亚洲栽培稻5种不同生态类型与籼粳间的杂种F1育性研究

为全面系统地研究亚洲栽培稻种内不同生态类型材料与籼、粳亚种杂交育性，对19份包含亚洲栽培稻种内5种不同生态类型品种与云南温带粳品种滇粳优1号及国际水稻所选育的籼稻品种IR64的杂种F₁育性进行分析.结果表明，亚洲栽培稻不同类型中，温带粳与籼亚种间普遍存在杂种不育，温带粳与热带粳间、籼籼部分组合也存在杂种不育，Aus类型与籼、粳亚种的杂种不育程度与品种相关.研究结果为系统研究亚洲栽培稻不同生态类型与籼、粳间杂种不育奠定了基础，为认识亚洲栽培稻的分化提供证据，并为利用亚洲栽培稻种内丰富的遗传变异来培育新品种、新组合提供理论指导.     

籼-粳稻特异插入／缺失分子标记揭示的稻属植物遗传分化

籼-粳分化在亚洲栽培稻(Oryza sativa L.)的驯化过程中非常重要,但其分化机制仍不清楚.有的学者认为水稻籼-粳分化是亚洲栽培稻在驯化过程中适应不同生境的结果,也有人认为籼-粳分化在水稻的野生祖先种中就已经存在.为了研究普通野生稻的籼-粳分化,并解析稻属(Oryza)植物的籼-粳遗传变异,利用34对籼-粳特异插入/缺失分子标记(InDel)引物,研究了50份典型籼稻(O.sativa L.subsp.Indica Kato)和粳稻(O.sativa L.subsp.japonica Kato)样本以及来源于35个国家的348份稻属其他种材料.结果表明,亚洲栽培稻存在明显的籼-粳分化,普通野生稻复合体(O.rufipogon complex)中存在"偏籼"和"偏粳"类型,而稻属的其他种均未出现籼-粳分化,普通野生稻复合体中"偏籼"和"偏粳"类型的地理分布格局与籼稻和粳稻的地理分布格局相吻合.考虑到大部分普通野生稻复合体的样本取自邻近有栽培稻种植的普通野生稻群体,推测得出部分普通野生稻样本中表现出的"偏籼"和"偏粳"类型可能是栽培稻的籼稻品种和粳稻品种在普通野生稻的长期协同进化过程中基因交流的结果.     

石峡遗址M104古稻稃壳印痕研究

水稻稃壳双峰乳突,纵列横行排列有序,是鉴别籼、粳亚种的主要性状之一;在缺乏古稻谷的情况下,稻壳印痕也不失一种鉴别古稻的方法。用此方法对100片石峡遗址M104古稻稃壳双峰乳突印痕的研究,确认石峡古稻是一个处于籼、粳分化,属性模糊的杂合种群,是古稻演化历史阶段的反映。石峡古稻所呈现的籼、粳模糊性和分化状态特征,表明石峡古稻是我国栽培稻起源与发展史上承上启下的重要一环。     

生态条件对籼粳稻杂交后代亚种特性与经济性状的影响

以两个典型的籼粳交重组自交系F6群体(RILSA:中优早8号/丰锦、RILSB:七山占/秋光)为试材,分别在四川和辽宁同年种植,研究亚种特性和经济性状变化规律及其相互关系的结果表明,不同生态条件下籼粳交后代的亚种特性和经济性状都发生了显著的变化.生态条件对程氏指数和构成性状的影响因群体和性状而异,RILSA群体在四川和辽宁都呈偏粳分布,RILSB群体在辽宁接近正态分布,在四川呈偏粳分布,总体上四川表现比辽宁更加偏粳.与辽宁相比,四川穗数、结实率和千粒重降低,因而产量极显著下降,而穗粒数在不同地区和不同群体间差异不显著.程氏指数和构成性状与产量和经济性状有正相关的趋势,据此进行人工选择可能是北方稻区籼粳稻杂交育成品种仍然基本保持粳亚种特性的重要原因.此外,还讨论了籼粳分化与生态条件的关系等相关问题.     

籼稻93-11和粳稻日本晴DNA插入缺失差异片段揭示的水稻籼-粳分化

碱基的插入/缺失(InDel)引起DNA序列变化并形成了DNA片段长度多态,可以用作遗传标记.为了评价基于籼稻93-11和粳稻日本晴全基因组序列比对获得的差异片段而设计的InDel引物在鉴定籼稻和粳稻两种生态型以及研究稻属不同物种之间亲缘关系的意义,采用45对InDel引物,对来自亚洲10个国家的49份籼稻、43份粳稻品种和24份野生稻进行了检验.结果表明,其中41对InDel引物鉴定籼稻或粳稻品种的准确率高于80%.主成分分析散点图显示:籼稻与粳稻存在明显的遗传分化;含AA基因组的野生稻物种与籼稻品种存在较近的亲缘关系;非AA基因组的野生稻物种不存在明显的籼-粳分化.并且证明了基于籼稻93-11和粳稻日本晴全基因组序列比对获得的InDel差异片段设计的引物可以用于栽培稻籼稻和粳稻品种的鉴定以及籼-粳分化问题的研究,及探索稻属不同物种间的亲缘关系.     

云南改良稻及元阳地方稻磷高效研究

 在土壤有效磷质量比分别为0.26mg·kg^-1和80.0mg·kg^-1条件下对云南111份改良稻和77份元阳地方稻进行形态性状与程氏指数间的磷高效研究,以明确籼粳分化和米色与磷高效间相互关系,对比改良稻和地方稻间磷高效差异.结果表明:①相对地上干物质量、相对地下干物质量、相对总干物质量和相对分蘖力4个耐低磷指标性状可作为水稻耐低磷能力强弱评价的理想指标,能明确反映出磷高效能力强弱;②元阳地方稻各亚种间平均耐低磷能力均明显高于云南改良稻,且差异明显,4个耐低磷指标性状平均值为籼>偏籼>偏粳>粳,无色米>有色米,耐低磷能力随程氏指数的增大呈递减趋势;③总体而言,程氏指数和耐低磷指标性状间多数呈显著负相关,磷高效特性和籼粳分化密切相关.     

籼稻和粳稻的高效分子鉴定方法及其在水稻育种和进化研究中的意义

为了建立准确高效的籼稻和粳稻鉴定方法,对基于籼稻(93-11)和粳稻(日本晴)的全基因组DNA序列比对而获得的45个特异插入/缺失(InDcl)位点进行了实验验证.以包括93-11和日本睛在内的44个典型籼稻和典型粳稻品种为实验群体,用45对InDel引物对上述水稻品种的DNA样品进行了PCR扩增和聚丙烯酰胺凝胶电泳,获得了多态的电泳条带.对获得的各InDel位点的基因型数据矩阵进行了中性检测(neutrality test),确定了与栽培稻的籼、粳遗传分化密切相关的34个InDel位点.进一步对来自亚洲11个国家的栽培稻品种和来源不同12个野生稻物种的PCR产物和电泳结果的读取和分析,计算这些栽培稻品种和野生稻DNA样品在这34个InDel位点上的籼型或粳型基因频率,最终确定了不同样品的籼、粳特性.该籼稻和粳稻鉴定方法被称为"InDel分子指数法".与传统基于形态特征鉴定栽培稻籼、粳特性的"程氏指数法"相比,该方法不仅能够准确鉴定籼稻和粳稻,而且还具有更快捷、简便和高效的特点.另外,InDel分子指数法还可以用于野生稻样品的籼、粳特性鉴定,扩大了被检测样品的范围,具有广阔的应用前景.InDel分子指数法的建立为栽培稻育种过程中正确选用籼稻或粳稻种质资源提供了新的技术方法,也为栽培稻的起源、籼-粳遗传分化、以及籼稻和粳稻在驯化过程中如何适应地理环境变化提供了新的研究思路.     

基于ISSR指纹标记的亚洲栽培稻与野生稻遗传多样性分析

为评价野生稻与亚洲栽培稻的遗传多样性及其变异关系,21对SSR引物被用于研究广泛地理分布的17份野生稻(其中11份O. rufipogon和6份O. nivara)和22份亚洲栽培稻(13份indica和9份japonica)样本.271条多态性片段被扩增出,占总扩增片段的96.10%.野生稻多态性片段的百分比(平均达64.30%)及Nei's遗传多样性值(GD)明显高于亚洲栽培稻,表明野生稻比亚洲栽培稻具更丰富的遗传多样性.UPGMA聚类分析显示野生稻的两个类群(O. rufipogon和O. nivara)关系密切,但在遗传上存在明显的分化,支持其作为两个独立物种的分类观点.野生稻中籼粳分化不很明显,亚洲栽培稻的籼粳亚种分化明显.亚洲栽培稻与多年生普通野生稻(O. rufipogon)关系更为密切,符合异源起源的遗传分化模式.     

籼粳我稻米品质性状恙农艺性状之间的遗传协方差分析

用２个籼型裟光温敏核不育系和７个粳型广亲和品种为材料，按ＮＣⅡ设计配制杂交组合，获得亲本、Ｆ１及Ｆ２代的籽粒群体，对８个主要品质性状和６个农艺性状进行测定，按混合线性模型的分析方法对品质性状与农艺之间的遗传方差进行了研究，结果表明：在籼粳亚种间杂交组合中，稻为理化品质性状与植株农艺性状之间遗传协方差大都不直接加性／母体加性协方差，外观品质性状与农艺性状之间的遗传协方差则主要是直接加性／母体加性协方     

籼稻和粳稻的高效分子鉴定方法及其在水稻育种和进化研究中的意义

为了建立准确高效的籼稻和粳稻鉴定方法,对基于籼稻（93—11）和粳稻（日本睛）的全基因组DNA序列比对而获得的45个特异插入／缺失（InDel）位点进行了实验验证．以包括93—11和日本晴在内的44个典型籼稻和典型粳稻品种为实验群体,用45对InDel引物对上述水稻品种的DNA样品进行了PCR扩增和聚丙烯酰胺凝胶电泳,获得了多态的电泳条带．对获得的各InDel位点的基因型数据矩阵进行了中性检测（neutrality test）,确定了与栽培稻的籼、粳遗传分化密切相关的34个InDel位点．进一步对来自亚洲11个国家的栽培稻品种和来源不同12个野生稻物种的PCR产物和电泳结果的读取和分析,计算这些栽培稻品种和野生稻DNA样品在这34个InDel位点上的籼型或粳型基因频率,最终确定了不同样品的籼、粳特性．该籼稻和粳稻鉴定方法被称为“InDel分子指数法99与传统基于形态特征鉴定栽培稻籼、粳特性的“程氏指数法”相比,该方法不仅能够准确鉴定籼稻和粳稻,而且还具有更快捷、简便和高效的特点．另外,InDel分子指数法还可以用于野生稻样品的籼、粳特性鉴定,扩大了被检测样品的范围,具有广阔的应用前景．InDel分子指数法的建立为栽培稻育种过程中正确选用籼稻或粳稻种质资源提供了新的技术方法,也为栽培稻的起源、籼-粳遗传分化、以及籼稻和粳稻在驯化过程中如何适应地理环境变化提供了新的研究思路．     

Spacer length variation in rice 5S rRNA genes revealed by polymerase chain reaction

Polymerase chain reaction (PCR) was used to amplify 5S rRNA spacer from wild rice (Oryza rufipogon and O. nivara) and cultivated rice (indica and japonica varieties of O. sativa L). The results show that there is spacer length variation within and between species, and the typical indica and japonica varieties have their unique banding patterns of amplified 5S rRNA spacers, whereas intermediate showed no specific amplification profile of spacer regions. The 5S rRNA genes in intermediate are either identical with that of indica variety or that of japonica variety. These data suggest that the spacer length polymorphisms can be used to distinguish between closely ralated species and subspecies. Supported by the National Natural Science Foundation of China Yi Qingming, born in Apr. 1938, Professor     

Efficient indica and japonica rice identification based on the InDel molecular method: Its implication in rice breeding and evolutionary research

An efficient molecular method for the accurate and efficient identification of indica and japonica rice was created based on the poly-morphisms of insertion/deletion (InDel) DNA fragments obtained from the basic local alignment search tool (BLAST) to the entire genomic sequences of indica (93-11) and japonica rice (Nipponbare). The 45 InDel loci were validated experimentally by the polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE) in 44 typical indica and japonica rice varieties, including 93-11 and Nipponbare. A neutrality test of the data matrix generated from electrophoretic banding patterns of various InDel loci indicated that 34 InDel loci were strongly associated with the differentiation of indica and japonica rice. More extensive analyses involving cultivated rice varieties from 11 Asian countries, and 12 wild Oryza species with various origins confirmed that indica and japonica characteristics could accurately be determined via calculating the average frequency of indica- or japonica-specific alleles on different InDel loci across the rice genome. This method was named as the "InDel molecular index" that combines molecular and statistical methods in determining the indica and japonica characteristics of rice varieties. Compared with the traditional methods based essentially on morphology, the InDel molecular index provides a very accurate, rapid, simple, and efficient method for identifying indica and japonica rice. In addition, the InDel index can be used to determine indica or japonica characteristics of wild Oryza species, which largely extends the utility of this method. The InDel molecular index provides a new tool for the effective selection of appropriate indica or japonica rice germplasm in rice breeding. It also offers a novel model for the study of the origin, evolution, and genetic differentiation of indica and japonica rice adapted to various environmental changes.     

水稻花药发育的细胞形态学精细分期

本研究通过观察籼型杂交水稻"川优9527"的花粉和药室壁的发育,对幼穗花药从雄蕊原基形成至花粉成熟的全过程进行了分期。选用不同发育时期的水稻幼穗(长度约1.0~10.0 mm),以石蜡切片技术分别制作其花药装片,置于光学显微镜(N ikon DS-R i1-U2)下观察并拍照。根据细胞及组织形态特征的显著变化,将花药的发育进程精细划分为:孢原细胞期、二层药室壁期、三层药室壁期、花粉母细胞形成期(以上4期对应小孢子母细胞形成期)、花粉母细胞减数分裂初期、花粉母细胞减数分裂中期、花粉母细胞二分体时期、花粉母细胞四分体时期(以上4期对应小孢子母细胞减数分裂期)、小孢子早期、小孢子中期、小孢子晚期、二胞花粉时期、成熟花粉时期等13个阶段。     

控制水稻剑叶形态相关性状的数量基因位点(QTL)的定位

水稻籽粒中一半以上的碳水化合物来自剑叶的光合作用，剑叶形态改良一直是水稻株型育种的一个重要目标．利用一个日本主要种植的粳稻品种越光（轮回亲本）和一个印度的籼稻品种Kasalath杂交产生的回交重组自交系群体（backcross recombinant inbred lines，BILs）对剑叶形态中的3个主要性状（剑叶长、叶宽以及其叶面积）进行了相关分析及其数量基因位点（quantitative trait loci，QTL）的定位．研究表明，控制剑叶形态的3个主要性状间存在极显著的正相关，并检测到影响3个性状的8个QTL，分布在第1，3，4，6条染色体上，贡献率介于4．94％～22．07％，其中第4染色体上C1016标记和第6染色体上C556标记附近的共有6个QTL，其两侧的紧密分子标记在水稻株型分子育种上具有一定应用价值．     

An active DNA transposon family in rice

The publication of draft sequences for the two subspecies of Oryza sativa (rice), japonica (cv. Nipponbare) and indica (cv. 93-11), provides a unique opportunity to study the dynamics of transposable elements in this important crop plant. Here we report the use of these sequences in a computational approach to identify the first active DNA transposons from rice and the first active miniature inverted-repeat transposable element (MITE) from any organism. A sequence classified as a Tourist-like MITE of 430 base pairs, called miniature Ping (mPing), was present in about 70 copies in Nipponbare and in about 14 copies in 93-11. These mPing elements, which are all nearly identical, transpose actively in an indica cell-culture line. Database searches identified a family of related transposase-encoding elements (called Pong), which also transpose actively in the same cells. Virtually all new insertions of mPing and Pong elements were into low-copy regions of the rice genome. Since the domestication of rice mPing MITEs have been amplified preferentially in cultivars adapted to environmental extremes-a situation that is reminiscent of the genomic shock theory for transposon activation.     

Production of bacterial blight resistant lines from somatic hybridization between Oryza sativa L. and Oryza meyeriana L

Novel bacterial blight (BB) resistance gene(s) for rice was (were) introduced into a cultivated japonica rice variety Oryza sativa (cv. 8411), via somatic hybridization using the wild rice Oryza meyeriana as the donor of the resistance gene(s). Twenty-nine progenies of somatically hybridized plants were obtained. Seven somatically hybridized plants and their parents were used for AFLP (amplified fragment length polymorphism) analysis using 8 primer pairs. Results confirmed that these plants were somatic hybrids containing the characteristic bands of both parents. The morphology of the regenerated rice showed characters of both O.sativa and O.meyeriana. Two somatic hybrids showed highest BB resistance and the other 8 plants showed moderate resistance. The new germplasms with highest resistance have been used in the rice breeding program for the improvement of bacterial blight resistance.     

农杆菌介导转化水稻成熟胚影响因素研究

实验对蜀恢881、蜀恢527、中花9号等7个水稻品种的成熟胚为受体材料，研究了影响农杆菌转化水稻频率的几个重要因素，实验结果表明：CC培养基是籼稻成熟胚愈伤组织的最适诱导培养基，而NB培养基是粳稻品种成熟胚的最适诱导培养基；诱导培养基中添加ABA并不能显著提高籼稻和粳稻成熟胚愈伤组织的诱导率，但在ABA为3mg／LN可以明显改善愈伤质量；适当的低温预处理可以不同程度提高诱导率；农杆菌菌株EHA105对水稻愈伤组织的转化能力优于LBA4404和AGL1；在分化时，采用适宜的激素配比有利于提高分化率。     

不同水稻品种种子发芽耐盐性研究

以江苏籼稻9311、湖南粳稻HN-1、豫粳6号、豫粳8号4个水稻品种为实验材料,进行种子发芽耐盐性比较实验,并分别测其种子的发芽率、发芽指数、活力指数、苗长、电导率以及POD(过氧化物酶)比活力,分析不同品种间幼苗耐盐性的差别.结果表明:4个品种中江苏籼稻9311的耐盐性最强,豫粳8号次之,再次为湖南粳稻HN-1,而豫粳6号最弱.     

Identification of salt-tolerance QTL in rice (Oryza sativa L.)

Quantitative trait loci (QTLs) controlling salt-tolerance at the seedling stage in rice (Oryza sativa L.) were identified by interval mapping (SIM) and composite interval mapping (CIM) using a doubled haploid population ZJDH and its high resolution genetic linkage map. The population was derived from an inter-subspecific cross between an indica variety Zhaiyeqing8 (ZYQ8) and a japonica variety Jingxi17 (JX17). Analysis of survival days of seedlings treated with 0.7% NaCI revealed that a major salt-tolerance quantitative trait locus (QTL), Std, was present between markers RG612 and C131 on chromosome 1 when using both MAPMAKER/QTL 1.1 and PLABQTL 1.0 (SIM). Its allele which contributes to salt-tolerance was from ZYQ8. In addition, seven more QTLs which give additive effect on salt-tolerance are identified when using PLABQTL (CIM), and most of them were from JX17.