flanking regions of the S13, S12, S21RNase genes with a length of 854 bp, 1 448 bp and 1 137 bp were successfully isolated from ‘Jinhua ’and ‘Maogong’(Pyrus pyrifolia) and ‘Yali’(Pyrus bretschneideri) genomic DNA, the GenBank numbers are HM047239, HM047240 and HM047241. The core promoter regions and some upstream regulatory elements in the three fragments were analyzed using PLACE and PlantCARE software. It is found that all of these genes have the putative TATA box and CAAT box, and the promoters were affected by a variety of conditions as light, ABA, salicylic acid. Alignment analysis for promoter sequences of S13, S12, S21 with 5′flanking sequences of S2, S3, S4, S5 revealed a homologous region of about 200 bp in the upstream sequences of the TATA box in SRNase promoters. Phylogenetic tree constructed by MEG 4.0 suggests that the divergence of SRNase gene was formed before the differentiation of subfamilies.
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参考文献
[1]王绪,邓俭英,方锋学,等. ISSR分子标记技术及其在园艺作物中的应用[J]. 广西农业科 学,2007,38(4):371-374.
[2]Sato Y, Kurihara A, Ogata T, et al. Mode of selfcompatibility inheritance in Japanese pear[J]. J Jpn Soc Hort Sci, 1988, Supp(2): 76-77.
[3]Liu Y G, Whittier R F. Thermal asymmetric interlaced PCR: automatable amplification and sequencing of insert end fragments from P1 and YAC clones for chromosome walking [J]. Genomics, 1995, 25: 674-681.
[4]Liu Y G, Mitsukawa N, Oosumi T, et al. Efficient isolation and mapping of Arabidopsis thaliana TDNA insert junctions by thermal asymmetric interlaced PCR[J]. Plant, 1995, 8: 457-463.
[5]Tsugeki R, Kochieva E Z, Fedoroff N V. A transposon insertion in the Arabidopsis SSR16 gene causes an embryodefective lethal mutation[J]. The Plant Journal, 1996, 10 (3): 479-489.
[6]乌云塔娜. 中国白梨自交不亲和基因的分离鉴定[D]. 长沙:中南林业科技大学,2003.
[7]Thompson J D, Gibson T J, Plewniak F, et al. The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research, 1997, 25: 4876-4882.
[8]Tamura K, Dudley J, Nei M, et al. MEGA4: molecular evolutionary genetics analysis (MEGA)software version 4.0[J]. Molecular Biology and Evolution, 2007: 1596-1599.
[9]Higo K, Ugawa Y, Iwamoto M, et al. Plant cisacting regulatory DNA elements(PLACE) database[J]. Nucl Acids Res, 1999, 27: 297-300.
[10]Lescot M, Déhais P, Thijs G, et al. PlantCARE,a database of plant cisacting regulatory elements and a portal to tools for in silico analysis of promoter sequences [J]. Nucl Acids Res, 2002, 30: 325-327.
[11]Ushijima K, Sassa H, Hirano H. Characterization of the flanking regions of the S RNase genes of Japanese pear(Pyrus serotina) and apple(Malus×domestica)[J]. Gene, 1998, 211: 159-167.
[12]Norioka N, Katayama H, Matsuki T, et al. Sequence comparison of the 5′ flanking regions of Japanese pear(Pyrus pyrifolia Naki)SRNases associated with gametophytic selfincompatibility[J]. Sex Plant, 2001, 13: 289-291.
[13]Dissanayake DMRKK, Norioka S, Norioka N, et al. Cisregulatory elements for pistil specific expression in SRNase promoter region of Japanese pear(Pyrus pyrifolia Nakai)[J]. Acta Hort, 2002, 587: 459-465.
[14]Kaufmann H, Salamini F, Thompson R D. Sequence variability and gene structure at the selfincompatibility locus of Solanum tuberosum[J]. Mol Gen Genet, 1991, 226:4 57 -466.
[15]Coleman C E, Kao Th. The 5′ flanking regions of two Petunia inflata S alleles are heterogeneous and repetitive sequences[J]. Plant Mol Biol, 1992, 18: 499-511.
[16]Chung I K, Lee S Y, Ito T, et al. The 5′ flanking sequences of two S alleles in Lycopersicon peruvianum are highly heterologous but contain short blocks of homologous sequences[J]. Plant Cell Physiol, 1995, 36: 1621-1627.
[17]Matton D P, Mau S L, Okamoto S, et al. The Slocus of Nicotiana alata: genomic organization and sequence analysis of two SRNase alleles[J]. Plant Mol Biol, 1995, 28: 847-858.
[18]Morgan D R, Soltis D E, Robertson K R. Systematic and evolutionary implications of rbcL sequence variation in Rosaceae[J]. Am J Bot, 1994, 81: 890-903.
[19]Prell H. Das problem der unfruchtbarkeit[J]. Nature Wochschr, N F, 1921, 20: 440- 446. (责任编辑郑琰燚)第34卷第4期2010年7月南京林业大学学报(自然科学版)Journal of Nanjing Forestry University (Natural Science Edition)Vol.34, No.4Jul., 2010
基金
收稿日期：2009-10-21修回日期：2010-04-26基金项目：湖南省科技计划项 目(K0904005-21)；湖南省教育厅科学研究项目(5014)；长沙市科技局科研项目(101-4586)作者简介 ：乌云塔娜(1975—)，副教授。Email: tanatana@sina.com。引文格式：乌云塔娜,刘学英,李振 国. 中国梨SRNase基因启动子的TAILPCR克隆及功能预测[J]. 南京林业大学学报:自然科学 版,2010,34(4):21-25.
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