[1]周晓婴,付三雄,陈松,等.甘蓝型油菜CRABS CLAW基因克隆及其RNA干扰载体的构建[J].江苏农业学报,2015,(04):737-742.[doi:10.3969/j.issn.1000-4440.2015.04.005]
　ZHOU Xiao-ying,FU San-xiong,CHEN Song,et al.Cloning of CRABS CLAW gene from Brassica napus and construction of its RNA interference vector[J].,2015,(04):737-742.[doi:10.3969/j.issn.1000-4440.2015.04.005]
点击复制

甘蓝型油菜CRABS CLAW基因克隆及其RNA干扰载体的构建()
分享到：

江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年04期
页码:
737-742
栏目:
遗传育种·生理生化
出版日期:
2015-08-31

文章信息/Info

Title:
Cloning of CRABS CLAW gene from Brassica napus and construction of its RNA interference vector
作者:
周晓婴123; 付三雄123; 陈松123; 张超3; 戚存扣123
(1.国家油菜改良中心南京分中心，江苏南京210014;2.农业部长江下游棉花与油菜重点实验室，江苏南京210014;3.江苏省农业科学院经济作物研究所，江苏南京210014）
Author(s):
ZHOU Xiao-ying123; FU San-xiong123; CHEN Song123; ZHANG Chao3; QI Cun-kou123
(1.Nanjing Sub-center of National Center of Rapeseed Crop Improvement /Key Laboratory of Cotton and Rapeseed, Nanjing 210014, China;2.Ministry of Agriculture /Institute of Industrial Crops, Nanjing 210014, China;3. Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China)
关键词:
油菜; CRABS CLAW基因; RNA干扰载体 
Keywords:
Brassica napus L.; CRABS CLAW gene; RNA interference vector
分类号:
S565.4
DOI:
10.3969/j.issn.1000-4440.2015.04.005
文献标志码:
A
摘要:
CRABS CLAW（CRC）转录因子是YABBY基因家族成员，在植物花器官发育过程中起重要作用。为进一步研究CRC转录因子在调控油菜花器官发育过程中的作用,本研究以甘蓝型油菜品种宁油10号花蕾RNA为材料,通过反转录PCR克隆到1个580 bp长度的CRC基因，并利用pHurricane中间载体构建CRC基因的反向重复序列表达框，将CRC基因片段以正向的方式连接在1个可剪切的内含子的5′末端，以反向的方式连接到该内含子的3′末端；然后将CaMV35S启动子序列和CRC基因的反向重复表达框再克隆到植物双元载体pCAMBIAI1390的pUC18多克隆位点，构建了干扰表达载体pA6-CRCi，经过酶切鉴定和测序分析，所构建的载体正确。
Abstract:
CRABS CLAW (CRC) is a member of the YABBYA transcription factor gene family and plays an important role in the development of plant flower organ. In order to study the CRC transcription factor’s function in regulating the rape flower organ development, a 580-bp segment was cloned by RT-PCR from the total RNA of rapeseed flower organ. An inverted repeated expression cassette was firstly constructed by cloning the 580-bp segment of CRC gene into an intermediate pHurrican vector. The fragment was then ligated to a spliceable fad2 intron sequence. Finally, both the inverted repeat cassette and CaMV35S promoter were inserted into the multiple clone site of binary vector pCAMBIA1390. The ihpRNA expression vector was named pA6-CRCi, and the construction of which was further confirmed by the digestion of restriction enzymes and sequencing.

参考文献/References:


[1]胡青,高述民,李凤兰.植物雌性不育的研究进展[J].北京林业大学学报,2004,26(1):87-91.
[2]陈新军,戚存扣,张洁夫，等.油菜(Brassica napus L.)雌性不育突变体FS-M1的生物学特性研究[J].中国油料作物学报,2003,25（3）:12-15．
[3]陈新军,戚存扣,张洁夫，等.甘蓝型油菜雌性不育突变体FS-M1花器官形态结构研究[J].作物学报,2005,31（9）:1239-1240.
[4]李春宏,付三雄,陈新军，等.甘蓝型油菜雌性不育突变体M7乳突细胞的细胞学观察[J].植物学报, 2012,47（1）:36-43.
[5]凌定厚,马镇荣,陈梅芳，等.起源于体细胞培养的籼稻雌性不育突变[J].遗传学报,1991,18(5):446-451.
[6]丁坤善,郑彩霞,包仁艳，等.油松雌性不育系球果蛋白质双向电泳技术的建立[J].植物学通报,2005,22(2):190-197.
[7]窦秉德,张新玲,马林，等.普通小麦中一种雌性不育现象的观察[J].作物学报,2001,27(6):1013-1016.
[8]赵团结,盖钧镒.大豆叶与花形态异常、雌性不育突变体NJS-10H的发现[J].大豆科学,2005，24(1):1-4.
[9]张静文,金 樑,冯光辉，等.苜蓿雌性不育发生机制及其分子调控研究进展[J].云南农业大学学报,2011,26(4):560-566.
[10]STORT．Sterility barriers in some artificial F1 orchid hybrids：female sterility[J]. Rev Bras Genet，1987，10(1)：109-118.
[11]刘永胜,周开达,阴国大，等.水稻籼梗杂种雌性不育的细胞学初步观察[J]．实验生物学报,1993,26(1):95-99.
[12]LILLECRAPP A M,WALLWORK M A,SEDGLEY M. Female and male sterility cause low fruit set in a clone of the“Trevatt”variety of apricot(Prunus armeniaca) [J].Scientia Horticulturae Amsterdam,1999,82(3-4):255-263.
[13]RIM Y W,BEUSELINCK P R,MCGRAW R L,et a1.Megagametophyte development in Lotus corniculatus  L. coninrbricensis and their protoplast fusion hybrid[J].Lotus Newsletter， 1998,19:45-47.
[14]OUYANG X Z.Ultrastructure and AcPase ultracytochemical localization of the abortive functional megaspores in intersubspecific F1 hybrid rice (Oryza sativa)[J]．Chinese Rice Research Newsletter,1996,10（4）:2-3.
[15]张宪省,马小杰.胚珠发育的分子基础[J].植物学通报,1998,15 (3):1-5.
[16]PENMESTA R V,COOK D R．Production and characterization of diverse developmental mutants of Medicago truncatula[J].Plant Physiology,2000,123 (4):1387-1397.
[17]LANG J D,RAY S,RAY A.Sinl，a mutation afecting female fertility in Arabidopsis,interacts with mod1, its recessive modifier[J]. Genetics,1994,137(4):1101-1110.
[18]TIAN L,CHEN Z J.Blocking histone deacetylation in Arabidopsis induces pleiotuopic effects on plant gene regulation and development[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98(1):200-205.
[19]CAPOMACCIO S,BARONE P，REALE L,et a1.Isolation of genes from female sterile flowers in M edicago satiu [J].Sexual Plant Reproduction,2009,22:97-107.
[20]CARAPETIAN J,KNOWLES P F.Genetic linkage between the trigenic male-female sterility and oil quality alleles in saflower[J].Crop Science,1993,33(2):239-242.
[21]GALLAGHER T L,GASSER C S.Independence and interaction of regions of the INNER NO OUTER protein in growth control during ovule development[J]. Plant Physiology，2008,147: 306-315.
[22]TANAKAA W，TORIBAA T，OHMORIA Y,et a1. The YABBY gene TONGARI-BOUSHI1 is involved in lateral organ development and maintenance of meristem organization in the rice spikelet[J].The Plant Cell,2012,24: 80-95.
[23]ALVAREZ J，SMYTH D R. CRABS CLAW and SPATULA，two Arabidopsis genes that control carpel development in parallel with AGAMOUS[J]. Development,1999,26:2377-2388.
[24]LEE J Y，BAUM S F，ALVAREZ J， et al. Activation of CRABS CLAW in the nectaries and carpels of Arabidopsis [J]. The Plant Cell,2005,17:25-36.
[25]FOURQUIN C，VINAUGER-DOUARD M，FOGLIANI B,et a1. Evidence that CRABS CLAW and TOUSLED have conserved their roles in carpel development since the ancestor of the extant angiosperms[J]. PNAS USA,2005,102:4649-4654.
[26]GROSZMANN M,BYLSTRA Y,LAMPUGNANI E R,et a1. Regulation of tissue-specific expression of SPATULA，a bHLH gene involved in carpel development，seedling germination，and lateral organ growth in Arabidopsi [J]. Journal of Experimental Botany,2010,61:1495-1508.
[27]GROSZMANN M,PAICU T,SMYTH D R. Functional domains of SPATULA,a bHLH transcription factor involved in carpel and fruit development in Arabidopsis[J].Plant J，2008,55:40-52.
[28]GOIZ, JOHN F, HUDSON, et al. Signaling in plant lateral organ development[J]. The Plant Cell，2002(Suppl):277-288.
[29]李春宏,付三雄,戚存扣.应用基因芯片分析甘蓝型油菜柱头特异表达基因[J].植物学报,2014,49(3):246-253.
[30]陈松,张洁夫,陈锋,等.甘蓝型油菜种子特异性表达fad2基因的ihpRNA载体构建[J].中国油料作物学报,2006,28(3):25l-256.
[31]付三雄,张超,陈松,等.甘蓝型油菜丙酮酸激酶基因RNA干扰载体的构建[J].江苏农业学报,2013,29(1):20-27.〖ZK）〗〖FL）〗

相似文献/References:

[1]付三雄,周晓婴,张 维,等.种植密度和施氮量对油菜产量、品质及机收性状的影响[J].江苏农业学报,2016,(03):548.[doi:10.3969/j.issn.1000-4440.2016.03.010]
　FU San-xiong,ZHOU Xiao-ying,ZHANG Wei,et al.Influences of planting density and nitrogen application rate on yield and quality and mechine harvest traits of rapeseed(Brassica napus L.)[J].,2016,(04):548.[doi:10.3969/j.issn.1000-4440.2016.03.010]
[2]陈魏涛,曹宏鑫,张保军,等.氮素营养诊断技术及其在油菜上的应用研究进展[J].江苏农业学报,2016,(04):953.[doi:10.3969/j.issn.100-4440.2016.04.038]
　CHEN Wei-tao,CAO Hong-xin,ZHANG Bao-jun,et al.Research progresses in nitrogen diagnosis technology and its application in rapeseed[J].,2016,(04):953.[doi:10.3969/j.issn.100-4440.2016.04.038]
[3]熊洁,邹晓芬,邹小云,等.干旱胁迫对不同基因型油菜农艺性状和产量的影响[J].江苏农业学报,2015,(03):494.[doi:10.3969/j.issn.1000-4440.2015.03.005]
　XIONG Jie,ZOU Xiao-fen,ZOU Xiao-yun,et al.Effects of drought stress on agronomic traits and yield of different rapeseed genotypes[J].,2015,(04):494.[doi:10.3969/j.issn.1000-4440.2015.03.005]
[4]熊洁,李书宇,邹晓芬,等.轻简化育苗移栽方式对油菜生长发育和产量的影响[J].江苏农业学报,2015,(02):317.[doi:10.3969/j.issn.1000-4440.2015.02.015]
　XIONG Jie,LI Shu-yu,ZOU Xiao-fen,et al.Effects of simplified seedling and transplanting patterns on growth and development and yield of rapeseed[J].,2015,(04):317.[doi:10.3969/j.issn.1000-4440.2015.02.015]
[5]李茹,陈国奇,张玉华,等.油菜和小麦秸秆水浸提液对千金子种子萌发和幼苗生长的影响及其应用[J].江苏农业学报,2018,(02):293.[doi:doi:10.3969/j.issn.1000-4440.2018.02.010]
　LI Ru,CHEN Guo-qi,ZHANG Yu-hua,et al.Influences of oilseed rape and wheat aquatic straw extract on Leptochloa chinensis seed germination and seedling growth, and the application potential[J].,2018,(04):293.[doi:doi:10.3969/j.issn.1000-4440.2018.02.010]
[6]葛道阔,曹宏鑫,杨余旺,等.干旱胁迫下油菜栽培模拟优化决策系统（Rape-CSODS）的订正及其检验[J].江苏农业学报,2019,(01):56.[doi:doi:10.3969/j.issn.1000-4440.2019.01.008]
　GE Dao-kuo,CAO Hong-xin,YANG Yu-wang,et al.Modification and verification of Rape-CSODS under drought stress[J].,2019,(04):56.[doi:doi:10.3969/j.issn.1000-4440.2019.01.008]
[7]岳绪国,葛永申,景德道,等.移栽密度和方式对不同类型油菜品种产量及构成的调控效应[J].江苏农业学报,2019,(01):63.[doi:doi:10.3969/j.issn.1000-4440.2019.01.009]
　YUE Xu-guo,GE Yong-shen,JING De-dao,et al.Effects of density and transplanting method on yield and yield components of different types of rapeseed[J].,2019,(04):63.[doi:doi:10.3969/j.issn.1000-4440.2019.01.009]
[8]程丽,胡茂龙,浦惠明,等.M342抗除草剂基因CAPS标记的开发与应用[J].江苏农业学报,2019,(02):241.[doi:doi:10.3969/j.issn.1000-4440.2019.02.001]
　CHENG Li,HU Mao-long,PU Hui-ming,et al.Development and application of the CAPS marker for herbicide-resistant gene in M342[J].,2019,(04):241.[doi:doi:10.3969/j.issn.1000-4440.2019.02.001]
[9]万倩,曹宏鑫,葛道阔,等.不同施肥量下花期渍水胁迫对油菜籽粒充实过程的定量影响[J].江苏农业学报,2020,(05):1144.[doi:doi:10.3969/j.issn.1000-4440.2020.05.011]
　WAN Qian,CAO Hong-xin,GE Dao-kuo,et al.Quantitative effects of waterlogging stress at florescence stage on the grain filling process of rape under different fertilization amounts[J].,2020,(04):1144.[doi:doi:10.3969/j.issn.1000-4440.2020.05.011]
[10]张伟欣,吴茜,孙传亮,等.油菜株型模拟研究进展[J].江苏农业学报,2022,38(02):549.[doi:doi:10.3969/j.issn.1000-4440.2022.02.031]
　ZHANG Wei-xin,WU Qian,SUN Chuan-liang,et al.Research progress on plant type simulation of rapeseed[J].,2022,38(04):549.[doi:doi:10.3969/j.issn.1000-4440.2022.02.031]

备注/Memo

备注/Memo:
收稿日期：2015-02-02 基金项目：江苏省农业科技自主创新基金项目[CX(11)4011];江苏省自然科学基金项目(BK2011668);国家“948”项目(2011-G23) 作者简介：周晓婴（1983-），女，福建宁德人，硕士，助理研究员，主要从事油菜遗传育种研究。（Tel）025-84390364；（E-mail）beibeinv@hotmail.com 通讯作者：陈松, （Tel）025-84390370；（E-mail）chensong1963@126.com
更新日期/Last Update: 2015-08-31