抗稻瘟病基因Pib启动子内YTCANTYY暗诱导分子元件功能的转基因验证
|
摘要
启动子是位于结构基因5′端上游区的调控序列,在转录水平控制着植物基因表达的时空性。研究植物抗病基因启动子,分析启动区内分子元件的功能对了解植物抗病基因的表达特异性有很重要的意义。不少与启动和诱导活性有关的分子元件在启动区内是多拷贝的。用逐段敲除这些功能性分子元件的启动子缺失体来进行转基因分析是了解这些分子元件的生物学功能的重要方法。
Pib基因属于NBS-LRR抗病基因家族,是目前为数不多已克隆的抗稻瘟病主效基因之一。研究报道表明,pib基因表达不受病原菌诱导,而黑暗、温度等环境和一些化学物质却能有效地诱导pib基因的表达。研究结果还显示,将不同长短的pib结构基因序列片段置于35S驱动下,它们各自的转基因植株表现出对稻瘟病生理小种的不同抗性,而pib结构基因上游的启动区与gus基因融合构建的转基因试验也证实了pib启动子具有很强的黑暗诱导活性。由于水稻的根是处于24h黑暗环境,而地上部器官处于光/暗循环交替之下,这就使得pib基因的抗病性表达具有了特殊的器官特异性和昼夜节律性。
为了探讨pib启动子这种环境条件诱导性的分子结构和分子机制,笔者用PLACE数据库对pib启动区核酸序列进行了分析,该启动区内有各种定义为启动相关分子元件近百个,除了TATA、CAAT等启动子共有元件外还有数十个不同类型的诱导性元件。其中被定义为光响应相关的顺式作用元件YTCANTYY以6个拷贝形式散布在启动子序列内。为了验证该启动子的暗诱导特性是否与此分子元件有关,这个分子元件又是怎样影响该启动子的暗诱导性的,笔者构建了不同长度、即含有不同数目的YTCANTYY元件的水稻pib基因启动子的5′端缺失体驱动gus基因的双元载体,用对其水稻转基因植株的gus基因表达分析的结果来研究该pib启动子中暗诱导分子元件的生物学功能及其对暗诱导的响应。
1.用PCR扩增并测序获得4个不同长度的pib启动子片段,各片段之间的区别仅在于5'端的缺失各不相同。4个片段的3'端均终止于碱基2(ATG信号的G),分别为:5'缺失体启动子1 PCR产物长222bp,为-219～2bp,无YTCANTYY元件,5'缺失体启动子2PCR产物长569bp,为-566～2bp,含1个YTCANTYY元件(-220～-227bp处),5'缺失体启动子3 PCR产物长1911bp,为-1908～2bp,含3个YTCANTYY元件(-220～-227,-567～-574,-728～-735bp处),全长启动子PCR产物长3575bp,为-3572～2bp,含6个YTCANTYY元件(-220～-227,-567～-574,-728～-735,-1909～-1916,-3042～-3049,-3382～-3389 bp处)。801(-219～2bp)、802(-566～2bp)、803(-1908～2bp)和804(-3572～2bp)分别连接进pCAMBIA1301中gus因前,取代原来的CaMV35S启动子,构建成双元载体pNAR801、pNAR802、pNAR803和pNAR804。重组双元质粒经过酶切验证,用农杆菌冻融转化法转化入农杆菌菌株EHA101中。
2.利用农杆菌介导的方法,将上述pNAR801、pNAR802、pNAR803、pNAR804重组质粒转化粳稻品种R109,同时以CaMV35S启动子驱动gus基因的pCAMBIA1301作对照。共获得136株再生植株,经PCR筛选确定得到96个阳性的转基因水稻植株,阳性率70.6%。通过Southern blot检测也表明目的基因已经整合到水稻基因组中。
3.对上述不同长度5′端缺失的pib启动子驱动gus基因的水稻转基因植株,进行Gus组织化学检测和荧光定量分析,系统研究了pib启动子中分子元件YTCANTYY拷贝数目与该启动子启动活性和暗诱导性的关系。Gus组织化学分析结果表明,含6个、3个、1个拷贝的pib启动子的转基因水稻愈伤在暗处理后均能在X-gluc溶液中显示不同深浅的Gus兰色,而6个拷贝完全缺失的启动子残端的水稻愈伤在X-gluc溶液中不显现Gus兰色。荧光定量分析结果表明,pib启动子序列具有很强的器官特异性,即便是6个暗诱导分子元件都被敲除的3'残端,其转基因植株仍保持根部启动活性高于地上部份的趋势。但其启动活性和暗诱导性随启动子缺失片段的缩短即暗诱导分子元件拷贝数增加而提高。这些结果表明,YTCANTYY在pib启动子序列中是一个暗诱导的功能性分子元件,他能赋予启动子的暗诱导性,起码在6个拷贝以内,pib启动子的暗诱导活性是与其拷贝数目呈正相关,各个YTCANTYY分子元件的启动活性可能是累加的。
4.利用pib基因启动区与gus构建的报告基因载体pNAR604得到的T_3转基因水稻,继续对pib基因启动子诱导性进行研究,再次证实pib启动子活性受黑暗和温度影响,无论诱导与否,在根中的Gus活性都远高于叶片具有强烈的器官特异性,pib启动子活性无论是在根还是叶都具有明显的昼夜节律性,白昼低夜晚高,以15:00活性水平为最低,03:00活性水平为最高。这些结果表明,pib基因表达的起始或抗稻瘟病产物的合成地在根系,而不是植株叶、茎等稻瘟病发病部位。也就是说pib基因抗稻瘟病功能性产物的合成起始地与抗病的靶点分离。据此,我们推断,pib基因编码的抗稻瘟病产物是在根部起始合成,抗病基因产物在夜间由根系输送到植株地上的靶组织,实现其抗病的功能。
As a important regulatory element of 5'upstream regions of structure gene, promoter controls gene expression characteristic of space and time in transcriptional level. Study on plant promoters of disease resistance and analysis of regulatory element's function in promoter region are the very significance to expression characteristic of disease resistance gene. The transformation analysis by 5'deletion promoter fragments is the important method to understand the biological function of these molecular elements.
Pib gene is the first major blast resistant gene cloned from rice by mapping clone technique and which is belongs to NBS-LRR resistant disease gene family. Previous studies show that the expression of pib gene was induced by darkness and chemistry factors but not pathogen. Our experiment results also indicated that different length fragments of pib structure gene sequence under the drive of 35S promoter, their To transgenic rice plants showed different resistances to rice blast pathogen races. Our results indicated that the pib promoter has the activity of darkness inducing in the rice transgenic experiments with the constructs of pib promoter-gus fusion. In natural environment, only root of rice plant could enjoy full darkness but the above ground organs are in the condition of light or dark cycle alternative, which offer the expression of pib gene the special organ specificity and diurnal rhythm.
For studying the molecular structure and mechanism of environmental condition inducing, we analysis this promoter region by using PLACE database. The results show that there are about hundred of molecular regulatory element, for activation in this region, except of TATA、CAAT commonly elements, there are six copies of YTCANTYY element defined for light-response of cis-regluatory element. To verifying whether the dark-inducing property of the pib promoter is relate to the YTCANTYY element and how this molecular element affect the dark-inducing property? By using transgenic rice plants of 5'deletion pib promoter fragment-gus constructions, the relationships between the copy number of molecular element YTCANTYY in pib promoter and activity of pib promoter and its darkness inducing attribute were systematically analyzed.
1. Four different regions of pib gene promoter were obtained by PCR, the difference among each fragments only lengths in 5'deletion, the 3' downstream of four fragments all terminate to base 2(base G of signal ATG). The PCR products of the first 5'deletion promoter is 222bp which is without YTCANTYY element; the PCR product of the second 5'deletion promoter is 569bp which contains one YTCANTYY element (-220～-227bp); the PCR products of the third 5'deletion promoter is 1911bp which contain three YTCANTYY element (-220～-227,-567～-574,-728～-735bp); the PCR products of the full length promoter is 3575bp which contain six YTCANTYY element (-220～-227bp, -567～-574bp, -728～-735, -1909～-1916, -3042～-3049, -3382～-3389bp), these fragments were ligated into the binary vector pCAMBIA1301 to substitute the CaMV35S promoter which is upstream of gus. The recombined plasmids are pNAR801, pNAR802, pNAR803 and pNAR804. All recombined binary vectors were confirmed by restricted enzyme digestion and transferred into Agrobacterium tumefaciens strain EHA101 by freeze-thaw method.
2. By Agrobacterium-mediated method, pNAR801, pNAR802, pNAR803, and pNAR804 were transferred into japonica rice R109. pCAMBIA1301 was also transferred at the same time. Total 136 independent transformed plants were got on regeneration medium. PCR analysis revealed that 96 plants were positive and the transform frequency is 70.6%. PCR and Southern blot tests confirmed the integration of foreign gene.
3. By using transgenic rice plants of 5'deletion pib promoter fragment-gus constructions, the relationships between the copy number of molecular element YTCANTYY in pib promoter and activity of pib promoter and its darkness inducing attribute were systematically analyzed. The results from Gus histochemistry detection revealed, after darkness inducing the transgenic rice calli of pib promoter fragments carrying 6, 3 or 1 copy of YTCANTYY motif were appeared different degree Gus blue in X-gluc solution, but the 5'deletion pib promoter fragment with no YTCANTYY element was unable to drive gus gene to show any Gus blue in X-gluc solution. Fluorescence quantitative analysis results showed that pib promoter sequence had strong organ specificity, even though all 6 darkness inducing elements were knocked out, its transgenic rice plants still kept the tendency of promoter activity in root higher than that of above ground parts of plants. But the activity of promotion and darkness inducing were increased with the increase of promoter fragments in length and with increase of darkness inducing element copy. These results indicated that YTCANTYY in pib promoter is a functional motif for darkness inducing and in other words this motif confer the darkness inducing of pib promoter. At least within 6 copies, the activity of darkness inducing was positively correlated with the copy number of YTCANTYY element. And the activity for each individual YTCANTYY element seemed to additive.
4. Using transgenic rice progenies of pib promoter-gus recombined plasmid, the inducing activities of pib promoter were analyzed. The fluorescence quantitative analysis of Gus activity in T_3 transgenic rice plants confirmed that 24h darkness treatment and temperature could influence promotion activity of pib promoter. Gus activity in root was always higher than leaf and they have strong organ specificity. Gus activity both in root and leaf of the transgenic rice plants all exhibited a clear diurnal rhythm, the Gus activity in daytime was at low level (15:00 was lowest) and high level was in night (3:00 was highest). These experimental results of Gus activity in the transgenic rice plants indicated the blast resistance production of pib structure gene was mainly synthesize in root and which was separated from infection position of rice blast. We infer that, the disease resistance product of pib structure gene is synthesized in root, and then transported to the target tissue of aboveground in the night to perform the function of blast resistance.
Pib基因属于NBS-LRR抗病基因家族,是目前为数不多已克隆的抗稻瘟病主效基因之一。研究报道表明,pib基因表达不受病原菌诱导,而黑暗、温度等环境和一些化学物质却能有效地诱导pib基因的表达。研究结果还显示,将不同长短的pib结构基因序列片段置于35S驱动下,它们各自的转基因植株表现出对稻瘟病生理小种的不同抗性,而pib结构基因上游的启动区与gus基因融合构建的转基因试验也证实了pib启动子具有很强的黑暗诱导活性。由于水稻的根是处于24h黑暗环境,而地上部器官处于光/暗循环交替之下,这就使得pib基因的抗病性表达具有了特殊的器官特异性和昼夜节律性。
为了探讨pib启动子这种环境条件诱导性的分子结构和分子机制,笔者用PLACE数据库对pib启动区核酸序列进行了分析,该启动区内有各种定义为启动相关分子元件近百个,除了TATA、CAAT等启动子共有元件外还有数十个不同类型的诱导性元件。其中被定义为光响应相关的顺式作用元件YTCANTYY以6个拷贝形式散布在启动子序列内。为了验证该启动子的暗诱导特性是否与此分子元件有关,这个分子元件又是怎样影响该启动子的暗诱导性的,笔者构建了不同长度、即含有不同数目的YTCANTYY元件的水稻pib基因启动子的5′端缺失体驱动gus基因的双元载体,用对其水稻转基因植株的gus基因表达分析的结果来研究该pib启动子中暗诱导分子元件的生物学功能及其对暗诱导的响应。
1.用PCR扩增并测序获得4个不同长度的pib启动子片段,各片段之间的区别仅在于5'端的缺失各不相同。4个片段的3'端均终止于碱基2(ATG信号的G),分别为:5'缺失体启动子1 PCR产物长222bp,为-219～2bp,无YTCANTYY元件,5'缺失体启动子2PCR产物长569bp,为-566～2bp,含1个YTCANTYY元件(-220～-227bp处),5'缺失体启动子3 PCR产物长1911bp,为-1908～2bp,含3个YTCANTYY元件(-220～-227,-567～-574,-728～-735bp处),全长启动子PCR产物长3575bp,为-3572～2bp,含6个YTCANTYY元件(-220～-227,-567～-574,-728～-735,-1909～-1916,-3042～-3049,-3382～-3389 bp处)。801(-219～2bp)、802(-566～2bp)、803(-1908～2bp)和804(-3572～2bp)分别连接进pCAMBIA1301中gus因前,取代原来的CaMV35S启动子,构建成双元载体pNAR801、pNAR802、pNAR803和pNAR804。重组双元质粒经过酶切验证,用农杆菌冻融转化法转化入农杆菌菌株EHA101中。
2.利用农杆菌介导的方法,将上述pNAR801、pNAR802、pNAR803、pNAR804重组质粒转化粳稻品种R109,同时以CaMV35S启动子驱动gus基因的pCAMBIA1301作对照。共获得136株再生植株,经PCR筛选确定得到96个阳性的转基因水稻植株,阳性率70.6%。通过Southern blot检测也表明目的基因已经整合到水稻基因组中。
3.对上述不同长度5′端缺失的pib启动子驱动gus基因的水稻转基因植株,进行Gus组织化学检测和荧光定量分析,系统研究了pib启动子中分子元件YTCANTYY拷贝数目与该启动子启动活性和暗诱导性的关系。Gus组织化学分析结果表明,含6个、3个、1个拷贝的pib启动子的转基因水稻愈伤在暗处理后均能在X-gluc溶液中显示不同深浅的Gus兰色,而6个拷贝完全缺失的启动子残端的水稻愈伤在X-gluc溶液中不显现Gus兰色。荧光定量分析结果表明,pib启动子序列具有很强的器官特异性,即便是6个暗诱导分子元件都被敲除的3'残端,其转基因植株仍保持根部启动活性高于地上部份的趋势。但其启动活性和暗诱导性随启动子缺失片段的缩短即暗诱导分子元件拷贝数增加而提高。这些结果表明,YTCANTYY在pib启动子序列中是一个暗诱导的功能性分子元件,他能赋予启动子的暗诱导性,起码在6个拷贝以内,pib启动子的暗诱导活性是与其拷贝数目呈正相关,各个YTCANTYY分子元件的启动活性可能是累加的。
4.利用pib基因启动区与gus构建的报告基因载体pNAR604得到的T_3转基因水稻,继续对pib基因启动子诱导性进行研究,再次证实pib启动子活性受黑暗和温度影响,无论诱导与否,在根中的Gus活性都远高于叶片具有强烈的器官特异性,pib启动子活性无论是在根还是叶都具有明显的昼夜节律性,白昼低夜晚高,以15:00活性水平为最低,03:00活性水平为最高。这些结果表明,pib基因表达的起始或抗稻瘟病产物的合成地在根系,而不是植株叶、茎等稻瘟病发病部位。也就是说pib基因抗稻瘟病功能性产物的合成起始地与抗病的靶点分离。据此,我们推断,pib基因编码的抗稻瘟病产物是在根部起始合成,抗病基因产物在夜间由根系输送到植株地上的靶组织,实现其抗病的功能。
As a important regulatory element of 5'upstream regions of structure gene, promoter controls gene expression characteristic of space and time in transcriptional level. Study on plant promoters of disease resistance and analysis of regulatory element's function in promoter region are the very significance to expression characteristic of disease resistance gene. The transformation analysis by 5'deletion promoter fragments is the important method to understand the biological function of these molecular elements.
Pib gene is the first major blast resistant gene cloned from rice by mapping clone technique and which is belongs to NBS-LRR resistant disease gene family. Previous studies show that the expression of pib gene was induced by darkness and chemistry factors but not pathogen. Our experiment results also indicated that different length fragments of pib structure gene sequence under the drive of 35S promoter, their To transgenic rice plants showed different resistances to rice blast pathogen races. Our results indicated that the pib promoter has the activity of darkness inducing in the rice transgenic experiments with the constructs of pib promoter-gus fusion. In natural environment, only root of rice plant could enjoy full darkness but the above ground organs are in the condition of light or dark cycle alternative, which offer the expression of pib gene the special organ specificity and diurnal rhythm.
For studying the molecular structure and mechanism of environmental condition inducing, we analysis this promoter region by using PLACE database. The results show that there are about hundred of molecular regulatory element, for activation in this region, except of TATA、CAAT commonly elements, there are six copies of YTCANTYY element defined for light-response of cis-regluatory element. To verifying whether the dark-inducing property of the pib promoter is relate to the YTCANTYY element and how this molecular element affect the dark-inducing property? By using transgenic rice plants of 5'deletion pib promoter fragment-gus constructions, the relationships between the copy number of molecular element YTCANTYY in pib promoter and activity of pib promoter and its darkness inducing attribute were systematically analyzed.
1. Four different regions of pib gene promoter were obtained by PCR, the difference among each fragments only lengths in 5'deletion, the 3' downstream of four fragments all terminate to base 2(base G of signal ATG). The PCR products of the first 5'deletion promoter is 222bp which is without YTCANTYY element; the PCR product of the second 5'deletion promoter is 569bp which contains one YTCANTYY element (-220～-227bp); the PCR products of the third 5'deletion promoter is 1911bp which contain three YTCANTYY element (-220～-227,-567～-574,-728～-735bp); the PCR products of the full length promoter is 3575bp which contain six YTCANTYY element (-220～-227bp, -567～-574bp, -728～-735, -1909～-1916, -3042～-3049, -3382～-3389bp), these fragments were ligated into the binary vector pCAMBIA1301 to substitute the CaMV35S promoter which is upstream of gus. The recombined plasmids are pNAR801, pNAR802, pNAR803 and pNAR804. All recombined binary vectors were confirmed by restricted enzyme digestion and transferred into Agrobacterium tumefaciens strain EHA101 by freeze-thaw method.
2. By Agrobacterium-mediated method, pNAR801, pNAR802, pNAR803, and pNAR804 were transferred into japonica rice R109. pCAMBIA1301 was also transferred at the same time. Total 136 independent transformed plants were got on regeneration medium. PCR analysis revealed that 96 plants were positive and the transform frequency is 70.6%. PCR and Southern blot tests confirmed the integration of foreign gene.
3. By using transgenic rice plants of 5'deletion pib promoter fragment-gus constructions, the relationships between the copy number of molecular element YTCANTYY in pib promoter and activity of pib promoter and its darkness inducing attribute were systematically analyzed. The results from Gus histochemistry detection revealed, after darkness inducing the transgenic rice calli of pib promoter fragments carrying 6, 3 or 1 copy of YTCANTYY motif were appeared different degree Gus blue in X-gluc solution, but the 5'deletion pib promoter fragment with no YTCANTYY element was unable to drive gus gene to show any Gus blue in X-gluc solution. Fluorescence quantitative analysis results showed that pib promoter sequence had strong organ specificity, even though all 6 darkness inducing elements were knocked out, its transgenic rice plants still kept the tendency of promoter activity in root higher than that of above ground parts of plants. But the activity of promotion and darkness inducing were increased with the increase of promoter fragments in length and with increase of darkness inducing element copy. These results indicated that YTCANTYY in pib promoter is a functional motif for darkness inducing and in other words this motif confer the darkness inducing of pib promoter. At least within 6 copies, the activity of darkness inducing was positively correlated with the copy number of YTCANTYY element. And the activity for each individual YTCANTYY element seemed to additive.
4. Using transgenic rice progenies of pib promoter-gus recombined plasmid, the inducing activities of pib promoter were analyzed. The fluorescence quantitative analysis of Gus activity in T_3 transgenic rice plants confirmed that 24h darkness treatment and temperature could influence promotion activity of pib promoter. Gus activity in root was always higher than leaf and they have strong organ specificity. Gus activity both in root and leaf of the transgenic rice plants all exhibited a clear diurnal rhythm, the Gus activity in daytime was at low level (15:00 was lowest) and high level was in night (3:00 was highest). These experimental results of Gus activity in the transgenic rice plants indicated the blast resistance production of pib structure gene was mainly synthesize in root and which was separated from infection position of rice blast. We infer that, the disease resistance product of pib structure gene is synthesized in root, and then transported to the target tissue of aboveground in the night to perform the function of blast resistance.
引文
董继新,董海淘,李德葆.水稻抗瘟性研究进展.农业生物技术学报,2000,8:99-102.
丁克坚,檀根甲,吴坚.稻瘟病为害损失研究.植物保护学报,1999,26(1):61-64.
邓晓梅,杨竹平.粳稻幼胚转化系统的建立及除草剂抗性植株的获得.上海农业学报,1998,4(2):29-34.
戴顺洪,李良材,丁月云,等.水稻基因枪法多基因转化研究.遗传学报,1998,25 4):345-350.
段晓岚,陈善葆.DNA导入水稻引起性状变异.中国农业科学,1985,18(3):6-9.
付永彩,月云,刘新仿,等.抑制衰老的嵌合基因在水稻中的转化.科学通报,1998,43(18):1963-1967.
郭岩,张莉,肖岗,等.甜菜碱醛脱氢酶在水稻中的表达及转基因植株的耐盐性研究.中国科学(C 辑),1997,27(2):151-155.
黄大年,朱冰,杨炜,等.抗菌肽B基因导入水稻及转基因植株的鉴定.中国科学(C辑),1997,27(1):55-62.
胡廷章,罗凯,甘丽萍,石汝杰.植物基因启动子的类型及其应用.湖北农业科学,2007,46(1):149-151
李婵娟,杨世湖,武亮,等.Pib基因启动子及其诱导活性初探.遗传,2006,28(6):689-694
李一醌,王金发.高等植物启动子研究进展.植物学通报,1998,15(增刊):1-6.
李杰,张福城,王文泉,等.高等植物启动子的研究进展.生物技术通讯,2006,17(4):658-661
刘后利.植物育种研究进展.北京:农业出版社,1993.
李宝健,欧阳学智,许耀.应用农杆菌Ti质粒系统将外源基因转入籼稻细胞研究.中国科学(B辑),1990,20(2):144-149.
李宝健,曾庆平.植物生物技术原理与方法.湖南:湖南科学技术出版社,1990.201-218.
凌定厚,陶利珍,马镇荣,等.以基因枪介导获转ps1-barnase基因的工程雄性不育水稻植株.遗传学报,1998,25(5):433-442.
卢泳全,吴为人.水稻遗传转化研究进展.福建农林大学学报(自然科学版),2003,32(1):27-31.
凌忠专,潘庆华.主要农作物抗病性遗传研究进展.南京:江苏科学技术出版社,1990,1111-1151.
刘国权等.水稻抗稻瘟病研究进展与对策.中国农学通报,2004,20(1):211-214.
林福呈,李德葆.稻瘟病菌侵入机理.微生物学通报,1997,24(3):167-170.
李落叶,井金学.稻瘟病抗性基因的分子定位及克隆.中国农学通报,2006,22(1):49-53.
李宁,樊守金,张增艳.植物抗病相关启动子及其研究进展.植物遗传资源学报,2007,8(2):234-239.
雷财林,王久林,凌忠专.云南粳稻品种毫变—1和扎缅尼—1抗稻瘟病基因分析.中国农业科 学,1995,28(4):66-71.
雷财林,王久林,毛世宏,等.籼稻品种窄叶青8号抗稻瘟病基因分析.遗传学报,1997,24(1):36-41.
却志群.水稻稻瘟病抗病基因的克隆.安徽农业科学,2007,35(25):7792-7793,7806.
苏金,Jargolli J,吴乃虎,等.在转基因植物中实现外源基因最佳表达的途径.生物工程进展,1999,19(4):3-6.
吴建利,庄杰云,李德葆,郑康乐.水稻对稻瘟病抗性的分子生物学研究进展.中国水稻科学,1999,13(2):123-128.
吴建利,柴荣耀,樊叶杨,等.抗稻瘟病水稻材料谷梅2号中主效抗稻瘟病基因的成簇分布.中国水稻科学,2004,18(6):567-569.
万丙良,张献龙.基因枪与水稻遗传改良.生命的化学,1998,18(6):41-42.
王葳.转基因植物研究概况.生物技术通报,1994(6):6-7.
王关林,方宏筠.物基因工程原理与技术.北京:中国科学出版社,1998.
许明辉,唐祚舜,谭亚玲,田颖川,李成云,张树华,陈正华,田文忠.几丁质酶-葡聚糖酶双价基因导入滇型杂交稻恢复系提高稻瘟病抗性的研究.遗传学报.2003,30(4):330-334.
向匈朝,李平,王世全.水稻基因工程育种及其进展.西南科技大学学报,2004,19(2):107-113.
许新萍,胡明,卫剑文,等.用基因枪转化水稻获得转基因植株.中山大学学报(自然科学版),1997,36(6):25-29.
杨慧,杨世湖,王庆,等.pib基因编码区功能的转基因初步鉴定.南京农业大学学报,2006,29(3):1-5.
杨剑波,吴李君,吴家道,等.应用低能离子介导法获得水稻转基因植株.科学通报,1994,39(16):1530-1534.
张春晓,王文棋,蒋湘宁,陈雪.植物基因启动子研究进展.遗传学报,2004,31(12):1455-1464.
张锦花,施碧红,赵明富.水稻稻瘟病抗性基因研究概况.生物技术通报,2007,2:17-22.
周建明,朱群,白永延.稻瘟病菌侵染水稻的机理.植物生理学通讯,1999,35(1):49-44.
朱献丰.水稻稻瘟病菌研究进展.江西农业学报,2002,14(3):51-55.
郑宏红,戴顺洪,何锶洁,等.籼稻基因枪转化的研究.遗传学报,996,23(4):286-292.
周文忠,杨金水,王蕴珠.用一种新型基因枪将外源基因转入水稻细胞.实验生物学报,1996,29(1):89-93.
朱祯,李向辉,孙勇如,等.转基因水稻植株再生及外源人a-干扰素cDNA的表达.中国科学(B 辑),1992,22(2):149-155.
Arondel V,Lemieux B,Hwang I,et al.Map-based cloning of a gene controlling omega-3 fatty acid desaturation in Arabidopsis.Science,1992,258:1353-1355.
Benoit S P and Normand B.5' deletion analysis of the potato starch phosphorylase gene:An upstream sequence definesdistal regulatory elements and a proximal organ-dependent promoter.Plant Science,1995,110:193-203
Bradford M M.A rapid and sensitive method after the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.AnalBiochem,1976,72(7):248-254.
Bryan GT,Wu KS,Farrall L,et al.A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta.Plant Cell,2000,12:2033-2045.
Berruyer R,Adreit H,Milazzo J,et al.Identification and fine mapping of Pi33,the rice resistance gene corresponding to the Magnaporthe grisea avirulence gene ACE1.Theor Appl Genet,2003,107:1139-1147.
Chen DH,Delavina M,Inukai T,et al.Molecular mapping of the blast resistance gene,Pi44(t),in a line derived from a durably resistant rice cultivar.Theor Appl Genet,1999,98:1046-1053.
Chen S,Wang L,Que Z,et al.Genetic and physical mapping of Pi37(t),a new gene conferring resistance to rice blast in the famous cultivar St.No.1.Theoretical and Applied Genetics,2005,111(8):1563-1570.
Chen X,Shang J,Chen D,et al.AB-lectin receptor kinase gene conferring rice blast resistance.Plant J,2006,46(5):794-804.
Christou P,Ford T L,Kofrom M.Production of transgenic rice from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos.Bio-Technology,1991,9:957-962.
Chan M T,Chang H H,Ho S L,et al.Agrobacterium mediated production of transgenic rice plants expressing achimeric amylase promoter glucuronidase gene.Plant Mol Biol,1993,22:491-506.
Christou P,Ford T L,Kofrom M.Production of transgenic rice(Oryza sativa L.) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zypotic embryos.Bio(o|¨) Technol,1991,9:957-962.
Cao J,Duan X,Mcelory D,et al.Regeneration of herbicide resistant transgenic rice plants follow in gmicropro-jectile-mediated transformation of suspension culture cells.Plant Cell Rep,1992,11:586-591.
Dellaporta S L,Wood J,Hicks J B.A plant DNA mini preparation:version Ⅱ.Plant Mol Biol Rep.1983,1:19-21.
Dickinson C D,Evans R P,Nielsen N C.RY repeats are conserved in the 5'-flanking regions of legume seed- protein genes.Nucleic Acids Res,1988,16(1):371.
Dorbe M F,Truong H N and Francoise D V.Deletion analysis of the tobacco Nii1 promoter in Arabidopsis thaliana.Plant Science,1998,139:13971-82.
Edwards J W,Walker E L,Coruzzi G M.Cell-specific expression in transgenic plants reveals nonoverlapping roles for chloroplast and cytosolic glutamine synthetase.Proc Natl Acad Sci USA,1990,87(9):3459-3463.
Fujiwara T,Beachy R N.Tissue-specific and temporal regulation of a beta- conglycin in gene:roles of the RY repeat and other cis-acting elements.Plant Mol Biol,1994,24(2):261-272.
Fukuoka S,Okuno K.QTL analysis and mapping of pi21,a recessive gene for field resistance to rice blast in Japanese upland rice.Theor Appl Genet,2001,103:185-190.
Flor H.The current status of the gene-for-gene concept.Annu Rev Phytopathol,1971,9:275-296.
Guo Y,Lian G H,Bernsm W.Laser-mediated gene transfer in rice.Plant Physiol,1995,93:19-24.
Giraudat J,Haugebm,Valon C,et al.Isolation of the Arabidopsis ABI3 gene by positional cloning.Plant Cell,1992,4:1251-1261.
Higo K,Ugawa Y,Lwamoto M,Korenaga K.Plant cis-acting regulatory DNA elements (PLACE)database.Nucleic Acids Res,1999,27:297-300.
Hiei Y,Ohta S T,Kumashiro T.Efficient transformation of rice(Oryza sativa L.) mediated by A grobacterium and sequence analysis of the boundaries of the T-DNA.Plant J,1994,6:271-282
Haley J,Bogorad L.Alternative promoters are used for genes with in maize chloroplast polycistronic transcription units.Plant Cell,1990,2(4):323-333.
Jfferson R A.Assaying chimeric genes in plants:the gus gene fusion system.Plant Mol Biol Rep,1987,5(4):387-405.
Jfferson R A.GUS fusions:β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.The Embo J,1987,6(3):3901-3907.
Jiang H,Qin H M and Tian Y C.Cloning of a promoter fragment of bark storage protein gene from populus deltoids and its function in transgeneic tobacco plants.Scientia Silvae Sinicae,1999,35:46-50.
Joshl C P.An inspection of the domain between putative TATA box and translation start site in 79 plant genes.Nucleic Acids Research,1987,15:6643-6653.
Keddie J S,Tsiantis M,Piffanelli P,et al.A seed-specific brassica napusoleos in promoter interacts with a G- box- specific protein and may be bi- directional.Plant Mol Biol,1994,24(2):327-340.
Keller B and Lamb C J.Specific expression of a novel cell wall hydroxyproline-rich glycoprotein gene in lateral root initiation.Genes and Development,1989,3:1639-46.
Kiyo sawa S.In Rice Breeding.IRRI,1972,1203-225.
Kosugi S,Suzuka I,Ohashi Y.Two of three promoter elements identified in a rice gene for proliferating cell nuclear antigen are essential for meristematic tissue-specific expression.Plant J,1995,7(6):877-886.
Lagrange T,Franzetti B,Axelos M,et al.Structure and expression of the nuclear gene coding for the chloroplast ribosomal protein L21:developmental regulation of a housekeeping gene by alternative promoters.Mol Cell Biol,1993,13(4):2614-2622.
Lumbreras V,Canpos N,Boronat A.The use of an alternative promoter in the arabidopsis thaliana HMG1 gene generates an mRNA that encodes a novel 3-hydroxy-3-methylglutaryleoenzyme a reductaseisoform with an extended N-terminal region,Plant J,1995,8(4):541-549.
LiYi-Kun,Wang Jin-Fa.Advances of the studies on plant promoter.Chinese Bullentin of Botany,1998,15(Suppl):1-6.
Li Z,Upadnyaya NM,Meena S,Comparison of promoters and selectable marker genes for use in indica rice transformation.Molecular breeding.1997,3:1-14.
Li Y F,Zhu R,Xie L X,and Xu P L.Inducible expression of a promoter of the gene encoding barley b-l,3-glucanase iso-enzyme GⅢ in transgenie flee.Journal of Plant Physiology and Molecular Biology,2005,31:143-148.
Liu B,Zhang S,Zhu X,et al.Candidate defense genes as predictors of quantitative blast resistance in rice.Molecular Plant-Microbe Interactions,2004,17(10):1146-1152.
Liu Y H,Jia S R.Vascular specific promoters and cis-regulatory elements.Chinese Journal of Biotechnology,2003,19:131-135.
Li X,Ning Sh B,Song Y Ch,et al.Comparative physical localization of rice Pi-b gene and its linked RFLP markers in Oryza sativa O.officinalis and Zea mays.Acta Botanica Sinica,2002,44(1):49-54.
Liu X Q,Wang L,Chen S,et al.Genetic and physical mapping of Pi36(t),a new rice blast resistance gene located on rice chromosome 8.Molecular Genetics and Genomics,2005,274(4):394-401.
Liu G,Lu G,Zeng L,Wang G L.Two broad-spectrum blast resistance genes,Pi9(t) and Pi2(t) are physically linked on rice chromosome 6.Mol Genet Gen,2002,267:472-480.
Maria-Jesus C,Diane L,Kathleen MB.Activity of a maize ubiquitin promoter in transgenic rice.Plant Mol Biol,1993,23:567-581.
Martin G B,Brommonschenkel S H,Chunwongse J,et al.Map-based cloning of a protein kinase gene conferring disease resistance in tomato.Science,1993,262:1432-1436.
Michael Carey S T S.Transcriptional Regulation in Eukaryotes Concepts,Strategies,and Techniques.Clod Spring Harbor Laboratory Press,2001.
Maier R G,Brown J W S,Toloczyki C.Binding of a nuclear factor to a consensus sequence in the 5'-flanking region of zein genes from maize.EMBO J,1987,6:17-22.
Mao Z C,Yu Q J,Zeng W,Guo J Y and Hu Y L.Effects of ipt gene expression driven by fruit-specific promoter on development intransgenic tomato plants.Chinese Science Bulletin,2002,47:444-448.
Murray M G,Thompson W F.Rapid isolation of high molecular weight plant DNA.Nucl Acids Res,1980,8:4321-4325.
Nakamural M,Tatsuhiko Tsunoda T,Obokatal J.Photosynthesis nuclear genes Generally lack TATA-boxes:a tobacco photosystem Ⅰ gene responds to light through an initiator.The Plant Journal,2002,29(1),1-10.
Nakazono M,Itadani H,Wakasugi T,et al.The rps3-rpl16-nad3-rps12gene cluster in rice mitochondrial DNA is transcribed from alternative promoters.Curr Genet,1995,27(2):184-189.
Pan Q,Hu Z,Takatoshi T,et al.Fine Mapping of the Blast Resistance Gene Pi15,Linked to Pii,on Rice Chromosome 9.Acta Botanica Sinica,2003,45(7):871-877.
Puzio P S,Lausen J,Heinen P and Grtmdler F M W.Promoter analysis of pyk20,a gene from Arabidopsis thaliana.Plant Science,2000,157:245-25.
Qu S H,Liu G F,Zhou B,et al.The broad spectrum blast resistance gene Pi9 encodes an NBS-LRR protein and is a member of a multi-gene family in rice.Genetic,2006,172:1901-1914.
Rao Z,Wu J,Zhuang J,et al.Genetic Dissections of Partial Resistances to Leaf and Neck Blast in Rice(Oryza sativa L.).Acta Genetica Sinica,2005,32(6):556-565.
Rainerid M,Boyyino P,Gordon M P,et al.Agrobacterium-mediated transformation of rice(Oryza sativa L.).Bio(o|¨)Technol,1990,8:33-38.
Sallaud C,Lorieux M,Roumen E,et al.Identification of five new blast resistance genes in the highly blast-resistant rice variety IR64 using a QTL mapping strategy.Theoretical and Applied Genetics,2003,106(5):794-803.
Schernthaner J P,Matzke M A,Matzke A J M.Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants.EMBO J,1988,7:1249-1256.
Simpson S D,Nakashima K,Narusaka Y,Seki M,Shinozaki K,Sugita M and Gruissem W.
Developmental,organ-specific,and light-dependent expression of the tomato ribulose-1,5-bisphosphate carboxylase small subunit gene family.Proceedings of the National Academy of Sciences of the USA,1987,84:7104-710.
Sun X L,Cao Y L,Yang Z F,et al.Xa26,a gene conferring resistance to Xanthomonas oryzae pv.oryzae in rice,encodes an LRR receptor kinase-like protein.Plant J,2004,37:517-527.
Song W Y,Wang G L,Chen L L,et al.A receptor kinase-like protein encoded by the rice disease resistance gene,Xa2.Science,1995,270:1804-1806.
Toriyama K,Arimoto Y,Uchmiya H,et al.Transgenic rice plants after direct gene transfer into protoplast.Bio(o|¨) Technology,1988,6:1072-1074.
Tsuchiya T,Toriyama K,Nasrallah M E,et al.Isolation of genes abundantly expressed in rice anthers at the microsporostage.Plant Mol Biol,1992,20:1189-1193.
Uchimiya H,Fushim I T,Hashimoto H,et al.Expression of a foreign gene in callus derived from DNA-treated protoplasts of rice(Oryza sativa L.),Mol Gen Genet,1986,204:204-207.
Wang Z X,Yano M,Yanamouchi U,et al.The Pib gene for rice blast resistance belongs to the nucleotide binding and leucine rich repeat class of plant disease resistance genes.Plant J,1999,19:55-64.
Wang Z X,Yanamouchi U,Yano M,et al.Expression of the Pi-b rice-blast-resistance gene family is up-regulated by environmental conditions favoring infection and by chemical signals that trigger secondary plant defenses.Plant Molecular Biology,2001,47:653-661.
Yoshimura S,Yamanouchi U,Katayose Y,et al.Expression of Xa1,abacterial blight-resistance gene in rice,is induced by bacterial inoculation.Proc Natl Acad Sci USA,1998,95:1663-1668.
Zang W,David M,Ray W.Analysis of rice Actl 5'region activity in transgenic rice plants.Plant Cell,1991,3:1155-1165.
Zhuang J,Ma W,Wu J,et al.Mapping of leaf and neck blast resistance genes with resistance gene analog,RAPD and RFLP in rice.Euphytica,2002,128(3):363-370.
Zhu Q,Lamb C.TATA box and initiator functions in the accurate transcription of a plant minimal promoter in vitro.Plant Cell,1995,7(10):1681-1689.
Zhang H M,Yang H,Rech E L,et al.Transgenic rice plant produced by electro-poration-mediated plasmid up-take into protoplasts.Plant Cell Rep,1988,7:379-384.
Zhan G W,Wu R.Efficient regeneration of transgenic plant from rice protoplasts and correctly regulated expression of the foreign gene in the plant.Theor Appl Genet,1988,76:835-840.
Zhang S Z,Yang B P and Liu F H.Cloning and sequence analysis of a flower-specific expression promoter PchsA.Journal of Agricultural Biotechnology,2002,10:116-119.
丁克坚,檀根甲,吴坚.稻瘟病为害损失研究.植物保护学报,1999,26(1):61-64.
邓晓梅,杨竹平.粳稻幼胚转化系统的建立及除草剂抗性植株的获得.上海农业学报,1998,4(2):29-34.
戴顺洪,李良材,丁月云,等.水稻基因枪法多基因转化研究.遗传学报,1998,25 4):345-350.
段晓岚,陈善葆.DNA导入水稻引起性状变异.中国农业科学,1985,18(3):6-9.
付永彩,月云,刘新仿,等.抑制衰老的嵌合基因在水稻中的转化.科学通报,1998,43(18):1963-1967.
郭岩,张莉,肖岗,等.甜菜碱醛脱氢酶在水稻中的表达及转基因植株的耐盐性研究.中国科学(C 辑),1997,27(2):151-155.
黄大年,朱冰,杨炜,等.抗菌肽B基因导入水稻及转基因植株的鉴定.中国科学(C辑),1997,27(1):55-62.
胡廷章,罗凯,甘丽萍,石汝杰.植物基因启动子的类型及其应用.湖北农业科学,2007,46(1):149-151
李婵娟,杨世湖,武亮,等.Pib基因启动子及其诱导活性初探.遗传,2006,28(6):689-694
李一醌,王金发.高等植物启动子研究进展.植物学通报,1998,15(增刊):1-6.
李杰,张福城,王文泉,等.高等植物启动子的研究进展.生物技术通讯,2006,17(4):658-661
刘后利.植物育种研究进展.北京:农业出版社,1993.
李宝健,欧阳学智,许耀.应用农杆菌Ti质粒系统将外源基因转入籼稻细胞研究.中国科学(B辑),1990,20(2):144-149.
李宝健,曾庆平.植物生物技术原理与方法.湖南:湖南科学技术出版社,1990.201-218.
凌定厚,陶利珍,马镇荣,等.以基因枪介导获转ps1-barnase基因的工程雄性不育水稻植株.遗传学报,1998,25(5):433-442.
卢泳全,吴为人.水稻遗传转化研究进展.福建农林大学学报(自然科学版),2003,32(1):27-31.
凌忠专,潘庆华.主要农作物抗病性遗传研究进展.南京:江苏科学技术出版社,1990,1111-1151.
刘国权等.水稻抗稻瘟病研究进展与对策.中国农学通报,2004,20(1):211-214.
林福呈,李德葆.稻瘟病菌侵入机理.微生物学通报,1997,24(3):167-170.
李落叶,井金学.稻瘟病抗性基因的分子定位及克隆.中国农学通报,2006,22(1):49-53.
李宁,樊守金,张增艳.植物抗病相关启动子及其研究进展.植物遗传资源学报,2007,8(2):234-239.
雷财林,王久林,凌忠专.云南粳稻品种毫变—1和扎缅尼—1抗稻瘟病基因分析.中国农业科 学,1995,28(4):66-71.
雷财林,王久林,毛世宏,等.籼稻品种窄叶青8号抗稻瘟病基因分析.遗传学报,1997,24(1):36-41.
却志群.水稻稻瘟病抗病基因的克隆.安徽农业科学,2007,35(25):7792-7793,7806.
苏金,Jargolli J,吴乃虎,等.在转基因植物中实现外源基因最佳表达的途径.生物工程进展,1999,19(4):3-6.
吴建利,庄杰云,李德葆,郑康乐.水稻对稻瘟病抗性的分子生物学研究进展.中国水稻科学,1999,13(2):123-128.
吴建利,柴荣耀,樊叶杨,等.抗稻瘟病水稻材料谷梅2号中主效抗稻瘟病基因的成簇分布.中国水稻科学,2004,18(6):567-569.
万丙良,张献龙.基因枪与水稻遗传改良.生命的化学,1998,18(6):41-42.
王葳.转基因植物研究概况.生物技术通报,1994(6):6-7.
王关林,方宏筠.物基因工程原理与技术.北京:中国科学出版社,1998.
许明辉,唐祚舜,谭亚玲,田颖川,李成云,张树华,陈正华,田文忠.几丁质酶-葡聚糖酶双价基因导入滇型杂交稻恢复系提高稻瘟病抗性的研究.遗传学报.2003,30(4):330-334.
向匈朝,李平,王世全.水稻基因工程育种及其进展.西南科技大学学报,2004,19(2):107-113.
许新萍,胡明,卫剑文,等.用基因枪转化水稻获得转基因植株.中山大学学报(自然科学版),1997,36(6):25-29.
杨慧,杨世湖,王庆,等.pib基因编码区功能的转基因初步鉴定.南京农业大学学报,2006,29(3):1-5.
杨剑波,吴李君,吴家道,等.应用低能离子介导法获得水稻转基因植株.科学通报,1994,39(16):1530-1534.
张春晓,王文棋,蒋湘宁,陈雪.植物基因启动子研究进展.遗传学报,2004,31(12):1455-1464.
张锦花,施碧红,赵明富.水稻稻瘟病抗性基因研究概况.生物技术通报,2007,2:17-22.
周建明,朱群,白永延.稻瘟病菌侵染水稻的机理.植物生理学通讯,1999,35(1):49-44.
朱献丰.水稻稻瘟病菌研究进展.江西农业学报,2002,14(3):51-55.
郑宏红,戴顺洪,何锶洁,等.籼稻基因枪转化的研究.遗传学报,996,23(4):286-292.
周文忠,杨金水,王蕴珠.用一种新型基因枪将外源基因转入水稻细胞.实验生物学报,1996,29(1):89-93.
朱祯,李向辉,孙勇如,等.转基因水稻植株再生及外源人a-干扰素cDNA的表达.中国科学(B 辑),1992,22(2):149-155.
Arondel V,Lemieux B,Hwang I,et al.Map-based cloning of a gene controlling omega-3 fatty acid desaturation in Arabidopsis.Science,1992,258:1353-1355.
Benoit S P and Normand B.5' deletion analysis of the potato starch phosphorylase gene:An upstream sequence definesdistal regulatory elements and a proximal organ-dependent promoter.Plant Science,1995,110:193-203
Bradford M M.A rapid and sensitive method after the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.AnalBiochem,1976,72(7):248-254.
Bryan GT,Wu KS,Farrall L,et al.A single amino acid difference distinguishes resistant and susceptible alleles of the rice blast resistance gene Pi-ta.Plant Cell,2000,12:2033-2045.
Berruyer R,Adreit H,Milazzo J,et al.Identification and fine mapping of Pi33,the rice resistance gene corresponding to the Magnaporthe grisea avirulence gene ACE1.Theor Appl Genet,2003,107:1139-1147.
Chen DH,Delavina M,Inukai T,et al.Molecular mapping of the blast resistance gene,Pi44(t),in a line derived from a durably resistant rice cultivar.Theor Appl Genet,1999,98:1046-1053.
Chen S,Wang L,Que Z,et al.Genetic and physical mapping of Pi37(t),a new gene conferring resistance to rice blast in the famous cultivar St.No.1.Theoretical and Applied Genetics,2005,111(8):1563-1570.
Chen X,Shang J,Chen D,et al.AB-lectin receptor kinase gene conferring rice blast resistance.Plant J,2006,46(5):794-804.
Christou P,Ford T L,Kofrom M.Production of transgenic rice from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos.Bio-Technology,1991,9:957-962.
Chan M T,Chang H H,Ho S L,et al.Agrobacterium mediated production of transgenic rice plants expressing achimeric amylase promoter glucuronidase gene.Plant Mol Biol,1993,22:491-506.
Christou P,Ford T L,Kofrom M.Production of transgenic rice(Oryza sativa L.) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zypotic embryos.Bio(o|¨) Technol,1991,9:957-962.
Cao J,Duan X,Mcelory D,et al.Regeneration of herbicide resistant transgenic rice plants follow in gmicropro-jectile-mediated transformation of suspension culture cells.Plant Cell Rep,1992,11:586-591.
Dellaporta S L,Wood J,Hicks J B.A plant DNA mini preparation:version Ⅱ.Plant Mol Biol Rep.1983,1:19-21.
Dickinson C D,Evans R P,Nielsen N C.RY repeats are conserved in the 5'-flanking regions of legume seed- protein genes.Nucleic Acids Res,1988,16(1):371.
Dorbe M F,Truong H N and Francoise D V.Deletion analysis of the tobacco Nii1 promoter in Arabidopsis thaliana.Plant Science,1998,139:13971-82.
Edwards J W,Walker E L,Coruzzi G M.Cell-specific expression in transgenic plants reveals nonoverlapping roles for chloroplast and cytosolic glutamine synthetase.Proc Natl Acad Sci USA,1990,87(9):3459-3463.
Fujiwara T,Beachy R N.Tissue-specific and temporal regulation of a beta- conglycin in gene:roles of the RY repeat and other cis-acting elements.Plant Mol Biol,1994,24(2):261-272.
Fukuoka S,Okuno K.QTL analysis and mapping of pi21,a recessive gene for field resistance to rice blast in Japanese upland rice.Theor Appl Genet,2001,103:185-190.
Flor H.The current status of the gene-for-gene concept.Annu Rev Phytopathol,1971,9:275-296.
Guo Y,Lian G H,Bernsm W.Laser-mediated gene transfer in rice.Plant Physiol,1995,93:19-24.
Giraudat J,Haugebm,Valon C,et al.Isolation of the Arabidopsis ABI3 gene by positional cloning.Plant Cell,1992,4:1251-1261.
Higo K,Ugawa Y,Lwamoto M,Korenaga K.Plant cis-acting regulatory DNA elements (PLACE)database.Nucleic Acids Res,1999,27:297-300.
Hiei Y,Ohta S T,Kumashiro T.Efficient transformation of rice(Oryza sativa L.) mediated by A grobacterium and sequence analysis of the boundaries of the T-DNA.Plant J,1994,6:271-282
Haley J,Bogorad L.Alternative promoters are used for genes with in maize chloroplast polycistronic transcription units.Plant Cell,1990,2(4):323-333.
Jfferson R A.Assaying chimeric genes in plants:the gus gene fusion system.Plant Mol Biol Rep,1987,5(4):387-405.
Jfferson R A.GUS fusions:β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants.The Embo J,1987,6(3):3901-3907.
Jiang H,Qin H M and Tian Y C.Cloning of a promoter fragment of bark storage protein gene from populus deltoids and its function in transgeneic tobacco plants.Scientia Silvae Sinicae,1999,35:46-50.
Joshl C P.An inspection of the domain between putative TATA box and translation start site in 79 plant genes.Nucleic Acids Research,1987,15:6643-6653.
Keddie J S,Tsiantis M,Piffanelli P,et al.A seed-specific brassica napusoleos in promoter interacts with a G- box- specific protein and may be bi- directional.Plant Mol Biol,1994,24(2):327-340.
Keller B and Lamb C J.Specific expression of a novel cell wall hydroxyproline-rich glycoprotein gene in lateral root initiation.Genes and Development,1989,3:1639-46.
Kiyo sawa S.In Rice Breeding.IRRI,1972,1203-225.
Kosugi S,Suzuka I,Ohashi Y.Two of three promoter elements identified in a rice gene for proliferating cell nuclear antigen are essential for meristematic tissue-specific expression.Plant J,1995,7(6):877-886.
Lagrange T,Franzetti B,Axelos M,et al.Structure and expression of the nuclear gene coding for the chloroplast ribosomal protein L21:developmental regulation of a housekeeping gene by alternative promoters.Mol Cell Biol,1993,13(4):2614-2622.
Lumbreras V,Canpos N,Boronat A.The use of an alternative promoter in the arabidopsis thaliana HMG1 gene generates an mRNA that encodes a novel 3-hydroxy-3-methylglutaryleoenzyme a reductaseisoform with an extended N-terminal region,Plant J,1995,8(4):541-549.
LiYi-Kun,Wang Jin-Fa.Advances of the studies on plant promoter.Chinese Bullentin of Botany,1998,15(Suppl):1-6.
Li Z,Upadnyaya NM,Meena S,Comparison of promoters and selectable marker genes for use in indica rice transformation.Molecular breeding.1997,3:1-14.
Li Y F,Zhu R,Xie L X,and Xu P L.Inducible expression of a promoter of the gene encoding barley b-l,3-glucanase iso-enzyme GⅢ in transgenie flee.Journal of Plant Physiology and Molecular Biology,2005,31:143-148.
Liu B,Zhang S,Zhu X,et al.Candidate defense genes as predictors of quantitative blast resistance in rice.Molecular Plant-Microbe Interactions,2004,17(10):1146-1152.
Liu Y H,Jia S R.Vascular specific promoters and cis-regulatory elements.Chinese Journal of Biotechnology,2003,19:131-135.
Li X,Ning Sh B,Song Y Ch,et al.Comparative physical localization of rice Pi-b gene and its linked RFLP markers in Oryza sativa O.officinalis and Zea mays.Acta Botanica Sinica,2002,44(1):49-54.
Liu X Q,Wang L,Chen S,et al.Genetic and physical mapping of Pi36(t),a new rice blast resistance gene located on rice chromosome 8.Molecular Genetics and Genomics,2005,274(4):394-401.
Liu G,Lu G,Zeng L,Wang G L.Two broad-spectrum blast resistance genes,Pi9(t) and Pi2(t) are physically linked on rice chromosome 6.Mol Genet Gen,2002,267:472-480.
Maria-Jesus C,Diane L,Kathleen MB.Activity of a maize ubiquitin promoter in transgenic rice.Plant Mol Biol,1993,23:567-581.
Martin G B,Brommonschenkel S H,Chunwongse J,et al.Map-based cloning of a protein kinase gene conferring disease resistance in tomato.Science,1993,262:1432-1436.
Michael Carey S T S.Transcriptional Regulation in Eukaryotes Concepts,Strategies,and Techniques.Clod Spring Harbor Laboratory Press,2001.
Maier R G,Brown J W S,Toloczyki C.Binding of a nuclear factor to a consensus sequence in the 5'-flanking region of zein genes from maize.EMBO J,1987,6:17-22.
Mao Z C,Yu Q J,Zeng W,Guo J Y and Hu Y L.Effects of ipt gene expression driven by fruit-specific promoter on development intransgenic tomato plants.Chinese Science Bulletin,2002,47:444-448.
Murray M G,Thompson W F.Rapid isolation of high molecular weight plant DNA.Nucl Acids Res,1980,8:4321-4325.
Nakamural M,Tatsuhiko Tsunoda T,Obokatal J.Photosynthesis nuclear genes Generally lack TATA-boxes:a tobacco photosystem Ⅰ gene responds to light through an initiator.The Plant Journal,2002,29(1),1-10.
Nakazono M,Itadani H,Wakasugi T,et al.The rps3-rpl16-nad3-rps12gene cluster in rice mitochondrial DNA is transcribed from alternative promoters.Curr Genet,1995,27(2):184-189.
Pan Q,Hu Z,Takatoshi T,et al.Fine Mapping of the Blast Resistance Gene Pi15,Linked to Pii,on Rice Chromosome 9.Acta Botanica Sinica,2003,45(7):871-877.
Puzio P S,Lausen J,Heinen P and Grtmdler F M W.Promoter analysis of pyk20,a gene from Arabidopsis thaliana.Plant Science,2000,157:245-25.
Qu S H,Liu G F,Zhou B,et al.The broad spectrum blast resistance gene Pi9 encodes an NBS-LRR protein and is a member of a multi-gene family in rice.Genetic,2006,172:1901-1914.
Rao Z,Wu J,Zhuang J,et al.Genetic Dissections of Partial Resistances to Leaf and Neck Blast in Rice(Oryza sativa L.).Acta Genetica Sinica,2005,32(6):556-565.
Rainerid M,Boyyino P,Gordon M P,et al.Agrobacterium-mediated transformation of rice(Oryza sativa L.).Bio(o|¨)Technol,1990,8:33-38.
Sallaud C,Lorieux M,Roumen E,et al.Identification of five new blast resistance genes in the highly blast-resistant rice variety IR64 using a QTL mapping strategy.Theoretical and Applied Genetics,2003,106(5):794-803.
Schernthaner J P,Matzke M A,Matzke A J M.Endosperm-specific activity of a zein gene promoter in transgenic tobacco plants.EMBO J,1988,7:1249-1256.
Simpson S D,Nakashima K,Narusaka Y,Seki M,Shinozaki K,Sugita M and Gruissem W.
Developmental,organ-specific,and light-dependent expression of the tomato ribulose-1,5-bisphosphate carboxylase small subunit gene family.Proceedings of the National Academy of Sciences of the USA,1987,84:7104-710.
Sun X L,Cao Y L,Yang Z F,et al.Xa26,a gene conferring resistance to Xanthomonas oryzae pv.oryzae in rice,encodes an LRR receptor kinase-like protein.Plant J,2004,37:517-527.
Song W Y,Wang G L,Chen L L,et al.A receptor kinase-like protein encoded by the rice disease resistance gene,Xa2.Science,1995,270:1804-1806.
Toriyama K,Arimoto Y,Uchmiya H,et al.Transgenic rice plants after direct gene transfer into protoplast.Bio(o|¨) Technology,1988,6:1072-1074.
Tsuchiya T,Toriyama K,Nasrallah M E,et al.Isolation of genes abundantly expressed in rice anthers at the microsporostage.Plant Mol Biol,1992,20:1189-1193.
Uchimiya H,Fushim I T,Hashimoto H,et al.Expression of a foreign gene in callus derived from DNA-treated protoplasts of rice(Oryza sativa L.),Mol Gen Genet,1986,204:204-207.
Wang Z X,Yano M,Yanamouchi U,et al.The Pib gene for rice blast resistance belongs to the nucleotide binding and leucine rich repeat class of plant disease resistance genes.Plant J,1999,19:55-64.
Wang Z X,Yanamouchi U,Yano M,et al.Expression of the Pi-b rice-blast-resistance gene family is up-regulated by environmental conditions favoring infection and by chemical signals that trigger secondary plant defenses.Plant Molecular Biology,2001,47:653-661.
Yoshimura S,Yamanouchi U,Katayose Y,et al.Expression of Xa1,abacterial blight-resistance gene in rice,is induced by bacterial inoculation.Proc Natl Acad Sci USA,1998,95:1663-1668.
Zang W,David M,Ray W.Analysis of rice Actl 5'region activity in transgenic rice plants.Plant Cell,1991,3:1155-1165.
Zhuang J,Ma W,Wu J,et al.Mapping of leaf and neck blast resistance genes with resistance gene analog,RAPD and RFLP in rice.Euphytica,2002,128(3):363-370.
Zhu Q,Lamb C.TATA box and initiator functions in the accurate transcription of a plant minimal promoter in vitro.Plant Cell,1995,7(10):1681-1689.
Zhang H M,Yang H,Rech E L,et al.Transgenic rice plant produced by electro-poration-mediated plasmid up-take into protoplasts.Plant Cell Rep,1988,7:379-384.
Zhan G W,Wu R.Efficient regeneration of transgenic plant from rice protoplasts and correctly regulated expression of the foreign gene in the plant.Theor Appl Genet,1988,76:835-840.
Zhang S Z,Yang B P and Liu F H.Cloning and sequence analysis of a flower-specific expression promoter PchsA.Journal of Agricultural Biotechnology,2002,10:116-119.