水稻OsHKT基因表达模式分析

【目的】OsHKT(High-affinity K~+transporter)是与水稻耐盐胁迫有关的一类Na~+或K~+转运体或Na~+-K~+共转运体,对水稻体内Na~+再循环,维持植株地上部特别是叶片中的低Na~+浓度和低Na~+/K~+比具有重要作用。本研究的目的是分析OsHKT基因家族表达的组织特异性、昼夜节律性以及盐、ABA对其表达的影响。【方法】利用荧光定量PCR技术分析OsHKT家族基因的表达模式。【结果】OsHKT基因家族中不同成员的表达具有明显的组织特异性,OsHKT1;1、OsHKT1;3、OsHKT2;3以及OsHKT2;4主要在水稻的叶片中表达,其余基因在根中表达水平较高。分析盐处理后OsHKT家族基因在水稻根和叶中的表达模式,结果表明OsHKT家族基因的表达均受盐胁迫的影响,但是不同OsHKT家族成员对盐胁迫反应不同。盐处理后OsHKT1;5、Os HKT2;1和OsHKT2;2基因在根和叶中的表达模式是一致的,而其他家族成员在根和叶中存在差异。所有OsHKT基因的表达在根或叶片中均受ABA调控,其中只有OsHKT1;3和OsHKT1;5在根和叶片中表达模式相似。光周期表达模式结果表明,OsHKT基因家族成员的表达具有相似的昼夜节律性。【结论】本研究结果表明水稻OsHKT家族不同成员的表达具有组织特异性和相似的昼夜节律性,并且不同成员对盐胁迫和ABA应答反应不同。这种表达模式可能与水稻在不同环境下的盐胁迫反应相适应。本研究结果为进一步揭示OsHKT基因的作用机制奠定了基础。     

太湖流域粳稻地方品种韭菜青的苗期耐盐性遗传分析

为了研究太湖流域强耐盐粳稻地方品种韭菜青的苗期耐盐性遗传机理,配制了韭菜青/IR26籼粳杂交组合,在盐胁迫条件下,运用剥蘖分株法鉴定了P[sub]1[/sub]、P[sub]2[/sub]、F[sub]1[/sub]和F[sub]2[/sub]苗期的盐害级别和根系Na+/K+两个耐盐相关指标。结果表明,在0.8%的NaCl胁迫下,亲本的两个耐盐相关指标有明显的差异,F[sub]1[/sub]均表现出超亲优势,F[sub]2[/sub]呈正态混合分布。采用主基因 多基因遗传模型进行遗传分析,发现韭菜青苗期的盐害级别和根系Na[sup]+[/sup]/K[sup]+[/sup]均由2对主效基因控制,并存在多基因修饰。盐害级别的主基因遗传率仅为16.10%,而根系Na[sup]+[/sup]/K[sup]+[/sup]的主基因遗传率高达50.78%,两个指标的主基因加多基因共同解释的遗传率都在70%以上。     

Monoclonal Antibody Production and Immunolocalization of a Salinity Stress-Related Protein in Rice (Oryza sativa)

Among various physiological responses to salt stress, the synthesis of a lectin-related protein of 14.5 kDa was observed in rice plants (Oryza sativa L.) under the treatment of 170 mmol/L NaCl. In order to better understand the role of the SALT protein in the physiological processes involving salinity, it was immunolocalized in mesophilic cells of leaf sheath and blade of a rice variety IAC-4440 following monoclonal antibodies produced by hybridome culture technique. This variety turned out to be an excellent model for that purpose, since it accumulates SALT protein even in absence of salt treatment and it has been classified as moderately sensitive to salinity and a superior grain producer. This feature was relevant for this work since it allowed the use of plants without the deleterious effects caused by salinity. Immunocytochemistry assays revealed that the SALT protein is located in the stroma of chloroplasts under non-stressing condition. Since the chloroplast is the main target affected by salinity and considering that the SALT protein does not present any apparent signal peptide for organelle localization, its lectin-like activity seems to play an important role in the establishment of stable complexes, either to other proteins or to oligosaccharides that are translocated to the chloroplast.     

黄淮海稻区水稻品种的穗颈瘟抗性

[目的]为明确黄淮海稻区育成水稻品种的稻瘟病抗性及其基因型,[方法]收集了来自本稻区5个不同地区的水稻品种,连续两年通过稻瘟病菌接种鉴定和田间病圃自然诱发对供试品种进行了稻瘟病抗性鉴定,并利用4个抗病基因Pita, Pib, Pi54和Pikm的分子标记,对水稻品种基因型进行了统计分析,并以抗性等级为表型值对品种资源进行聚类分析。[结果]黄淮海稻区水稻品种发病率较高,分别达 85.2%和 65.9%。Pi54和Pib检出率分别达 84%、65%。同时抗性聚类分析表明Pi54和Pib在本稻区的抗性贡献率却很低。[结论]黄淮海稻区水稻品种稻瘟病抗性弱,携带的抗瘟基因的抗性在不断减弱或丧失,需加强稻瘟病主效抗性的发掘、开发与利用。     

黄淮海稻区水稻品种的穗颈瘟抗性

【目的】为明确黄淮海稻区育成水稻品种的稻瘟病抗性及其基因型，【方法】收集了来自本稻区5个不同地区的水稻品种，连续两年通过稻瘟病菌接种鉴定和田间病圃自然诱发对供试品种进行了稻瘟病抗性鉴定，并利用4个抗病基因Pita, Pib, Pi54和Pikm的分子标记，对水稻品种基因型进行了统计分析，并以抗性等级为表型值对品种资源进行聚类分析。【结果】黄淮海稻区水稻品种发病率较高，分别达85.2%和65.9%。Pi54和Pib检出率分别达84%、65%。同时抗性聚类分析表明Pi54和Pib在本稻区的抗性贡献率却很低。【结论】黄淮海稻区水稻品种稻瘟病抗性弱，携带的抗瘟基因的抗性在不断减弱或丧失，需加强稻瘟病主效抗性的发掘、开发与利用。     

Systematic Identification of Rice ABC1 Gene Family and Its Response to Abiotic Stress

Members of the activity of bc1 complex (ABC1) family are protein kinases that are widely found in prokaryotes and eukaryotes. Previous studies showed that several plant ABC1 genes participated in the abiotic stress response. Here, we present the systematic identification of rice and Arabidopsis ABC1 genes and the expression analysis of rice ABC1 genes. A total of 15 and 17 ABC1 genes from the rice and Arabidopsis genomes, respectively, were identified using a bioinformatics approach. Phylogenetic analyses o...     

Differential Proteins Expressed in Rice Leaves and Grains in Response to Salinity and Exogenous Spermidine Treatments

Exogenous application of spermidine(Spd) has been reported to modulate physiological processes and alleviate salt-induced damage to growth and productivity of several plants including rice. Employing a proteomic approach, we aimed at identifying rice leaf and grain proteins differentially expressing under salt stress, and in response to Spd prior to Na Cl treatment. A total of 9 and 20 differentially expressed protein spots were identified in the leaves of salt-tolerant(Pokkali) and saltsensitive(KDML105) rice cultivars, respectively. Differential proteins common to both cultivars included a photosynthetic light reaction protein(oxygen-evolving complex protein 1), enzymes of Calvin cycle and glycolysis(fructose-bisphosphate aldolase and triose-phosphate isomerase), malate dehydrogenase, superoxide dismutase and a hypothetical protein(Os I_18213). Most proteins were present at higher intensities in Pokkali leaves. The photosynthetic oxygen-evolving enhancer protein 2 was detected only in Pokkali and was up-regulated by salt-stress and further enhanced by Spd treatment. All three spots identified as superoxide dismutase in KDML105 were up-regulated by Na Cl but down-regulated when treated with Spd prior to Na Cl, indicating that Spd acted directly as antioxidants. Important differential stress proteins detected in mature grains of both rice cultivars were late embryogenesis abundant proteins with protective roles and an antioxidant protein, 1-Cys-peroxiredoxin. Higher salt tolerance of Pokkali partly resulted from higher intensities and more responsiveness of the proteins relating to photosynthesis light reactions, energy metabolism, antioxidant enzymes in the leaves, and stress proteins with protective roles in the grains.     

外源物质对水稻盐胁迫缓解效应研究进展

盐胁迫是影响水稻生长发育和产量的主要非生物因素之一,外源物质能有效缓解盐对水稻的毒害作用并增加水稻产量。本文主要综述了不同外源物质(生长素、赤霉素、细胞分裂素、脱落酸、乙烯、褪黑素、水杨酸、多胺、油菜素类固醇、茉莉酸类外源生长调节物质以及钙、硅离子类外源物)对盐胁迫下水稻生长的不同调控机理及其缓解效应,并进一步对外源物质在水稻耐盐上的利用与研究工作进行了展望,以期为改善盐渍土环境下水稻的生长发育以及提高产量和品质提供理论依据。     

Cloning and Expression Analysis of OsNADPH1 Gene from Rice in Drought Stress Response

Drought resistance in rice has long been one of the important scientific research programs in cereal crops [1-3]. At present, the finding of novel drought genes is common in rice genetic resource development and varietal improvement. According to the action modes, the drought resistance genes can be divided into two classes [4]. The first class constitutes functional genes coding for such enzymes as late embryogenesis abundant protein (LEA) [5], △1-pyrroline-5-carboxylate synthetase (P5CS) …     

水稻盐胁迫应答的分子机制

水稻是一种对盐胁迫中度敏感的作物,它的耐盐性反应是个复杂的过程,包括对外界盐信号的感知和传递、特异转录因子的激活和下游胁迫应答基因的表达。综述了负责维持和重建离子平衡的Na+转运蛋白、转录调控因子和渗透调节基因的功能。它们的作用结果,或通过将盐离子隔离进液泡,或排出体外,或在胞质中积累大量渗透调节物质,最终使得植株表现耐盐性。     

水稻ABC1基因家族的鉴定及在非生物胁迫下的表达分析

ABC1(Activity of bc1 complex)家族属于蛋白质激酶家族,其成员普遍存在于原核和真核生物中.已有研究表明,几个植物ABC1基因参与非生物胁迫应答.为了解ABC1基因在水稻中的结构和功能,采用生物信息学方法分别在水稻和拟南芥上鉴定出15个和17个ABC1基因,并进行了系统发育和表达分析.结果表明,该家族在单、双子叶植物分离之前就已经发生了分化,其基本特征已经形成;单、双子叶植物分离之后,该家族在水稻和拟南芥中均以物种特异的方式进行了扩增.内含子/外显子结构分析显示多数直系同源基因之间外显子大小接近,而内含子差别较大,水稻含有更多大的内含子;内含子获得是近期伴随水稻ABC1家族进化的重要事件.多序列比对显示,ABC1结构域具有1个保守的氨基酸片段和4个保守的氨基酸残基.在线亚细胞定位预测9个水稻ABC1蛋白定位在叶绿体上.实时定量RT-PCR分析表明,水稻ABC1基因主要在叶片中表达,并且受多种非生物胁迫因素包括H2O2、脱落酸、低温、干旱、黑暗和高盐的调控.说明水稻ABC1家族不仅在逆境胁迫应答中发挥重要作用,可能还与水稻特定的生理过程有关.     

盐胁迫对粳稻品种生长和生理特性的影响

以津原85、金穗26、隆粳27、辽粳763为试验材料,研究了不同盐胁迫浓度对这4个品种生长和抗逆性的影响,以期为盐碱地适宜水稻品种的筛选提供理论参考.结果 表明,盐胁迫显著抑制水稻幼苗的生长,金穗26受到的影响最小.在盐胁迫下,津原85离子选择吸收性较强,有利于提高耐盐性.盐胁迫对水稻的主要伤害是破坏膜质结构,进而影响...     

水稻抗感纹枯病品种Ospgip1基因的表达特征

 从水稻基因组DNA中扩增出1 184 bp的Ospgip1基因片段。该片段包含930 bp的完整开放阅读框,终止密码子为TAA,无内含子。RT PCR结果表明,Ospgip1基因在水稻抗、感纹枯病品种中均能表达,但不同生育期、不同部位表达量有差异。实时PCR结果显示,抗病品种YSBR1和中感品种Jasmine 85中Ospgip1的表达量要明显高于感病品种Lemont;稻苗黄化处理后,无论是抗病品种还是感病品种的Ospgip1表达量均显著提高;纹枯病菌侵染使得抗病品种Ospgip1表达量大大增加,而对感病品种影响不大。     

利用CRISPR/Cas9技术编辑AFP1基因提高水稻耐逆性

目的 为鉴定水稻AFP1在非生物胁迫响应中的作用,创制非生物胁迫抗性的水稻新材料。方法 以优异籼稻恢复系华占为转化受体,利用CRISPR/Cas9技术创制afp1突变体,并对afp1突变体的耐逆性进行初步鉴定。结果 AFP1靶点1和靶点2的编辑效率分别为66.67%和75.00%。所有突变株系中,突变类型仅有插入和缺失突变,90%突变株系的突变长度为小片段突变（<5bp）。获得了6种无转基因成分的afp1纯合突变体。正常条件下,afp1突变体株高和结实率降低,有效分蘖增加,穗长显著升高,单株产量在-4.06%和11.75%之间变化。和野生型相比,afp1突变体的ABA敏感性和叶片水分散失率降低,耐干旱、热和渗透胁迫能力提高。结论 编辑AFP1基因可提高水稻多种非生物胁迫抗性。     

大田全生育期盐水灌溉胁迫筛选水稻耐盐恢复系

 2009-2010年在浙江省三门县沿海滩涂采用0.5%盐浓度海水进行全生育期灌溉的方法,以胁迫后植株存活率、单株结实率和单株粒重为指标对明恢86背景的49个回交导入系群体进行全生育期耐盐性评价。结果表明,不同群体的田间耐盐性存在很大差异,粳稻供体导入系群体的田间耐盐性总体好于籼稻供体的导入系群体,而且在同一亚种内不同供体品种的导入系群体的耐盐性差异明显。其中,沈农265、Y134、早籼14和Gayabyeo 为供体的4个导入系群体的耐盐株系的比率较高,分别为10.04%、6.98%、11.83%和7.9%。耐盐重复鉴定表明,大田全生育期耐盐性鉴定结果具有较高的可靠性。在常规栽培条件下对丰产性较好的耐盐恢复系与协青早A测交种进行鉴定,筛选出单株穗数、每穗总粒数和千粒重优势明显,单株产量比生产上对照组合显著增加的杂交组合6个。研究表明,利用大田全生育期盐胁迫筛选耐盐恢复系,不仅可以选育到在常规栽培条件下的高产组合,而且将为高产耐盐杂交新组合的选育创造有利条件。     

Identification and Cloning of Tillering-Related Genes OsMAX1 in Rice

Tillering is an important agronomic trait which has a direct impact on plant type and grain yield. Strigolactones are a class of important phytohormones regulating rice tillering. ATMAX1 is an important gene involved in strigolactone biosynthesis through encoding the protein P450 in Arabidopsis. Based on sequence BLASTp, we identified five homologous genes of ATMAX1 in rice, i.e., OsMAX1a, OsMAX1b, OsMAX1c, OsMAX1d and OsMAX1e. Among them, OsMAX1a and OsMAX1e showed stable and high expression in rice tissues. In addition, we observed that Os MAX1a and OsMAX1e can rescue the branched phenotype and the influences caused by MAX1 mutation in Arabidopsis. Moreover, the expression of OsMAX1a and OsMAX1e can respond to phosphate deficiency and different phytohormones, especially GR24, a strigolactone analogue. Therefore, it is concluded that Os MAX1a and OsMAX1e are involved in the biosynthesis of strigolactones and regulated rice tillering.     

水稻耐盐性数量性状位点的初步检测

用RFLP分析技术和分蘖株系法对由耐盐性品种Pokkali和盐敏感品种Peta配制的BC1(Peta/Pokkali∥Peta)群体分别检测水稻苗期和成熟期耐盐性数量性状位点(QTLs)。表型鉴定在含(处理)或不含(对照)60 mol/m3 NaCl的营养液中进行,苗期观测盐害级别、苗Na+含量和鲜重/干重比值3项指标,成熟期测定10种农艺性状处理与对照的相对值。从水稻12条染色体上筛选出43个多态性标记,对上述指标分别作点分析,共检出15个连锁标记。连锁标记的分布特点显示,在研究所涉及的基因组范围内存在4个影响苗期耐盐性的QTL,其增效等位基因均来自耐盐品种Pokkali;影响成熟期耐盐性的QTL分布于7条染色体的1或2个连锁区间上,其有利基因来自双亲;RG678和RZ400B~RZ792附近的2个QTL在全生育期都能表达较强耐盐性。     

水稻耐盐分子机制与育种研究进展

盐碱土壤对水稻不同生长发育时期均形成盐胁迫,进而导致水稻产量降低。耐盐水稻选育是提高水稻产量的有效途径。本文分析了盐胁迫对水稻不同发育时期农艺性状的影响,并从渗透调节、离子应答、激素调控和活性氧清除四个方面综述了近年来水稻耐盐分子机制的研究进展。最后,本文总结了水稻耐盐育种现状,并对耐盐水稻新品种的选育和推广进行了展望。     

一个新的水稻液泡膜Na+/H+逆向转运蛋白基因的克隆及表达特征

通过RT PCR技术从水稻幼苗组织中克隆了一个新的Na+/H+逆向转运蛋白基因OsNHX2,它编码一条长544氨基酸的多肽。OsNHX2与水稻和拟南芥的液泡膜Na+/H+逆向转运蛋白OsNHX1、AtNHX1氨基酸序列一致性分别为78%和77%。基因结构分析表明OsNHX2与OsNHX1具有类似的外显子和内含子构成,但不同于拟南芥AtNHX1,表明OsNHX2与OsNHX1可能起源于单双子叶植物分化后的水稻染色体复制。半定量RT PCR方法检测了OsNHX2与OsNHX1在水稻耐盐品种韭菜青与盐敏感品种IR28的地上部与根部的表达。结果表明：耐盐品种韭菜青的OsNHX2与OsNHX1基因在盐胁迫下表达持续增强,而在盐敏感品种IR28的地上部,OsNHX2基因在盐胁迫处理后1 h表达增强后立即减弱,根部的表达则基本不变,而IR28地上部与根部OsNHX1则在盐胁迫处理初期表达增强,随后即减弱。研究结果表明水稻两个液泡膜Na+/H+逆向转运蛋白基因OsNHX2、OsNHX1在盐敏感程度不同的水稻品种中表达有所不同,液泡膜Na+/H+逆向转运蛋白基因的转录调控可能是决定水稻耐盐能力的一个重要因素。