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1.
Summary Two alleles of the structural gene prxA from Petunia, prxA6 and prxA7, could be identified by their differential temporal expression. The alleles prxA6 and prxA7 code for peroxidases with a similar electrophoretic mobility as the products of the previously described alleles prxA1 and prxA5, respectively. The former two alleles differ in that they have a different temporal expression with regard to the temporal expression of the allele prxA2. Crossing experiments indicated that the mutations involved are (cisacting) internal site mutations. In the case of the allele prxA6, the experiments indicated a difference with respect to the allele prxA1 in responsiveness to the action of a trans-acting factor.  相似文献   

2.
A genomic library was prepared from Populus kitakamiensis and screened with the cDNA for an anionic peroxidase from P. kitakamiensis. One genomic clone was isolated that contained two tandemly oriented genes for anionic peroxidases, prxA3a and prxA4a. Both genes consisted of four exons and three introns; the introns had consensus nucleotides, namely, GT and AG, at their 5 and 3 ends, respectively. The prxA3a and prxA4a genes encoded 347 and 343 amino acid residues, respectively, including putative signal sequences at the amino-termini. Putative promoters and polyadenylation signals were found in the flanking regions of both genes. The sequence of the coding region of prxA3a was completely identical to that of the cDNA clone pA3, whereas the sequence of the coding region of prxA4a was only 73% identical to that of the cDNA clone pA3. Northern blot analysis showed that the patterns of expression of the mRNAs that corresponded to prxA3a and prxA4a differed in stems of P. kitakamiensis.  相似文献   

3.
Summary The structural geneprxD inPetunia codes for a slow moving anodic peroxidase whose activity is sensitive to high concentrations of hydrogen peroxide. The PRXd enzyme could be found in mature and old leaf and stem tissue of full-grown flowering plants. PRXd was found to be absent in tissues from flower corolla and root. The geneprxD is the fourth gene that codes for peroxidases in leaf and stem. Two mobility variants of the PRXd enzyme have been found among our inbred lines using starch gel system II electrophoresis. The geneprxD could be located on chromosome III by a four-point-cross involving the genesprxA, prxD, Mf1 andHt1. The order of the genes established is:Ht1 — Mf1 — prxD — prxA.  相似文献   

4.
Summary As detected by starch gel electrophoresis, the fast moving anodal group of peroxidase isoenzymes, the PRXa complex, of a Petunia homozygous for the encoding gene can be made up of one to four bands, depending on the tissue sampled, the age of the tissue and of the plant, and the genetic background. Additional evidence is presented showing that the PRXa complex is encoded by one structural gene, prxA, rather than by tandem duplicated genes. On the basis of electrophoretic variation in Petunia hybrida and related species, five prxA alleles were found. A prxA internal site mutation was found recognized by the absence of recombination between the mutation that affected the temporal programme of the gene and the mutation that altered the mobility of the enzyme. By a three-point test, the gene prxA was located on chromosome III and found to be linked to the genes Mf1 and Ht1 in the order prxA-Mf1-Ht1. The construction of a trisomic III triply heterozygous for prxA confirmed the location of prxA.  相似文献   

5.
It is generally accepted that peroxidases catalyze the final step in the biosynthesis of lignin. In this study, to examine how expression of prxA3a, a gene for an anionic peroxidase, might be related to lignification in plant tissues, we produced transgenic tobacco plants that harbored a gene for β-glucuronidase (GUS) fused to the prxA3a promoter. Histochemical staining for GUS activity indicated that the prxA3a promoter was active mainly in the lignifying cells of stem tissues. Further, to examine the effects of suppressing the expression of prxA3a, we transferred an antisense prxA3a gene construct into the original host, hybrid aspen (Populus sieboldii ×P. gradidentata), under the control of the original promoter of the prxA3a gene. Eleven transformed aspens were obtained and characterized, and the stable integration of the antisense construct was confirmed by PCR and Southern blotting analysis in all these lines. Assays of enzymatic activity showed that both total peroxidase activity and acidic peroxidase activity were lower in most transgenic lines than in the control plants. In addition, the reduction of peroxidase activity was associated with lower lignin content and modified lignin composition. Transgenic lines with the highest reduction of peroxidase activity displayed a higher syringyl/vanillin (S/V) ratio and a lower S+V yield, mainly because of a decreased amount of V units. Thus, our results indicate that prxA3a is involved in the lignification of xylem tissue and that the down-regulation of anionic peroxidase alters both lignin content and composition in hybrid aspen.  相似文献   

6.
Two nitrogen-deregulated mutants of Phanerochaete chrysosporium, der8-2 and der8-5, were isolated by subjecting wild type conidia to gamma irradiation, plating on Poly-R medium containing high levels of nitrogen, and identifying colonies that are able to decolorize Poly-R. The mutants showed high levels of ligninolytic activity (14C-synthetic lignin 14CO2), and lignin peroxidase, manganese peroxidase and glucose oxidase activities in both low nitrogen (2.4 mM) and high nitrogen (24 mM) media. The wild type on the otherhand displayed these activities in low nitrogen medium but showed little or no activities in high nitrogen medium. Fast protein liquid chromatographic analyses showed that the wild type as well as the der mutants produce three major lignin peroxidase peaks (designated L1, L2 and L3) with lignin peroxidase activity in low nitrogen medium. Furthermore, in low nitrogen medium, mutant der8-5 produced up to fourfold greater lignin peroxidase activity than that produced by the wild type. In high nitrogen medium, the wild type produced no detectable lignin peroxidase peaks whereas the mutants produced peaks L1 and L2, but not L3, and a new lignin peroxidase protein peak designated LN. Mutants der8-2 and der8-5 also produced high levels of glucose oxidase, an enzyme known to be associated with secondary metabolism and an important source of H2O2 in ligninolytic cultures, both in low and high nitrogen media. In contrast, the wild type produced high levels of glucose oxidase in low nitrogen medium and only trace amounts of this enzyme in high nitrogen medium. The results of this study indicate that the der mutants are nitrogen-deregulated for the production of a set of secondary metabolic activities associated with lignin degradation such as lignin peroxidases, manganese peroxidases and glucose oxidase.  相似文献   

7.
Low-molecular-weight glutenin subunits (LMW-GS) have great effect on wheat processing quality, but were numerous and difficult to dissect by SDS-PAGE. The development of functional markers may be the most effective way for a clear discrimination of different LMW-GS genes. In the present study, three different approaches were used to identify SNPs of different genes at Glu-D3 and Glu-B3 loci in bread wheat for the development of six STS markers (3 for Glu-D3 and 3 for Glu-B3 genes) that were validated with distinguished wheat cultivars. Firstly, seven LMW-GS gene sequences ( AY585350, AY585354, AY585355, AY585356, AY585349, AY585351 and AY585353 ) from Aegilops tauschii, the diploid donor of the D-genome of bread wheat, were chosen to design seven pairs of AS-PCR primers for Glu-D3 genes. By amplifying the corresponding genes from five bread wheat cultivars with different Glu-D3 alleles (a, b, c, d and e) and Ae. tauschii, a primer set, S13F2/S13R1, specific to the gene AY585356, was found to be positive to cultivars with alleles Glu-D3c and d. Nevertheless, the other five pairs of primers designed from AY585350, AY585349, AY585353, AY585354 and AY585355, respectively, did not produce specific PCR products to the cultivars tested. Secondly, all the PCR products from the five primer sets without specific characteristics were sequenced and an SNP from the gene AY585350 was detected in the cultivar Hartog, which resulted in the second STS marker S1F1/S1R3 specific to the allelic variant of AY585350. Thirdly, three Glu-D3 sequences (AB062851, AB062865 and AB062872) and three Glu-B3 sequences (AB062852, AB062853 and AB062860) defined by Ikeda et al. (2002) were chosen to query wheat EST and NR databases, and DNA markers were developed based on the putative SNPs among the sequences. Using this approach, four STS markers were developed and validated with 16-19 bread wheat cultivars. The primer set T1F4/T1R1 was also a Glu-D3 gene-specific marker for AB062872, while T2F2/T2R2, T5F3/T5R1 and T13F4/T13R3 were all Glu-B3 gene specific markers for AB062852, BF293671 and AY831800, respectively. The chromosomal locations of the six markers were verified by amplifying the genomic DNA of Ae. tauschii (DD), T. monococcum (AA) and T. turgidum (AABB) entries, as well as Chinese Spring and its group 1 chromosome nulli-tetrasomic lines. The results are useful to discriminate the corresponding Glu-D3 and Glu-B3 genes in wheat breeding programs.  相似文献   

8.
Ketan Nangka, the donor of wide compatibility genes, showed sterility when crossed to Tuanguzao, a landrace rice from Yunnan province, China. Genetic and cytological analyses revealed that the semi-sterility was primarily caused by partial abortion of the embryo sac. Genome-wide analysis of the linkage map constructed from the backcross population of Tuanguzao/Ketan Nangka//Ketan Nangka identified two independent loci responsible for the hybrid sterility located on chromosomes 2 and 5, which explained 18.6 and 20.1% of phenotypic variance, respectively. The gene on chromosome 5 mapped to the previously reported sterility gene S31(t), while the gene on chromosome 2, a new hybrid sterility gene, was tentatively designated as S32(t). The BC1F2 was developed for further confirmation and fine mapping of S32(t). The gene S32(t) was precisely mapped to the same region as that detected in the BC1F1 but its position was narrowed down to an interval of about 1.9 cM between markers RM236 and RM12475. By assaying the recombinant events in the BC1F2, S32(t) was further narrowed down to a 64 kb region on the same PAC clone. Sequence analysis of this fragment revealed seven predicted open reading frames, four of which encoded known proteins and three encoded putative proteins. Further analyses showed that wide-compatibility variety Dular had neutral alleles at loci S31(t) and S32(t) that can overcome the sterilities caused by these two genes. These results are useful for map-based cloning of S32(t) and for marker-assisted transferring of the neutral allele in hybrid rice breeding.  相似文献   

9.
Summary By starch gel electrophoresis three mobility variants of a cathodic moving doublet of bands, encoded by the structural gene prxC, were detected in all organs of flowering petunias. In root tissue two of the variants showed a lower electrophoretic mobility than in other organs. During development of flower buds the PRXc enzymes showed an increase in mobility. The gene prxC was located on chromosome IV by showing linkage to the genes An3 and Dw1, by trisomic segregation, and by the construction of triply heterozygous trisomics IV. The gene order on chromosome IV is B1-An3/Dw1-prxC. It was concluded that the temporal programming difference in the expression of the alleles prxC2 and prxC3 is caused by internal site mutation. Analysis of progeny obtained by crossing of lines to the trisomic IV with genotype prxC1/C1/C2 showed differential expression of the two prxC1 alleles of the trisomic IV.  相似文献   

10.
Summary The expressed activity in pollen and stigma was determined for both S alleles of sixteen S-alíele heterozygous genotypes and for one of the two S alleles of two additional heterozygotes. Activities were measured using pollen tube penetration and seed set data from reciprocal crosses between each S-allele heterozygote and its two corresponding S-allele homozygotes.In pollen the S-allele activities ranged from zero to 100% inhibition of pollen tube penetration and seed set, and in the stigma they ranged from 8 to 100% inhibition. Of the sixty-eight S-allele activities measured, thirty-three (48%) were 90 to 100% inhibition, nine (13%) were 80 to 89% inhibition and one to five were within each ten-unit range below 80% inhibition.In an S-allele heterozygote, each subset of two S alleles had an activity for each allele in both pollen and stigma which was highly repeatable among duplicate pollinations within and among successive years. Each subset of two S alleles had a specific S-allele interaction in the pollen, and the same or another specific interaction in the stigma. In pairings with six other S alleles, allele S 2 had four calculated levels of activity in pollen that ranged from 88 to 94%, and five levels in the stigmas between 15 and 94%. When paired in a heterozygote, alleles S 3 and S 5 had activities ranging between 42 and 59%, representing mutual weakening of S-allele activity. Also, heterozygote S 15 S 3 had pollen activities, respectively, of 25 and 6%, i.e. mutual weakening in the pollen.These results indicate that in heterozygous combination with a series of other S alleles, each S-allele may have activity in pollen and also in stigma that potentially is between zero and 100% inhibition. They further indicate that the defined sexual-organ X S-allele-interaction Types I, II, III and IV are extremes; all intermediate variations including complete weakening of both alleles are possible. Recessiveness is weakening of the activity of but one of the two S alleles. The pollen tube penetrations into the style and seed set were highly correlated.Department of Plant Breeding and Biometry Paper No. 683  相似文献   

11.
Puroindolines, the tryptophan-rich proteins controlling grain hardness in wheat, appeared as two pairs of 13 kDa polypeptides in the Acid-PAGE (A-PAGE) and two-dimensional A-PAGE×SDS-PAGE patterns of starch-granule proteins from wild allotetraploid wheat Aegilops ventricosa Tausch. (2n = 4x = 28, genomes DvDvNvNv). Puroindoline pair a1 + a2 reacted strongly with an antiserum specific for puroindoline-a from common wheat (Triticum aestivum L.), whereas puroindoline pair b1 + b2 exhibited A-PAGE relative mobilities similar to that of puroindoline-b in Aegilops tauschii (Coss.), the D-genome donor to both common wheat and Ae. ventricosa. Puroindolines a2 and b1 were found to be encoded by alleles Pina-D1a and Pinb-D1h on chromosome 5Dv, respectively, whereas puroindolines a1 and b2 were assumed to be under the genetic control of chromosome 5Nv. Puroindoline a1 encoded by the novel Pina-N1a allele exhibited a high level of amino acid variation with respect to puroindoline-a. On the other hand, the tryptophan-rich region of puroindoline b2 encoded by allele Pinb-N1a showed a sequence change from lysine-42 to arginine, with no effect on the amount of protein b2 accumulated on the starch granules. A partial duplication of the pin-B gene (Pinb-relic) was identified about 1100 bp downstream from Pinb-D1 on chromosome 5Dv. The present findings are the first evidence of a tetraploid wheat species in which four puroindoline genes are expressed. The potential of Ae. ventricosa as a source of genes that may be used to modulate endosperm texture and other valuable traits in cultivated wheat species is discussed.  相似文献   

12.
Summary Transposable element Activator (Ac) induced wild-type stable revertants, derived from McClintock's Dissociation (Ds) insertion shrunken (sh) mutant sh-m5933, have been examined for sucrose synthases, SS1 and SS2, encoded by the revertant (Sh) locus and the non-allelic gene Sus (previously designated as Ss2), respectively. A structurally normal Sh locus has been previously described in these revertants. Immuno-blot (Western) and Southern hybridization analyses reported here identify one of the nine alleles, Sh-r5, as unique for several features. It showed altered tissue specificity, as the SS1 protein encoded by the Sh-r5 allele was readily detectable in the immature embryo which is otherwise characterized by the Sus expression only. The level of Sh-r5 expression at the protein and enzyme level was marked by endosperm specific SS1 abundance and a significant down-regulation in the embryo similar to the standard Sh and Sus loci in endosperm and embryo, respectively. We infer that tissue specific levels of gene expression among maize Ss genes is significantly determined by trans-regulatory factors present in these two tissues. The Sh-r5 strain also exhibited a complete loss of the Sus expression in all tissues tested in the plant. Lack of any detectable phenotypic abnormality in the Sh-r5 strain due to the loss of SS2 protein indicated that either the SS2 protein is nonessential or that the two SS isozymes are functionally compensatory. Genomic filter hybridizations with the Sus cDNA clone indicated that the Sus locus in the Sh-r5 strain was not deleted and was, in fact, unique among these revertants. Together, these data provide an unusual insight into the regulation and function of the two SS isozymes in the maize plant.  相似文献   

13.
Summary Allelic complexity is a key feature of self-incompatibility (S) loci in gametophytic plants. We describe in this report the allelic diversity and gene structure of the S locus in Solanum tuberosum revealed by the isolation and characterization of genomic and cDNA clones encoding S-associated major pistil proteins from three alleles (S 1, S r1, S 2). Genomic clones encoding the S1 and S2 proteins provide evidence for a simple gene structure: Two exons are separated by a small intron of 113 (S 1) and 117 by (S 2). Protein sequences deduced from cDNA clones encoding S1 and Sr1 proteins show 95% homology. 15 of the 25 residues that differ between these S 1and S r1alleles are clustered in a short hypervariable protein segment (amino acid positions 44–68), which corresponds in the genomic clones to DNA sequences flanking the single intron. In contrast, these alleles are only 66% homologous to the S 2allele, with the residues that differ between the alleles being scattered throughout the sequence. DNA crosshybridization experiments identify a minimum of three classes of potato S alleles: one class contains the alleles S 1, S r1and S 3, the second class S 2and an allele of the cultivar Roxy, and the third class contains at present only S 4. It is proposed that these classes reflect the origin of the S alleles from a few ancestral S sequence types.  相似文献   

14.
Summary Multigene families are a ubiquitous feature of eukaryotes; however, their presence in Saccharomyces is more limited. The MAL multigene family is comprised of five unlined loci, MAL1, MAL2, MAL3, MAL4 and MAL6, any one of which is sufficient for yeast to metabolize maltose. A cloned MAL6 locus was used as a probe to facilitate the cloning of the other four functional loci as well as two partially active alleles of MAL1. Each locus could be characterized as a cluster of three genes, MALR (regulatory), MALT (maltose transport or permease) and MALS (structural or maltase), encoded by a total of about 7 kb of DNA; however, homologous sequences at each locus extend beyond the coding regions. Our results indicate that there is extensive homology among the MAL loci, especially within their maltase genes. The greatest sequence diversity occurs in their regulatory gene regions. Southern cross analyses of the cloned MAL loci indicate a single duplication of the MAL6R-homologous sequences upstream of the MAL6R gene as well as an extensive duplication of more than 10 kb at the MAL3 locus. The large repeat at the MAL3 locus results in the presence of four copies of MAL3R-homologous sequences and two copies of MAL3T-homologous sequences at that locus. Two naturally occurring inactive alleles of MAL1 show a deletion or divergence of their MALR sequences. The significance of these repeats in the evolution of the MAL multigene family is discussed.  相似文献   

15.
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.  相似文献   

16.
Cell cycle control in the fission yeastSchizosaccharomyces pombe involves interplay amongst a number of regulatory molecules, including thecdc2, cdc13, cdc25, weel, andmik1 gene products. Cdc2, Cdc13, and Cdc25 act as positive regulators of cell cycle progression at the G2/M boundary, while Wee1 and Mik1 play a negative regulatory role. Here, we have screened for suppressors of the lethal premature entry into mitosis, termed mitotic catastrophe, which results from simultaneous loss of function of both Wee1 and Mik1. Through such a screen, we hoped to identify additional components of the cell cycle regulatory network, and/or G2/M-specific substrates of Cdc2. Although we did not identify such molecules, we isolated a number of alleles of bothcdc2 andcdc13, including a novel wee allele ofcdc2, cdc2-5w. Here, we characterizecdc2-5w and two alleles ofcdc13, which have implications for the understanding of details of the interactions amongst Cdc2, Cdc13, and Wee1.  相似文献   

17.
Notch-mediated lateral inhibition has been reported to regulate auditory hair cell and supporting cell development from common precursors. While the Notch effector genes Hes1, Hes5 and Hey1 are expressed in the developing cochlea, inactivation of either of them causes only mild abnormality, suggesting their functional redundancy. To explore the roles of Hes/Hey genes in cochlear development, we examined compound heterozygous or homozygous mutant mice that lacked Hes1, Hes5 and Hey1 alleles. We found that a reduction in Hes/Hey gene dosage led to graded increase of hair cell formation. However, if at least one allele of Hes1, Hes5 or Hey1 was intact, excessive hair cells were accompanied by overproduction of supporting cells, suggesting that the hair cell increase does not occur at the expense of supporting cells, and that each Hes/Hey gene functions to induce supporting cells. By contrast, when all alleles of Hes1, Hes5 and Hey1 were inactivated, the number of hair cells increased more drastically, whereas that of supporting cells was unchanged compared with control, suggesting that supporting cell formation was balanced by their overproduction and fate conversion into hair cells. The increase of the cell numbers seemed to occur after the prosensory domain formation in the mutants because the proliferation state and the size of the prosensory domain were not affected. Thus, Hes1, Hes5 and Hey1 cooperatively inhibit hair cell formation, and one allele of Hes1, Hes5 or Hey1 is sufficient for supporting cell production probably by lateral inhibition in the sensory epithelium. Strikingly, Hes/Hey mutations lead to disorganized cell alignment and polarity and to hearing loss despite hair cell overproduction. These results suggest that Hes/Hey gene dosage is essential not only for generation of appropriate numbers of hair cells and supporting cells by controlling cell proliferation and lateral inhibition but also for the hearing ability by regulating the cell alignment and polarity.  相似文献   

18.
Plants are continually exposed to external and internal DNA-damaging agents. Although lesions can be removed by different repair processes, damages often remain in the DNA during replication. Synthesis of template damages requires the replacement of replicative enzymes by translesion synthesis polymerases, which are able to perform DNA synthesis opposite specific lesions. These proteins, in contrast to replicative polymerases, operate at low processivity and fidelity. DNA polymerase η and Rev 1 are two proteins found in eukaryotes that are involved in translesion DNA synthesis. In Arabidopsis, DNA polymerase η and Rev 1 are encoded by AtPOLH and AtREV1 genes, respectively. Transgenic plants over-expressing AtPOLH showed increased resistance to ultraviolet light. Only plants with moderate AtREV1 over-expression were obtained, indicating that this enzyme could be toxic at high levels. Transgenic plants that over-expressed or disrupted AtREV1 showed reduced germination percentage, but the former exhibited a higher stem growth rate than the wild type during development.  相似文献   

19.
Summary Embryo and endosperm peroxidases from dry mature seeds of three subspecies of tetraploid wheat (Triticum turgidum L.) were subjected to genetic analysis. The inheritance of eight isozymes (embryo isozymes a2, d1, d2, e and f; and endosperm isozymes b, d and 4) were studied in F2's obtained from different wheat accessions. Simple monogenic inheritance producing three banded: one null segregation and two epistatic segregations (97 and 151) were found. In the case of isozymes b, d and 4, monogenic or epistatic segregation depended on the F2 analyzed. Segregation data indicated that at least 9 different loci would determine the peroxidase isozymes of tetraploid wheat seed, all the loci studied containing null alleles. Furthermore, several loci determining embryo peroxidases were noticed to be mutually linked. All these data are discussed in context of the inheritance of seed peroxidases in hexaploid wheat and rye.  相似文献   

20.
Many bacteria adapt to microoxic conditions by synthesizing a particular cytochrome c oxidase (cbb 3) complex with a high affinity for O2, encoded by the ccoNOQP operon. A survey of genome databases indicates that ccoNOQP sequences are widespread in all sub-branches of Proteobacteria but otherwise are found only in bacteria of the CFB group (Cytophaga, Flexibacter, Bacteroides). Our analysis of available genome sequences suggests four major strategies of regulating ccoNOQP expression in response to O2. The most widespread strategy involves direct regulation by the O2-responsive protein Fnr. The second strategy involves an O2-insensitive paralogue of Fnr, FixK, whose expression is regulated by the O2-responding FixLJ two-component system. A third strategy of mixed regulation operates in bacteria carrying both fnr and fixLJ-fixK genes. Another, not yet identified, strategy is likely to operate in the -Proteobacteria Helicobacter pylori and Campylobacter jejuni which lack fnr and fixLJ-fixK genes. The FixLJ strategy appears specific for the -subclass of Proteobacteria but is not restricted to rhizobia in which it was originally discovered.  相似文献   

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