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Artykuły w czasopismach na temat "Two loci segregating"
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Echt, C. S., K. K. Kidwell, S. J. Knapp, T. C. Osborn i T. J. McCoy. "Linkage mapping in diploid alfalfa (Medicago sativa)". Genome 37, nr 1 (1.02.1994): 61–71. http://dx.doi.org/10.1139/g94-008.
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A genome map of cultivated alfalfa was constructed using segregating restriction fragment length polymorphisms (RFLPs) and random amplified polymorphic DNAs (RAPDs) in a diploid backcross population generated from noninbred parents. Among the 153 loci scored in 87 progeny, four segregation ratios were observed for codominant and dominant markers: 1:1, 1:2:1, 1:1:1:1, and 3:1. Deviations from expected Mendelian ratios (p < 0.05) were observed for 34% of the loci studied. A genome map was assembled from two separate linkage maps, each constructed from a subset of the segregation data. One linkage map was constructed from 46 RFLP and 40 RAPD markers segregating 1:1 from the F1 parent of the backcross and the other linkage map was constructed from 33 RFLP and 28 RAPD markers segregating 1:1 from the recurrent parent. Sixteen loci with alleles segregating 1:1 from both parents were used as locus bridges to align individual linkage groups between the two maps. The combined use of RFLPs and RAPDs was an effective method for developing an alfalfa genome map.Key words: genome mapping, RAPD, RFLP, locus bridges.
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Verhaegen, Daniel, i Christophe Plomion. "Genetic mapping in Eucalyptus urophylla and Eucalyptus grandis using RAPD markers". Genome 39, nr 6 (1.12.1996): 1051–61. http://dx.doi.org/10.1139/g96-132.
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Two single-tree linkage maps were constructed for Eucalyptus urophylla and Eucalyptus grandis, based on the segregation of 480 random amplified polymorphic DNA (RAPD) markers in a F1 interspecific progeny. A mixture of three types of single-locus segregations were observed: 244 1:1 female, 211 1:1 male, and 25 markers common to both, segregating 3:1. Markers segregating in the 1:1 ratio (testcross loci) were used to establish separate maternal and paternal maps, while markers segregating in the 3:1 ratio were used to identify homology between linkage groups of the two species maps. An average of 2.8 polymorphic loci were mapped for each arbitrary decamer primer used in the polymerase chain reaction. The mean interval size beween framework markers on the maps was 14 cM. The maps comprised 269 markers covering 1331 cM and 236 markers covering 1415 cM, in 11 linkage groups, for E. urophylla (2n = 2x = 22) and E. grandis (2n = 2x = 22), respectively. A comparative mapping analysis with two other E. urophylla and E. grandis linkage maps showed that RAPDs could be reliable markers for establishing a consensus species map. RAPD markers were automatically and quantitatively scored with an imaging analyzer. They were classified into four categories based on their optical density. A fragment intensity threshold is proposed to optimize the selection of reliable RAPD markers for future mapping experiments. Key words : genetic linkage map, Eucalyptus urophylla, Eucalyptus grandis, random amplified polymorphic DNA, RAPD, automated data collection.
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Wang, Xindan, i David J. Sherratt. "Independent Segregation of the Two Arms of the Escherichia coli ori Region Requires neither RNA Synthesis nor MreB Dynamics". Journal of Bacteriology 192, nr 23 (1.10.2010): 6143–53. http://dx.doi.org/10.1128/jb.00861-10.
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ABSTRACT The mechanism of Escherichia coli chromosome segregation remains elusive. We present results on the simultaneous tracking of segregation of multiple loci in the ori region of the chromosome in cells growing under conditions in which a single round of replication is initiated and completed in the same generation. Loci segregated as expected for progressive replication-segregation from oriC, with markers placed symmetrically on either side of oriC segregating to opposite cell halves at the same time, showing that sister locus cohesion in the origin region is local rather than extensive. We were unable to observe any influence on segregation of the proposed centromeric site, migS, or indeed any other potential cis-acting element on either replication arm (replichore) in the AB1157 genetic background. Site-specific inhibition of replication close to oriC on one replichore did not prevent segregation of loci on the other replichore. Inhibition of RNA synthesis and inhibition of the dynamic polymerization of the actin homolog MreB did not affect ori and bulk chromosome segregation.
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Foolad, M. R. "Unilateral Incompatibility as a Major Cause of Skewed Segregation in the Cross between Lycopersicon esculentum and L. pennellii". HortScience 31, nr 4 (sierpień 1996): 625d—625. http://dx.doi.org/10.21273/hortsci.31.4.625d.
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Skewed segregations are frequent events in segregating populations derived from different interspecific crosses in tomato. To determine a basis for skewed segregations in the progeny of the cross between Lycopersicon esculentum and L. pennellii, monogenic segregations of 16 isozyme loci were analyzed in an F2 and two backcross populations of this cross. In the F2, nine loci mapping to chromosomes 1, 2, 4, 9, 10, and 12 exhibited skewed segregations and in all cases there was an excess of L. pennellii homozygotes. The genotypic frequencies at all but one locus were at Hardy–Weinberg equilibria. In the backcross populations, all except two loci exhibited normal Mendelian segregations. No postzygotic selection model could statistically or biologically explain the observed segregation patterns. A prezygotic selection model, assuming selective elimination of the male gametophytes during pollen function (i.e., from pollination to karyogamy) adequately explained the observed segregations in all three populations. The direction of the skewed segregations in the F2 was consistent with that expected based on the effects of unilateral incompatibility reactions between the two species. In addition, the chromosomal locations of five of the nine markers that exhibited skewed segregations coincided with the locations of several known compatibility-related genes in tomato. Multigenic unilateral incompatibility reactions between L. esculentum pollen and the stigma or style of L. pennellii (or its hybrid derivatives) are suggested to be the major cause of the skewed segregations in the F2 progeny of this cross.
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Groover, A., M. Devey, T. Fiddler, J. Lee, R. Megraw, T. Mitchel-Olds, B. Sherman, S. Vujcic, C. Williams i D. Neale. "Identification of quantitative trait loci influencing wood specific gravity in an outbred pedigree of loblolly pine." Genetics 138, nr 4 (1.12.1994): 1293–300. http://dx.doi.org/10.1093/genetics/138.4.1293.
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Abstract We report the identification of quantitative trait loci (QTL) influencing wood specific gravity (WSG) in an outbred pedigree of loblolly pine (Pinus taeda L.). QTL mapping in an outcrossing species is complicated by the presence of multiple alleles (> 2) at QTL and marker loci. Multiple alleles at QTL allow the examination of interaction among alleles at QTL (deviation from additive gene action). Restriction fragment length polymorphism (RFLP) marker genotypes and wood specific gravity phenotypes were determined for 177 progeny. Two RFLP linkage maps were constructed, representing maternal and paternal parent gamete segregations as inferred from diploid progeny RFLP genotypes. RFLP loci segregating for multiple alleles were vital for aligning the two maps. Each RFLP locus was assayed for cosegregation with WSG QTL using analysis of variance (ANOVA). Five regions of the genome contained one or more RFLP loci showing differences in mean WSG at or below the P = 0.05 level for progeny as grouped by RFLP genotype. One region contained a marker locus (S6a) whose QTL-associated effects were highly significant (P > 0.0002). Marker S6a segregated for multiple alleles, a prerequisite for determining the number of alleles segregating at the linked QTL and analyzing the interactions among QTL alleles. The QTL associated with marker S6a appeared to be segregating for multiple alleles which interacted with each other and with environments. No evidence for digenic epistasis was found among the five QTL.
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Kianian, Shahryar F., i Carlos F. Quiros. "Genetic analysis of major multigene families in Brassica oleracea and related species". Genome 35, nr 3 (1.06.1992): 516–27. http://dx.doi.org/10.1139/g92-076.
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Sequences homologous to rRNA, napin, cruciferin, self-incompatibility, isocitrate lyase, and malate synthase were mapped to the Brassica oleracea genome. Four segregating populations were used to disclose possible distortions in segregation and linkage ratios while maximizing detectable polymorphism. rRNA mapped to three unlinked loci, which reside on different chromosomes. Certain restriction fragment length polymorphism variations detected by the rDNA probe reflect changes in the number of intergenic spacer subrepeats, the size of which was estimated to be about 300 base pairs. All five napin, three self-incompatibility, and two isocitrate lyase loci mapped in linkage clusters, while those of cruciferin and malate synthase (two loci each) were independent.Key words: Brassica oleracea, RFLP analysis, linkage analysis, multigene family.
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Jarrell, David C., i Mikeal L. Roose. "A GENETIC MAP OF CITRUS BASED ON ISOZYMES AND RFLPS." HortScience 25, nr 9 (wrzesień 1990): 1154b—1154. http://dx.doi.org/10.21273/hortsci.25.9.1154b.
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We report a preliminary genetic map of citrus based on segregation of 8 isozyme and at least 33 RFLP loci. The segregating population consisted of 60 plants from a cross of two citrus rootstock, `Sacaton' citrumelo × `Troyer' citrange. This cross represents an intergeneric F2 since `Sacaton' is Citrus paradisi (grapefruit) × Poncirus trifoliata (trifoliate orange) and `Troyer' is C. sinensis (sweet orange) × P. trifoliata. RFLPs were identified using anonymous probes from both cDNA and genomic DNA libraries of citrus. About 20% of the loci deviated significantly from Mendelian segregation. Two-point linkage analysis identified 8 linkage groups in which pairs of loci were within 30 centimorgans. This suggests that we have markers on most of the 9 chromosomes of Citrus. A map based on multipoint linkage estimates will be reported. Evidence for structural rearrangements between Citrus and Poncirus and extension of the map to additional marker and disease resistance loci will be discussed.
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Beaver, J. A., i A. F. Iezzoni. "Allozyme Inheritance in Tetraploid Sour Cherry (Prunus cerasus L.)". Journal of the American Society for Horticultural Science 118, nr 6 (listopad 1993): 873–77. http://dx.doi.org/10.21273/jashs.118.6.873.
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Inheritance for seven enzyme loci was determined in seeds produced from crosses and self-pollinations involving four sour cherry parents and one open-pollinated ground cherry (P. fruticosa Pall.) parent. Segregation data were used to identify allozymes and determine whether sour cherry is a naturally occurring allo- or autotetraploid. Three allozymes were identified at the 6-Pgd-1 locus, and two were identified at each of the following loci: Pgi-2, Lap-1, Adh-1, Idh-2, Pgm-2, and 6-Pgd-2. Segregating allozyme patterns for the diagnostic loci Idh-2, Pgm-2, 6-Pgd-1, and 6-Pgd-2 tit disomic inheritance models and thus confirmed the allotetraploid hypothesis for sour cherry. Chi-square tests of independence between loci indicated that Pgi-2, Adh-1, Idh-2, 6-Pgd-1, and 6-Pgd-2 were not linked.
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Landry, Benoit S., Rick V. Kesseli, Barry Farrara i Richard W. Michelmore. "A Genetic Map of Lettuce (Lactuca sativa L.) With Restriction Fragment Length Polymorphism, Isozyme, Disease Resistance and Morphological Markers". Genetics 116, nr 2 (1.06.1987): 331–37. http://dx.doi.org/10.1093/genetics/116.2.331.
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ABSTRACT A detailed linkage map of lettuce was constructed using 53 genetic markers including 41 restriction fragment length polymorphism (RFLP) loci, five downy mildew resistance genes, four isozyme loci and three morphological markers. The genetic markers were distributed into nine linkage groups and cover 404 cM which may be 25-30% of the lettuce genome. The majority (31 of 34) of the RFLP probes detected single segregating loci, although seven of these may have been homologous to further monomorphic loci. When several loci were detected by a single probe, the loci were generally linked, suggesting tandem duplications. One probe, however, detected loci in three linkage groups suggesting translocations. The five downy mildew resistance genes (Dm1, Dm3, Dm4, Dm5/8 and Dm13), segregating in the Calmar × Kordaat cross, represented each of the four resistance gene linkage groups. Dm5/8 is flanked by two cDNA loci, each located 10 cM away. These flanking markers will be used to study the source of variation in downy mildew genes and are also part our strategy to clone resistance genes.
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Ritter, E., C. Gebhardt i F. Salamini. "Estimation of recombination frequencies and construction of RFLP linkage maps in plants from crosses between heterozygous parents." Genetics 125, nr 3 (1.07.1990): 645–54. http://dx.doi.org/10.1093/genetics/125.3.645.
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Abstract The construction of a restriction fragment length polymorphism (RFLP) linkage map is based on the estimation of recombination frequencies between genetic loci and on the determination of the linear order of loci in linkage groups. RFLP loci can be identified as segregations of singular or allelic DNA-restriction fragments. From crosses between heterozygous individuals several allele (fragment) configurations are possible, and this leads to a set of formulas for the evaluation of p, the recombination frequency between two loci. Tables and figures are presented illustrating a general outline of gene mapping using heterozygous populations. The method encompasses as special cases the mapping of loci from segregating populations of pure lines. Formulas for deriving the recombination frequencies and information functions are given for different fragment configurations. Information functions derived for relevant configurations are also compared. A procedure for map construction is presented, as it has been applied to RFLP mapping in an allogamous crop.
Rozprawy doktorskie na temat "Two loci segregating"
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Zhu, Guohua. "Ascertainment in two-phase sampling designs for segregation and linkage analysis /". Connect to text online, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=case1112844349.
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Thesis (Ph. D.)--Case Western Reserve University, 2005.[School of Medicine] Department of Epidemiology and Biostatistics. Includes bibliographical references. Available online via OhioLINK's ETD Center.
Części książek na temat "Two loci segregating"
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Raemisch, Rick. "Colorado Ends Prolonged, Indeterminate Solitary Confinement". W Solitary Confinement, 311–24. Oxford University Press, 2019. http://dx.doi.org/10.1093/oso/9780190947927.003.0018.
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When did it become okay to lock someone in a cell that is the size of a parking space, twenty-three hours per day, seven days a week for decades? When did it become okay to lock someone who is mentally ill in a cell the size of a parking space, twenty-three hours per day for years, and let the demons chase him or her around in the cell? And when did it ever become okay to take someone who had spent years in segregation and release them directly to the community? If our mission is public safety, and it is, why are we sending people back to society worse than when they came in? Ninety-five percent of those incarcerated in Colorado will return to their community. In September of 2017, the Colorado Department of Corrections abolished the use of administrative segregation, or as it is now nationally termed by corrections, extended restrictive housing. Currently, Colorado is the only state in the United States that has abolished this practice. In order to see where we are, it is important to see where we were, and the journey that brought us here.
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Ponmalar A, Anand J, Dharshini S, Aishwariya K, Mahalakshmi S i Abinaya S. "Smartphone Controlled Fingerprint Door Look System (SCFDLS)". W Advances in Parallel Computing Technologies and Applications. IOS Press, 2021. http://dx.doi.org/10.3233/apc210164.
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Currently, the entire globe is infected with COVID 19, and everyone is doing everything they can do to avoid contracting this serious sickness by adhering to social segregation, wearing coverings, using credit-only exchanges, and refraining from contacting anything to prevent the spread of germs. With the advancement of technology, traditional locks are becoming relics of the past, while new biometric-based locks and Radio Frequency Identification (RFID)-based locks are becoming increasingly in common. The unique finger impression based locks, as well as participation record-keeping devices, are used in the vast majority of offices and universities, however due to the pandemic, it is no longer appropriate to do so. As a result, the proposed system use a solenoid lock to make the door lock. Aside from the equipment, a mobile application is used to inspect and double-check the finger impression and send the confirmation, as well as an id, to Arduino through Bluetooth. The entry way lock will be bolted and opened using the smartphone fingerprint sensor.
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Abdallah, Sarah B., i Thomas V. Fernandez. "Genetic Susceptibility in Tourette Syndrome". W Tourette Syndrome, redaktorzy Sarah B. Abdallah i Thomas V. Fernandez, 125–36. Wyd. 2. Oxford University Press, 2022. http://dx.doi.org/10.1093/med/9780197543214.003.0009.
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Abstract Early twin and family studies point to a significant genetic contribution to Tourette syndrome (TS). Informed by early segregation analyses in TS families pointing to a single-gene autosomal dominant inheritance pattern with partial penetrance, initial efforts at gene discovery in TS utilized parametric linkage analysis in large multigenerational families but failed to identify a single specific genetic locus. Later segregation analyses supported the current characterization of TS as a complex, genetically heterogeneous disorder. Nonparametric linkage analyses have yet to identify common TS risk alleles. Candidate gene association studies in TS have not yielded significant reproducible findings. Genome-wide association studies (GWAS) have proven valuable for identifying and replicating loci for common complex traits and disorders across the medical field. A GWAS meta-analysis of almost 5,000 cases identified a significant locus in FLT3. Polygenic risk scores, calculated from the most recent TS GWAS summary statistics, have shown correlation with tic severity and affected status in independent samples. There has been an increasing effort to evaluate the contribution of rare allele variants toward TS. Recent studies have found a greater burden of rare copy number variants in TS cases; the largest study to date identified NRXN1 and CNTN6 as TS risk genes. Identifying rare de novo single nucleotide variants and indels in more than 800 parent–child trios has so far identified six likely TS risk genes (WWC1, CELSR3, OPA1, NIPBL, FN1, and FBN2), which present an enriched function for cell polarity.
Streszczenia konferencji na temat "Two loci segregating"
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Bernardi, F., G. Marchetti, F. Vannini, L. Felloni, F. Panicucci i F. Conconi. "SPORADISM INVESTIGATION AND CARRIER DETECTION IN HAEMOPHILIA A BY RFLP ANALYSIS". W XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644011.
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Restriction fragment lenght polymorphisms (RFLPs)analysis has been employed for carrier detection andfor sporadism study in Haemophilia A. Three RFLPs, one intragenic in FC8 (647/BcII) and two with close linkage to Haemophilia A at DXS52 (Stl4/Taql) and DXS15 (DX13/BgIII), were used.In 20 families 29 carrier status determinations havebeen performed.In order to investigate sporadicity and to estimate the sex ratio of mutation rates directely, 17 families with isolated cases of haemophilia A were studied.In eight out of the 17 families the RFLPsanalysis excluded the carrier status of the maternalgrandmothers.Since by hemostatic studies the eight mothers of the propositi were shown to be haemophilia carriers, the origin of the newly mutated genes was inferred from the RFLP patterns: six haemophilic genes derive from the normal maternal grandfathers and two from the maternal grandmothers.Possible recombinations between FVIII locus and the extragenic RFLPs loci have to be considered; however the intragenic Bell RFLP is informative in five out of the eight families and the DXl3 and Stl4 patterns are concordant.The data indicate a higher mutation rate in males than in females gametes as previously suggested, althought not unanimously, by segregation analysis and coagulation studies. The RFLP analysis in a large number of families with isolated cases of haemophilia isnecessary to define the precise ratio of sex mutation rate for this disease.Work supported by P.F. Ing. Gen. Basi Mol. Mai. Ered. Contratto CNR N 8400877.
Raporty organizacyjne na temat "Two loci segregating"
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Gall, Graham A. E., Gideon Hulata, Eric M. Hallerman, Bernard May i Umiel Nakdimon. Creating and Characterizing Genetic Variation in Tilapia through the Creation of an Artificial Center of Origin. United States Department of Agriculture, luty 2000. http://dx.doi.org/10.32747/2000.7574344.bard.
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Five stocks of tilapia [oreochromis niloticus (on), red O. niloticus (ROn), O. aureus (Oa), O. mossambicus (Om), and Sarotherodon galilaeus (Sg)] were used to produce two-way (F1), three-way (3WC) and four-way crosses (4WC). Three 4WC groups, containing equal representation of all four species, formed the base population for a new synthetic stock, called an "artificial center of origin" (ACO). Four genomic maps were created using microsatellite and AFLP markers, two from a 3WC family [Om female and (Oa x ROn) male] and two from a 4WC family [(Om x Oas) females and (Sg x On) male]. Sixty-two loci segregating from the female parent of the 3WC mapped to 14 linkage groups while 214 loci from the male parent mapped to 24 linkage groups. Similarly, 131 loci segregating from the female parent of the 4WC mapped to 26 linkage groups and 118 loci from the male parent mapped to 25 linkage groups. Preliminary screening of an F2 and a 4WC family identified a number of loci associated with cold tolerance and body weight. These loci were clustered in a few linkage groups, suggesting they may be indicative of quantitative trait loci.
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Weller, Joel I., Harris A. Lewin i Micha Ron. Determination of Allele Frequencies for Quantitative Trait Loci in Commercial Animal Populations. United States Department of Agriculture, luty 2005. http://dx.doi.org/10.32747/2005.7586473.bard.
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Individual loci affecting economic traits in dairy cattle (ETL) have been detected via linkage to genetic markers by application of the granddaughter design in the US population and the daughter design in the Israeli population. From these analyses it is not possible to determine allelic frequencies in the population at large, or whether the same alleles are segregating in different families. We proposed to answer this question by application of the "modified granddaughter design", in which granddaughters with a common maternal grandsire are both genotyped and analyzed for the economic traits. The objectives of the proposal were: 1) to fine map three segregating ETL previously detected by a daughter design analysis of the Israeli dairy cattle population; 2) to determine the effects of ETL alleles in different families relative to the population mean; 3) for each ETL, to determine the number of alleles and allele frequencies. The ETL on Bostaurusautosome (BT A) 6 chiefly affecting protein concentration was localized to a 4 cM chromosomal segment centered on the microsatellite BM143 by the daughter design. The modified granddaughter design was applied to a single family. The frequency of the allele increasing protein percent was estimated at 0.63+0.06. The hypothesis of equal allelic frequencies was rejected at p<0.05. Segregation of this ETL in the Israeli population was confirmed. The genes IBSP, SPP1, and LAP3 located adjacent to BM143 in the whole genome cattle- human comparative map were used as anchors for the human genome sequence and bovine BAC clones. Fifteen genes within 2 cM upstream of BM143 were located in the orthologous syntenic groups on HSA4q22 and HSA4p15. Only a single gene, SLIT2, was located within 2 cM downstream of BM143 in the orthologous HSA4p15 region. The order of these genes, as derived from physical mapping of BAC end sequences, was identical to the order within the orthologous syntenic groups on HSA4: FAM13A1, HERC3. CEB1, FLJ20637, PP2C-like, ABCG2, PKD2. SPP, MEP, IBSP, LAP3, EG1. KIAA1276, HCAPG, MLR1, BM143, and SLIT2. Four hundred and twenty AI bulls with genetic evaluations were genotyped for 12 SNPs identified in 10 of these genes, and for BM143. Seven SNPs displayed highly significant linkage disequilibrium effects on protein percentage (P<0.000l) with the greatest effect for SPP1. None of SNP genotypes for two sires heterozygous for the ETL, and six sires homozygous for the ETL completely corresponded to the causative mutation. The expression of SPP 1 and ABCG2 in the mammary gland corresponded to the lactation curve, as determined by microarray and QPCR assays, but not in the liver. Anti-sense SPP1 transgenic mice displayed abnormal mammary gland differentiation and milk secretion. Thus SPP 1 is a prime candidate gene for this ETL. We confirmed that DGAT1 is the ETL segregating on BTA 14 that chiefly effects fat concentration, and that the polymorphism is due to a missense mutation in an exon. Four hundred Israeli Holstein bulls were genotyped for this polymorphism, and the change in allelic frequency over the last 20 years was monitored.
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Veilleux, Richard E., Jossi Hillel, A. Raymond Miller i David Levy. Molecular Analysis by SSR of Genes Associated with Alkaloid Synthesis in a Segregating Monoploid Potato Family. United States Department of Agriculture, maj 1994. http://dx.doi.org/10.32747/1994.7570550.bard.
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More than 15,000 anthers of an interspecific hybrid (CP2) between two diploid (2n=2x=24) potato species, Solanum chacoense (weedy) and S. phureja (cultivated), were cultured to generate a family of monoploid (haploid, 2n-1x=12) plants. Of 260 regenerated plants, 34 were monoploid, 210 diploid and 16 tetraploid. SSR analysis revealed that six monoploids were genetically identical and 14 diploids were homozygous, thus limiting the population to 42 (28 monoploids and 14 homozygous diploids). New microsatellite loci were developed for potato from database sequences (15), a conventional genomic library (6), an enriched library (18) and tomato (11). Of these, 13 were polymorphic in the CP2 family and 11 were used to study genetic segregatin. Four of 11 exhibited skewed segregation in the monoploid family. Seven of 18 microsatellite markers were polymorphic and informative on a set of 12 tetraploid potato cultivars. Acetylleptinidine (ALD) is the aglycone of leptines, a natural defense against insects, especially the highly destructuve Colorado potato beetle. ALD is absend in S. phureja but highly expressed in the S. chacoense parent of CP2. A backcross population between CP2 and tis S. phureja parent was used to examine segregation for ALD. Bulks of 10 backcross individuals that expressed ALD and 10 that did not were used to identify putative RAPD markers associatd with the trait. Of 80 primers tested, one putative marker amplified by OPQ02 was present in eight of ten individuals comprising the high bulk and absent in all 10 individuals comprising the low bulk. This is a putative marker for ALD expression in potato.
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Guy, Charles, Gozal Ben-Hayyim, Gloria Moore, Doron Holland i Yuval Eshdat. Common Mechanisms of Response to the Stresses of High Salinity and Low Temperature and Genetic Mapping of Stress Tolerance Loci in Citrus. United States Department of Agriculture, maj 1995. http://dx.doi.org/10.32747/1995.7613013.bard.
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The objectives that were outlined in our original proposal have largely been achieved or will be so by the end of the project in February 1995 with one exception; that of mapping cold tolerance loci based on the segregation of tolerance in the BC1 progeny population. Briefly, our goals were to 1) construct a densely populated linkage map of the citrus genome: 2) map loci important in cold and/or salt stress tolerance; and 3) characterize the expression of genes responsive to cold land salt stress. As can be seen by the preceding listing of accomplishments, our original objectives A and B have been realized, objective C has been partially tested, objective D has been completed, and work on objectives E and F will be completed by the end of 1995. Although we have yet to map any loci that contribute to an ability of citrus to maintain growth when irrigated with saline water, our very encouraging results from the 1993 experiment provides us with considerable hope that 1994's much more comprehensive and better controlled experiment will yield the desired results once the data has been fully analyzed. Part of our optimism derives from the findings that loci for growth are closely linked with loci associated with foliar Cl- and Na+ accumulation patterns under non-salinization conditions. In the 1994 experiment, if ion exclusion or sequestration traits are segregating in the population, the experimental design will permit their resolution. Our fortunes with respect to cold tolerance is another situation. In three attempts to quantitatively characterize cold tolerance as an LT50, the results have been too variable and the incremental differences between sensitive and tolerant too small to use for mapping. To adequately determine the LT50 requires many plants, many more than we have been able to generate in the time and space available by making cuttings from small greenhouse-grown stock plants. As it has turned out, with citrus, to prepare enough plants needed to be successful in this objective would have required extensive facilities for both growing and testing hardiness which simply were not available at University of Florida. The large populations necessary to overcome the variability we encountered was unanticipated and unforeseeable at the project's outset. In spite of the setbacks, this project, when it is finally complete will be exceedingly successful. Listing of Accomplishments During the funded interval we have accomplished the following objectives: Developed a reasonably high density linkage map for citrus - mapped the loci for two cold responsive genes that were cloned from Poncirus - mapped the loci for csa, the salt responsive gene for glutathione peroxidase, and ccr a circadian rhythm gene from citrus - identified loci that confer parental derived specific DNA methylation patterns in the Citrus X Poncirus cross - mapped 5 loci that determine shoot vigor - mapped 2 loci that influence leaf Na+ accumulation patterns under non-saline conditions in the BC1 population - mapped 3 loci that influence leaf Na+ accumulation paterns during salt sress - mapped 2 loci that control leaf Cl- accumulation patterns under non-saline conditions - mapped a locus that controls leaf Cl- accumulation patterns during salt stress Screened the BC1 population for growth reduction during salinization (controls and salinized), and cold tolerance - determined population variation for shoot/root ratio of Na+ and Cl- - determined levels for 12 inorganic nutrient elements in an effort to examine the influence of salinization on ion content with emphasis on foliar responses - collected data on ion distribution to reveal patterns of exclusion/sequestration/ accumulation - analyzed relationships between ion content and growth Characterization of gene expression in response to salt or cold stress - cloned the gene for the salt responsive protein csa, identified it as glutathione peroxidase, determined the potential target substrate from enzymatic studies - cloned two other genes responsive to salt stress, one for the citrus homologue of a Lea5, and the other for an "oleosin" like gene - cold regulated (cor) genes belonging to five hybridization classes were isolated from Poncirus, two belonged to the group 2 Lea superfamily of stress proteins, the others show no significant homology to other known sequences - the expression of csa during cold acclimation was examined, and the expression of some of the cor genes were examined in response to salt stress - the influence of salinization on cold tolerance has been examined with seedling populations - conducted protein blot studies for expression of cold stress proteins during salt stress and vice versa
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Sherman, Amir, Rebecca Grumet, Ron Ophir, Nurit Katzir i Yiqun Weng. Whole genome approach for genetic analysis in cucumber: Fruit size as a test case. United States Department of Agriculture, grudzień 2013. http://dx.doi.org/10.32747/2013.7594399.bard.
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The Cucurbitaceae family includes a broad array of economically and nutritionally important crop species that are consumed as vegetables, staple starches and desserts. Fruit of these species, and types within species, exhibit extensive diversity as evidenced by variation in size, shape, color, flavor, and others. Fruit size and shape are critical quality determinants that delineate uses and market classes and are key traits under selection in breeding programs. However, the underlying genetic bases for variation in fruit size remain to be determined. A few species the Cucurbitaceae family were sequenced during the time of this project (cucumber was already sequenced when the project started watermelon and melon sequence became available during the project) but functional genomic tools are still missing. This research program had three major goals: 1. Develop whole genome cucumber and melon SNP arrays. 2. Develop and characterize cucumber populations segregating for fruit size. 3. Combine genomic tools, segregating populations, and phenotypic characterization to identify loci associated with fruit size. As suggested by the reviewers the work concentrated mostly in cucumber and not both in cucumber and melon. In order to develop a SNP (single nucleotide polymorphism) array for cucumber, available and newly generated sequence from two cucumber cultivars with extreme differences in shape and size, pickling GY14 and Chinese long 9930, were analyzed for variation (SNPs). A large set of high quality SNPs was discovered between the two parents of the RILs population (GY14 and 9930) and used to design a custom SNP array with 35000 SNPs using Agilent technology. The array was validated using 9930, Gy14 and F1 progeny of the two parents. Several mapping populations were developed for linkage mapping of quantitative trait loci (QTL) for fruit size These includes 145 F3 families and 150 recombinant inbred line (RILs F7 or F8 (Gy14 X 9930) and third population contained 450 F2 plants from a cross between Gy14 and a wild plant from India. The main population that was used in this study is the RILs population of Gy14 X 9930. Phenotypic and morphological analyses of 9930, Gy14, and their segregating F2 and RIL progeny indicated that several, likely independent, factors influence cucumber fruit size and shape, including factors that act both pre-anthesis and post-pollination. These include: amount, rate, duration, and plane of cell division pre- and post-anthesis and orientation of cell expansion. Analysis of F2 and RIL progeny indicated that factors influencing fruit length were largely determined pre-anthesis, while fruit diameter was more strongly influenced by environment and growth factors post-anthesis. These results suggest involvement of multiple genetically segregating factors expected to map independently onto the cucumber genome. Using the SNP array and the phenotypic data two major QTLs for fruit size of cucumber were mapped in very high accuracy (around 300 Kb) with large set of markers that should facilitate identification and cloning of major genes that contribute to fruit size in cucumber. In addition, a highly accurate haplotype map of all RILS was created to allow fine mapping of other traits segregating in this population. A detailed cucumber genetic map with 6000 markers was also established (currently the most detailed genetic map of cucumber). The integration of genetics physiology and genomic approaches in this project yielded new major infrastructure tools that can be used for understanding fruit size and many other traits of importance in cucumber. The SNP array and genetic population with an ultra-fine map can be used for future breeding efforts, high resolution mapping and cloning of traits of interest that segregate in this population. The genetic map that was developed can be used for other breeding efforts in other populations. The study of fruit development that was done during this project will be important in dissecting function of genes that that contribute to the fruit size QTLs. The SNP array can be used as tool for mapping different traits in cucumber. The development of the tools and knowledge will thus promote genetic improvement of cucumber and related cucurbits.
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Ginzberg, Idit, i Walter De Jong. Molecular genetic and anatomical characterization of potato tuber skin appearance. United States Department of Agriculture, wrzesień 2008. http://dx.doi.org/10.32747/2008.7587733.bard.
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Potato (Solanum tuberosum L.) skin is composed of suberized phellem cells, the outer component of the tuber periderm. The focus of the proposed research was to apply genomic approaches to identify genes that control tuber skin appearance - smooth and shiny skin is highly preferred by the customers while russeted/netted skin potatoes are rejected. The breeding program (at Cornell University) seeks to develop smooth-skin varieties but has encountered frequent difficulties as inheritance of russeting involves complementary action by independently segregating genes, where a dominant allele at each locus is required for any degree of skin russeting. On the other hand, smooth-skin varieties frequently develop unsightly russeting in response to stress conditions, mainly high soil temperatures. Breeding programs in Israel aimed towards the improvement of heat tolerant varieties include skin quality as one of the desired characteristics. At the initiation of the present project it was unclear whether heat induced russeting and genetically inherited russeting share the same genes and biosynthesis pathways. Nevertheless, it has been suggested that russeting might result from increased periderm thickness, from strong cohesion between peridermal cells that prevents the outer layers from sloughing off, or from altered suberization processes in the skin. Hence, the original objectives were to conduct anatomical study of russet skin development, to isolate skin and russeting specific genes, to map the loci that determine the russet trait, and to compare with map locations the candidate russet specific genes, as well as to identify marker alleles that associated with russet loci. Anatomical studies suggested that russet may evolve from cracking at the outer layers of the skin, probably when skin development doesn’t meet the tuber expansion rate. Twodimensional gel electrophoresis and transcript profiling (cDNA chip, potato functional genomic project) indicated that in comparison to the parenchyma tissue, the skin is enriched with proteins/genes that are involved in the plant's responses to biotic and abiotic stresses and further expand the concept of the skin as a protective tissue containing an array of plantdefense components. The proteomes of skin from heat stressed tubers and native skin didn’t differ significantly, while transcript profiling indicated heat-related increase in three major functional groups: transcription factors, stress response and protein degradation. Exceptional was ACC synthase isogene with 4.6 fold increased level in the heat stressed skin. Russeting was mapped to two loci: rusB on chromosome 4 and rusC on chromosome 11; both required for russeting. No evidence was found for a third locus rusA that was previously proposed to be required for russeting. In an effort to find a link between the russeting character and the heat-induced russeting an attempt was made to map five genes that were found in the microarray experiment to be highly induced in the skin under heat stress in the segregating russet population. Only one gene was polymorphic; however it was localized to chromosome 2, so cannot correspond to rusB or rusC. Evaluation of AFLP markers tightly linked to rusB and rusC showed that these specific alleles are not associated with russeting in unrelated germplasm, and thus are not useful for MAS per se. To develop markers useful in applied breeding, it will be necessary to screen alleles of additional tightly linked loci, as well as to identify additional russet (heat-induced and/or native) related genes.
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Fridman, Eyal, Jianming Yu i Rivka Elbaum. Combining diversity within Sorghum bicolor for genomic and fine mapping of intra-allelic interactions underlying heterosis. United States Department of Agriculture, styczeń 2012. http://dx.doi.org/10.32747/2012.7597925.bard.
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Heterosis, the enigmatic phenomenon in which whole genome heterozygous hybrids demonstrate superior fitness compared to their homozygous parents, is the main cornerstone of modern crop plant breeding. One explanation for this non-additive inheritance of hybrids is interaction of alleles within the same locus. This proposal aims at screening, identifying and investigating heterosis trait loci (HTL) for different yield traits by implementing a novel integrated mapping approach in Sorghum bicolor as a model for other crop plants. Originally, the general goal of this research was to perform a genetic dissection of heterosis in a diallel built from a set of Sorghum bicolor inbred lines. This was conducted by implementing a novel computational algorithm which aims at associating between specific heterozygosity found among hybrids with heterotic variation for different agronomic traits. The initial goals of the research are: (i) Perform genotype by sequencing (GBS) of the founder lines (ii) To evaluate the heterotic variation found in the diallel by performing field trails and measurements in the field (iii) To perform QTL analysis for identifying heterotic trait loci (HTL) (iv) to validate candidate HTL by testing the quantitative mode of inheritance in F2 populations, and (v) To identify candidate HTL in NAM founder lines and fine map these loci by test-cross selected RIL derived from these founders. The genetic mapping was initially achieved with app. 100 SSR markers, and later the founder lines were genotyped by sequencing. In addition to the original proposed research we have added two additional populations that were utilized to further develop the HTL mapping approach; (1) A diallel of budding yeast (Saccharomyces cerevisiae) that was tested for heterosis of doubling time, and (2) a recombinant inbred line population of Sorghum bicolor that allowed testing in the field and in more depth the contribution of heterosis to plant height, as well as to achieve novel simulation for predicting dominant and additive effects in tightly linked loci on pseudooverdominance. There are several conclusions relevant to crop plants in general and to sorghum breeding and biology in particular: (i) heterosis for reproductive (1), vegetative (2) and metabolic phenotypes is predominantly achieved via dominance complementation. (ii) most loci that seems to be inherited as overdominant are in fact achieving superior phenotype of the heterozygous due to linkage in repulsion, namely by pseudooverdominant mechanism. Our computer simulations show that such repulsion linkage could influence QTL detection and estimation of effect in segregating populations. (iii) A new height QTL (qHT7.1) was identified near the genomic region harboring the known auxin transporter Dw3 in sorghum, and its genetic dissection in RIL population demonstrated that it affects both the upper and lower parts of the plant, whereas Dw3 affects only the part below the flag leaf. (iv) HTL mapping for grain nitrogen content in sorghum grains has identified several candidate genes that regulate this trait, including several putative nitrate transporters and a transcription factor belonging to the no-apical meristem (NAC)-like large gene family. This activity was combined with another BARD-funded project in which several de-novo mutants in this gene were identified for functional analysis.
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Weller, Joel I., Ignacy Misztal i Micha Ron. Optimization of methodology for genomic selection of moderate and large dairy cattle populations. United States Department of Agriculture, marzec 2015. http://dx.doi.org/10.32747/2015.7594404.bard.
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The main objectives of this research was to detect the specific polymorphisms responsible for observed quantitative trait loci and develop optimal strategies for genomic evaluations and selection for moderate (Israel) and large (US) dairy cattle populations. A joint evaluation using all phenotypic, pedigree, and genomic data is the optimal strategy. The specific objectives were: 1) to apply strategies for determination of the causative polymorphisms based on the “a posteriori granddaughter design” (APGD), 2) to develop methods to derive unbiased estimates of gene effects derived from SNP chips analyses, 3) to derive optimal single-stage methods to estimate breeding values of animals based on marker, phenotypic and pedigree data, 4) to extend these methods to multi-trait genetic evaluations and 5) to evaluate the results of long-term genomic selection, as compared to traditional selection. Nearly all of these objectives were met. The major achievements were: The APGD and the modified granddaughter designs were applied to the US Holstein population, and regions harboring segregating quantitative trait loci (QTL) were identified for all economic traits of interest. The APGD was able to find segregating QTL for all the economic traits analyzed, and confidence intervals for QTL location ranged from ~5 to 35 million base pairs. Genomic estimated breeding values (GEBV) for milk production traits in the Israeli Holstein population were computed by the single-step method and compared to results for the two-step method. The single-step method was extended to derive GEBV for multi-parity evaluation. Long-term analysis of genomic selection demonstrated that inclusion of pedigree data from previous generations may result in less accurate GEBV. Major conclusions are: Predictions using single-step genomic best linear unbiased prediction (GBLUP) were the least biased, and that method appears to be the best tool for genomic evaluation of a small population, as it automatically accounts for parental index and allows for inclusion of female genomic information without additional steps. None of the methods applied to the Israeli Holstein population were able to derive GEBV for young bulls that were significantly better than parent averages. Thus we confirm previous studies that the main limiting factor for the accuracy of GEBV is the number of bulls with genotypes and progeny tests. Although 36 of the grandsires included in the APGD were genotyped for the BovineHDBeadChip, which includes 777,000 SNPs, we were not able to determine the causative polymorphism for any of the detected QTL. The number of valid unique markers on the BovineHDBeadChip is not sufficient for a reasonable probability to find the causative polymorphisms. Complete resequencing of the genome of approximately 50 bulls will be required, but this could not be accomplished within the framework of the current project due to funding constraints. Inclusion of pedigree data from older generations in the derivation of GEBV may result is less accurate evaluations.
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Fallik, Elazar, Robert Joly, Ilan Paran i Matthew A. Jenks. Study of the Physiological, Molecular and Genetic Factors Associated with Postharvest Water Loss in Pepper Fruit. United States Department of Agriculture, grudzień 2012. http://dx.doi.org/10.32747/2012.7593392.bard.
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The fruit of pepper (Capsicum annuum) commonly wilts (or shrivels) during postharvest storage due to rapid water loss, a condition that greatly reduces its shelf life and market value. The fact that pepper fruit are hollow, and thus have limited water content, only exacerbates this problem in pepper. The collaborators on this project completed research whose findings provided new insight into the genetic, physiological, and biochemical basis for water loss from the fruits of pepper (Capsicum annuum and related Capsicum species). Well-defined genetic populations of pepper were used in this study, the first being a series of backcross F₁ and segregating F₂, F₃, and F₄ populations derived from two original parents selected for having dramatic differences in fruit water loss rate (very high and very low water loss). The secondly population utilized in these studies was a collection of 50 accessions representing world diversity in both species and cultivar types. We found that an unexpectedly large amount of variation was present in both fruit wax and cutin composition in these collections. In addition, our studies revealed significant correlations between the chemical composition of both the fruit cuticular waxes and cutin monomers with fruit water loss rate. Among the most significant were that high alkane content in fruit waxes conferred low fruit water loss rates and low permeability in fruit cuticles. In contrast, high amounts of terpenoids (plus steroidal compounds) were associated with very high fruit water loss and cuticle permeability. These results are consistent with our models that the simple straight chain alkanes pack closely together in the cuticle membrane and obstruct water diffusion, whereas lipids with more complex 3-dimensional structure (such as terpenoids) do not pack so closely, and thus increase the diffusion pathways. The backcross segregating populations were used to map quantitative trait loci (QTLs) associated with water loss (using DART markers, Diversity Arrays Technology LTD). These studies resulted in identification of two linked QTLs on pepper’s chromosome 10. Although the exact genetic or physiological basis for these QTLs function in water loss is unknown, the genotypic contribution in studies of near-isogenic lines selected from these backcross populations reveals a strong association between certain wax compounds, the free fatty acids and iso-alkanes. There was also a lesser association between the water loss QTLs with both fruit firmness and total soluble sugars. Results of these analyses have revealed especially strong genetic linkages between fruit water loss, cuticle composition, and two QTLs on chromosome 10. These findings lead us to further speculate that genes located at or near these QTLs have a strong influence on cuticle lipids that impact water loss rate (and possibly, whether directly or indirectly, other traits like fruit firmness and sugar content). The QTL markers identified in these studies will be valuable in the breeding programs of scientists seeking to select for low water loss, long lasting fruits, of pepper, and likely the fruits of related commodities. Further work with these newly developed genetic resources should ultimately lead to the discovery of the genes controlling these fruit characteristics, allowing for the use of transgenic breeding approaches toward the improvement of fruit postharvest shelf life.
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Abbott, Albert G., Doron Holland, Douglas Bielenberg i Gregory Reighard. Structural and Functional Genomic Approaches for Marking and Identifying Genes that Control Chilling Requirement in Apricot and Peach Trees. United States Department of Agriculture, wrzesień 2009. http://dx.doi.org/10.32747/2009.7591742.bard.
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Structural and functional genomic approaches for marking and identifying genes that control chilling requirement in apricot and peach trees. Specific aims: 1) Identify and characterize the genetic nature of chilling requirement for flowering and dormancy break of vegetative shoots in Prunusgermplasm through the utilization of existing apricot (NeweYa'ar Research Center, ARO) and peach (Clemson University) genetic mapping populations; 2) Use molecular genetic mapping techniques to identify markers flanking genomic regions controlling chilling; 3) Comparatively map the regions controlling chilling requirement in apricot and peach and locate important genomic regions influencing chilling requirement on the Prunus functional genomic database as an initial step for identification of candidate genes; 4) Develop from the functional genomics database a set of markers facilitating the development of cultivars with optimized chilling requirements for improved and sustained fruit production in warm-winter environments. Dormant apricot (prunus armeniaca L.) and peach [Prunus persica (L.) Batsch] trees require sustained exposure to low, near freezing, temperatures before vigorous floral and vegetative bud break is possible after the resumption of warm temperatures in the spring. The duration of chilling required (the chilling requirement, CR) is determined by the climatic adaptation of the particular cultivar, thus limiting its geographic distribution. This limitation is particularly evident when attempting to introduce superior cultivars to regions with very warm winter temperatures, such as Israel and the coastal southern United States. The physiological mechanism of CR is not understood and although breeding programs deliberately manipulate CR in apricot and peach crosses, robust closely associated markers to the trait are currently not available. We used segregating populations of apricot (100 Fl individuals, NeweYa'ar Research Center, ARO) and peach (378 F2 individuals, Clemson University) to discover several discreet genomic loci that regulate CR and blooming date. We used the extensive genomic/genetic resources available for Prunus to successfully combine our apricot and peach genetic data and identify five QTL with strong effects that are conserved between species as well as several QTL that are unique to each species. We have identified markers in the key major QTL regions for testing in breeding programs which we are carrying out currently; we have identified an initial set of candidate genes using the peach physical/transcriptome map and whole peach genome sequences and we are testing these currently to identify key target genes for manipulation in breeding programs. Our collaborative work to date has demonstrated the following: 1) CR in peach and apricot is predominantly controlled by a limited number ofQTL loci, seven detected in a peach F2 derived map comprising 65% of the character and 12 in an apricot Fl map comprising 71.6% and 55.6% of the trait in the Perfection and A. 1740 parental maps, respectively and that peach and apricot appear in our initial maps to share five genomic intervals containing potentially common QTL. 2) Application of common anchor markers of the Prunus/peach, physical/genetic map resources has allowed us not only to identify the shared intervals but also to have immediately available some putative candidate gene information from these intervals, the EVG region on LG1 in peach the TALY 1 region in apricot on LG2 in peach; and several others involved in vernalization pathways (LGI and LG7). 3) Mapped BACcontigs are easily defined from the complete physical map resources in peach through the common SSR markers that anchor our CR maps in the two species, 4) Sequences of BACs in these regions can be easily mined for additional polymorphic markers to use in MAS applications.