Genetic Diversity of Rice Landraces from Lowland and Upland Accessions of China

Genetic diversity of rice landraces from lowland and upland accessions of China was investigated using 66 polymorphic simple sequence repeat (SSR) markers.The total number of alleles detected from all 324 tested accessions was 555 with an average allele number (Na) of 8.409 per locus,the average effective number of alleles (Ne) of 3.574 and the average Shannon’s information index (I) of 1.378.The genetic diversity was higher for the indica landraces compared to the japonica landraces,and the upland landraces were more genetically diverse than the lowland landraces.The SSR markers,RM72,RM232,RM219,RM241,RM224 and RM3 showed the highest rates of polymorphism and these SSR markers were suitable to assess the genetic diversity of rice germplasm resources.A dendrogram of 324 accessions of lowland and upland landraces showed that all rice accessions were mainly subdivided into two groups,japonica and indica,with some being intermediate.The distribution of lowland and upland landraces among the japonica and indica rice groups was distinct,with obvious differentiation between the lowland and upland landraces in japonica rice,but no such clear distinction in indica rice.     

利用微卫星标记比较云南元阳哈尼梯田两个不同时期种植的水稻地方品种的遗传多样性

 为了揭示云南元阳哈尼梯田两个不同时期种植的水稻地方品种的遗传多样性,以元阳哈尼梯田过去种植（20世纪70年代收集于元阳县,现保存于种质库）的72个和当前种植的76个品种为材料,采用48个SSR分子标记进行多态性、遗传相似性和聚类分析。平均每个标记检测到的等位基因数、有效等位基因数、位点多态信息含量和基因型多样性指标在过去与当前的品种间,差异均未达显著水平。共有24个SSR标记的52个等位基因位点在过去与当前种植的品种中存在差异,其中,有18个SSR标记的32个等位基因位点在当前的品种中未检测到,同样,有16个SSR标记的20个等位基因位点在过去的品种中未检测到。基于48个SSR分子标记的平均遗传相似性系数,当前种植的品种为03340,过去为03313,过去与当前的品种分别在遗传相似性系数0203和0174处分为籼、粳两个亚种群。表明元阳哈尼梯田水稻地方品种经过30多年的自然与人工选择,虽有部分等位基因位点发生了变异,但仍保留了原有的遗传多样性;元阳哈尼梯田水稻地方品种存在明显的籼、粳分化。元阳哈尼梯田及其水稻地方品种是研究稻作籼粳演化的理想基地和材料。     

贵州地方水稻品种的SSR遗传多样性分析

利用21对SSR引物对74份贵州"禾"水稻品种及6份国际上常用的典型籼稻和粳稻品种进行遗传多样性研究。共检测到92个等位基因,每个位点的等位基因数变幅为2~9个,平均4.381个;平均Shannon信息指数为0.935,变幅为0·2793~1.8732;期望杂合度为0.5145,范围在0.0988~0.8313;品种间遗传相似系数为0.64~0.98。UPGMA聚类分析表明,在相似系数为0.682处可将"禾"品种分为4大类,大部分材料被聚类到典型粳稻的附近,品种间的亲缘关系与地理来源关系不大;主坐标分析结果与UPGMA聚类结果基本吻合。SSR检测结果表明,贵州省水稻地方品种——"禾"的多样性程度较低,且大多数属于粳稻。     

Morphological and Molecular Characterization of Novel Salt-Tolerant Rice Germplasms from the Philippines and Bangladesh

To screen for new sources of salinity tolerance, 688 traditional rice varieties from the Philippines and Bangladesh were obtained, and their tolerance to hypersaline conditions at the seedling stage was examined. A total of 29 Philippine lines and 15 Bangladeshi lines were scored as salt-tolerant.Morphological assessment(plant height, biomass and Na-K ratio) revealed that among the 44 salt-tolerant accessions, Casibon, Kalagnon and Ikogan had significantly higher relative shoot length difference, relative shoot growth reduction and shoot Na-K ratio than the tolerant check FL478.Additionally, AC and Akundo exhibited significantly higher Na-K ratios than the other genotypes. The genetic diversity of the 44 genotypes was assessed using 34 simple sequence repeat markers. A total of 133 alleles were detected across all loci. Cluster analysis showed that AC, Akundo and Kuplod were clustered along with FL478, indicating a strong genetic relatedness between these genotypes. IR29(susceptible check) was singly separated. The haplotype analysis revealed that none of the 44 genotypes had a similar allele combination as FL478. These accessions are of interest since each genotype might be different from the classical salinity-tolerant Pokkali.     

Rice landrace diversity in Nepal: Variability of agro-morphological traits and SSR markers in landraces from a high-altitude site

Nepal has a wide diversity of rice landraces in all rice-growing areas from low to high altitude. A baseline survey in Jumla, a high-altitude site (2200–3000 m), identified rice landraces with different names that differed in phenotype and were adapted to the extreme high-altitude environment. Jumli Marshi was the most common traditional rice variety, accounting for 85% of samples collected. The genetic diversity of these landraces was assessed by agro-morphological variability and microsatellite marker polymorphism. Forty-two qualitative and quantitative traits and 39 microsatellite (simple sequence repeats, SSR) markers were assessed among accessions with 10 different names. The accessions showed low morphological diversity with an average Shannon Weaver diversity index of 0.23. Only 16 morphological traits showed significant variation among the accessions. Discriminant function analysis showed that only 36% of accessions could be clustered according to name by morphological traits. Only one SSR locus was polymorphic, distinguishing only one accession. The names used by farmers to describe the phenotypic characteristics of panicle and grains were inconsistent indicators of genetic identity. We conclude that the Jumla landrace population has a narrow genetic base. The diversity detected was sufficiently low for it to be possible that there was a single origin for all of the Jumla landraces.     

Genetic Diversity Analysis of Rice Germplasm in Tripura State of Northeast India Using Drought and Blast Linked Markers

We genotyped 74 rice germplasms including Tripura's local landraces, improved varieties, cultivars and breeding lines and other rice varieties using molecular markers for genetic diversity, drought QTLs, and blast resistance genes. The number of alleles per locus ranged from 2 to 5 with an average of 2.9. The polymorphic information content value per locus ranged from 0.059 (RM537) to 0.755 (RM252) with an average of 0.475. Cluster analysis based on 30 simple sequence repeat markers revealed 5 clusters and also indicated the presence of variability within the rice accessions. The drought QTL qDTY2.1 was found in 56.0% of germplasms and qDTY1.1 was detected in only 6.8% of the germplasms. Out of seven rice blast resistance genes screened, only two rice varieties, RCPL-1-82 and Buh Vubuk (Lubuk), were positive for four blast resistance genes while only Releng possessed two blast resistance genes. Among 74 rice germplasms, only three accessions, Releng, RCPL1-82 and Buh Vubuk (Lubuk), possessed both drought-related QTLs and blast resistance genes. Overall, the 74 indigenous rice genotypes showed low level of genetic diversity, which is in contrast to high level of genetic diversity among rice varieties in northeast India, where highlights the good farming practice, conservation of germplasms and the limitation of molecular markers employed in this study. The presence of both drought related QTLs and blast resistance genes in some of the germplasms can be useful in future breeding programmes.     

布朗族当前种植稻作地方品种的SSR位点多样性分析

 调查收集了80份布朗族当前仍在种植的稻作地方品种,利用分布于水稻12条染色体上的72对SSR引物,进行了遗传多样性分析。共检测出271个等位基因,平均每对引物3.764个;197个有效等位基因,平均每对引物2.739个;多态性信息含量变幅为0.163~0.827,平均为0.582;稀有等位基因数20个。云南省内3个不同地区布朗族种植的地方品种遗传多样性有差异,表现为勐海县>双江县>墨江县。地方品种的遗传相似系数变幅为0.111~0.875,平均为0.411;在相似系数0.266处可以将供试品种分为籼粳两大类,但不能再将布朗族同一集居区的地方品种完全聚成同一亚类,暗示着布朗族不同集居区的条件对地方品种进化的影响还比较有限。最后还就布朗族对地方品种多样性的贡献及可能的原因等进行了讨论分析,对如何防止地方品种收集中同名异种和同种异名问题发生提出了建议。     

Genetic Diversity and Allelic Frequency of Selected Thai and Exotic Rice Germplasm Using SSR Markers

A collection of 167 Thai and exotic rice accessions was subjected for evaluation of genetic diversity and assessment of relationship by simple sequence repeat(SSR) markers. Among a total of 49 SSR markers, 13 markers distributing over 12 rice chromosomes showed clear polymorphic band patterns, and they were selected for genetic assessment. A total of 110 alleles were detected with an average of 8.46 alleles per locus. The averages of gene diversity, heterozygosity and polymorphic information content were 0.59, 0.02 and 0.56, respectively. The unweighted-pair group method with arithmetic averages(UPGMA) clustering analysis was performed for genetic distance, and phylogenetic tree was constructed. The result showed that this rice collection was divided into two major groups, classified as japonica and indica subspecies. Within the japonica group, temperate japonica and tropical japonica subgroups can be clearly separated. Three-dimensional principal component analysis projection and model-based population structure analysis showed consistent clustering results with two major groups of UPGMA analysis, supporting the classification of japonica and indica subspecies. The indica allelic frequency was also investigated to provide an indicative guide for breeders to overcome the practical problems on sterility of inter-subspecies hybrid offspring. This rice collection and information obtained in this study will be useful for rice breeding programs.     

太湖流域粳稻地方品种的微卫星分析

 采用45对SSR引物对224份太湖流域粳稻地方品种进行遗传多样性分析,共检测到162个等位变异,每个位点等位基因数的变幅为2～7,平均为3.6。各位点Nei基因多样性指数变异较大,为0.009（RM169）～0.663（RM444）,平均为0.197。青稻、黄稻、红稻和白稻各传统生态型均具较低的遗传多样性。分子方差分析表明,SSR遗传变异绝大部分存在于传统生态型内。青稻与红稻间、青稻与白稻、红稻与白稻、红稻与黄稻间遗传分化显著。太湖流域粳稻地方品种SSR多样性较低,稀有类型等位基因较多。这对科学制定太湖粳稻地方资源的保护和利用策略具有指导意义。     

野生大豆芽期耐盐性状的关联分析

盐渍化是危害大豆生产的主要非生物胁迫因素之一。目前大豆耐盐性研究主要集中在栽培大豆的苗期耐盐性,而芽期耐盐性状的研究相对较少。野生大豆蕴含丰富的耐逆基因,是栽培大豆遗传改良的重要资源。为了研究野生大豆芽期耐盐性状的遗传机制,以113份野生大豆为试验材料,进行芽期耐盐性状的鉴定,结合群体的分子标记对包括2年平均值在内的3个环境下的3个耐盐指数进行全基因组关联分析,共检测到与野生大豆芽期耐盐相关的位点26个,6个SSR标记Satt521、Satt022、Satt239、Satt516、Satt251和Satt285在2个或3个环境下均被检测到,4个SSR标记Satt516、Satt251、Satt285和GMES4990与2个或3个耐盐指数显著相关。对这些SSR标记进行分析,挖掘了最优的等位基因及其载体材料。以上这些结果对于阐明野生大豆芽期耐盐性状的遗传机制,进一步发掘新的耐盐基因具有重要意义。同时也为栽培大豆遗传基础的拓宽、大豆耐盐分子标记辅助选择和分子设计育种等后续研究提供重要依据。     

An in planta, Agrobacterium-mediated transient gene expression method for inducing gene silencing in rice (Oryza sativa L.) leaves

Background

Although the genetic structure of rice germplasm has been characterized worldwide, few studies investigated germplasm from Thailand, the world’s largest exporter of rice. Thailand and the International Rice Research Institute (IRRI) have diverse collections of rice germplasm, which could be used to develop breeding lines with desirable traits. This study aimed to investigate the level of genetic diversity and structures of Thai and selected IRRI germplasm. Understanding the genetic structure and relationships among these germplasm will be useful for parent selection used in rice breeding programs.

Results

From the 98 InDel markers tested for single copy and polymorphism, 19 markers were used to evaluate 43 Thai and 57 IRRI germplasm, including improved cultivars, breeding lines, landraces, and 5 other Oryza species. The Thai accessions were selected from all rice ecologies such as irrigated, deep water, upland, and rainfed lowland ecosystems. The IRRI accessions were groups of germplasm having agronomic desirable traits, including temperature-sensitive genetic male sterility (TGMS), new plant type, early flowering, and biotic and abiotic stress resistances. Most of the InDel markers were genes with diverse functions. These markers produced the total of 127 alleles for all loci, with a mean of 6.68 alleles per locus, and a mean Polymorphic Information Content (PIC) of 0.440. Genetic diversity of Thai rice were 0.3665, 0.4479 and 0.3972 for improved cultivars, breeding lines, and landraces, respectively, while genetic diversity of IRRI improved and breeding lines were 0.3272 and 0.2970, respectively. Cluster, structure, and differentiation analyses showed six distinct groups: japonica, TGMS, deep-water, IRRI germplasm, Thai landraces and breeding lines, and other Oryza species.

Conclusions

Thai and IRRI germplasm were significantly different. Thus, they can be used to broaden the genetic base and trait improvements. Cluster, structure, and differentiation analyses showed concordant results having six distinct groups, in agreement with their development, and ecologies.     

Population Structure and Marker-Trait Association in Indigenous Aromatic Rice

Joha rice of Assam is a class of rice with aroma, good cooking qualities and excellent palatability, which are grown in different places of Assam with their adaptability to local situations. A total of 143 simple sequence repeat markers were used in this study to reveal sufficient genetic variations among the 40 Joha and 14 non-Joha rice genotypes. Polymorphism information content of these markers were from 0.17 to 0.88, where the average of 3.7 alleles were observed. The model-based population assignment and dendrogram analysis identified three distinct groups in the 54 rice accessions, which established a clear identity of Joha accessions from the Basmati accessions, indicating high diversity and strong population structure in Joha rice. Furthermore, a total of 29 significant marker-trait associations(P 0.05) for 10 characters were detected. The QTLs related with yield and grain quality can be used effectively in crop improvement programs and for further fine mapping and validation of specific genes to develop gene-based perfect markers in rice breeding and for mining of better alleles of these genes in Joha rice collections.     

利用SRAP分子标记分析47份杂草稻样品遗传多样性

 利用SRAP分子标记分析了采集于杂草稻多发地区的47份样本的遗传多样性,旨在从基因的开放式阅读框区域（ORFs）深入研究杂草稻的遗传背景,为进一步揭示其来源提供依据。结果表明,参试的我国杂草稻材料在110对引物组合中共扩增出845种类型的谱带,多态性谱带590种,多态率为6982%,平均每对引物可以检测到54条多态性谱带。说明SRAP分子标记对于杂草稻基因组开放式阅读框区域序列的多态性具有很高的检测效率,适用于遗传多样性研究。我国杂草稻材料Shannon Weaver多样性指数（I）为063,且南北方杂草稻多样性指数（I）相当,分别为063和062。聚类分析结果表明,南方收集的杂草稻与北方收集杂草稻群体间遗传分化较大,遗传差异明显。程氏指数判定结果也表明杂草稻呈南籼北粳的分布状态。     

基于SSR标记的云南地方稻种群体内遗传多样性分析

 用20对SSR引物对原产于云南的16份地方稻种和2份选育品种进行单个品种群体内的遗传多样性分析。结果表明,87.5%的地方品种群体内SSR多态性高于选育品种,而12.5%的地方品种群体内SSR标记多态性与选育品种相近。81.2%的地方稻种群体内的等位基因数(Na)和Nei基因遗传多样性指数(He)高于选育品种,而18.8%的地方品种群体内Na和He与选育品种相同或略小。水稻地方稻种群体内He的差异较大,其变异范围为0.0146～0.5117,其中,黄板所 1（1980年收集）、黄板所 2（2007年收集）、麻线谷 1（1980年收集）、麻线谷 2（2007年收集）的群体内遗传多样性较高,分别为0.2327、0.4214、0.5117和0.4489。87.5%地方品种的杂合度(Ho)明显高于选育品种;地方品种群体间遗传多样性差异很大,其中1/4的遗传差异来源于地方稻种群体内,差异极显著。RM333、RM257和RM180在供试云南地方稻种群体内的多态性、等位基因数、多样性指数和变异百分率均较高,适用于云南地方稻种群体内遗传多样性检测。     

亚洲栽培稻遗传变异分析最少SSR引物数的研究

以69份类型明确的亚洲栽培稻品种为材料,选用120对SSR引物,通过评估遗传多样性和群体结构,研究分析其遗传变异对SSR引物数的要求。结果表明:1)120对引物在69份试验材料中共检测到1256个等位变异,每个位点的等位基因数差异较大,变化范围为2~27,平均为10.5;平均Nei基因多样性指数变化范围为0.4058~0.9452,平均为0.7616;2)分析亚洲栽培稻遗传多样性时,至少需要72对SSR引物;3)分析亚洲栽培稻群体结构时所需最少引物数可降低至60对。     

Identification of Rice Accessions Associated with K~+/Na~+ Ratio and Salt Tolerance Based on Physiological and Molecular Responses

The key for rice plant survival under Na Cl salt stress is maintaining a high K~+/Na~+ ratio in its cells. Selection for salt tolerance rice genotypes based on phenotypic performance alone will delay in progress in breeding. Use of molecular markers in tandem with physiological studies will help in better identification of salt tolerant rice accessions. Eight rice accessions along with the check Dongjin were screened using 1/2 Yoshida solution with 50 mmol/L NaCl at the seedling stage. The accessions IT001158, IT246674, IT260533 and IT291341 were classified as salt tolerant based on their K~+/Na~+ ratios. Seventeen SSR markers reported to be associated with K~+/Na~+ ratio were used to screen the accessions. Five SSR markers(RM8053, RM345, RM318, RM253 and RM7075) could differentiate accessions classified based on their K~+/Na~+ ratios. Banding pattern of the accessions was scored compared to the banding pattern of Dongjin. The study differentiated accessions based on their association of K~+/Na~+ ratio with molecular markers which are very reliable. These markers can play a significant role in screening large set of rice germplasms for salt tolerance and also help in identification of high-yielding varieties with better salt tolerance. The salt tolerant accessions can be taken forward into developing better varieties by conventional breeding and exploring genes for salt tolerance.     

Comparative Analysis of Genetic Diversity and Structure in Rice Using ILP and SSR Markers

Genetic diversity of 36 rice entries from the United States Department of Agriculture(USDA)rice collection was assessed using 103 ILP(intron length polymorphism)and 54 SSR(simple sequence repeats)markers.A total of 236 and 332 alleles were detected by the ILP and SSR markers,respectively.On average,the SSR markers produced higher polymorphism information content value and number of alleles than the ILP markers.Whereas the Nei's genetic distance measured using the SSR markers was much higher than that measured using the ILP markers.Mantel's test indicated that there was a statistically significant correlation(r=0.827,P0.001)between the two marker systems.UPGMA clustering based on the ILP and SSR markers resulted in consensus dendrograms.The cophenetic correlation coefficient(r=0.918,0.878 and 0.924,P0.001 for the ILP,SSR and combined markers,respectively)showed a highly accurate dendrogram represented the genetic distance among these entries.The 36 entries were divided into four groups.Four African Oryza glaberrima accessions were clustered within a distinct group(Ⅰ),and the remaining entries were separated into three groups(Ⅱ,Ⅲ and IV).All the entries could be also clustered into two main groups:One was composed of Ⅲ and IV,considered as indica group, and the other was composed of Ⅰ(O.glaberrima)and Ⅱ(japonica-like).Model-based cluster analysis revealed that the japonica-like group maintained very pure ancestry while the indica group shared mixed ancestry,especially for group Ⅲ, which had seven admixtures sharing from 19.5%to 30.0%of ancestry with group Ⅳ(based on SSR markers).It is suggested that ILP and SSR markers could be very useful for the genetic study and breeding in rice.     

Haplotyping of Rice Genotypes Using Simple Sequence Repeat Markers Associated with Salt Tolerance

Salt stress is a major problem in most of the rice growing areas in the world. A major QTL Saltol associated with salt tolerance at the seedling stage has been mapped on chromosome 1 in rice. This study aimed to characterize the haplotype diversity at Saltol and additional QTLs associated with salt tolerance. Salt tolerance at the seedling stage was assessed in 54 rice genotypes in the scale of 1 to 9 score at EC = 10 d Sm-1 under controlled environmental conditions. Seven new breeding lines including three KMR3/O. rufipogon introgression lines showed similar salt tolerant ability as FL478 and can be good sources of new genes/alleles for salt tolerance. Simple sequence repeat(SSR) marker RM289 showed only two alleles and RM8094 showed seven alleles. Polymorphic information content value varied from 0.55 for RM289 to 0.99 for RM8094 and RM493. Based on 14 SSR markers, the 54 lines were clearly separated into two major clusters. Fourteen haplotypes were identified based on Saltol linked markers with FL478 as the reference. Alleles of RM8094 and RM3412 can discriminate between the salt tolerant and susceptible genotypes clearly and hence can be useful in marker-assisted selection at the seedling stage. Other markers RM10720 on chromosome 1 and RM149 and RM264 on chromosome 8 can also distinguish tolerant and susceptible lines but with lesser stringency.     

大豆耐盐性种质的分子标记辅助鉴定及其利用研究

大豆耐盐种质的鉴定对于促进大豆耐盐育种具有重要作用。本文利用已获得的大豆耐盐性基因的共显性PCR标记，对选自于国家作物种质库的59份耐盐和盐敏感种质加以鉴定，同时进行了田间耐盐性重复鉴定，并对种质库耐盐性记载结果、田间耐盐性重复鉴定结果与分子标记鉴定结果进行比较，从中选出田间耐盐性重复鉴定结果与分子标记鉴定结果一致的耐盐种质42份，分析了这些种质间的遗传多样性，为大豆处质资源的改良及耐盐遗传育种中的亲本选配提供了理论依据，同时对分子标记应用于种质鉴定和育种实践进行了有益的尝试。     

Inheritance and QTL Mapping of Salt Tolerance in Rice

An F2 population derived from the cross between Jiucaiqing (japonica) and IR36 (indica) was used to analyze the inheritance of salt tolerance in rice by genetic model of major-genes plus polygenes, and to map the corresponding QTLs by SSR molecular markers. Rice plants of P1, P2, F1 and F2 at 5- to 6- leaf stage were treated under 140 mmol/L NaCI for 10 days. Three indices representing the ability of salt tolerance of rice seedlings were measured, including salt tolerance rating (STR), Na^ /K^ ratio in roots and dry matter weight of shoots (DWS). STR, Na^ /K^ and DWS were all controlled by two major genes with modification by polygenes. Heritability of these traits from major genes was 17.8, 53.3 and 52.3%, respectively. The linkage map constructed by 62 SSR molecular markers covered a total length of about 1 142 cM. There were three QTLs detected for STR located on chromosome 1, 5 and 9, two QTLs for DWS on chromosomes 8 and 9, and two QTLs for Na^ /K^ on chromosomes 2 and 6, one on each chromosome respectively. Single QTL accounted for 6.7 to 19.3% of phenotypic variation. Identification method of salt tolerance in rice and breeding of rice varieties with salt tolerance based on molecular markers assisted selection had been discussed.