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1.
Gametophytic self-incompatibility (GSI) in sweet cherry is determined by a locus S with multiple alleles. In the style, the S-locus codifies for an allele-specific ribonuclease (S-RNase) that is involved in the rejection of pollen that carries the same S allele. In this work we report the cloning and genomic DNA sequence analysis including the 5 flanking regions of four S-RNases of sweet cherry (Prunus avium L., Rosaceae). DNA from the cultivars Ferrovia, Pico Colorado, Taleguera Brillante and Vittoria was amplified through PCR using primers designed in the conserved sequences of sweet cherry S-RNases. Two alleles were amplified for each cultivar and three of them correspond to three new S-alleles named S 23 , S 24 and S 25 present in 'Pico Colorado', 'Vittoria' and 'Taleguera Brillante' respectively. To confirm the identity of the amplified fragments, the genomic DNA of these three putative S-RNases and the allele S 12 amplified in the cultivar Ferrovia were cloned and sequenced. The nucleotide and deduced amino-acid sequences obtained contained the structural features of rosaceous S-RNases. The isolation of the 5-flanking sequences of these four S-RNases revealed a conserved putative TATA box and high similarity among them downstream from that sequence. However, similarity was low compared with the 5-flanking regions of S-RNases from the Maloideae. S 6 - and S 24 -RNase sequences are highly similar, and most amino-acid substitutions among these two RNases occur outside the rosaceous hypervariable region (RHV), but within another highly variable region. The confirmation of the different specificity of these two S-RNases would help elucidate which regions of the S-RNase sequences play a role in S-pollen specific recognition.Communicated by H.F. Linskens  相似文献   

2.
Nicotiana tabacum and Nicotiana alata plants were transformed with genomic clones of two S-RNase alleles from N. alata. Neither the S 2 clone, with 1.6 kb of 5 sequence, nor the S 6 clone, with 2.8 kb of 5 sequence, were expressed at detectable levels in transgenic N. tabacum plants. In N. alata, expression of the S 2 clone was not detected, however the S 6 clone was expressed (at low levels) in three out of four transgenic plants. An S 6-promoter-GUS fusion gene was also expressed in transgenic N. alata but not N. tabacum. Although endogenous S-RNase genes are expressed exclusively in floral pistils, the GUS fusion was expressed in both styles and leaves.  相似文献   

3.
The rate of CO2- and p-benzoquione-dependent photosynthetic O2 evolution by Anabaena variabilis cells remained unaltered and the rate of O2 uptake observed after switching off the light (endogenous respiration) was enhanced by a factor of 6–8 when the O2 concentration was increased from 200 to 400 M. Photosystem-I-linked O2 uptake and respiration of the cells incubated with ascorbate and N,N,NN-tetramethyl-p-phenylenediamine was not appreciable influenced by the O2 concentration. 2-Iodo-6-isopropyl-3-methyl-2,4,4-trinitrodiphenyl ether, blocking electron transfer at the plastoquinone level, suppressed O2 evolution and had no influence on endogenous respiration. 2-n-Heptyl-4-hydroxyquinoline-N-oxide, an inhibitor of electron transfer between photosystems II and I, as well as the cytochrome-oxidase inhibitors N 3 - , CN- and NH2OH, caused a 35–50% retardation of endogenous respiration and blocked photosynthetic O2 evolution. The molar ratio of cytochromes b6, f, c-553, aa3 and photosystem-I reaction centers in the isolated membranes equalled approx. 2:1:2:0.7:2. It is inferred that endogenous respiration of A. variabilis cells is inhibited by the light-induced electron flow through both photosystems at the level of the plastoquinone-plastocyanin-oxidoreductase complex.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - DNP-INT 2-iodo-6-isopropyl-3-methyl-2,4,4-trinitrodiphenyl ether - Hepes 4-(2-hydroxyethyl)-1-piperazine ethansulfonic acid - TMPD N,N,NN-tetramethyl-p-phenylenediamine  相似文献   

4.
The seeds of 19 sunflower species were compared on the basis of their protein contents and the relative proportions of their protein fractions. The globulin content varied from 50% to about 70% and the albumin content from 18% to 35% according to the species. The level of intermediateMr polypeptides showed a great variability (9.6 to 24.3%). Comparative studies onMr polymorphism were carried out by means of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of non reduced and/or reduced samples using both mono- and bidimensional procedures. Polypeptide constituents of helianthinin were compared including both number and molecular size (cultivatedH. annuus was used as a standard). Studies focused on differences observed between the major two (Mr 38 000), (Mr 32 000) and (Mr 25 500), (21 000) polypeptides families constituting the main A, B, and C subunits. and polypeptides analyses permit to discriminate easilyH. petiolaris from the other species. Charge polymorphism was studied using isoelectric focusing (IEF) and IEF-PAGE in mono and bidimensional procedures in the presence or absence of 2-mercaptoethanol (2-ME). Only a specific 4 polypeptide enables an easy discrimination betweenH. petiolaris and all the other species. Detailed nomenclature of the , and , polypeptides constituting the different helianthinin globulin subunits is given via the results of pI andMr analyses. Monodimensional IEF patterns of the more basic albumins (pI > 8.0) appear to provide a more valuable approach to identifying specific protein markers.  相似文献   

5.
The S incompatibility system of apple was confirmed through the application of the gene Got-1 for glutamate oxaloacetate transaminase as a marker for the S locus. The 11S alleles proposed by Kobel et al. (1939) were confirmed through anomalous segregations for Got-1 observed in 14 semi-compatible crosses and regular segregations observed in 2 fully compatible crosses. The S allele genotypes of Idared (S 3 S 7), Cox (S 5 S 9) and Fiesta (S 3 S 5) were determined and found to fall within the original series. By associating parental incompatibility genotypes with the segregation of Got-1 alleles, we were able to deduce the coupling of S and Got-1 alleles in 9 varieties.  相似文献   

6.
Summary In flower extracts of defined genotypes of Petunia hybrida, an enzyme activity was demonstrated which catalyses the hydroxylation of naringenin and dihydrokaempferol in the 3-position. Similar to the flavonoid 3-hydroxylases of other plants, the enzyme activity was found to be localized in the microsomal fraction and the reaction required NADPH as cofactor. A strict correlation was found between 3-hydroxylase activity and the gene Ht1, which is known to be involved in the hydroxylation of flavonoids in the 3-position in Petunia. Thus, the introduction of the 3-hydroxyl group is clearly achieved by hydroxylation of C15-intermediates, and the concomitant occurrence of the 3,4-hydroxylated flavonoids quercetin and cyanidin (paeonidin) in the presence of the functional allele Ht1 is due to the action of one specific hydroxylase catalysing the hydroxylation of common precursors for both flavonols and anthocyanins.  相似文献   

7.
An abnormal, structurally modified, kinetically stable S2-state has been reported to be induced when Photosystem II was treated with NaCl-EGTA (or EDTA) in the light or with pH in darkness, both are assumed to release functional Ca2+. In order to compare the mechanism of induction of modified S2-state between the two treatments, effects of illumination during or before low pH-treatment on formation of the abnormal S2-state were investigated by means of thermoluminescence measurements and low temperature EPR spectroscopy. Following results have been obtained: Flash illumination during low pH-treatment did not practically induce the abnormal S2-state, whereas preflash illumination given immediately before low pH-treatment efficiently induced the abnormal S2-state, and its amplitude showed a period-four oscillation on varying the preflash number with maxima at the second and sixth flashes. The abnormal S2-state thus induced by preflashes was identical with the modified S2-state that could be induced in dark-low pH-treated Photosystem II by excitation at 0°C after neutralization to pH 6.5. It was inferred that in low pH-treatment, modified S2-state can be formed from both S2- and S3-states, but its yield from the latter is much higher than from the former, consistent with the early results by Boussac et al. obtained for NaCl-EGTA-light or NaCl-citrate-light treatment.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - EDTA ethylenediaminetetraactate - EGTA ethylene glycol bis(-aminoethylether)-N,N,N,N-tetraacetic acid - Mes 2-(N-morpholino)ethanesulfonic acid - PS II Photosystem II  相似文献   

8.
9.
We have investigated the effect of some metabolic drugs, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), 2,4-dinitrophenol (DNP), sodium azide (NaN3), on the photobehavior of single cells of Euglena gracilis, in order to clarify the relevance of different metabolic pathways in the process of photoperception and sensory transduction in this alga. The results obtained show that the photophobic response of Euglena is not affected by the action of these drugs. This suggests that neither the photosynthetic process nor oxidative phosphorylation play a significant role in the phenomenon of photosensory transduction in Euglena.List of Abbreviations DNP 2,4-dinitrophenol - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - PSI Photosystem I - PSII Photosystem II  相似文献   

10.
The seed storage globulins from sixHelianthus and four hybrids were studied using mono and bidimensional gel SDS electrophoresis (+ 2 mercaptoethanol). The polypeptide composition of each subunit was determined. Different pairs are specifically expressed according to the species studied. Three typical patterns were discriminated. All the studied species exhibit five subunits: two of them are expressed in all the species (11 and 22). The subunit corresponding to the 11 pair is present inH. petiolaris and in the three populations ofH. annuus studied. The 2b2 pair is common toH. annuus andH. argophyllus. H. petiolaris presents two specific 2a2 and 44 pairs andH. annuus a specific 33 pair. InH. argophyllus 11 33 or 44 are never observed but are replaced by 13 and 31 pairs. Some globulins, poorly represented, are of forms but present chains of higher molecular weights (in the range 54–56 kDa). Expressing variations in the banding patterns between these species by the use of a similarity index reveals complete identity between the three populations ofH. annuus. Identity between the twoH. petiolaris studied is also observed.H. annuus andH. argophyllus appear to be closer to each other thanH. petiolaris concerning the seed storage globulins.  相似文献   

11.
The interaction of 3,5-bis-O-(,,,-tetrafluoropyrid--yl)thymidine with various nucleophilic reagents was studied to evaluate the possibility of molecular design of new types of nucleic acid analogues using S NAr reactions. The reactions with morpholine and sodium azide led to the introduction of one and two nucleophilic residues into each of the polyfluorinated pyridine rings. The nucleophilic polycondensation with bifunctional reagents ethylenediamine and hexamethylenediamine depended on the nature of nucleophile and reaction conditions and resulted in the formation of supramolecules containing about five or more than 20 pyrimidine bases.  相似文献   

12.
Resistance to verticillium wilt, a vascular disease causing yield losses in many crops, is conferred in tomato by a single dominant allele, Ve. A population segregating for the Ve allele was generated using near-isogenic tomato lines. Analysis of the parental tomato DNA using the polymerase chain reaction and 400 random primers, each 10 deoxyribonucleotides in length, produced 1,880 amplified DNA fragments. Of the four polymorphisms observed between the resistant and susceptible parental genotypes, only one was linked to the Ve gene. No recombination was observed between this DNA marker and the Ve locus, indicating that the linkage is less than 3.5±2.7 cM. The marker detected both the susceptible and resistant alleles, producing amplified DNA fragments of approximately 1,300 and 1,350 bp, respectively. The sequence of the primer, determined from cloned amplified products, was 5 CTCACATGCA 3 instead of the expected 5 CTCACATGCC 3. The marker will be of value to tomato breeding programs because of the tight linkage, Codominant nature, and analytical procedure utilized.  相似文献   

13.
2D NMR spectroscopy and J coupling constant analysis are applied to resolve the structure of two photoproducts of thymidylyl-(35)-thymidine. These products are cyclobutane type thymine dimers possessing the cis-syn (the predominant one) and trans-syn geometry. The cis-syn is formed in an ANTI-ANTI conformation about the N-glycosyl linkages and resembles the normal base-stacked configuration. The glycosidic conformation in solution of the 5 terminal fragment differs from the crystal in which the less common SYN conformation is observed. In this isomer only the sugar pucker of the 3 terminal fragment is changed substantially with respect to the dinucleotide. The trans-syn isomer is formed in a SYN-ANTI glycosidic conformation. In this isomer the sugar puckers of both deoxyribose rings are affected and a preference for a pure 2-endo conformation is observed.Abbreviations dTpdT 2-deoxythymidylyl-(35)-2-deoxythymidine - dTp[]dT cyclobutane type photodimers of dTpdT - dTp- and dTp[]- their 5' terminal fragments (fragment A) - -pdT and-[]pdT their 3 terminal fragments (fragment B) - RP-HPLC reversed-phase high-performance liquid chromatography - COSY two-dimensional correlated spectroscopy - 2D NOE two-dimensional nuclear Overhauser spectroscopy  相似文献   

14.
Summary The development of a technique for the identification of S alleles involved in self-incompatibility in Brassica oleracea which is based on the polymerase chain reaction (PCR) amplification of genomic DNA followed by restriction analysis is described. Primers homologous to conserved regions near to the 5 and 3 ends of the S coding sequence were used to amplify a number of members of the S multigene family. However, by designing a selective primer and using a higher temperature for annealing in the PCR, we were able to amplify certain members from the multigene family preferentially. These were considered to be the S-locus glycoprotein genes (SLG), since the patterns of restriction bands of the PCR products were shown to correspond to those of the SLG where sequence data were available. DNA samples from plants with certain S alleles were found not to amplify efficiently using the selective primers and high annealing temperature. This property, however, could be used as a means of distinguishing plants homozygous for these S alleles, as was demonstrated by an examination of a small F2 population that was segregating for the S5 and S29 alleles. In the investigation of the F2 population, it was found that preferential amplification of one of the alleles of the heterozygotes occurred when the consensus primers were used in the PCR. However, by using different primers, homologous to another region of the S sequence, we were able to amplify both alleles of the heterozygotes equally. The genotypes of the plants were determined by restriction analysis of PCR products and agreed with results based on pollen-tube growth tests.  相似文献   

15.
Egbert  K.J.  Martin  C.E. 《Photosynthetica》2002,40(1):35-39
Four fluorescence parameters [Fv/Fm = the intrinsic efficiency of energy conversion via photosystem 2 (PS2); Fv/Fm= the efficiency of energy conversion via PS2 in the light; P = fraction of absorbed radiant energy utilized for photosynthesis; and D = fraction of absorbed radiant energy dissipated as heat] were measured on leaves of seven species of succulents having epidermal windows. While the function of leaf windows has reportedly been to increase absorption of radiant energy and, hence, the rate of photosynthesis in these species, recent evidence indicates that this translucent portion of epidermal tissue, lacking chlorophyll, may also result in photoinhibition in these species, especially for those with growth habits aboveground. Species with aboveground and belowground growth habits were compared with their leaf windows covered with reflective tape and with windows unobstructed. Results showed no increase in photoinhibition for these species resulting from the radiant energy penetrating the window tissue. Although the efficiency of the photosynthetic mechanism was not significantly influenced by the additional radiant energy provided by the window for individual species, there were significant differences in the efficiencies of radiant energy capture (Fv/Fm) and utilization (P) between the two growth habits. Species with an aboveground growth habit were less efficient in radiant energy utilization compared with the species having a belowground growth habit.  相似文献   

16.
Using primary cultures of gill pavement cells from freshwater rainbow trout, a method is described for achieving confluent monolayers of the cells on glass coverslips. A continuous record of intracellular pH was obtained by loading the cells with the pH-sensitive flourescent dye 2,7-bis(2-carboxyethyl)-5(6)-carboxyfluorescein and mounting the coverslips in the flowthrough cuvette of a spectrofluorimeter. Experiments were performed in HEPES-buffered media nominally free of HCO3. Resting intracellular pH (7.43 at extracellular pH=7.70) was insensitive to the removal of Cl or the application of 4-acetamido-4-isothiocyanatostilbene-2,2-disulfonic acid (0.1 mmol·l–1), but fell by about 0.3 units when Na+ was removed or in the presence of amiloride (0.2 mmol·l–1). Exposure to elevated ammonia (ammonia prepulse; 30 mmol·l–1 as NH4Cl for 6–9 min) produced an increase in intracellular pH (to about 8.1) followed by a slow decay, and washout of the pulse caused intracellular pH to fall to about 6.5. Intracellular non-HCO 3 buffer capacity was about 13.4 slykes. Rapid recovery of intracellular pH from intracellular acidosis induced by ammonia prepulse was inhibited more than 80% in Na+-free conditions or in the presence of amiloride (0.2 mmol·l–1). Neither bafilomycin A1 (3 mol·l–1) nor Cl removal altered the intracellular pH recovery rate. The K m for Na+ of the intracellular pH recovery mechanism was 8.3 mmol·l–1, and the rate constant at V max was 0.008·s–1 (equivalent to 5.60 mmol H+·l–1 cell water·min–1), which was achieved at external Na+ levels from 25 to 140 mmol·l–1. We conclude that intracellular pH in cultured gill pavement cells in HEPES-buffered, HCO 3 -free media, both at rest and during acidosis, is regulated by a Na+/H+ antiport and not by anion-dependent mechanisms or a vacuolar H+-ATPase.Abbreviations BCECF 2,7-bis(2-carboxyethyl)-5(6)-carboxy-fluorescein - BCECF/AM 2,7-bis(2-carboxyethyl)-5(6)-carboxy-fluorescein, acetoxymethylester - Cholin-Cl choline chloride - DMSO dimethyl sulfoxide - EDTA ethylene diamine tetra-acetic acid - FBS foetal bovine serum - H + -ATPase Proton-dependent adenosine triphosphatase - HEPES N-[2-hydroxyethyl]piperazine-N[2-ethanesulfonic acid] - pH i intracellular pH - pH e extracellular pH - PBS phosphate-buffered saline - SITS 4-acetamido-4-isothiocyanatostilbene-2,2-disulfonic acid  相似文献   

17.
A potent toxin with phospholipase A2 (PLA2) and hemolytic activity in vitro was purified from the Russell's viper venom of eastern India (RVV-EI). The purified protein (RVV-PFIIc) of 15.3 kDa molecular weight, and a lethal toxicity dose (LD50 i.p.) of 0.1 mg/kg body weight, was the most toxic PLA2 so far reported from the Indian subcontinent. The material also possessed anticoagulant activity as it enhanced the prothrombin induced plasma clotting time in vitro. The PLA2 toxin (RVV-PFIIc) was shown to be different from other PLA2s of RVV in respect to one or more of these parameters e.g. molecular weight, isoelectric pH, in vivo toxicity, specific activity of the enzyme and certain other biological activities. The first 19 amino terminal sequence (NLFQFAEMIVKMTGKEAVH) of RVV-PFIIc showed variable degree of homology (42.1–94.7%) with those of other RVV-PLA2s described in the literature. Antisera raised against RVV-EI or RVV-PFIIc, though completely neutralized the in vivo lethal toxicity of RVV-EI or RVV-PFIIc, failed to inhibit their PLA2 activity in vitro thereby suggesting that in vivo toxicity and in vitro activity of the enzyme may not be directly related. Apart from RVV-PFIIc, at least two other PLA2 isozymes were found to be present in RVV-EI that were distinct from RVV-PFIIc in respect to their molecular, biological as well as serological properties. The significance of these and related data in antivenom therapy is discussed.  相似文献   

18.
Summary The nuclear 18 S, 5.8 S and 25 S ribosomal RNA genes (rDNA) of Cucumis sativus (cucumber) occur in at least four different repeat types of 10.2, 10.5, 11.5, and 12.5 kb in length. The intergenic spacer of these repeats has been cloned and characterized with respect to sequence organization. The spacer structure is very unusual compared to those of other eukaryotes. Duplicated regions of 197 bp and 311 bp containing part of the 3 end of the 25 S rRNA coding region and approximately 470 bp of 25 S rRNA flanking sequences occur in the intergenic spacer. The data from sequence analysis suggest that these duplications originate from recombination events in which DNA sequences of the original rDNA spacer were paired with sequences of the 25 S rRNA coding region. The duplicated 3ends of the 25 S rRNA are separated from each other mostly by a tandemly repeated 30 bp element showing a high GC-content of 87.5%. In addition, another tandemly repeated sequence of 90 bp was found downstream of the 3flanking sequences of the 25 S rRNA coding region. These results suggest that rRNA coding sequences can be involved in the generation of rDNA spacer sequences by unequal crossing over.  相似文献   

19.
Summary In hybrid breeding programs, testcross evaluation of lines can be done during the early stages of selfing (early testing) or delayed until the lines are near-homozygous. To evaluate the usefulness of early testing, the expected genetic and phenotypic correlations between testcross performance at different selfing generations were examined. The genetic correlation (r GnGn ) between testcross performance of S n and S n , (n>n) individuals or lines is equal to the square root of the ratio of their testcross genetic variances, and it is a function of the inbreeding coefficients (F) at the two selfing generations, i.e., r GnGn=[(1+F n )/(1+F n )]0.5. The genetic correlation between testcross performance of lines and their directly descended homozygous (n=) lines is 0.71 for S1; 0.87 for S2, 0.93 for S3, 0.97 for S4, 0.98 for S5, and 0.99 for S5 lines. The effectiveness of early testing is limited mainly by nongenetic effects. The square root of testcross heritability at generation n sets the upper limit on the correlation between phenotypic value at generation n and genotypic value at homozygosity. The probabilities of correctly retaining S n individuals or lines that have superior testcross performance at homozygosity (n=) indicate that early testing should be effective in identifying lines with above- and below-average combining ability. However, the risk of losing lines with superior combining ability is high if strong (best 10%) selection pressure is applied during early testing. If only a small proportion of lines is retained based on testcross performance and/or if the heritability of the trait is low, selfing for two or three generations prior to testcrossing may be desirable to increase the likelihood of retaining lines that perform well at homozygosity. The theoretical results in this study support the testcross evaluation procedures for grain yield used by most maize (Zea mays L.) breeders.A contribution from Limagrain Genetics, a Groupe Limagrain company  相似文献   

20.
Summary The nucleotide sequence of cytoplasmic 5S ribosomal RNA fromEuglena gracilis has been determined to be: G- A C -G-U-A-C-G-G-C-C-A-U-A-C-U-A-C-C-G-G-G-A-A-U-A-C-A-C-C-U-G-A-A-C-C-C-G--U-C-G-A-U-U-U-C-A-G-A-A-G-U-U-A-A-G-C-C-G-G-G-U-U-A-G-G-C-C-C-A-G-U-U-A-G-U-A-C-U-G-A-G-U-G-G-G-C-G-A-C-C-A-C-U-U-G-G-G-A-A-C-A-C-U-G-G-G-U-G-C-U-G-U-A-C-G-C-U-Up. This RNA is 119 nucleotides long and the sequence of a probable tRNA-binding site is GAUU (position 41–44 from the 5-terminus), which is the same as that of a trypanosoma species,Crithidia fasci-culata. TheEuglena 5S rRNA has a pseudouridine residue at position 38 and 3-terminus is phosphorylated. The 5S rRNA sequence ofEuglena resembles those of several other protozoa and higher animals rather than plants.On leave from Department of Zoology, Hiroshima University, Hiroshima, Japan  相似文献   

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