云南栽培稻种SSR 遗传多样性比较

采用64个SSR标记对96份云南水稻(Oryz a sativa)地方品种和选育品种的遗传多样性进行比较分析。结果发现64个标记都具有多态性, 共检测到741个等位基因, 每个多态性位点检测到的等位基因数为2-29个, 平均11.57个; Nei基因多样性指数(He)范围在0.345(RM321)-0.932(RM1)之间, 平均为0.56。水稻品种的遗传多样性并非按地理位置均匀分布, 而是在相 似系数为0.17的水平上明显分为2个不同类群, 即籼稻类群和粳稻类群, 且籼粳亚种间的SSR多样性差异不明显, 籼稻平均等位基因数(Ap)和Nei基因多样性指数(Ap=10.6, He=0.46)与粳稻品种(Ap=10.7, He=0.48)十分接近, 可能与这些品种间存在一定频率的基因交流有关。糯稻和非糯稻在籼稻群和粳稻群中都有表现, 没有特别的分布规律。云南栽培稻选育品种与地方稻亲缘关系较近, 其遗传基础可能来源于云南水稻地方品种。本研究结果表明, SSR标记能较好地区分云南栽培稻品种, 且云南水稻地方品种遗传多样性丰富, 存在大量的优质性状可供育种实践选择。     

云南栽培稻种SSR遗传多样性比较

采用64个SSR标记对96份云南水稻（Oryza sativa）地方品种和选育品种的遗传多样性进行比较分析。结果发现64个标记都具有多态性,共检测到741个等位基因,每个多态性位点检测到的等位基因数为2—29个,平均11．57个：Nei基因多样性指数（He）范围在0．345（RM321）-0．932（RM1）之间,平均为0．56。水稻品种的遗传多样性并非按地理位置均匀分布,而是在相似系数为0．17的水平上明显分为2个不同类群,即籼稻类群和粳稻类群,且籼粳亚种间的SSR多样性差异不明显,籼稻平均等位基因数（Ap）和Nei基因多样性指数（Ap=10．6,He=0．46）与粳稻品种（Ap=10．7,He=0．48）十分接近,可能与这些品种间存在一定频率的基因交流有关。糯稻和非糯稻在籼稻群和粳稻群中都有表现,没有特别的分布规律。云南栽培稻选育品种与地方稻亲缘关系较近,其遗传基础可能来源于云南水稻地方品种。本研究结果表明,SSR标记能较好地区分云南栽培稻品种,且云南水稻地方品种遗传多样性丰富,存在大量的优质性状可供育种实践选择。     

云南地方稻核心种质耐低磷特性研究

1引言磷是作物的必需性营养元素,人类将面临农业可持续发展与磷素资源严重短缺的矛盾[17].土壤普查结果表明,全国有59%以上的土壤缺磷[1].为了提高作物产量,生产中通过施用磷肥来解决土壤缺磷的问题.由于磷肥利用率低,不仅增加生产成本,还带来资源短缺、环境污染及食品安全等诸多问题[21,26].发掘水稻利用磷的内在潜力,培育磷高效利用新品种,可减少磷的施用量.近年来,野生或地方稻绿色基因发掘及改良进展显著,如资源高效利用[22~24]、高产基因[3,20,27]和抗性基因[2,13]等,利用植物基因型间磷元素利用效率的差异,筛选和培育磷高效基因型作物…     

云南地方稻waxy基因序列多样性分析

利用特异引物Wx-F/AG-2对来源于云南省16个州市64个县的252份地方稻种waxy基因中含有微卫星序列(CT)n和第一内含子的序列进行PCR扩增并测序。结果表明在268个碱基的序列内检测到4个变异位点:第一内含子上游56位处(CT)n存在(CT)10、(CT)11、(CT)12、(CT)14、(CT)16、(CT)17、(CT)18、(CT)19、(CT)20、(CT)21等10种变异;(CT)11、(CT)18、(CT)17、(CT)12、(CT)10等5种单倍型是云南优势类型,合计占供试材料的92.85%;籼稻以(CT)10、(CT)11、(CT)12为主,而粳稻以(CT)11、(CT)12、(CT)17和(CT)18为主;第一内含子+1位存在G/T变异,81.75%品种是G,T只出现在(CT)16、(CT)17、(CT)18和(CT)20的品种中,其频率在籼稻中为13.48%,粳稻20.86%,水稻16.17%,陆稻22.35%,粘稻10.47%,糯稻42.62%。+86-88位存在ATA/GTA/A--3种变异;+128位处存在(AATT)5和(AATT)6 2种变异。根据这4个变异位点,可将252个云南地方稻种归为16种单倍型,其中Wx4(32.54%)、Wx9(13.89%)、Wx12(12.7%)、Wx5(12.3%)、Wx1(8.33%)、Wx11(7.94%)是主要类型,合计87.7%,其他类型频率较低。籼/粳亚种、水/陆稻和粘/糯中存在单倍型种类和单倍型频率两方面的差异,籼稻/粳稻、水/陆稻和粘/糯稻各亚种或生态型均有独享的单倍型,共享单倍型频率也存在差异,表明亚种间或生态型间发生了一定的遗传分化。单倍型地理分布分析表明,临沧、普洱单倍型种类最丰富,以之为中心向外扩展,单倍型种类有减少的趋势,第一内含子+1位的T主要分布在临沧、普洱、西双版纳、德宏等南部地区。本研究揭示了云南地方稻种群体waxy基因的变异和分布特点。     

不同类型特种稻种质营养及功能性成分含量的差异

本研究以具有有色种皮、巨胚、甜味、香味、糯性等单一特殊性状或2个以上特殊性状聚合于一体的新创制特种稻种质39份和1份白米种质(对照)为试验材料,进行了黑米、黑褐米、红米、香糯米、黑巨胚糯米、红巨胚糯米、巨胚糯米、黑甜米、红甜米、白甜米等不同类型特种稻种质的营养及功能性成分含量的差异评价。结果表明,供试特种稻类型在大部分氨基酸含量和矿质元素含量上与白米差异不显著,只在个别氨基酸和矿质元素含量上与白米呈显著或极显著差异。黑褐米、红米、黑巨胚糯米、红巨胚糯米、巨胚糯米、红甜米和白甜米等7种类型的赖氨酸含量显著或极显著高于白米,高2.91%~24.68%;黑米、黑褐米、红米、香糯米、黑巨胚糯米和红甜米等6种类型的铁含量显著或极显著高于白米,高17.62%~68.09%;黑褐米、红米、黑巨胚糯米、红巨胚糯米、黑甜米、红甜米、白甜米等7种类型的钙含量显著或极显著高于白米,高23.56%~49.46%;黑米、黑褐米、红米、黑甜米、红甜米、白甜米等6种类型的锌含量显著或极显著高于白米,高12.21%~55.87%。由此表明,具有有色种皮、巨胚、甜味、香味、糯性等单一特殊性状或2~3个特殊性状的聚合对赖氨酸含量与铁、钙和锌含量的提高方面具有一定的增加效应,认为在今后以赖氨酸、铁、钙和锌含量为目标性状的功能性水稻育种中,多个特殊性状的聚合将是增加上述功能性成分含量的有效途径之一。通过鉴定评价,从创新种质中还筛选出一些功能性成分含量相对较高的优异种质,白甜米1553和红巨胚糯米1476的赖氨酸含量较高,比白米分别高29.37%和23.42%;红米1439和红米1440的铁含量较高,比白米分别高99.05%和80.00%;黑甜米1511和黑甜米1515的硒含量较高,比白米分别高194.14%和136.48%;白甜米1551和香糯米1446的γ-氨基丁酸含量较高,比白米分别高14.56%和11.83%;黑巨胚糯米1464和黑米1432的花色苷含量较高,比供试18份有色稻米的平均值分别高253.23%和248.83%。这些新创制的功能性成分含量较高的水稻种质有待于今后在育种、生态适应性鉴定与产业化中进一步得到利用。     

水稻种质资源芽期耐冷性的鉴定与评价

对新收集的879份水稻种质资源进行了水稻芽期耐冷性的鉴定与评价,从中筛选出芽期耐冷性极强的水稻种质资源39份,均为贵州地方粳稻品种.供试粳稻品种61.23%具有强芽期耐冷性(3级),而籼稻品种只有13.77%具有强芽期耐冷性(3级 ),粳稻品种的芽期耐冷性显著强于籼稻品种.无论籼稻,还是粳稻,贵州地方稻种的芽期耐冷性显著强于来自其他地区的品种,从贵州地方稻种中挖掘极强耐冷种质资源的潜力较大 .     

Genetic diversity analysis of traditional and improved Indonesian rice (Oryza sativa L.) germplasm using microsatellite markers

The archipelago of Indonesia has a long history of rice production across a broad range of rice-growing environments resulting in a diverse array of local Indonesian rice varieties. Although some have been incorporated into modern breeding programs, the vast majority of these landraces remain untapped. To better understand this rich source of genetic diversity we have characterized 330 rice accessions, including 246 Indonesian landraces and 63 Indonesian improved cultivars, using 30 fluorescently-labeled microsatellite markers. The landraces were selected across 21 provinces and include representatives of the classical subpopulations of cere, bulu, and gundil rices. A total of 394 alleles were detected at the 30 simple sequence repeat loci, with an average number of 13 alleles per locus across all accessions, and an average polymorphism information content value of 0.66. Genetic diversity analysis characterized the Indonesian landraces as 68% indica and 32% tropical japonica, with an indica gene diversity of 0.53 and a tropical japonica gene diversity of 0.56, and a Fst of 0.38 between the two groups. All of the improved varieties sampled were indica, and had an average gene diversity of 0.46. A set of high quality Indonesian varieties, including Rojolele, formed a separate cluster within the tropical japonicas. This germplasm presents a valuable source of diversity for future breeding and association mapping efforts.     

Genetic Variation and Indica Japonica Differentiation in Yunnan Indigenous Rice

Yunnan, in Southwest China, has long been recognized as part of a center of diversity for Asia cultivated rice (Oryza sativa L. ). The authors surveyed DNA restriction fragment length polymorphisms (RFLPs) in a sample of 87 accessions from Yunnan indigenous rice with 7 single copy probes. The results ihdicated that both indica and japonica rice are genetically highly diverse. The number of alleles and the level of genic diversity in japonica rice were higher than those in indica rice. The results also showed that indica and japonica rices were clearly differentiated and the extent of differentiation varies with chromosomal regions as represented by the RFLP markers. Authors' results appeared to be in favor of the diphyletic hypothesis concerning the origin of cultivated rice.     

中国粳稻地方品种孕穗期耐冷性评价及聚类分析

用原产于中国18个省的329份粳稻地方品种为材料,分析了自然低温和冷水胁迫下不同省份粳稻地方品种孕穗期耐冷性及主要农艺性状表型差异和聚类特点。结果表明,在自然低温和冷水胁迫下各省份粳稻地方品种的孕穗期耐冷性状及主要农艺性状有明显的差异。在自然低温和冷水胁迫下,天津、四川和台湾品种的结实率及冷水反应指数均较高,表现较强的孕穗期耐冷性和迟钝的冷水反应。云南品种在自然低温下表现为较强的孕穗期耐冷性（结实率）,而在冷水胁迫下播种至抽穗天数和株高的冷水反应指数较高,表现为迟钝的冷水反应。从总体趋势上看,自然低温和冷水胁迫下,除个别省份外,纬度相对较高的北方省份品种的孕穗期耐冷性（结实率）强于纬度相对较低的南方省份品种。此外,在自然低温和冷水胁迫下,各省粳稻地方品种的聚类结果总体上与各省品种的地理位置及其耐冷性有密切的联系,而在冷水胁迫下品种的聚类结果与品种地理位置的关系比自然低温下更为密切。     

Genetic structure and phylogeography of rice landraces in Yunnan, China, revealed by SSR.

Yunnan Province is one of the largest centers of genetic diversity of Oryza sativa L. in China, and in the world. Using a genetically representative core collection of 692 rice landraces from Yunnan, the genetic structure, differentiation, and geographic diversity of this rice germplasm were analyzed. The accessions were classified into different populations, according to the model-based structure analysis. Model-based populations were characterized through the reconstruction of a neighbor-joining tree, principal coordinate analysis, and the estimation of morphologic and SSR variations. Finally, the distribution of genetic diversity and differentiation among districts were studied. Seven model-based populations were identified on the basis of the structure analysis. This classification was partly consistent with Ting's 5-level taxonomic system. Differentiation between 2 rice subspecies is the most apparent, with a clearer differentiation between soil-watery ecotypes in japonica than in indica; however, differentiation among seasonal ecotypes in indica is clearer than in japonica. Cropping system and man-made restricted-growth environments could be considered to be the main forces driving the intraspecific differentiation of cultivated rice. It has been suggested that, because it possesses the highest genetic diversity and all the necessary conditions as a center of genetic diversity, the southwest region of Yunnan, encompassing Simao, Lincang, and Xishuangbanna districts, is the center of genetic diversity of Yunnan rice landraces.     

Diversity in the Content of Some Nutritional Components in Husked Seeds of Three Wild Rice Species and Rice Varieties in Yunnan Province of China

In addition to rice (Oryza sativa L.) cultivars, there are three wild rice species, namely O.rufipogon Griff, O. officinalis Wall and O. granulata Baill, in Yunnan Province, China. Each species has different subtypes and ecological distributions. Yunnan wild rice species are excellent genetic resources for developing new rice cultivars. The nutritional components of the husked seeds of wild rice have not been investigated thus far. Herein, we report on the contents of total protein, starch, amylose, 17 amino acids, and five macro and five trace mineral elements in husked seeds from three wild rice species and six O. sativa cultivars. The mean (± SD) protein content in the husked rice of O. rufipogon, O. officinalis, and O. granulata was (14.5 ± 0.6)%, (16.3 ± 1. 1)%, and (15.3 ± 0.5)%, respectively. O. officinalis Ⅲ originating from Gengma had the highest protein content (19.3%). In contrast, the average protein content of six O. sativa cultivars was only 9.15%. The total content of 17 amino acids of three wild rice species was 30%-50% higher than that of the six cultivars. Tyrosine, lysine, and valine content in the three wild rice species was 34%-209% higher than that of the cultivars. However, the difference in total starch content among different O. sativa varieties or types of wild rice species was very small. The average amylose content of O. rufipogon, O. officinalis,and O. granulata was 12.0%, 9.7%, and 11.3%, respectively, much lower than that of the indica and japonica varieties (14.37%-17.17%) but much higher than that of the glutinous rice cultivars (3.89%). The sulfur, phosphorus, magnesium, zinc, and ferrite content in the three wild rice species was 30%-158% higher than that of the six cultivars. The considerable difference in some nutritional components among wild rice species and O. sativa cultivars represents a wide biodiversity of Yunnan Oryza species. Based on the results of the present study, it is predicted that some good genetic traits, especially high protein and ideal amylose content, of Yunnan wild rice species may be useful in improving the nutritional value of rice. This is the first report regarding the amino acid, mineral element, protein and amylose content of husked seeds of some Yunnan wild rice species that have important genetic characteristics for rice quality and nutritional value.     

云南普通野生稻和地方稻WRKY45基因的克隆及转化

以云南地方稻‘三磅七十箩’(SB70)和景洪普通野生稻为材料,根据GenBank中水稻WRKY45基因序列设计引物,进行PCR扩增。结果表明,经测序得到目的基因长约1.3kb,推导的氨基酸具有WRKY蛋白典型的保守区域WRKYGQK。经BLAST分析,与GenBank中不同栽培稻WRKY45基因的相似性在85%以上,但也存在一些核苷酸差异,这些差异可能导致其调控抗逆能力更强。构建该基因的表达载体pCAMBIA1300,重组克隆后通过农杆菌介导法转入云南栽培粳稻‘云资粳41号’中,经PCR鉴定获得24株转基因阳性植株。     

Genetic Identification of Quantitative Trait Loci for Contents of Mineral Nutrients in Rice Grain

In present study, Fe, Zn, Mn, Cu, Ca, Mg, P and K contents of 85 Introgression linee (ILs) derived from a cross between an elite indica cultivar Teqing and the wild rice (Oryza rufipogon) were measured by inductively coupled argon plasma (ICAP) spectrometry. Substantial variation was observed for all traits and most of the mineral elements were significantly positive correlated or independent except for Fe with Cu. A total of 31 putative quantitative trait loci (QTLs) were detected for these eight mineral elements by single point analysis. Wild rice (O. rufipogon) contributed favorable alleles for most of the QTLs (26 QTLs), and chromosomes 1, 9 and 12 exhibited 14 QTLs (45%) for these traits. One major effect of QTL for zinc content accounted for the largest proportion of phenotypic variation (11%-19%) was detected near the simple sequence repeats marker RM152 on chromosome 8. The co-locations of QTLs for some mineral elements observed in this mapping population suggested the relationship was at a molecular level among these traits and could be helpful for simultaneous improvement of these traits in rice grain by marker assisted selection.     

利用基因组数据分析水稻基因顺式作用元件

为了研究籼粳亚种基因调控序列的总体特性,我们利用籼粳稻以及拟南芥基因组和全长mRNA序列获取了大量高可信度的调控序列,通过这些序列,分析了水稻基因调控序列顺式作用元件(信号)的数量、分布以及与GC含量的关系等．研究结果表明:一些信号在水稻基因调控序列中发生显著的数量变化,同时一些信号数量在水稻与拟南芥基因间存在明显差异, 这说明这两种单双子叶植物间信号的使用上存在偏好,同时水稻不同类型基因以及特有与非特有基因间在信号的使用上也存在差异．这些差异信号的分布直接导致了调控序列GC含量的波动．本研究没有发现水稻籼粳两个亚种间在调控序列方面(顺式调节因子和GC含量等)存在明显差异．     

The Effects of Kinetin and Gibberellin on the Germination of Dehusked Seeds of Indica and Japonica Rice (Oryza sativaL.) under Anaerobic and Aerobic Conditions

The effects of kinetin and gibberellin were examined under anaerobicconditions (0% oxygen) and aerobic conditions (20% oxygen) onthe germination of dehusked seeds of indica and japonica ricecultivars that had been harvested at different times duringthe formation of seeds. Surjamkhi was used as a representativeof deep dormant indica cultivars and Assam IV as a less dormantindica cultivar. Sasanishiki was used as the japonica rice cultivar.Both phytohormones were applied at a concentration of 10^-3Mwhichproved to have the greatest stimulatory effect in preliminarywork at concentrations of 10^-3–10^-5M. Under aerobic conditions,inhibition of germination by dehusking of Sasanishiki seedsthat had been harvested either 30 or 60 d after anthesis wasovercome by kinetin and all seeds germinated. Complete germinationinduced by kinetin under aerobic conditions was also achievedwith the dehusked seeds of the indica rice cultivar Assam IVthat had been harvested on two occasions and of Surjamkhi thathad been harvested 28 d after anthesis. In contrast, germinationof dehusked japonica seeds stimulated by anaerobiosis was inhibitedby kinetin. The stimulatory effects of gibberellin on the germinationof indica and japonica rice seeds were observed under aerobicand anaerobic conditions. Under anaerobic conditions, the responsesof dehusked indica and japonica rice seeds to kinetin and gibberellindiffered, being negative with kinetin and positive with gibberellin.Under aerobic conditions, the stimulatory effects of kinetinon germination of dehusked seeds were greater than those ofgibberellin. Thus, treatment with kinetin appears to be usefulfor breaking the considerable dormancy commonly observed inthe dehusked seeds of indica rice. Mechanisms are proposed toexplain the stimulatory effects of these phytohormones on thegermination of dehusked seeds of indica and japonica rice underaerobic and anaerobic conditions. Rice; Oryza sativaL.; seed germination; dehusking treatment; gibberellin; indica; japonica; kinetin; oxygen; dormancy; germination inhibition; seed formation     

RAPD Determinations on the Genetic Differentiation of Chinese Common Wild Rice

Ninety accessions which included Chinese common wild rice (Oryza rufipogon) from 8 provinces and traditional cultivars from lower and middle basins of Yangtze River, southeast of China and Yunnan Province as well as some commercial varieties were analyzed by RAPD with 24 primers. A scattered figure suggesting the indica-japonica and wild-domestication differentiations among 90 rice accessions was generated based on RAPD data. The results indicated that Chinese common wild rice, indica and japonica accessions were divided into 3 groups respectively. Chinese common wild rice were somewhat closer to the japonica type than the indica type.     

元阳3个长期连续栽培水稻地方品种内部遗传异质性分析

水稻地方品种是稻种资源的重要组成部分,遗传多样性十分丰富,具有推广改良品种所缺少或没有的优质种质,是水稻育种和稻种起源、进化研究不可缺少的过渡材料。目前,对水稻地方品种间遗传多样性研究较多,而对其内部异质性研究甚微。本研究用24对微卫星(SSR)引物对云南元阳梯田3个栽种历史悠久的水稻地方品种的内部遗传异质性进行了分析。共检测出117个等位基因,香农指数为红脚老粳居群(0.5911)>白脚老粳A居群(0.4875)>月亮谷居群(0.3070)。结果显示:3个地方品种的内部遗传异质性丰富,且遗传异质性主要得益于个体间,而非居群间。     

Molecular evaluation of genetic diversity and association studies in rice (<Emphasis Type="BoldItalic">Oryza sativa</Emphasis> L.)

In the present study, we tested rice genotypes that included un(der)exploited landraces of Tamil Nadu along with indica and japonica test cultivars to ascertain their genetic diversity structure. Highly polymorphic microsatellite markers were used for generating marker segregation data. A novel measure, allele discrimination index, was used to determine subpopulation differentiation power of each marker. Phenotypic data were collected for yield and component traits. Pattern of molecular differentiation separated indica and japonica genotypes; indica genotypes had two subpopulations within. Landraces were found to have indica genome, but formed a separate subgroup with low linkage disequilibrium. The landraces further separated into distinct group in both hierarchical clustering analysis using neighbour-joining method as well as in the model based population structure analysis. Japonica and the remaining indica cultivars formed two other distinct groups. Linkage disequilibrium observed in the whole population was considerably reduced in subpopulations. Low linkage disequilibrium of landforms suggests their narrow adaptation in local geographical niche. Many population specific alleles could be identified particularly for japonica cultivars and landraces. Association analysis revealed nine marker-trait associations with three agronomic traits, of which 67% were previously reported. Although the testing landraces together with known cultivars had permitted genomewide association mapping, the experiment offers scope to study more landraces collected from the entire geographical region for drawing more reliable information.