不同杂种优势群玉米子粒脱水速率分析

采用烘干法测定170份玉米自交系的子粒的脱水速率及其相关性状,利用覆盖玉米全基因组的210对SSR标记对试验材料进行全基因组扫描,通过Structure V2.3.4软件揭示其群体结构。对不同杂种优势群平均子粒脱水速率进行方差分析,并筛选出各个类群中子粒脱水速率快的自交系。结果表明:子粒脱水速率在不同自交系间存在显著差异,与穗轴和子粒的含水率等性状间存在显著相关性。试验共筛选到子粒脱水速率大于1%的自交系20个;授粉后40 d子粒含水率低于21%的自交系10个。参试自交系分成P、旅大红骨、瑞德、兰卡斯特和唐四平头5个杂种优势群;授粉后40 d子粒脱水速率依次是瑞德群0.92%、兰卡斯特群0.85%、旅大红骨群0.82%、混合群0.80%、P群0.76%、唐四平头群0.56%。本文旨在通过研究不同杂种优势群玉米自交系子粒脱水速率的特性,筛选子粒脱水速率快的自交系,为选育适应机械化作业的玉米杂交种提供借鉴。     

基于荧光SSR标记的玉米自交系遗传结构解析

利用多重SSR-PCR荧光标记检测技术,对本单位自育和引进的259份玉米自交系进行遗传多样性和亲缘关系分析,并以11个归属于不同杂种优势群的代表自交系为参照进行群体结构分析。结果显示,50对SSR引物共检测到214个等位基因变异,基因多态性指数的平均值为0.50,每个SSR标记的多态性信息量(PIC)平均值为0.44。用UPGMA方法将270份自交系划分成3大类群,进一步进行数值化杂种优势群分析,以明确所用种质资源的杂优类群利用方向,为玉米自交系的有效利用和品种的选育提供依据。     

120份欧美玉米自交系的遗传多样性分析

为了拓宽黄淮海区玉米自交系的遗传基础,加快欧美优异种质的融入与利用,本研究利用SSR分子标记对120份来自美国和塞尔维亚及2份中国的玉米自交系进行遗传多样性和聚类分析。结果表明:29个多态性SSR标记共检测到115个等位位点,平均3.97个,位点多态性信息指数(PIC)平均为0.50,较好地揭示了自交系间的遗传多样性;观测杂合度(Ho)仅为0.03,表明参试自交系遗传稳定、纯合度高;美国SS、美国NSS、塞尔维亚和中国骨干自交系4个群之间相比,美国NSS群的等位位点数(3.55)、Shannon信息指数(0.93)最高,而塞尔维亚群的有效等位位点数(2.37)最高,表明美国NSS和塞尔维亚自交系群比其他两个群遗传多样性高;4个自交系群间的遗传距离介于0.1403~0.4695之间,美国NSS群与美国SS群、塞尔维亚群之间较小(0.1419,0.1403),与中国骨干自交系群之间最大(0.4695),4个群的遗传一致度介于0.6253~0.8691之间,美国NSS群与美国SS、塞尔维亚两个群之间的遗传一致度较高,表明美国与塞尔维亚自交系之间基因交流频繁,亲缘关系较近;聚类分析将122份玉米自交系分为9大主要类群,美国SS种质、NSS种质自交系被明显的区分开,并且SS种质被分为2个主要类群(Ⅰ和Ⅸ),NSS种质被分为6个主要类群(Ⅱ-Ⅶ),来自塞尔维亚的材料分散在美国NSS种质类群。本研究结果为来自欧美的自交系在玉米育种中合理利用提供可靠依据。     

180份玉米自交系亲缘关系的分子评价

利用覆盖玉米全基因组的22对SSR引物,对180份玉米自交系的亲缘关系进行分子评价.结果显示:22对SSR引物共检测到129个等位基因,每一位点平均等位基因数5.9,变幅2～13;平均基因多样性指数和平均多态性信息含量分别为0.583和0.528.基于模型的群体结构分析将所有材料分为5个类群,与国内自交系划分的杂种优势...     

玉米耐盐种质筛选及群体遗传结构分析

本研究采用盆栽法评价了157份玉米自交系的苗期耐盐性,并利用115对SSR标记解析了耐盐自交系的群体遗传结构。结果表明,T5V、N1026、农大1145及4S等10份玉米自交系为高耐盐玉米种质;处理后10d株高、地上鲜重、地下鲜重、地上干重、地下干重及存活率可作为玉米苗期耐盐鉴定的重要指标;利用SSR分子标记,结合系谱资料,将157份自交系划为6个类群,其中具有通系5血缘（Ⅰ类群）、泰国糯玉米种质血缘（Ⅲ类群）及旅大红骨、黄早四等血缘（Ⅵ类群）的自交系耐盐性较强,是开展玉米耐盐育种的重要种质类群。本研究筛选到的耐盐种质将为玉米耐盐遗传机制研究、玉米种质遗传改良及耐盐分子育种提供优异的基因资源。     

玉米抗腐霉茎腐病种质标记基因型鉴定与遗传多样性分析

腐霉茎腐病(Pythium stalk rot)是玉米生产上的重要病害。本研究利用14个与8个抗玉米茎腐病基因连锁的分子标记对196份抗腐霉茎腐病玉米种质进行抗病标记基因型鉴定,并采用42对多态性SSR标记对54份抗病自交系进行遗传多样性分析,以期阐明玉米抗腐霉茎腐病种质的标记基因型和遗传背景,并为资源的有效利用、新基因的挖掘和杂种优势模式确定提供参考信息。14个与抗病基因连锁的分子标记将196份抗性种质鉴定为128种标记基因型,表明存在多样的抗性基因组合方式。191份种质获得与齐319、X178或1145中一个或多个的相同的扩增,表明97.45%种质可能含有与3个抗玉米茎腐病材料相同的抗病基因;粤61、郑653、赤L136、白53和18--14共5份种质均未扩增出与齐319、X178和1145相同的标记基因型,可能携带其他抗茎腐病基因;遗传背景相近的抗性种质分属不同的标记基因型,表明抗病种质携带的抗病基因可能在育种选择中发生了分离。42对多态性SSR引物在54份抗病材料中共检测出119个等位基因(Na),多态位点百分率(PPB)为99.17%,平均有效等位基因数(Ne)为1.7070,平均Nei′s基因多样性(H)为0.3999,平均Shannon′s信息指数(I)为0.5844,平均多态信息含量(PIC)为0.5527,变幅为0.2061～0.7844;通过UPGMA聚类分析,54份抗病材料被划分为2个类群,共6个亚群中,分别是旅大红骨亚群、BSSS亚群、塘四平头亚群、PA亚群、PB亚群、Lan亚群,表现出较高的遗传多样性。结果表明,我国6个杂种优势群中均含有较为丰富的抗腐霉茎腐病种质资源,其中PA亚群包含的抗病种质最多。     

我国玉米自交系茎秆性状多样性分析

研究我国玉米自交系茎秆性状特征及其多样性,是培育宜机收玉米品种的重要前提。本研究以兰卡斯特、PB、四平头、旅大红骨和瑞德五大主要类群70份主要玉米自交系为材料,调查12个茎秆相关性状(茎高、穗位高、穗位系数、茎节数、穗位节、穗节系数、穗茎长、穗茎粗、茎鲜重、茎干重、含糖量和含水量),分析性状相关性和类群多样性。结果表明,我国地方种质四平头和旅大红骨茎秆性状表型变异丰富;灌浆期玉米茎秆含水量比较稳定;玉米植株高度与茎节长度显著相关;玉米雌、雄穗节之间的节间数比较恒定;玉米茎秆含糖量与茎节长度、茎粗、果穗着生位置有关;有效降低穗位高度应从降低果穗着生节入手;类群茎秆特征鲜明:兰卡斯特茎节较少,瑞德茎秆较粗,PB茎秆较细,旅大红骨茎秆较粗、茎节较短,四平头植株较矮、茎秆含糖量较低、干物质含量较低;兰卡斯特×四平头和兰卡斯特×PB类群间存在较强的生物量及籽粒产量杂种优势;挖掘和利用茎节较长、穗位较低的玉米地方种质是我国宜机收玉米育种的技术途径。本研究结果对玉米育种具有重要指导意义。     

喀斯特高海拔山区玉米骨干自交系遗传多样性RAPD标记分析

以代表我国玉米6个主要杂种优势群旅大红骨、Lancaster、Reid、苏湾、墨白和塘四平头的标准测验种丹340、自330、7922、苏37、449、黄早四和适应我国喀斯特高海拔山区的玉米骨干自交系为材料,利用RAPD标记对其进行遗传多样性和杂种优势群划分.从90个随机引物中筛选出21个多态性好的引物扩增材料,共产生146条谱带,其中124条谱带有多态性,占86.3%,说明喀斯特高海拔山区的玉米骨干自交系具有较丰富的遗传多样性.通过UPGMA聚类分析,以遗传相似系数为0.632,可将我国喀斯特高海拔山区玉米种质资源的16个骨干自交系划分为5个类群.     

不同杂种优势群玉米籽粒脱水速率分析

研究不同杂种优势群玉米自交系籽粒脱水速率的特性,筛选脱水速率快的自交系,为选育适应机械化作业的玉米杂交种提供借鉴。本试验采用烘干测定173份玉米自交系的籽粒的脱水速率及其相关性状,利用覆盖玉米全基因组的210对SSR标记对实验材料进行全基因组扫描,通过Structure V2.3.4 软件揭示其群体结构。对不同杂种优势群平均籽粒脱水速率进行方差分析,并筛选出各个群中籽粒灌浆速率快的自交系。研究结果如下：籽粒脱水速率在不同自交系间存在显著差异,与苞叶、穗轴及籽粒的含水率等性状间存在显著相关性。试验共筛选到脱水速率大于1%的自交系20个;授粉后40天时籽粒的含水率低于21%的自交系10个。参试自交系分成P、旅大红骨、瑞德、兰卡斯特和塘四平头5个杂种优势群;授粉后40天脱水速率依次是Reid群0.92%、Lancaster群0.85%、旅大红骨群0.82%、混合群0.80%、P群0.76%、塘四平头群0.56%。     

推测187份玉米自交系基因组血统与分子亲缘关系

为提高育种效率以及开展重要 QTL 和关键基因关联性分析研究, 以 187 份生产上重要玉米自交系为材料, 以 70 个均匀分布于全基因组简单重复序列(SSR)基因座鉴定出的290个等位基因多态性为分析数据, 采用联合连锁位点与混合模型分析, 推测这些自交系的基因组血缘构成以及分子亲缘关系, 并分析了全基因连锁不平衡。当亚群数目 K=5 时, 导致似然值 P 明显下降, 亚群数据 K > 6 时, 似然值 P 没有明显上升, 表明群体结构的亚群数 K 最佳推测为 6。六个亚群分别为 PA、BSSS (含 Reid)、PB、兰卡斯特 (Lancaster)、旅大红骨 (旅大红骨及其衍生系)、四平头 (唐四平头及其衍生系)。亚群间的Kullback-Leibler 距离自 0.13 至 1.06 不等, 平均为 0.599, 各亚群间区分度较好。全基因组连锁不平衡(LD)分析表明: 与四平头种质类群内相比, 遗传基础宽泛种质的基因组内存在 LD"区块"(LD block)少且小, 对重要 QTLs 与基因的关联性分析可以避免假阳性。本研究群体结构与基因组构成分析数据为这些材料育种应用与改良提供了重要信息, 也为基于这些材料的关联性分析奠定了分析基础。     

Analysis of genetic diversity and relationships among waxy maize inbred lines in Korea using SSR markers

Information regarding the genetic diversity and genetic relationships among elite inbred lines is necessary to improve new cultivars in maize breeding programs. In this study, genetic diversity and genetic relationships were investigated among 84 waxy maize inbred lines using 50 SSR markers. A total of 269 alleles were identified at all the loci with an average of 5.38 and a range between 2 and 13 alleles per locus. The gene diversity values varied from 0.383 to 0.923 with an average of 0.641. The cluster tree generated using the described SSR markers recognized two major groups at 32% genetic similarity. Group I included 33 inbred lines while group II included 51 inbred lines. The clustering patterns of most of the waxy maize inbred lines did not clearly agree with their source, pedigree or geographic location. The average GS among all inbred lines was 35.7 ± 10.8. Analysis of waxy maize inbred lines collected from Korea and China at 50 SSR loci revealed higher values of average number of alleles (4.9) and gene diversity (0.638) in Korean inbred lines as compared to Chinese inbred lines (3.5 and 0.563, respectively). The information obtained from the present studies would be very useful for maize breeding programs in Korea.     

Microsatellite marker-based genetic diversity assessment among exotic and native maize inbred lines of Bangladesh

Hybrid development is basically dependent on the variability among available genetic resources. Polymorphism among the maize inbreds is essentially needed for maize hybridization. This study aimed at the assessment of diversity among 22 maize inbreds by 18 microsatellite markers. The study identified 187 alleles at 18 SSR loci. The amplified allele frequency per microsatellite locus was 10.4 and the highest allele per locus was 17 in SSR primer pair phi026. SSR primer set p-umc1292, phi074 and phi090 showed the lowest 6 alleles per genotype per locus. The locus phi026 showed the highest degree of gene diversity (0.92), and the locus p-umc1292 had the lowest of gene diversity (0.77) with a mean value of 0.862 among the microsatellites. At each site, the most prevalent allele varied between 0.14 (bnlg371) and 0.36. (p-umc1292). At any given locus, an average of 0.22 out of the 22 selected maize inbred lines had a common major allele. The average value of the polymorphic information content (PIC) was 0.85, within the range of 0.74 at the lowest to 0.92 at the highest. The higher PIC values of phi026 and nc013 established them to be the best markers for maize inbred lines. The UPGMA clustering generated seven distinct groups having 12.5% of similarity coefficient. The results revealed that inbred lines E10, E27, E19, E34, E35, E4, E43, E28, E11, E21, E17, E38, E25, E34, E14, E16, E39 and E3 were more diversified. These lines are promising to be used as parent materials for hybrid maize development in the future.     

Genetic diversity of African maize inbred lines revealed by SSR markers

Knowledge of genetic diversity (GD) and relationships among maize inbred lines is indispensable in a breeding program. Our objectives were to (1) investigate the level of genetic diversity among maize inbred lines and (2) assess their genetic structures by applying simple sequence repeat (SSR) markers. Fifty-six highland and mid-altitude maize inbred lines obtained from CIMMYT programs in Ethiopia and Zimbabwe were genotyped using 27 SSR loci. All of the genotypes studied could unequivocally be distinguished with the combination of the SSRs used. In total, 104 SSR alleles were identified, with a mean of 3.85 alleles per locus. The average polymorphism information content (PIC) was 0.58. GD expressed as Euclidean distance, varied from 0.28 to 0.73 with an average of 0.59. Cluster analysis using unweighted pair group method with arithmetic average (UPGMA) suggested five groups among the inbred lines. Most of the inbred lines adapted to the highlands and the mid-altitudes were positioned in different clusters with a few discrepancies. The pattern of groupings of the inbred lines was mostly consistent with available pedigree information. The variability detected using SSR markers could potentially contribute towards effective utilization of the inbred lines for the exploitation of heterosis and formation of genetically diverse source populations in Ethiopian maize improvement programs.     

Selection of high heterozygosity popcorn varieties in Brazil based on SSR markers

We analyzed genetic structure and diversity among eight populations of popcorn, using SSR loci as genetic markers. Our objectives were to select SSR loci that could be used to estimate genetic diversity within popcorn populations, and to analyze the genetic structure of promising populations with high levels of heterozygosity that could be used in breeding programs. Fifty-seven alleles (3.7 alleles per locus) were detected; the highest effective number of alleles (4.21) and the highest gene diversity (0.763) were found for the Umc2226 locus. A very high level of population differentiation was found (F(ST) = 0.3664), with F(ST) for each locus ranging from 0.1029 (Umc1664) to 0.6010 (Umc2350). This analysis allowed us to identify SSR loci with high levels of heterozygosity and heterozygous varieties, which could be selected for production of inbred lines and for developing new cultivars.     

PCR-multiplexes for a genome-wide framework of simple sequence repeat marker loci in cultivated sunflower

Simple sequence repeat (SSR) and other DNA sequence-tagged site markers can be genotyped more rapidly and cost efficiently by simultaneously amplifying multiple loci (multiplex PCR). The development of PCR-multiplexes for a nearly genome-wide framework of 78 SSR marker loci in cultivated sunflower ( Helianthus annuus L.) is described herein. The most outstanding single-locus SSR markers in the public collection (300 out of 1,089) were identified and screened for polymorphisms among 24 elite inbred lines, preparatory to selecting SSR markers for testing in multiplex PCRs. The selected SSR markers produced robust PCR products, amplified a single locus each, were polymorphic among elite inbred lines (minimum, mean and maximum heterozygosities were 0.08, 0.53 and 0.85, respectively), and supply a dense genome-wide framework of predominantly or completely codominant, single-locus DNA markers for molecular breeding and genomics research in sunflower. Thirteen six-locus multiplex PCRs were developed for 78 SSR marker loci strategically positioned throughout the sunflower genome (three to five per linkage group) by identifying compatible SSR primer combinations and optimizing multiplex PCR protocols. The multiplexed SSR markers, when coupled with 17 complementary SSR marker loci, create a 'standard genotyping' set ideal for first-pass scans of the genome, as are often needed when screening bulked-segregant DNA samples or mapping phenotypic trait loci. The minimum, mean and maximum heterozygosities of the multiplexed SSR markers were 0.38, 0.62 and 0.83, respectively. The PCR-multiplexes increase genotyping throughput, reduce reagent costs, and are ideal for repetitive genotyping applications where common sets of SSR marker loci are required or advantageous.     

Assessment of genetic variation in tomato （Solanum lycopersicum L.） inbred lines using SSR molecular markers

A study was conducted to determine the genetic diversity of 39 determinate and indeterminate tomato inbred lines collected from China, Japan, S. Korea, and USA. Using 35 SSR polymorphic markers, a total of 150 alleles were found with moderate levels of diversity, and a high number of unique alleles existing in these tomato lines. The mean number of alleles per locus was 4.3 and the average polymorphism information content (PIC) was 0.31. Unweighted Pair Group Method with Arithmetic Mean (UPGMA) clustering at genetic similarity value of 0.85 grouped the inbred lines into four groups, where one USA cultivar formed a separate and more distant cluster. The most similar inbred lines are from USA, both with determinate type, whereas the most different lines are from USA (Us-16) and Japan (Ja-2) with determinate and indeterminate growth habit, respectively. Clustering was consistent with the known information regarding geographical location and growth habit. The genetic distance information reported in this study might be used by breeders when planning future crosses among these inbred lines.     

Identification of unique alleles and assessment of genetic diversity of rabi sorghum accessions using simple sequence repeat markers

Genetic diversity among 42 sorghum accessions representing landraces (19), advanced breeding lines (16), local cultivars (2) and release varieties (5) with 30 simple sequence repeat (SSR) markers revealed 7.6 mean number of alleles per locus showing 93.3% polymorphism and an average polymorphism information content of 0.78 which range from 0.22 (Xtxp12) and 0.91(Xtxp321). The average heterozygosity and effective number of alleles per locus were 0.8 and 6.65 respectively. Cluster analysis based on microsatellite allelic diversity clearly demarcated the accessions into ten clusters. A total of 24 unique alleles were obtained from seven SSR loci in 23 accessions in a size range of 110–380 bp; these unique alleles may serve as diagnostic tools for particular region of the genome of respective genotypes. Selected SSR markers from different linkage groups provided an accurate way of determining genetic diversity at the molecular level.     

Allelic diversity of simple sequence repeats among elite inbred lines of cultivated sunflower.

Simple sequence repeat (SSR) markers were developed for cultivated sunflower (Helianthus annuus L.) from the DNA sequences of 970 clones isolated from genomic DNA libraries enriched for (CA)n,, (CT)n, (CAA)n, (CATA)n, or (GATA)n. The clones harbored 632 SSRs, of which 259 were unique. SSR markers were developed for 130 unique SSRs by designing and testing primers for 171 unique SSRs. Of the total, 74 SSR markers were polymorphic when screened for length polymorphisms among 16 elite inbred lines. The mean number of alleles per locus was 3.7 for dinucleotide, 3.6 for trinucleotide, and 9.5 for tetranucleotide repeats and the mean polymorphic information content (PIC) scores were 0.53 for dinucleotide, 0.53 for trinucleotide, and 0.83 for tetranucleotide repeats. Cluster analyses uncovered patterns of genetic diversity concordant with patterns produced by RFLP fingerprinting. SSRs were found to be slightly more polymorphic than RFLPs. Several individual SSRs were significantly more polymorphic than RFLP and other DNA markers in sunflower (20% of the polymorphic SSR markers had PIC scores ranging from 0.70 to 0.93). The newly developed SSRs greatly increase the supply of sequence-based DNA markers for DNA fingerprinting, genetic mapping, and molecular breeding in sunflower; however, several hundred additional SSR markers are needed to routinely construct complete genetic maps and saturate the genome.     

Simple sequence repeat map of the sunflower genome

Several independent molecular genetic linkage maps of varying density and completeness have been constructed for cultivated sunflower ( Helianthus annuus L.). Because of the dearth of sequence and probe-specific DNA markers in the public domain, the various genetic maps of sunflower have not been integrated and a single reference map has not emerged. Moreover, comparisons between maps have been confounded by multiple linkage group nomenclatures and the lack of common DNA markers. The goal of the present research was to construct a dense molecular genetic linkage map for sunflower using simple sequence repeat (SSR) markers. First, 879 SSR markers were developed by identifying 1,093 unique SSR sequences in the DNA sequences of 2,033 clones isolated from genomic DNA libraries enriched for (AC)(n) or (AG)(n) and screening 1,000 SSR primer pairs; 579 of the newly developed SSR markers (65.9% of the total) were polymorphic among four elite inbred lines (RHA280, RHA801, PHA and PHB). The genetic map was constructed using 94 RHA280 x RHA801 F(7) recombinant inbred lines (RILs) and 408 polymorphic SSR markers (462 SSR marker loci segregated in the mapping population). Of the latter, 459 coalesced into 17 linkage groups presumably corresponding to the 17 chromosomes in the haploid sunflower genome ( x = 17). The map was 1,368.3-cM long and had a mean density of 3.1 cM per locus. The SSR markers described herein supply a critical mass of DNA markers for constructing genetic maps of sunflower and create the basis for unifying and cross-referencing the multitude of genetic maps developed for wild and cultivated sunflowers.     

Cytomorphological and molecular diversity in backcross-derived inbred lines of sunflower (Helianthus annuus L.).

A set of 250 distinct, stable, and uniform backcross-derived inbred lines were developed in sunflower through 5 interspecific cross combinations involving 4 wild diploid annual species (Helianthus argophyllus, H. petiolaris, H. annuus, and H. debilis). The presence of the wild-species genome in these inbred lines was confirmed through higher chromosome associations (tri- and quadrivalents) at diakinesis. Maximum structural rearrangements of chromosomes were observed in lines derived from H. petiolaris. Forty morphologically diverse inbred lines along with 2 controls were subjected to measurements of phenotypic and genetic distance using 118 simple sequence repeat (SSR) markers of known map location. A total of 204 alleles were identified and the number of alleles per locus varied between 2 and 5. There were 46 unique alleles and the number of unique alleles was highest in the lines derived from the cross involving H. petiolaris. The polymorphism information content (PIC) values ranged from 0.05 to 0.575. The pair-wise comparison values based on genetic dissimilarity estimates computed using molecular marker data varied between 0.143 and 0.486 among the 42 lines. The results indicate that the sunflower gene pool could benefit from introgression of novel alleles from the latent genetic diversity present in the wild species and particularly through exploitation of the diploid annual H. petiolaris.