Gary E. Hart

Characteristics, linkage-map positions, and allelic differentiation of Sorghum bicolor (L.) Moench DNA simple-sequence repeats (SSRs).

Abstract Fifty one clones isolated from a size-fractionated genomic DNA library of Sorghum bicolor (L.) Moench, that had been probed with four radiolabeled di- and tri-nucleotide oligomers, were sequenced. Fifty of the clones contained one or more simple-sequence repeats (SSRs) [72% of which were (AG/TC)n SSRs] and, following analysis of the clones, polymerase-chain-reaction primer sets that amplify 38 unique SSR loci were developed. Genotyping of the 38 loci in 18 sorghum accessions, including the parents of a recombinant inbred (RI) mapping population, revealed polymorphism at 36 of the loci among the 18 accessions and at 31 of the loci (not including null alleles at two loci) between the parents of the RI population. All of the latter 31 loci were mapped. The genotypes at 17-mapped SSR loci were assayed in 190 S. bicolor accessions in order to determine δ*T, the estimated level of allelic differentiation (the estimated probability that two members of a population, chosen at random and without replacement, differ in allelic composition), at each of the loci. The mean δ*Tvalue determined for S. bicolor overall was 0.89, the range of mean δ*T values for ten S. bicolor races was from 0.88 to 0.83, and the range of mean δ*T values for ten working groups (= sub-races) of the race caudatum, with only two exceptions, was from 0.87 to 0.79. The lowest δ*T values for six of the loci among the ten race-caudatum working groups ranged from 0.86 to 0.70; thus, the probability that different alleles will be present at one or more of these loci in two accessions chosen at random from a working group is > 0.996 when three of the loci are genotyped, and > 0.9999 when all six of the loci are genotyped. The results of this study confirm that most S. bicolor SSR loci are sufficiently polymorphic to be useful in marker- assisted selection programs and they indicate that the levels of polymorphism at some loci are high enough to allow the vast majority of S. bicolor accessions, even accessions within working groups, to be distinguished from one another by determining the genotypes at a small number, perhaps as few as a half-dozen, SSR loci.

Chromosomal location and evolution of isozyme structural genes in hexaploid wheat

SummaryThe genetic control of the production of multiple forms of lipoxygenase, endopeptidase, acid phosphatase, aminopeptidase, and alcohol dehydrogenase in hexaploid wheat has been studied using the zymogram technique. Aneuploid strains which encompass a range of from zero to four doses of each chromosome in the complement were analysed. The localisation of 22 structural genes to specific chromosome arms is reported. Extensive inter-genomic variation between homoeologous isozyme structural genes has been detected. The results of this study are consistent with the hypothesis that many of the duplicated structural genes of this species have diverged in structure and function subsequent to the origin of the polyploid wheats.

Alignment of genetic maps and QTLs between inter- and intra-specific sorghum populations

To increase the value of associated molecular tools and also to begin to explore the degree to which interspecific and intraspecific genetic variation in Sorghum is attributable to corresponding genetic loci, we have aligned genetic maps derived from two sorghum populations that share one common parent (Sorghum bicolor L. Moench accession BTx623) but differ in morphological and evolutionarily distant alternate parents (S. propinquum or S. bicolor accession IS3620C). A total of 106 well-distributed DNA markers provide for map alignment, revealing only six nominal differences in marker order that are readily explained by sampling variation or mapping of paralogous loci. We also report a total of 61 new QTLs detected from 17 traits in these crosses. Among eight corresponding traits (some new, some previously published) that could be directly compared between the two maps, QTLs for two (tiller height and tiller number) were found to correspond in a non-random manner (P<0.05). For several other traits, correspondence of subsets of QTLs narrowly missed statistical significance. In particular, several QTLs for leaf senescence were near loci previously mapped for ‘stay-green’ that have been implicated by others in drought tolerance. These data provide strong validation for the value of molecular tools developed in the interspecific cross for utilization in cultivated sorghum, and begin to separate QTLs that distinguish among Sorghum species from those that are informative within the cultigen (S. bicolor).

Genetic mapping of Sorghum bicolor (L.) Moench QTLs that control variation in tillering and other morphological characters

Abstract Grain yield of Sorghum bicolor (L.) Moench is significantly influenced by genetically controlled variation in the number of tillers, plant height, time of anthesis, and various other morphological and physiological characters. In this study, a minimum of 27 unique QTLs that control variation in nine morphological traits, including the presence versus the absence and the height of basal tillers, were mapped, and the percentage of additive genetic variance explained by the QTLs was determined in a population of 137 recombinant inbred lines in two environments. Four QTLs explained from 86.3% to 48.9% (depending upon the environment) of the additive genetic variance in the number of basal tillers with heads, and seven QTLs explained from 85.9% to 47.9% of the additive genetic variance in panicle width. It is unlikely that different alleles were segregating in the mapping population at any of the major dwarfing loci, but five QTLs that explained from 65.8% to 52.0% of the additive genetic variance in main-culm height were mapped. QTLs controlling variation in height of the tallest basal tiller, number of basal tillers per basal-tillered plant, panicle length, leaf angle, maturity, and awn length also were mapped. Three or more QTLs were mapped in linkage groups A, E, G, and I, while none were mapped in linkage groups B and D. Several of the QTLs mapped in this study are likely candidates for marker-assisted selection in breeding programs.

Use of isozymes as chromosome markers in the isolation and characterization of wheat-barley chromosome addition lines.

The alcohol dehydrogenase (ADH), glutamic oxaloacetic transaminase (GOT), aminopeptidase (AMP), endopeptidase (EP), and esterase (EST) zymogram phenotypes of Chinese Spring wheat, Betzes barley, Chinese Spring-Betzes heptaploids, and a number of presumptive Betzes chromosome additions to Chinese Spring were determined. It was found that four disomic chromosome addition lines could be distinguished from one another and from the other three possible lines on the basis of the zymogram phenotypes of these isozymes. The structural gene Adh-H1 was located in Betzes chromosome 4, the genes Got-H2 and Amp-H1 in chromosome 6, and the gene Ep-H1 in chromosome 1. These gene locations provide evidence of homoeology between Betzes chromosomes 4, 6, and 1 and the Chinese Spring chromosomes of homoeologous groups 4, 6, and 7, respectively.

A RFLP linkage map of Sorghum bicolor (L.) Moench

A RFLP linkage map of sorghum composed principally of markers detected with sorghum low-copy-number nuclear DNA clones has been constructed. The map spans 1789 cMs and consists of 190 loci grouped into 14 linkage groups. The 10 largest linkage groups consist of from 10 to 24 markers and from 103 to 237 cMs, and the other 4 linkage groups consist of from 2 to 5 markers and from 7 to 62 cMs. The map was derived in Sorghum bicolor ssp. bicolor by analysis of a F2 population composed of 50 plants derived from a cross of IS 3620C, a guinea line, and BTx 623, an agronomically important inbred line derived from a cross between a zera zera (a caudatum-like sorghum) and an established kafir line. The restriction fragment length polymorphism (RFLP) frequency detected in this population using polymerase chain reaction (PCR)-amplifiable low-copy-number sorghum clones and five restriction enzymes was 51%. A minimal estimate of the number of clones that detect duplicate sequences is 11 %. Null alleles occurred at 13% of the mapped RFLP loci.

Chromosomal locations of eleven Elytrigia elongata (= Agropyron elongatum ) isozyme structural genes

The zymogram phenotypes of 11 enzymes were determined for 22 Triticum aestivum cv. Chinese Spring-Elytrigia elongata disomic and ditelosomic chromosome addition lines. Eleven isozyme structural genes were located in specific arms of six E. elongata chromosomes, as follows: Gpi-El in 1ES, Est-El in 3ES, Got-E3 in 3EL, Adh-El and Lpx-El in 4ES, Adh-E2 and Lpx-E2 in 5EL, Amp-El in 6Ea, Adh-E3 and Got-E2 in 6E/?, and Ep-El in 7EL. The E. elongata chromosomes present in five disomic addition lines have previously been designated IE, 2E, 4E, 6E, and 7E to indicate their homoeology with Chinese Spring chromosomes. The results of this study support these designations. The development of disomic putative 3E and 5E addition lines is reported. The added chromosomes designated IV, V, and VI that are present in three of the seven original disomic T. aestivum-E. elongata addition lines are translocated. Evidence that VL and VIL are opposite arms of 2E and that IV is partially homoeologous to 3E has been published. The results reported in this paper indicate that IVS = 3ES, IVL = 7EL, VS = 3ES, and VIS = 5ES and are consistent with VL and VIL being opposite arms of 2E. The synteny relationships of the 11 E. elongata isozyme genes identified in this study are fully consistent with those of homoeologous T. aestivum cv. Chinese Spring genes and thus provide evidence that the gene synteny groups which these two species inherited from their common ancestor are conserved. This study further documents the valuable role that studies of isozyme genes can play in the isolation, characterization, and maintenance of alien chromosomes, telosomes, and chromosomal segments in wheat strains.

RFLP-based assay of Sorghum bicolor (L.) Moench genetic diversity

Sixty-two single-copy sorghum DNA clones were used to compare restriction fragment patterns of 53 sorghum accessions from Africa, Asia and the United States. Included were accessions from five morphological races of the cultivated subspecies bicolor, and four races of the wild subspecies verticilliflorum. From two to twelve alleles were detected with each probe. There was greater nuclear diversity in the wild subspecies (255 alleles in ten accessions) than in the domestic accessions (236 alleles in 37 accessions). Overall, 204 of the 340 alleles (60%) that were detected occurred in both subspecies. Phylogenetic analysis using parsimony separated the subspecies into separate clusters, with one group of intermediate accessions. Though exceptions were common, especially for the race bicolor, accessions classified as the same morphological race tended to group together on the basis of RFLP similarities. Selection for traits such as forage quality may have led to accessions genetically more similar to other races being classified as bicolors, which have a loose, small-grained panicle similar to wild races. Population statistics, calculated using four nuclear and four cytoplasmic probes that detect two alleles each, revealed a low but significant amount of heterozygosity, and showed little differentiation in alleles in the wild and cultivated subspecies. Outcrossing with foreign pollen appears to have been more important than migration via seed dispersal as a mechanism for gene flow between the wild and domestic accessions included in this study.

GLUTAMATE OXALOACETATE TRANSAMINASE ISOZYMES OF TRITICUM: EVIDENCE FOR MULTIPLE SYSTEMS OF TRIPLICATE STRUCTURAL GENES IN HEXAPLOID WHEAT

ABSTRACT. The glutamate oxaloacetate transaminase (GOT) zymogram phenotypes of 54 wheat strains, each possessing a distinctive chromosomal constitution, were determined. Three of the several genetically independent GOT systems expressed were analyzed. The results obtained support the hypothesis that the active GOT isozymes of each system are dimers composed of the six possible combinations of subunits coded by triplicate structural genes. Sets of triplicate GOT structural genes were linked to the chromosome arms 3Aα, 3BL, and 3Dα, to 6Aα, 6BS, and 6Dα, and to 6Aβ, 6BL, and 6Dβ. Genes involved in the production of GOT isozymes were also located in the chromosomes of homoeologous group 7. Tissue specificity in the expression of the GOT systems was observed. The results of this study suggest that genetic regulation of the expression of the GOT isozymes occurs at the level of gene transcription and that the rate of transcription is the same for each copy present of each of the three possible members of the homoeologous sets of GOT structural genes.

Extended physical maps and a consensus physical map of the homoeologous group-6 chromosomes of wheat (Triticum aestivum L. em Thell.)

Abstract Extended physical maps of chromosomes 6A, 6B and 6D of common wheat (Triticum aestivum L. em Thell., 2n=6x=42, AABBDD) were constructed with 107 DNA clones and 45 homoeologous group-6 deletion lines. Two-hundred and ten RFLP loci were mapped, including three orthologous loci with each of 34 clones, two orthologous loci with each of 31 clones, one locus with 40 clones, two paralogous loci with one clone, and four loci, including three orthologs and one paralog, with one clone. Fifty five, 74 and 81 loci were mapped in 6A, 6B and 6D, respectively. The linear orders of the mapped orthologous loci in 6A, 6B and 6D appear to be identical and 65 loci were placed on a group-6 consensus physical map. Comparison of the consensus physical map with eight linkage maps of homoeologous group-6 chromosomes from six Triticeaespecies disclosed that the linear orders of the loci on the maps are largely, if not entirely, conserved. The relative distributions of loci on the physical and linkage maps differ markedly, however. On most of the linkage maps, the loci are either distributed relatively evenly or clustered around the centromere. In contrast, approximately 90% of the loci on the three physical maps are located either in the distal one-half or the distal two-thirds of the six chromosome arms and most of the loci are clustered in two or three segments in each chromosome.