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.     

Grouping of Micromonospora based on their cultural-morphologic features, antibiotic-sensitivity and formation of antibiotics

500 Micromonospora cultures were subdivided into nine groups on the basis of their cultural-morphological properties, the ability to produce antibiotics of certain chemical classes, and the sensitivity to 18 different antibiotics: aurantiaca (I), cinnamomea (II), cinnamomea-vinacea (III), cinnamomea-olivacea (IV), nigra (V), nigra-violacea (VI), lilacinescens (VII), coerulea (VIII) and brunnea (IX). Cultures belonging to groups I, II, III, V and VI are moderately sensitive to most of the antibiotics and often occur in natural substrates. Black Micromonospora cultures (groups V and VI) mostly produce aminoglycoside antibiotics while brown cultures (groups II and III) form macrolide antibiotics. Cultures belonging to groups IV, VII, VIII and IX have a higher sensitivity to most of the antibiotics and are rarely isolated from natural substrates. These cultures have a weak ability to produce antibiotics.     

基于DUS测试性状的玉米自交系形态多样性分析

利用DUS测试中的47个性状对15份普通玉米自交系和13份糯玉米骨干自交系进行了形态多样性研究。结果表明:本研究所选用的普通玉米自交系较糯玉米自交系的形态变异更为丰富;47个性状的Shannon-Weaver多样性指数存在较大差异。聚类分析表明,糯玉米自交系紧密地聚在一起,与普通玉米自交系存在较大形态差别。DUS测试性状涵盖的信息量大且受环境影响相对较小,可应用于玉米自交系的形态多样性分析。     

Identification of Genetic Differentiation between Waxy and Common Maize by SNP Genotyping

Waxy maize (Zea mays L. var. ceratina) is an important vegetable and economic crop that is thought to have originated from cultivated flint maize and most recently underwent divergence from common maize. In this study, a total of 110 waxy and 110 common maize inbred lines were genotyped with 3072 SNPs to evaluate the genetic diversity, population structure, and linkage disequilibrium decay as well as identify putative loci that are under positive selection. The results revealed abundant genetic diversity in the studied panel and that genetic diversity was much higher in common than in waxy maize germplasms. Principal coordinate analysis and neighbor-joining cluster analysis consistently classified the 220 accessions into two major groups and a mixed group with mixed ancestry. Subpopulation structure in both waxy and common maize sets were associated with the germplasm origin and corresponding heterotic groups. The LD decay distance (1500–2000 kb) in waxy maize was lower than that in common maize. Fourteen candidate loci were identified as under positive selection between waxy and common maize at the 99% confidence level. The information from this study can assist waxy maize breeders by enhancing parental line selection and breeding program design.     

Genome-Wide Comparative Analysis of Genetic Diversity of Regular and Specialty Maize Inbred Lines Through Genotyping by Target Sequencing (GBTS)

Specialty maize is widely used for human and animal consumption. It is therefore important to conduct a genome-wide comparison of regular and specialty maize. In the present study, a total of 20,000 SNP markers were generated using genotyping by target sequencing (GBTS) approach, of which 18,497 high-quality SNPs were used to explore the population structure and genetic diversity of both regular (163) and specialty (124) maize resources. Neighbor-joining clustering analysis classified the 287 tested maize inbred lines into three major groups, in which popcorn and other types of maize clustered into separate groups. The regular temperate maize was further divided into five heterotic subgroups, while the waxy maize and regular tropical maize were divided into two separate subgroups. However, the quality protein maize (QPM) could not be distinguished from regular tropical maize. The highest difference in allele frequency (P?<?0.05) was observed between popcorn and waxy maize (28.2%), while 26.0% SNPs between popcorn and regular tropical maize, and 25.4% SNPs between popcorn and regular temperate maize. The information generated in this study is helpful for incorporating identified genetic variation into hybrid breeding programs for specialty maize improvement and also for genome-wide association mapping in specialty maize.

     

The chemical synthesis of the anticodon loop of an eukaryotic initiator tRNA containing the hypermodified nucleoside N6-/N-threonylcarbonyl/-adenosine/t6A/1.

In this work, the first example of chemical synthesis of oligoribonucleotide containing the hypermodified nucleoside N6-/N-threonylcarbonyl/-adenosine /t6A/ is presented. Synthesis of the heptamer C-C-C-A-U-t6A-A IX, the sequence of which is related to the anticodon loop of the initiator tRNA from yellow lupine, was achieved by: /i/ phosphotriester block synthesis of suitably protected heptamer VI containing an adenosine unit with a free exo-NH2 group, /ii/ highly effective "one-flask" procedure for the transformation of the free exo-NH2 group of adenosine unit of heptamer VI into a N,N'-disubstituted urea system of t6A of heptamer VII /hypermodification/, and /iii/ final deprotection of VIII /32% total yield/ with the use of a new approach for simultaneous hydrogenolysis /PdO-hydrogen-pyridine/ of the p-nitrobenzyl group and 2,2,2-trichloroethyl groups from carboxyl function of t6A and internucleotide phosphates respectively.     

Paradoxical sleep deprivation, stress and emotionality in the rat

88 adult male rats were divided into 9 groups. Group I and II served as controls. The rats of group III were repeatedly aroused during 4 days at the very onset of each paradoxical sleep period by direct MRF stimulation. This deprivation reduced the daily amount of paradoxical sleep by 70%, while the slow wave sleep was reduced by 10% only. In group IV, the animals were given food and water for one hour a day only. Groups V and VI were subjected to immobilization and cold stress, respectively. Groups VII, VIII and IX were deprived of paradoxical sleep on platforms of 15, 11 and 6.5 cm in diameter, respectively. Stress was estimated by the classical Selye's triad: weight of adrenals and thymus and gastric ulceration. Emotionality was measured in the open field and also by self-stimulation of the lateral hypothalamus. Neither emotional behaviour disturbances nor stress features were found after paradoxical sleep deprivation in the group III. Moreover, this deprivation induced a slight, though significant, reduction in adrenals weight. Also, no changes in emotional behaviour were noted in the stress-exposed group V and VI. Only the interplay between REM-sleep deprivation and stress on the platforms in groups VII, VIII and especially IX led to a considerable shift in emotionality.     

新疆自然高温环境下玉米自交系开花期耐热性鉴定与评价

在新疆夏季自然高温环境下,以田间玉米空秆率、果穗结实率、相对结实率为主要评价指标,综合籽粒产量与穗部性状,对26份国内外玉米骨干自交系连续2年进行耐热性鉴定与评价,结合新疆当地高温干燥的气候特点,建立一套适宜玉米种质资源大田耐热性的鉴定、评价体系,为中国玉米耐热种质资源的遗传改良和新品种选育提供参考。结果表明:(1)不同基因型玉米自交系空秆率、果穗结穗率、相对结实率和耐热性差异较大,其中GW5F、GW4F、GW7F和PH6WC耐热性最强,在高温胁迫下空秆率最低,果穗结实率与相对结实率最高;其次为PHBA6、新自351等8份自交系,在高温胁迫下籽粒产量及综合表现较好;其余自交系对高温均表现敏感。(2)高温胁迫导致玉米穗部性状严重衰退,其中的穗重、穗行数和行粒数所受影响最大。(3)在年份或材料之间,玉米自交系籽粒产量与其主要农艺性状相关性差异较大,并以果穗重与籽粒产量相关性最高。(4)以相对结实率为主要指标通过层次聚类可将参试材料分为两大群,耐热性最强的GW5F、PH6WC、GW4M聚为第Ⅰ群,其余自交系聚为第Ⅱ群;第Ⅱ群中‘吉63’、Mo17等6份自交系可聚为第1亚群,PHBA6、LH82等6份自交系聚为第2亚群,郑58、新农育6390M等11份自交系聚为第3亚群。综合分析来看,该研究中表现极耐高温的4份自交系均为国外优异种质,可用于玉米耐高温基础研究和遗传改良;表现较强耐热性的8份品系多来自于新疆本地选育的材料,其遗传基础较为广泛,适应当地的高温环境,也是优异的耐热育种资源。     

Linkage groups inAscobolus immersus

Thirty-two genes (31 mutants and themt gene) have been mapped in 9 linkage groups. There are at least 28 loci distributed as follows: two loci each in groups I, IV, V, VII, VIII and IX; 3 in group VI; 8 or more loci in group II; and 5 loci in group III.Supported by Grant GB-3325 from the National Science Foundation.     

中国17个优良玉米自交系的分子标记杂合性及其与杂交种性状的关系研究

在玉米单交种育种中 ,鉴定高产杂交种和具有优良特性的自交系是一个重要的问题。研究以 1 7个优良玉米自交系为亲本 ,按照双列杂交配组合 ,利用 RAPD技术分析了 1 7个自交系的多态性以及 RAPD标记与 9个重要农艺性状 (包括产量 )的关系。基于 RAPD标记计算的相似系数聚类将 1 7个自交系分为 5个类群 ,经分析与系谱亲缘关系基本一致。杂交种性状及其特殊配合力与亲本间的遗传距离是高度相关的 ,与聚类前比较 ,聚类后平均遗传距离与平均产量、平均特殊配合力的相关系数显著提高 ,类间平均产量高于类内平均产量。RAPD技术可揭示优良玉米自交系的系谱亲缘关系 ,将自交系划分成不同的类群 ,从而为选择类间自交系杂交 ,进行亲本选配和分子标记辅助育种提供一种方法。     

New Proctolin Analogues: Synthesis and Biological Investigation in Insects

We have extended our work on structure/activity relationship studies of the neuropeptiden proctolin (H-Arg-Tyr-Leu-Pro-Thr-OH) by evaluating the effects of the following proctolin analogues: H-X¹-Tyr-Leu-Pro-Thr-OH, where X¹ = D-Arg (I), N-Me-Arg (II), Can (III), Orn(di-Me) (IV), Orn(iPr) (V), Lys(N, N-di-Me) (VI), Lys(iPr) (VII), Lys(Nic) (VIII) and D-Lys(Nic) (IX). In analogues I–IX, the N-terminal Arg residue was replaced by basic amino acid derivatives with peptides containing amino acid residues with an isosteric system on the back side chain relative to Arg (compounds III, V and VI) or homo-Arg (compound VII). Analogues I–IX were evaluated for myotropic activity on the in vitro heart preparation of Tenebrio molitor, whereas peptides II, V, and VII–IX were tested for contractile activity on the isolated foregut of locust Schistocerca gregaria. Peptide II and III showed full cardiotropic activity in T. molitor while peptides V and VII showed 40% and 15%, respectively, locust-gut contracting activity of proctolin.     

Genetic Diversity and Molecular Evolution of Chinese Waxy Maize Germplasm

Waxy maize (Zea mays L. var. certaina Kulesh), with many excellent characters in terms of starch composition and economic value, has grown in China for a long history and its production has increased dramatically in recent decades. However, the evolution and origin of waxy maize still remains unclear. We studied the genetic diversity of Chinese waxy maize including typical landraces and inbred lines by SSR analysis and the results showed a wide genetic diversity in the Chinese waxy maize germplasm. We analyzed the origin and evolution of waxy maize by sequencing 108 samples, and downloading 52 sequences from GenBank for the waxy locus in a number of accessions from genus Zea. A sharp reduction of nucleotide diversity and significant neutrality tests (Tajima’s D and Fu and Li’s F*) were observed at the waxy locus in Chinese waxy maize but not in nonglutinous maize. Phylogenetic analysis indicated that Chinese waxy maize originated from the cultivated flint maize and most of the modern waxy maize inbred lines showed a distinct independent origin and evolution process compared with the germplasm from Southwest China. The results indicated that an agronomic trait can be quickly improved to meet production demand by selection.     

Fine genetic characterization of elite maize germplasm using high-throughput SNP genotyping

To investigate the genetic structure of Chinese maize germplasm, the MaizeSNP50 BeadChip with 56,110 single nucleotide polymorphisms (SNPs) was used to genotype a collection of 367 inbred lines widely used in maize breeding of China. A total of 41,819 informative SNPs with minor allele number of more than 0.05 were used to estimate the genetic diversity, relatedness, and linkage disequilibrium (LD) decay. Totally 1,015 SNPs evenly distributed in the genome were selected randomly to evaluate the population structure of these accessions. Results showed that two main groups could be determined i.e., the introduced germplasm and the local germplasm. Further, five subgroups corresponding to different heterotic groups, that is, Reid Yellow Dent (Reid), Lancaster Sure Crop (Lancaster), P group (P), Tang Sipingtou (TSPT), and Tem-tropic I group (Tem-tropic I), were determined. The genetic diversity of within subgroups was highest in the Tem-Tropic I and lowest in the P. Most lines in this panel showed limited relatedness with each other. Comparisons of gene diversity showed that there existed some conservative genetic regions in specific subgroups across the ten chromosomes, i.e., seven in the Lancaster, seven in the Reid, six in the TSPT, five in the P, and two in the Tem-Tropical I. In addition, the results also revealed that there existed fifteen conservative regions transmitted from Huangzaosi, an important foundation parent, to its descendants. These are important for further studies since the outcomes may provide clues to understand why Huangzaosi could become a foundation parent in Chinese maize breeding. For the panel of 367 elite lines, average LD distance was 391 kb and varied among different chromosomes as well as in different genomic regions of one chromosome. This analysis uncovered a high natural genetic diversity in the elite maize inbred set, suggesting that the panel can be used in association study, esp. for temperate regions.     

New Proctolin Analogues: Synthesis and Biological Investigation in Insects

Summary We have extended our work on structure/activity relationship studies of the neuropeptiden proctolin (H-Arg-Tyr-Leu-Pro-Thr-OH) by evaluating the effects of the following proctolin analogues: H-X¹-Tyr-Leu-Pro-Thr-OH, where X¹=d-Arg(I),N-Me-Arg (II), Can (III), Orn(di-Me) (IV), Orn (iPr) (V), Lys(N, N-di-Me) (VI), Lys(iPr) (VII), Lys(Nic) (VIII) andd-Lys(Nic) (IX). In analogues I–IX, the N-terminal Arg residue was replaced by basic amino acid derivatives with peptides containing amino acid residues with an isosteric system on the back side chain relative to Arg (compounds III, V and VI) orhomo-Arg (compound VII). Analogues I–IX were evaluated for myotropic activity on thein vitro heart preparation ofTenebrio molitor, whereas peptides II, V, and VII–IX were tested for contractile activity on the isolated foregut of locustSchistocerca gregaria. Peptide II and III showed full cardiotropic activity inT. molitor while peptides V and VII showed 40% and 15%, respectively, locust-gut contracting activity of proctolin.     

Genetic distance of inbred lines and prediction of maize single-cross performance using RAPD markers

 To evaluate the genetic diversity of 18 maize inbred lines, and to determine the correlation between genetic distance and single-cross hybrid performance, we have used random amplified polymorphic DNA (RAPD), a PCR-based technique. Eight of these lines came from a Thai synthetic population (BR-105), and the others derived from a Brazilian composite population (BR-106). Thirty two different primers were used giving a total of 325 reproducible amplification products, 262 of them being polymorphic. Genetic divergence was determinated using Jaccard’s similarity coefficient, and a final dendrogram was constructed using an unweighted pair-group method with arithmetical averages (UPGMA). Cluster analysis divided the samples into three distinct groups (GI, GII and GIII) that were confirmed by principal-coordinate analysis. The genetic distances (GD) were correlated with important agronomic traits for single-cross hybrids and heterosis. No correlation was found when group division was not considered, but significant correlations were detected between GI×GII and GI×GIII GDs with their respective single-cross hybrid grain-yield values. Three groups were identified; that is, the BR-106 population was divided in two different groups and the BR-105 population remained mostly as one group. The results indicated that RAPD can be used as a tool for determining the extent of genetic diversity among tropical maize inbred lines, for allocating genotypes into different groups, and also to aid in the choice of the superior crosses to be made among maize inbred lines, so reducing the number of crosses required under field evaluation. Received: 24 May 1996 / Accepted: 22 November 1996     

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.     

Grouping of tropical mid-altitude maize inbred lines on the basis of yield data and molecular markers

The classification of maize inbred lines into heterotic groups is an important undertaking in hybrid breeding. The objectives of our research were to: (1) separate selected tropical mid-altitude maize inbred lines into heterotic groups based on grain yield data; (2) assess the genetic relationships among these inbred lines using amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) markers; (3) examine the consistency between yield-based and marker-based groupings of the inbred lines. Thirty-eight tropical mid-altitude maize inbred lines were crossed to two inbred line testers representing the flint and dent heterotic pattern, respectively. The resulting testcrosses were evaluated in a trial at three locations for 2 years. Significant general combining ability (GCA) and specific combining ability (SCA) effects for grain yield were detected among the inbred lines. The tester inbred lines classified 23 of the 38 tested inbred lines into two heterotic groups based on SCA effects and testcross mean grain yields. This grouping was not related to endosperm type of the inbred lines. The outstanding performance of testcrosses of the remaining 15 inbred lines indicates the presence of significant genetic diversity that may allow the assignment of the lines into more than two heterotic groups. Diversity analysis of the 40 maize inbred lines using AFLP and SSR markers found high levels of genetic diversity among these lines and subdivided them into two main groups with subdivision into sub-groups consistent with breeding history, origin and parentage of the lines. However, heterotic groups formed using yield-based combining ability were different from the groups established on the basis of molecular markers. Considering the diversity of the genetic backgrounds of the mid-altitude inbred lines, the marker-based grouping may serve as the basis to design and carry out combining ability studies in the field to establish clearly defined heterotic groups with a greater genetic similarity within groups.Communicated by H.H. Geiger     

Molecular characterization of global maize breeding germplasm based on genome-wide single nucleotide polymorphisms

Characterization of genetic diversity is of great value to assist breeders in parental line selection and breeding system design. We screened 770 maize inbred lines with 1,034 single nucleotide polymorphism (SNP) markers and identified 449 high-quality markers with no germplasm-specific biasing effects. Pairwise comparisons across three distinct sets of germplasm, CIMMYT (394), China (282), and Brazil (94), showed that the elite lines from these diverse breeding pools have been developed with only limited utilization of genetic diversity existing in the center of origin. Temperate and tropical/subtropical germplasm clearly clustered into two separate groups. The temperate germplasm could be further divided into six groups consistent with known heterotic patterns. The greatest genetic divergence was observed between temperate and tropical/subtropical lines, followed by the divergence between yellow and white kernel lines, whereas the least divergence was observed between dent and flint lines. Long-term selection for hybrid performance has contributed to significant allele differentiation between heterotic groups at 20% of the SNP loci. There appeared to be substantial levels of genetic variation between different breeding pools as revealed by missing and unique alleles. Two SNPs developed from the same candidate gene were associated with the divergence between two opposite Chinese heterotic groups. Associated allele frequency change at two SNPs and their allele missing in Brazilian germplasm indicated a linkage disequilibrium block of 142 kb. These results confirm the power of SNP markers for diversity analysis and provide a feasible approach to unique allele discovery and use in maize breeding programs.     

Monolayer studies on derivatives of sphinganine and 4t-sphingenine

The pressure-area isotherms (π-A-isotherms) at temperatures between 4° and 47°C of the following derivatives of sphinganine and 4t-sphingenine have been determined.N-palmitoyl-D,L-sphingenine (I), N-palmitoyl-diacetyl-D,L-sphingenine (II), N-acetyl-sphingenine (III), N-acetyl-D,L-erythro-sphinganine (IV), N-acetyl-D,L-threo-sphinganine (V), triacetyl-D,L-erythro-sphinganine (VI), N-acetyl-3-dehydrosphingenine (VII), N-acetyl-3-dehydro-D,L-sphinganine (VIII) and diacetyl-3-dehydro-D,L-sphinganine (IX).The phases of the monolayer films have been discussed. Compounds III, V, VI, VIII and IX form trilayers when their monolayers are compressed beyond the collapse point. The folding to a trilayer is only possible from the liquid expanded state of the monolayer. In addition the formation of trilayer films occurs only when the area of the hydrophilic group exceeds that of the alkane chain of the long chain base.     

Assessment of testcross performance and genetic diversity of yellow endosperm maize lines derived from adapted × exotic backcrosses

Introduction of exotic maize (Zea mays L.) into adapted tropical germplasm may enhance genetic variability and lead to greater progress from selection. The first objective of this study was to determine if yellow endosperm lines derived from adapted × exotic backcrosses contain exotic alleles that are superior to the recurrent adapted parental line for yield and other agronomic traits in tropical environments. Thirteen exotic yellow maize inbred lines were crossed to an adapted orange line (KUSR) and the F₁s were backcrossed to KUSR to generate the first backcrosses. Fifty BC₁F₄ lines derived from these backcrosses and the recurrent parent were crossed to a common inbred tester (L4001) to form testcrosses, which were evaluated at eight environments in Nigeria. Testcrosses of the BC-derived lines differed significantly for grain yield and other agronomic traits. Only two testcrosses yielded significantly less than L4001 × KUSR, with the best 15 testcrosses producing between 289 and 1,056 kg/ha more grain yield than L4001 × KUSR. The best testcrosses were similar to or better than L4001 × KUSR for other agronomic traits. The second objective of this study was to assess the extent of genetic diversity present among the BC-derived lines. We genotyped 46 BC-derived lines including KUSR and L4001 with 10 AFLP primer pairs and found 491 polymorphic fragments. The average allelic diversity of the lines was 0.30 ± 0.01. The genetic distance of each BC-derived line from KUSR ranged between 0.49 and 0.91. The average genetic distance for all pairs of the BC-derived lines was 0.68 ± 0.004, varying from 0.34 to 0.92. The increased grain yield and genetic diversity observed in these studies provide evidence that exotic germplasm can contribute new alleles to expand the genetic base of tropical maize and develop high-yielding hybrids.