Analysis of SSLP and Soluble Protein Contents in Leaves of Mutants Induced by High Pressure in Rice （Oryza sativa）

Rice variety Yuexiangzhan and its mutants induced by high pressure were studied using microsatcllite markers and soluble protein content analyses. Eleven of the 88 microsatellite primer pairs showed evident polymorphisms repeatedly, and the polymorphic frequencies were 3.4-11.3% between the mutants and Yuexiangzhan. The polymorphic markers were randomly located on chromosomes. The more similar the plant types of the mutants like their original variety, the less polymorphic loci were detected. In addition, there was variation in the soluble protein contents among the leaves of mutants, and the contents were significantly lower than those of the original variety.     

水稻高压变异材料的SSR标记和叶片可溶性蛋白质含量分析

应用微卫星标记技术和可溶性蛋白质含量测定,对水稻品种粤香占及其由高压引起的变异材料粤压1、2、3、4号进行分子生物学研究。结果表明,在选取的88对微卫星引物中有11对表现多态性,变异材料与原种的多态性频率介于3.4%~11.3%,多态性标记随机地分布于水稻染色体上,形态上与原种越相近的材料多态性位点数越少。此外,每一变异材料叶片可溶性蛋白质含量互不相同,均显著低于对照。     

Changes in Chlorophyll Fluorescence of Rice Mutants Induced by High Hydrostatic Pressure

Three mutants of rice (Oryza sativa L. ), Mutant 1, Mutant 2and Mutant 3, which were selected by high hydrostatic pressure (75 MPa), and their parent Yuexiangzhan were used to study the changes in chlorophyll fluorescence during different growth stages. In all the three mutants, the function of PSⅡ was improved, Fy/Fm ratio of mutants increased compared to their parent at tillering and heading stage, and φPS Ⅱ also improved except for Mutant 2 at heading stage. Similar to their parent, the mutants exhibited slight photoinhibition at noon and almost complete recovery to initial levels of 6:00 after 18:00 at heading stage. At milking stage, the photoinhibition in the mutants was obvious, and recovered rapidly compared to the parent. Yields of individual plant and grain/straw ratio were also higher in three mutants than the parent. Results indicated that characteristics of chlorophyll fluorescence in leaves of mutants and their photoinhibition in the field had changed. It is suggested that high hydrostatic pressure induction could be applied as a new effective approach in high-yield rice breeding in the future.     

Changes in Chlorophyll Fluorescence of Rice Mutants Induced by High Hydrostatic Pressure

Three mutants of rice (Oryza sativa L. ), Mutant 1, Mutant 2 and Mutant 3, which were selected by high hydrostatic pressure (75 MPa), and their parent Yuexiangzhan were used to study the changes in chlorophyll fluorescence during different growth stages. In all the three mutants, the function of PS Ⅱ was improved, Fv/ Fm ratio of mutants increased compared to their parent at tillering and heading stage, and ФPS Ⅱ also improved except for Mutant 2 at heading stage. Similar to their parent, the mutants exhibited slight photoinhibition at noon and almost complete recovery to initial levels of 6:00 after 18:00 at heading stage. At milking stage, the photoinhibition in the mutants was obvious, and recovered rapidly compared to the parent. Yields of individual plant and grain/straw ratio were also higher in three mutants than the parent. Results indicated that characteristics of chlorophyll fluorescence in leaves of mutants and their photoinhibition in the field had changed. It is suggested that high hydrostatic pressure induction could be applied as a new effective approach in high-yield rice breeding in the future.     

高静水压诱变水稻突变株系的叶绿素荧光特性变化

以经75 MPa高静水压处理后筛选得到的3个水稻突变株系及其亲本粤香占为材料,测定了田间生长条件下不同生育时期水稻剑叶的叶绿素荧光动力学参数。与对照亲本相比较,3个突变株系的Fv/Fm在分蘖期和抽穗期均增加,除抽穗期突变2的ΦPSⅡ略低外,其余的ΦPSⅡ都表现为增加,说明光系统Ⅱ的功能得到改善。在抽穗期,突变株系与亲本在中午高光强下受到的抑制相比,突变株系受到的抑制较小,而且在18:00以后几乎能恢复到早上6:00的水平;在灌浆期,突变株系受到的光抑制增加,但能够快速恢复。突变1、突变2和粤压1号单株产量比对照亲本分别提高268%、279%和188%。结果显示,高静水压处理萌动的水稻种子后筛选得到的突变株系叶片的叶绿素荧光特性及田间光抑制发生了改变,产量也明显提高。高静水压诱变可能成为一种筛选高产水稻品种的新途径之一。     

水稻中花11辐射突变体的分离与鉴定

利用伽玛射线对粳稻品种中花11进行辐照,以诱导广泛的形态突变体,为水稻功能基因组研究提供基础材料。辐射第1代单本移栽并实行单株收获;辐射第2代（M2）播种1万个家系,自苗期开始至成熟期各个生育阶段从中独立选获各类形态突变体540个;经M3复选获得包括叶、茎、穗和育性变化等各类突变体431份。统计结果表明,形态性状的突变频率为5.1%。关键词     

水稻粳型亲籼系S-b座位基因型鉴定

S-b是控制水稻籼粳亚种间杂种不育性的基因座位之一。在该座位,台中65的基因型为Sj/Sj,广陆矮4号的基因型为Si/Si。以台中65和广陆矮4号为遗传测验种,分别与4个粳型亲籼系配组,根据紧密连锁分子标记基因型在这些组合F2群体中的偏态分离程度, 测定了这4个粳型亲籼系在F1花粉不育性S-b基因座位的基因型,结果表明,G2416-3和G2417-2-1在S-b基因座位的基因型为Si/Si,而G2605和G3004-4的基因型为Sn/Sn。比较了不同粳型亲籼系S-b基因座位等位基因遗传效应的大小,结果表明G2416-3携带的Si基因与G2417-2-1携带的Si基因之间,以及G2605携带的Sn基因与G3004 4携带的Sn基因之间没有显著差异,而Si 与Sn之间遗传效应差异极显著。     

光照敏感型水稻红根突变体遗传分析及基因的分子定位

 在自然光条件下水培籼稻品种Nankinkodo中,发现根为红色的自然突变体,命名为HG1。水培条件下该突变体在光照强度大于29 μmol/（m2·s）的可见光下,根开始转红,在光照强度为180 μmol/（m2·s）的可见光下,呈鲜红,具有明显的光照敏感性。遗传分析表明,该突变体光照敏感性的红根性状由1对显性基因控制, 暂命名为Lsr。利用微卫星标记将Lsr基因定位在第4染色体上RM252与RM303之间,遗传距离分别为9.8 cM和6.4 cM, 这为Lsr基因的精细定位和克隆奠定了基础。     

一个水稻穗特异表达锌指蛋白基因的克隆与结构分析

利用生物信息学和RT PCR方法从水稻中克隆鉴定了一个新的具有TFⅢA型锌指结构的锌指蛋白基因,其开放阅读框为1092 bp,编码363个氨基酸残基。组织表达谱结果表明,该基因只在水稻幼穗组织中表达,在成株期的根、叶、茎以及花药中不表达,将其命名为OsZPT3 1。序列分析表明OsZPT3 1具有3个C2H2型锌指结构,在氨基酸的C端没有典型的转录抑制区域DLN box,但LXLXL的结构仍然表明OsZPT3 1可能是一个转录抑制因子。对OsZPT3 1启动子区域进行预测,结果发现3个MADS box转录因子识别位点,推测OsZPT3 1可能在MADS box转录因子的调节下,通过抑制下游基因的表达在水稻穗部器官的生长发育过程中发挥重要的调控作用。     

Traits Related to Chilling-Induced Photoinhibition in Leaves of indica and japonica Rice （Oryza sativa）

Physiological indices related to PS II photochemical efficiency (Fv/Fm) and membrane lipid peroxidation were measured in leaves of indica rice cv Shanyou 63 and japonica rice 9516 at different temperatures and light intensities for four days. No obvious changes in Fv/Fm and MDA were observed in both indica and japonica rice at moderate temperature and medium PFD, implying neither photoinhibition nor photooxidation happened in these cases. In indica rice either at medium temperature with higher PFD or at lower temperature with medium PFD Fv/Fm dropped obviously with no changes in MDA contents, and photoinhibition appeared while photooxidation did not occur. However, D1 protein, Fv/Fm, (A Z)/(A Z V), and SOD activities dropped, and O2 - production and MDA content increased accordingly, as well as both photoinhibition and photooxidation appeared in two rice varieties at lower temperature and higher PFD. Experiment with inhibitors at lower temperature and higher PFD showed that as compared with japonica rice the decrements appeared in D1 protein contents, SOD activities, and (A Z)/(A Z V) ratios, the xanthophyll cycle and non-photochemical quench (qN) were inhibited in a more degree, as well as increments of MDA content were greater, thus exhibiting more distinct photoinhibition and photooxidation in indica rice. It is suggested that Fv/Fm and membrane lipid peroxidation product-MDA were the key indices to predict and diagnose photooxidation.     

Traits Related to Chilling-Induced Photoinhibition in Leaves of indica and japonica Rice (Oryza sativa)

Physiological indices related to PS Ⅱ photochemical efficiency (Fv/Fm) and membrane lipid peroxidation were measured in leaves of indica rice cv Shanyou 63 and japonica rice 9516 at different temperatures and light intensities for four days. No obvious changes in Fv/Fm and MDA were observed in both indica and japonica rice at moderate temperature and medium PFD,implying neither photoinhibition nor photooxidation happened in these cases. In indica rice either at medium temperature with higher PFD or at lower temperature with medium PFD Fv/Fm dropped obviously with no changes in MDA contents, and photoinhibition appeared while photooxidation did not occur. However, D1 protein, Fv/Fm, (A Z)/(A Z V), and SOD activities dropped, and O2 -. production and MDA content increased accordingly, as well as both photoinhibition and photooxidation appeared in two rice varieties at lower temperature and higher PFD. Experiment with inhibitors at lower temperature and higher PFD showed that as compared with japonica rice the decrements appeared in D1 protein contents, SOD activities, and (A Z)/(A Z V) ratios, the xanthophyll cycle and non-photochemical quench (qN) were inhibited in a more degree, as well as increments of MDA content were greater, thus exhibiting more distinct photoinhibition and photooxidation in indica rice. It is suggested that Fv/Fm and membrane lipid peroxidation product-MDA were the key indices to predict and diagnose photooxidation.     

水稻斑点叶变异研究进展

大多数水稻斑点叶突变体是研究稻瘟病抗性、白叶枯病抗性以及植物程序性细胞死亡机理的理想材料。已经阐明了49份水稻斑点叶突变体的遗传模式,并成功克隆分离了若干斑点叶基因。综述了这些突变体的来源、遗传模式、基因定位和克隆,基因的功能以及它们对水稻主要病害的反应。     

BT型杂交粳稻育性及其三系的若干蛋白质标记

 用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）方法，对粳稻BT 细胞质雄性不育系六千辛A，保持系六千辛B，恢复系六千辛R、77302-1，以及杂交组合六千辛A/77302-1的F1和F2 种子的胚乳贮藏蛋白进行了分析。结果表明，在谷蛋白α3区域，恢复系有两条带α3a 和α3b,而六千辛A和六千辛B只有一条带α3。 F2代具有α3的种子和具有α3a 加α3b的种子1∶1分离。谷蛋白α4带的移动速率，恢复系比六千辛A快。把较快的α4带记为α4f。 F2代具有α4的种子和具有α4 加α4f的种子也是1∶1分离，与配子体不育类型的F2代花粉育性恢复基因分离比一致。系谱分析表明六千辛R中α3a 和α3b来源于IR8。六千辛A比六千辛B容易提取醇溶蛋白。     

测定水稻硅含量的一种简易方法

 介绍了一种简易的测量水稻硅含量的方法，其主要特点为准确、方便、能批量化测定水稻硅含量。以珍汕97B/密阳46重组自交系群体，两次重复，共498个株系的谷壳、剑叶和茎秆为材料，验证了该方法的可靠性。每份材料测定2份样品，两次测定结果达极显著正相关（P<0.01），线性相关系数分别为0.97954、0.97026和0.98848。取10份代表性样品进行了高温碱熔解法的测定，两种方法测定结果高度一致， r = 0.9993。     

Genetic Analysis and Mapping of a Thermo-sensitive White Stripe-Leaf Mutant at Seedling Stage in Rice (Oryza sativa)

A thermo-sensitive white stripe-leaf mutant(tws)was selected from the M2 progeny of a japonica variety,Jiahua 1, treated by 60 Coγ-radiation.In comparison with the wild type parent,the mutant displayed a phenotype of white stripe on the 3rd and 4th leaves,but began to turn normal green on the 5th leaf when grown at low temperatures(20°C and 24°C). Furthermore,the content of total chlorophyll showed an obvious decrease in the leaves with white stripe.These results suggest that the expression of the mutant trait was thermo-sensitive and correlated with the leaf age of seedlings.The genetic analysis indicated that the mutant trait was controlled by a single recessive nuclear gene,designated as tws.In addition,by using SSR markers and an F2 segregating population derived from the cross between the tws mutant and 9311, tws was mapped between the markers MM3907 and MM3928 with a physical distance of 86 kb on rice chromosome 4.     

萍乡显性核不育水稻幼穗发育时期可溶性蛋白质含量的变化研究

对萍乡显性核不育水稻纯合不育株、杂合不育株与隐性可育株3种不同基因型植株幼穗发育期的叶片、幼穗和花药中的可溶性蛋白质含量变化进行了比较分析。结果表明，显性核不育水稻3种不同基因型植株在幼穗发育时期叶片中的可溶性蛋白质含量差异不大，且变化趋势相同；不育株(纯合与杂合)幼穗中可溶性蛋白质的含量在雌雄蕊形成期与可育株基本相同，而在花粉母细胞形成期和花粉母细胞减数分裂期迅速下降；不育株(纯合与杂合)花药内可溶性蛋白质含量明显低于可育株，这是不育株花粉败育的一个重要生理特征。     

Transgenic Expression of the Recombinant Phytase in Rice （Oryza sativa）

In many plants, phytic acid (phytate, 1, 2, 3, 4, 5, 6-hexakisphosphate) is one of the main storage forms of phosphate. About 80% of phosphorus (P) in cereal plants, including rice is stored as phytic acid [1-2]. P in phytic acid can’t be utilized by monogastric animals including human, while it was estimated that only 1/3 of the total P in most of the vegetal feedstuff could be efficiently utilized by the livestock. Therefore, for animal feed with P supplementation is expected to meet the d…     

孕穗期水稻不同功能叶的发育蛋白质组学分析

 从蛋白质组学角度对杂交水稻汕优63孕穗期4个叶位功能叶片蛋白质组变化进行了研究,以揭示水稻孕穗期叶片蛋白质组的表达差异。4个不同时期功能叶片蛋白质经双向电泳得到分离,应用Imagemaster 2D Elite 5.0软件对所得到的双向电泳图谱进行分析比较,共得到差异表达蛋白质点23个,差异表达蛋白质点经质谱分析,有14个得到鉴定。对鉴定出的蛋白功能进行研究,结果表明:差异表达的蛋白质大部分(9个)为参与光合作用和呼吸作用的蛋白质;其次为与蛋白结构及功能调控相关的蛋白质(3个),表达量表现为随叶位上升而上调;其余为参与氨基酸代谢的蛋白质(2个),表达量在较低叶位叶片中无显著规律,在剑叶中的表达量显著上调。蛋白质是细胞功能执行者,受细胞功能或细胞环境变化的影响,因此,上述蛋白质发生变化的原因应与水稻不同叶位叶片生长发育特性有关。     

Morphological Structure and Genetic Mapping of New Leaf-Color Mutant Gene in Rice (Oryza sativa)

Leaf-color mutations are a widely-observed class of mutations, playing an important role in the study of chlorophyll biosynthesis and plant chloroplast structure, function, genetics and development. A naturally-occurring leaf-color rice mutant, Baihuaidao 7, was analyzed. Mutant plants typically exhibited a green-white-green leaf-color progression, but this phenotype was only expressed in the presence of a stress signal induced by mechanical scarification such as transplantation. Prior to the appearance of white leaves, mutant plant growth, leaf color, chlorophyll content, and chloroplast ultrastructure appeared to be identical to those of the wild type. After the changeover to white leaf color, an examination of the mutated leaves revealed a decrease in total chlorophyll, chlorophyll a, chlorophyll b, and carotenoid content, a reduction in the number of chloroplast grana lamella and grana, and a gradual degradation of the thylakoid lamellas. At maturity, the mutant plant was etiolated and dwarfed compared with wild-type plants. Genetic analysis indicated that the leaf mutant character is controlled by a recessive nuclear gene. Genetic mapping of the mutant gene was performed using an F2 population derived from a Baihuaidao 7 × Jiangxi 1587 cross. The mutant gene was mapped to rice chromosome 11, positioned between InDel markers L59.2-7 and L64.8-11, which are separated by approximately 740.5 kb. The mutant gene is believed to be a new leaf-color mutant gene in rice, and is tentatively designated as gwgl.     

Expression Profiles and Protein Complexes of Starch Biosynthetic Enzymes from White-Core and Waxy Mutants Induced from High Amylose Indica Rice

Physicochemical properties of endosperm starches in milled rice determine its cooking and eating quality. Amylose is synthesized by granule-bound starch synthase I (GBSSI), whilst amylopectin is synthesized by the synergistic activities of starch synthases (SSs), branching enzymes (BEs) and debranching enzymes (DBEs). However, the complexes formed by starch biosynthetic enzymes are not well characterized. Gene expression profiles and protein complexes were determined in white-core (GM645) and waxy (GM077) mutants derived from a high amylose indica rice Guangluai 4 (GLA4). In GM645, genes including AGPS1, GBSSI, SSIIa, BEI, BEIIa, BEIIb, PUL, ISA1 and SP were significantly downregulated during seed development. In GM077, the expression levels of AGPL2, AGPS1, AGPS2b, SSIIIa, BEI, PUL and ISA1 were significantly upregulated. Co-immunoprecipitation assays revealed interactions of SSs-BEs, SSs-PUL and BEs-PUL in developing seeds. However, weak SSI-SSIIa interaction was detected in GM077, whilst SSI-PUL interaction was absent. Weak interaction signals for SSI-SSIIa, SSIIa-BEI, SSIIa-BEIIb, BEI-BEIIb and SSI-BEI were also observed in GM645. These results suggest that the protein-protein interactions for starch biosynthesis are modified in mutants, which provides insight into the mechanisms of starch biosynthesis, particularly in indica rice.