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.     

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.     

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.     

溶出固形物对软米硬度和黏弹性的影响

以粳米和糯米为对照,研究了溶出固形物对软米硬度和黏弹性的影响。分析发现软米溶出固形物质量分数为11.85%,显著高于粳米和糯米(P<0.05),与弹性显著正相关(P<0.05)；与粳米和糯米相比,软米中的固形物直支链淀粉比最低,利于形成连续凝胶网结构。此外,软米溶出淀粉侧链中聚合度为6~12的短链A链比例最高,赋予其较好的黏弹性。质构和流变分析结果表明:除溶出固形物后,软米的硬度显著增加,黏硬比和弹性显著降低。扫描电子显微镜结果表明:除溶出固形物后米粒表面粗糙、凹凸不平,有明显的孔洞；软米表面所形成的渔网状孔洞状结构能够对饭粒起到支撑作用,使得整个饭粒弹性更好。溶出固形物含量、组成、分子结构及其在米饭表面分布的差异是软米较低硬度和较好黏弹性优势产生的主要原因。     

基于InDel分子标记的20个水稻品种的籼粳属性分析

水稻品种的籼粳属性分析是培育高产优质杂交水稻亲本选择的重要依据之一。本研究以93-11与日本晴分别为籼粳稻参考标准,利用均匀分布于水稻基因组12条染色体并具有较高籼粳特异性的37对In Del分子标记,对20份水稻育种材料进行In Del标记的籼粳属性分析。结果表明:所选取的20份水稻材料中,有8份为典型籼稻,3份为典型粳稻,4份为偏籼类型,另有5份为偏粳类型。NTsys聚类分析结果显示,在遗传相似系数为0.43处,20份水稻材料被分为两大类群;在遗传相似数为0.70处,所有水稻材料则被分为4个类群。     

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     

Genetic analysis and fine mapping of an enclosed panicle mutant esp2 in rice （Oryza sativa L.）

The phenomenon of panicle enclosure in rice is mainly caused by the shortening of uppermost internode.Elucidating the molecular mechanism of panicle enclosure will be helpful for solving the problem of panicle enclosure in male sterile lines and creating new germplasms in rice.We acquired a monogenic recessive enclosed panicle mutant,named as esp2 (enclosed shorter panicle 2),from the tissue culture progeny of indica rice cultivar Minghui-86.In the mutant,panicles were entirely enclosed by flag leaf sheaths and the uppermost internode was almost completely degenerated,but the other internodes did not have obvious changes in length.Genetic analysis indicated that the mutant phenotype was controlled by a recessive gene,which could be steadily inherited and was not affected by genetic background.Apparently,ESP2 is a key gene for the development of uppermost internode in rice.Using an F 2 population of a cross between esp2 and a japonica rice cultivar Xiushui-13 as well as SSR and InDel markers,we fine mapped ESP2 to a 14-kb region on the end of the short arm of chromosome 1.According to the rice genome sequence annotation,only one intact gene exists in this region,namely,a putative phosphatidylserine synthase gene.Sequencing analysis on the mutant and the wild type indicated that this gene was inserted by a 5287-bp retrotransposon sequence.Hence,we took this gene as a candidate of ESP2.The results of this study will facilitate the cloning and functional analysis of ESP2 gene.     

一个水稻白化致死突变体abl25鉴定及其基因定位

经Co60辐照的粳稻嘉花1号得到一个新的致死白化突变体albino lethal 25(abl25),该突变体从发芽至4叶期表现为白化苗,之后逐渐死亡.与野生型嘉花1号相比,abl25突变体的叶绿素含量和类胡萝卜素的含量大大降低,叶绿体结构不正常,说明其叶绿体发育受到严重阻碍,导致植物死亡.遗传分析表明:该突变体受一对隐性核基因(abl25)控制,进一步利用abl25与广占63S杂交的F2分离群体,将该突变体基因(abl25)定位于第2染色体上SSR标记RM424与Indel分子标记ID7330之间,随后利用新开发的分子标记和扩大群体将其定位在Indel分子标记ID9111和ID9261之间的150 kb内,发现abl25是一个新的水稻苗期白化致死基因.     

平安竹纤维形态变异特征及其成因分析

【目的】阐明矢竹矮秆稳定变异体——平安竹纤维形态学特征并初步阐释其变异机制。【方法】利用石蜡切片及数量统计学方法进行显微观测与数据分析; qRT-PCR分析基因相对表达量; UPLC/GC-MS法测定赤霉素含量。【结果】平安竹竹秆不同部位纤维长度、长宽比及壁腔比显著小于矢竹; 但其宽度与腔径值显著大于矢竹; 而其不同部位纤维壁厚值虽小于矢竹,但差异不显著。平安竹地下茎纤维在长度、长宽比、壁厚及壁腔比上显著小于矢竹,但其宽度与腔径值则显著大于矢竹。形态解剖学分析显示,平安竹地下茎纤维细胞形态变异出现于节间活跃生长阶段。赤霉素及细胞壁与细胞骨架相关基因表达量分析结果表明,赤霉素信号途径在平安竹节间生长起始阶段即受到抑制; 而其细胞壁与细胞骨架相关下游功能基因则在其伸长早期与活跃阶段显著低于矢竹; 赤霉素含量分析也证实各赤霉素含量早在平安竹节间伸长起始阶段就低于矢竹。【结论】赤霉素信号途径受抑导致其下游细胞壁与细胞骨架组织相关基因下调是最终导致平安竹纤维细胞生长异常的原因之一。     

利用分子标记分析22种上海市重要水稻10个食味品质相关基因的基因型

稻米食味品质改良是目前水稻育种的重要研究方向之一.明确不同水稻食味品质基因型,可为优质水稻育种研究的亲本选择提供依据.本研究利用分子标记检测技术,对22个目前上海市种植的重要常规水稻和杂交稻亲本食味品质基因的基因型进行检测.结果发现,"紫香糯861"水稻含有相对较多不良食味品质基因型.3种杂交稻父本,"湘晴"、"R44"和"繁14"的食味品质基因型比这3种杂交稻母本,"寒风A"、"秋风A"和"申9A"以及其他常规稻有欠缺.从蜡质基因多态性分析显示,在本研究选用的22种水稻中,2个为糯稻、20个为粳稻,但其中有4个为软米型粳稻;在16个非软米型粳稻中,"宝农34"、"金丰"和"银香18"是具有相对最好食味品质基因型的水稻品种.     

水稻栽培稻分类研究的现状与展望

简述了稻属、栽培稻种分类研究的现状,我国栽培稻种主要分类系统及籼、粳亚种的鉴别方法.全世界稻属植物的种共归纳为22个种,分属9个染色体组;栽培稻在亚种分级上主要可分为籼、粳两亚种或籼、粳和爪哇三个亚种,多数学者倾向于只分籼、粳两个亚种;亚种以下可分为生态群、生态型、品种或生态群、生态型、变种和品种,生态群、生态型的性状和指标不一,尚未统一;在分类方法上主要以形态特征,杂交亲和力,同工酶和植物学性状综合数值等来划分,而籼粳分子水平上的鉴别与分类也开始得到应用.栽培稻亚种间亲缘关系、亲和性及其与杂种优势的关系仍然是栽培稻分类研究的主要方向.     

两个水稻叶片反向卷曲突变体的表型及遗传分析

为了揭示水稻叶片卷曲的形态建成,分析了水稻Ac/Ds转座子插入突变体库中的2个叶片反向卷曲突变体的表型及组织形态.研究发现,与野生型平展叶片相比,内卷突变体和外卷突变体叶片中泡状细胞数目明显减少,这可能是造成水稻叶片卷曲的重要原因.另外,内卷突变体叶片主脉薄壁细胞少,薄壁细胞崩裂形成的气腔面积较大.对2个卷叶突变体的纤维素含量进行测定发现,内卷突变体叶片及茎杆中纤维素含量明显减少,而外卷突变体叶片及茎杆的纤维素含量与野生型没有显著差异,说明2个卷叶突变体叶片的卷曲可能由不同基因突变造成.分析了2个水稻卷叶突变体的遗传规律,结果表明,二者均由隐性单基因控制.     

分光光度法检测不同地区和类型稻种功能性成分的初步研究

 为研究不同地区和类型水稻品种糙米中γ-氨基丁酸和总黄酮等功能性成分,本实验采用分光光度法准确测定了中国、日本、韩国、国际稻作所和云南品种及云南野生稻γ-氨基丁酸和总黄酮的含量.结果表明:① 用分光光度法测定水稻籽粒中γ-氨基丁酸和总黄酮含量的方法是可靠的;② 不同地区和类型水稻品种功能成分含量存在差异;③ 相关分析表明,氨基丁酸与总黄酮的相关系数较高(r=0.575^**),在育种上可以实现多个相关性状间的遗传改良;④ 鉴定出墨江黄皮饵块谷、明恢86、墨江饵块谷、疣粒野生稻等18份高GABA和高总黄酮的水稻品种,可作为选育高黄酮和γ-氨基丁酸品种的亲本或直接作为功能稻开发利用.首次报道疣粒野生稻总黄酮质量分数达0.751%,在所有水稻品种中黄酮含量最高.     

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.     

Characteristics of photosynthesis in rice plants transformed with an antisense Rubisco activase gene

Transgenic rice plants with an antisense gene inserted via Agrobacterium tumefaciens were used to explore the impact of the reduction of Rubisco activase (RCA) on Rubisco and photosynthesis. In this study, transformants containing 15% to 35% wild type Rubisco activase were selected, which could survive in ambient CO2 concentration but grew slowly compared with wild type controls. Gas exchange measurements indicated that the rate of photosynthesis decreased sig-nificantly, while stomatal conductance and transpiration rate did not change; and that the intercellular CO2 concentration even increased. Rubisco determination showed that these plants had approximately twice as much Rubisco as the wild types,although they showed 70% lower rate of photosynthesis, whichRubsico activase and/or the reduction in carbamylation.was likely an acclimation response to the reduction in Rubsico activase and/or the reduction in carbamylation.     

水稻OsNAP基因的功能分析及其在育种价值方面的研究

NAC转录因子是植物特异的转录因子.本研究通过从水稻日本晴(Oryza sativa L.)的基因组中克隆到一个与拟南芥衰老相关的基因AtNAP(Atlg69490)的同源基因OsNAP.OsNAP的ORF为1 179bp,推测其编码蛋白含有392个氨基酸,等电点为8.55,分子质量为42.195ku.为进一步研究OsNAP的功能,利用显性抑制元件SRDX构建其pCAMBIA1304-35S:OsNAP-SRDX表达载体,通过农杆菌浸染方法将其转化水稻中花11品种.T1代转基因植株通过潮霉素、RealtimePCR、和Western blot鉴定,筛选出4个OsNAP高表达的阳性植株.在种子成熟期,观察到T1代转基因4号植株较野生型相比有明显的延缓衰老的表型,其他农艺性状如单株分蘖数、单株总粒数、单穗饱和穗粒数、千粒重、结实率等都优于野生型.实验证明抑制OsNAP基因的表达能够延缓水稻叶片衰老并可以提高产量,具有潜在的育种价值.     

野生稻材料中氨基酸、碳、氮含量分析及其在种质资源评价中的应用

 野生稻是宝贵的遗传资源库.云南有较丰富的野生稻资源,研究对云南及其它来源的野生稻和栽培稻茎、叶和稻米中氨基酸含量及碳、氮含量进行了分析.结果表明:所涉及的几种材料中茎叶内总氨基酸质量比差异较大,小粒野生稻最高56.8 mg/g,野生的元江普通野生稻最低22.8 mg/g.换算的C/N和蛋白质质量分数变化范围分别为12~38和6%~20%.对野生稻茎、叶和相应的稻米中氨基酸总量及氨基酸组成间的相关性进行了分析,发现两者具有显著相关性.并初步探讨了茎、叶中氨基酸含量在评价植物种质资源评价中的应用.     

利用广亲和品种提高“三系”杂交稻优势的探讨

对02428、cpslo17、培矮64在三系杂交稻中的利用进行分析,结果表明,亲和力高低依次为cpslo17、培矮64、02428。cpslo17、培矮64含有对籼粳不育系的恢复基因,02428含有对粳稻不育系的恢复基因。三者与BT型不育系的产量配合力都较好,杂种有明显的优势。单株产量主要受单株有效穗数和每穗粒数的影响。     

合浦绒螯蟹Eriocheir hepuensis 的形态学研究

通过野外调查和养殖试验观察确知合浦绒螯蟹的成体及各期幼体的形态构造均介于中华绒螯蟹和日本绒螯蟹之间,是绒螯蟹属的一个物种,故将其学名由“日本绒螯蟹合浦亚种”改为“合浦绒螯蟹”(Eriocheir hepuensis) ,该蟹个全大,生长快,繁殖力强,生长周期短,是一个优良的绒螯蟹新品种。     

北缘普通野生稻实验种群动态研究Ⅱ--生殖生长动态

以江西东乡和湖南茶陵普通野生稻北缘种群为材料,在繁育期对当地构建的普通野生稻种群实验小种群进行了连续的调查和测定,研究北缘普通野生稻在各种实验条件下有性生殖的生长动态,结果表明：(1)不同起始密度下普通野生稻各小种群的有性繁殖量有一定差异,较低的起始密度抽穗率低,而较高起始密度的种群有性繁殖的总量相对较高,到40枝／m^2时抽穗率增长到一个稳定水平．(2)放牧条件对普通野生稻的有性繁殖总量影响不大;而刈割则对普通野生稻的有性繁殖具有明显的抑制作用,在较轻度的切割条件下虽未使普通野生稻个体趋于灭绝,但对抽穗等有性繁殖指标来说则是灾难性的．(3)长期生长于过高的水深对北缘普通野生稻的有性繁殖也具有抑制作用．