Genetics of superoxide dismutase in apple

 Four zones of superoxide dismutase (SOD), activity were detected in apple. Genetic studies confirmed the presence of three genes Sod-1, Sod-3 and Sod-4 with three, two and two alleles respectively, including one null allele. One of these genes, Sod-1, was found to be loosely linked to Prx-2 and Prx-3. The Sod-1a allele predominated in all the three groups studied, cultivars, rootstocks and Malus species. The distinct achromatic bands produced by SOD after electrophoresis facilitate its important role with peroxidase (PRX), glutamate oxalacetate transaminase (GOT) and phosphogluconate dehydrogenase (PGD) in the discrimination of apple cultivars. Received: 22 November 1996/Accepted: 29 November 1996     

Genetics of leucine aminopeptidase in apple

Summary Six zones of LAP activity were detected in apples, some of them tissue specific. Genetic studies in four of them revealed the presence of four genes LAP-1, LAP-2, LAP-3 and LAP-4 with 4, 5, 4 and 4 alleles respectively including two null alleles. There were no big differences in allelic frequency within cultivars, selections, rootstocks and Malus species. Close linkage was found between LAP-2 and resistance to mildew derived from White Angel.     

Variability and genetics of spacer DNA sequences between the ribosomal-RNA genes of hexaploid wheat (Triticum aestivum)

Summary Using restriction enzyme digests of genomic DNA extracted from the leaves of 25 hexaploid wheat (Triticum aestivum L. em. Thell.) cultivars and their hybrids, restriction fragment length polymorphisms of the spacer DNA which separates the ribosomal-RNA genes have been examined. (From one to three thousand of these genes are borne on chromosomes 1B and 6B of hexaploid wheat). The data show that there are three distinct alleles of the 1B locus, designated Nor-B1a, Nor-B1b, and Nor-B1c, and at least five allelic variants of the 6B locus, designated Nor-B2a, Nor-B2b, Nor-B2c, Nor-B2d, and Nor-B2e. A further, previously reported allele on 6B has been named Nor-B2f. Chromosome 5D has only one allelic variant, Nor-D3. Whereas the major spacer variants of the 1B alleles apparently differ by the loss or gain of one or two of the 133 bp sub-repeat units within the spacer DNA, the 6B allelic variants show major differences in their compositions and lengths. This may be related to the greater number of rDNA repeat units at this locus. The practical implications of these differences and their application to wheat breeding are discussed.     

Isozyme gene markers in Vicia faba L.

Summary This study was conducted to assess the genetic basis of the variability observed for the glutamate oxaloacetate transaminase (GOT), Superoxide dismutase (SOD), esterase (EST), and malate dehydrogenase (MDH) isozyme systems in different open-pollinated Vicia faba varieties. Individual plants showing contrasting zymogram patterns were simultaneously selfed and cross-combined. Crossing was unsuccessful in producing progeny, and only selfed progenies were suitable for genetical analysis of isozyme variability. Three zones of GOT activity were made visible. The isozyme of GOT-2 and GOT-3 zones were dimeric and under the control of three alleles at the Got-2 locus and two alleles at the Got-3 locus, respectively. The isozymes of the GOT-1 zone did not show any variability. Three zones of SOD isozyme activity were made visible. The isozymes occurring in the SOD-1 (chloroplastic isozyme form) and SOD-2 (cytosol isozyme form) zones were dimeric and under the control of two alleles at the Sod-1 and Sod-2 loci. The isozyme visualized in the SOD-3 zone (mitochondrial isozyme form) were tetrameric and under the control of two alleles at the Sod-3 locus. Apparently the isozymes made visible in the most anodal esterase zones EST-1, EST-2, and EST-3 were monomeric, and the occurrence of two alleles at each of two different loci explained the variability observed in the EST-2 and EST-3 zones. For MDH, only two five-banded zymogram pattern types were found, and every selfed progeny showed only one of the two zymogram type, indicating that each individual possessed fixed alleles at the loci controlling MDH isozyme. Got-2, Got-3, Sod-1, Sod-2, and Sod-3 appear to be five new isozyme gene markers that can be useful in Vicia faba breeding for linkage study, varietal fingerprinting, outcrossing rate estimate, and indirect selection for quantitative characters.     

High molecular weight glutenin subunit in durum wheat (T. durum)

Summary The diversity of high molecular weight (HMW) glutenin subunits of 502 varieties of durum wheat (Triticum durum) from 23 countries was studied using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Twenty-nine types of patterns were observed with 18 mobility bands. A total of 18 alleles were identified by comparing the mobilities of their subunits to those previously found in hexaploid wheat (T. aestivum) and in Triticum turgidum var. dicoccum. Five new alleles were detected: two on the Glu A1 and three on the Glu B1 locus. Comparison of the frequency of alleles in the three species T. aestivum, T. dicoccum and T. durum was investigated. Significant differences exist between each of these species on the basis of the frequency distributions of their three and four common alleles at the Glu A1 and Glu B1 locus, respectively. The Glu B1c allele occuring very frequently in hexaploid wheats was not found in the two tetraploid species. More than 83% of the T. durum analysed were found to have the Glu A1c (null) allele.     

Constitutive expression of two apple (Malus x domestica Borkh.) homolog genes of LIKE HETEROCHROMATIN PROTEIN1 affects flowering time and whole-plant growth in transgenic Arabidopsis

Fruit trees, such as apple (Malus × domestica Borkh.), are woody perennial plants with a long juvenile phase. The biological analysis for the regulation of flowering time provides insights into the reduction of juvenile phase and the acceleration of breeding in fruit trees. In Arabidopsis, LIKE HETEROCHROMATIN PROTEIN1 (LHP1) is involved in epigenetic silencing of the target genes such as flowering genes. We isolated and characterized twin apple LHP1 homolog genes, MdLHP1a and MdLHP1b. These genes may have been generated as a result of ancient genome duplication. Although the putative MdLHP1 proteins showed lower similarity to any other known plant LHP1 homologs, a chromo domain, a chromo shadow domain, and the nuclear localization signal motifs were highly conserved among them. RT-PCR analysis showed that MdLHP1a and MdLHP1b were expressed constantly in developing shoot apices of apple trees throughout the growing season. Constitutive expression of MdLHP1a or MdLHP1b could compensate for the pleiotropic phenotype of lhp1/tfl2 mutant, suggesting that apple LHP1 homolog genes are involved in the regulation of flowering time and whole-plant growth. Based on these results, LHP1 homolog genes might have rapidly evolved among plant species, but the protein functions were conserved, at least between Arabidopsis and apple. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Allozyme Variation in a Threatened Freshwater Fish, Spotted Murrel (Channa punctatus) in a South Indian River System

Samples of the spotted murrel (Channa punctatus) were collected from three rivers of Tamil Nadu and Kerala. The allozyme variation of C. punctatus was investigated by polyacrylamide gel electrophoresis. Eighteen enzymes were detected, but only 10 (EST, PGM, G3PDH, G6PDH, SOD, GPI, ODH, GDH, XDH, and CK) showed consistent phenotypic variations. Allele frequencies were estimated at the 18 polymorphic loci representing 10 enzymes. Two rare alleles, EST-4*C and G6PDH-2*C, were noted in the Tamirabarani and Kallada populations but were absent in the Siruvani population. The allele frequencies of the Tamirabarani and Kallada populations were similar, except for a few loci. Among the three populations, the maximum genetic distance (0.026) and FST (0.203) were found between the geographically distant Siruvani and Kallada populations. Overall the study showed that among the three populations, the Tamirabarani and Kallada have similar genetic structures.     

Polymorphism and chromosomal location of endogenous α-amylase inhibitor genes in common wheat

Summary Polymorphism of an endogenous -amylase inhibitor in wheat was studied using iso-electric focusing followed by monoclonal antibody — based immunoblotting. Ten isoforms of the inhibitor detected in common wheat and its wild counterparts were assigned to five homoeologous loci. Three -amylase inhibitor loci (Isa-1) were identified in common wheat and located on the long arms of chromosomes 2A, 2B and 2D. In a sample of 27 bread wheats, eight durum wheats, and 12 diploid wheat relatives, amphiploids and triticales, a high resolution isoelectric-focusing separation demonstrated two active and one null allele at the Isa-A1, two alleles at the Isa-B1, one allele at the Isa-D1, four alleles at the Isa-S1, and one allele at the Isa-G1 locus. The most frequent electrophoretic pattern of common wheat cultivars consisted of two isoforms, encoded respectively by the Isa-B1b, Isa-D1 a alleles and the Isa-Alnull allele. All the durum wheats had only one inhibitor form controlled by allele Isa-B1b, which was accompanied by the null allele at the Isa-A1 locus.Contribution No. 210 of the Food Science Department, University of Manitoba     

Variation,geographical distribution and genetical analysis of esterase isozymes in foxtail millet,Setaria italica (L.) P. Beauv.

Summary The esterase isozymes of 432 strains of foxtail millet, Setaria italica (L.) P. Beauv., collected from different areas throughout Eurasia, were investigated by gel isoelectric focusing. Five phenotypes were recognized, based on the combination of five major activity bands. Cross experiments among different phenotypes revealed these isozymes to be controlled by two codominant alleles and a null allele on the locus, Est-1, and three codominant alleles on another independent locus, Est-2. On locus Est-1, 388 strains had Est-1 ^a, 41 had Est-1 ^b and three had Est-1 ^null alleles. Est-1 ^a was widely distributed throughout Eurasia, while the distribution of Est-1 ^b and Est-1 ^null was distinctly restricted. On locus Est-2, 417 strains had Est-2 ^a, nine had Est-2 ^b and six had Est-2 ^c alleles. Est-2 ^a was widely distributed throughout Asia to Czechoslovakia, but was not detected in the western part of Europe. Est-2 ^b was found in all of the strains from the western part of Europe and in one of the Indian strains. Est-2 ^c was rarely found in Japan and China. The distribution of Est-2 ^a and -2 ^b might indicate some degree of phylogenetic differentiation between the Asian and the European strains. Polymorphism in both loci was observed only in Chinese strains.Contribution No. 30 from the Plant Germ-plasm Institute, Faculty of Agriculture, Kyoto University, Kyoto, Japan     

Utilization of Different <Emphasis Type="Italic">Bmy1</Emphasis> Intron III Alleles for Predicting β-Amylase Activity and Thermostability in Wild and Cultivated Barley

The third intron of barley (Hordeum vulgare L.) β-amylase 1 (Bmy1) is extremely polymorphic. The use of specific insertion/deletions (indels) in the third intron as markers for cultivar development has been recommended based on associations with β-amylase activity and thermostability. The third intron of Bmy1 in 40 barley genotypes was sequenced and aligned with 15 Bmy1 intron III sequences from GenBank and four alleles (Bmy1.a, Bmy1.b, Bmy1.c, and Bmy1.d) were identified based on indels of 126, 38, 11, and 21 bp. β-Amylase activity and thermostability were assayed in 22 North American cultivars and 12 wild barley genotypes. Cultivars carrying the Bmy1.a and Bmy1.b alleles had β-amylase activity ranges calculated on a fresh weight (FW) basis of 1.8- and 1.5-fold, respectively, and thermostability ranges of 8.8- and 1.2-fold, respectively. β-Amylase activity calculated on a protein basis yielded a 2.4- and 1.4-fold range for Bmy1.a and Bmy1.b, respectively. Significantly different activities were observed in cultivars carrying either Bmy1.a or the Bmy1.b allele when calculated on a FW basis and the Bmy1.a allele when calculated on a protein basis. Significantly different thermostabilities were observed in cultivars carrying the Bmy1.a allele. Wild barleys were found to carry Bmy1.a, Bmy1.b, and Bmy1.c alleles with β-amylase activity ranges calculated on a FW basis of 1.7-, 1.7-, and 2.6-fold, respectively, and thermostability ranges of 1.3-, 1.4-, and 2.1-fold, respectively. β-Amylase activity measured on a protein basis identified a 1.3-, 1.4-, and 2.1-fold range for Bmy1.a, Bmy1.b, and Bmy1.c, respectively. Significantly different activities were found in genotypes with any of these three alleles when calculated on a FW basis yet only in those with the Bmy1.c allele when calculated on a protein basis. Significantly different thermostabilities in genotypes carrying either the Bmy1.b or Bmy1.c allele were observed. In the germplasm studied here, the Bmy1 intron III alleles are not reliable predictors of β-amylase activity and thermostability.     

Characterization of Esterases in a Brazilian Population of Zaprionus Indianus (Diptera: Drosophilidae)

The aim of this study was to characterize esterases in Zaprionus indianus, a drosophilid recently introduced into Brazil. A further aim was study the variation of activity of esterases in the presence of inhibitors and their expression according to sex, sexual activity and age of individual flies. Polymorphisms were detected in two esterase loci (Est-2 and Est-3) and monomorphisms in four others (Est-1, Est-4, Est-5 and Est-6). Biochemical tests using α- and β-naphthyl acetate and the inhibitors malathion, eserine sulphate and PMSF allowed us to classify EST-2 and EST-5 as β-esterases, both carboxyl-esterases, and EST-1, EST-3, EST-4 and EST-6 as α-esterases. EST-1 and EST-3 were classified as carboxyl-esterases and EST-4 and EST-6 as cholinesterases. EST-5 activity was more pronounced in males and EST-2 was restricted to them or to recently copulated females. EST-4, rarely detected, was not characterized. Based on their biochemical characteristics possible roles for these enzymes are suggested.     

Development of a multiallelic SCAR marker for the scab resistance gene Vr1/Vh4/Vx from R12740-7A apple and its utility for molecular breeding

A major scab resistance gene initially called Vr1 was identified in the apple cultivar “Regia” derived from the Malus scab resistance source R12740-7A (Russian seedling, RS). A codominant, multiallelic sequence characterized amplified region (SCAR) marker was developed from a random amplified polymorphic DNA marker identified by bulked-segregant analysis. Additional alleles of the AD13 marker locus proved to be informative for the analysis of genetic relationships within Malus including putative relatives of RS. Separate linkage maps were created for the two families derived from crosses with “Regia”. Using phenotypic data from the greenhouse scab tests, the recombination frequency between Vr1 and AD13-SCAR was between 6 and 17%. The Vr1 locus appeared to be closely linked to the Vx [Hemmat et al. J Am Soc Hortic Sci, 127:365–370, 2002], Vr2 [Patocchi et al. Theor Appl Genet, 109:1087–1092, 2004], and the Vh4 gene [Bus et al. Mol Breed, 15:103–116, 2005a]. Our linkage analysis of the molecular markers identified by Hemmat et al. [J Am Soc Hortic Sci, 127:365–370, 2002] for two scab resistance factors from RS (Vr and Vx) indicate that both genes are separated by a large distance on apple linkage group 2 [Boudichevskaia et al. Acta Hortic, 663:171–175, 2004]. This is in agreement with the results of Bus et al., [Mol Breed, 15:103–116, 2005a] who concluded that (1) the RS-derived gene Vh2 is identical to Vr, (2) the RS-derived gene Vh4 is identical to Vx and Vr1, (3) Vh2/Vr and Vh4/Vr1/Vx map on opposite sides of LG 2. One of our main goals was the verification of the Vr1-SCAR within a practical apple-breeding program. The utility of the AD13-SCAR was evident after 2 years under natural scab infection conditions in both families investigated. This is the first report about the confirmation of a molecular marker for a RS resistance factor in a 2-year field experiment. A multiplex polymerase chain reaction assay based on two codominant SCARs for Vf and Vr1 was tested in an apple progeny segregating for both genes. The result of the two-marker approach is discussed with respect to scab races, which are able to overcome the Vf resistance gene.     

Variations in biochemical parameters of Heterocapsa sp. and Olisthodiscus luteus grown in 12:12 light:dark cycles II. Changes in pigment composition

Photosynthetic pigment composition was studied in batch cultures of Heterocapsa sp. and Olisthodiscus luteus growing exponentially in a 12:12 light:dark cycle. Both species divided in the dark. The synthesis of pigments was continuous for both species. However for chlorophyll c and peridinin, in Heterocapsa sp., and chlorophyll c and fucoxanthin, in O. luteus, (pigments belonging to light harvesting complexes) the synthesis was significantly higher during the light period. Concentrations per total cell volume (TCV) of chlorophyll a, chlorophyll c, peridinin and diadinoxanthin in Heterocapsa sp., and chlorophyll a, chlorophyll c, fucoxanthin and violaxanthin in O. luteus, showed a maximum at the onset of light and decreased during the light period. The values of the chlorophyll a:chlorophyll c, chlorophyll a:peridinin and chlorophyll a:fucoxanthin ratios are compared with data reported in the literature.     

The genetic control of grain esterases in hexaploid wheat

Summary A comparison of EST-5 grain esterase phenotypes from wheat-alien amphiploid, addition and substitution genotypes, resolved by flat-bed isoelectric focusing identified homoeologous Est-5 loci on chromosome 3H of Hordeum vulgare, 3H^ch of H. chilense, 3S^b of Aegilops bicornis, 3S¹ of Ae. sharonensis and Ae. longissima and 6R of Secale cereale and 6R^m of S. montanum. The Est-5 genes in alien species provide evidence for chromosome homoeology with wheat.     

Genetic variation and hybridization in SwedishSchoenus (Cyperaceae)

Schoenus ferrugineus andS. nigricans have restricted distributions in Sweden and are almost exclusively confined to calcareous fen habitats. AtS. nigricans sites,S. ferrugineus is usually also present, and hybrids are frequently found. In this report, I used allozymes to estimate the amount of gene flow between the two species, and to compare the partitioning of genetic diversity in each of them. Thirteen loci were analysed at eight different enzyme systems. Seven loci were variable between or within the species. The two species had completely different alleles at two of the seven variable loci, whereas there was overlap at five loci. In all, 22 different alleles were found. Six of these alleles were confined toS. nigricans, and five alleles were confined toS. ferrugineus. Nei's genetic identity was 0.55.—InS. ferrugineus, three loci (23%) were polymorphic, and the average number of alleles per polymorphic locus was 2.0 (each polymorphic locus had two alleles). InS. nigricans, three loci (23%) were polymorphic, and the average number of alleles per polymorphic locus was 2.3.—The proportion of genetic diversity due to variation among sites (G _ST) was fairly similar in the two species, mean over loci = 0.12 inS. ferrugineus and 0.15 inS. nigricans. However, the proportion of genetic diversity due to variation among individuals within sites (G _IS) differed markedly between the two species, mean over loci = 0.54 inS. ferrugineus and 0.17 inS. nigricans. Accordingly, there was a much higher individual heterozygosity inS. nigricans than inS. ferrugineus. — Most hybrids were interpreted as F₁ hybrids. However, a small proportion, 0.5–1.6 %, were F_n hybrids or back-crosses.—On the Swedish mainland, all former occurrences ofS. nigricans are extinct, but viable hybrids are still present at a few sites in southernmost Sweden.     

Molecular cloning of the c2 locus of Zea mays, the gene coding for chalcone synthase

Summary The c2 locus of Zea mays, identified as one of the genes affecting anthocyanin biosynthesis, was cloned using the transposable element En (Spm) as a gene tag. The Spm element present at the c2 locus in the autonomously mutating c2-m1 line was isolated using En1 element specific probes. Sequences flanking the element were identified as c2 locus specific and were used to clone the nonautonomous c2-m2 and wild-type alleles. The cloning and analysis of a cDNA complementary to the c2 locus provided evidence that this gene encodes the enzyme chalcone synthase.     

多花海棠叶绿体基因组分析

多花海棠（Malus floribunda Siebold.）是世界范围内广泛栽培的苹果属物种,具有较高的观赏价值和育种意义。对其进行叶绿体基因组比较分析,有利于完善苹果属系统进化以及种质利用的研究内容。基于全基因组测序数据,组装获得一个完整的具有四分体结构的多花海棠叶绿体基因组。该基因组包括大单拷贝区（88 142 bp）、反向重复区B （26 353 bp）、小单拷贝区（19 189 bp）与反向重复区A （26 353 bp）,共计160 037 bp。多花海棠叶绿体全基因组共注释到111个基因,包括78个蛋白编码基因、29个tRNA基因和4个rRNA基因。此外,在其基因组中识别到大量的重复序列,与三叶海棠和变叶海棠略有差异。通过计算相对同义密码子使用度,发现其高频密码子共30种,并且密码子具有偏向A/T结尾的使用模式。种间序列比对、边界分析的结果表明,大单拷贝区序列变异较大,8种苹果属植物SC区与IR区扩张收缩情况整体上较为相似。基于叶绿体基因组序列的系统进化分析,将多花海棠、湖北海棠和变叶海棠聚为一类。多花海棠叶绿体基因组的研究可为今后遗传标记开发与种质资源利用等提供数据支持。     

Three types of polymorphisms in exon 14 in porcine Mx1 gene

Much is known about the antiviral activity of Mx proteins in species such as mouse and human. In the mouse, loss of resistibility to influenza virus has been shown to be due to specific polymorphisms in the Mx gene. This gene is therefore an interesting candidate gene for disease resistance in farm animals. The porcine Mx1 gene has already been identified and characterized based on its homology with mouse Mx1; however, until now no evidence of polymorphisms in the porcine gene has been reported. In this study, we have found two new polymorphisms in exon 14 of porcine Mx1 by DNA sequencing and confirmed their presence in different breeds, using polymerase chain reaction (PCR)–restriction fragment length polymorphisms (RFLP) with NarI and NaeI restriction enzymes. On the basis of the deduced amino acid sequence, one allele contains a deletion that may result in a frameshift to yield several amino acid substitutions and extension of the carboxyl terminal region of Mx1 protein. The deletion allele, Mx1 ^c, was found to be segregating in Landrace, Berkshire, Duroc, Hampshire, and Yucatan miniature pig. A second point mutation, Mx1 ^b, was detected in Meishan and two Vietnamese native pig breeds. All other breeds tested were fixed for the Mx1 ^a allele that is identical to the sequence reported previously. It will be interesting to determine if the Mx1 ^c deletion is associated with variation in resistance to the myxovirus family in the pig.     

Esterase-29 (ES-29): Biochemical characterization and control by two independent gene loci of a testosterone-dependent mouse serum esterase

Biochemistry and genetics of a testosterone-dependent murine serum esterase designated esterase-29 (ES-29) are described. The enzyme was identified after disc electrophoresis and subsequent staining for esterase using -naphthyl acetate as the substrate. It was inhibited by bis-p-nitrophenyl phosphate and was resistant top-chlorophenylsulphonate and hence was classified as carboxylesterase EC 3.1.1.1. The molecular mass was estimated to be about 130 kDa. It was shown that ES-29 is under the control of two independent genes. The first, termed Es-29, is suggested to be a structural locus, linked to the cluster-2 esterase loci on chromosome 8. Three alleles atEs-29, Es-29 ^a, Es-29^b, andEs-29 ^care distinguished, which determine absence (SEG/1), strong activity (BALB/cJ), and low activity (MOLH/Fre), respectively. The second locus, termedMse-1 (serum esterase modifying factor), was found to be closely linked toPre-2 on chromosome 12 and is suggested to be a modifying or regulatory gene. Two alleles were distinguished,Mse-1 ^a(BALB/cJ) andMse-1 ^m(MOL3/JA, CasBgr), which determine whether ES-29 appears as a single band or a double band, respectively.Mse-1 ^mis dominant toMse-1 ^a.This work was supported by the Deutsche Forschungsgemeinschaft. This is communication No. 70 of a research program devoted to the cellular distribution, genetics, and regulation of nonspecific esterases.