High-level Expression of Maize <Emphasis Type="Italic">γ</Emphasis>-zein Protein in Transgenic Soybean (<Emphasis Type="Italic">Glycine max</Emphasis>)

Primers were developed for 118 microsatellites isolated from grape (Vitis vinifera) genomic libraries enriched for (AC)n repeats. Only one microsatellite sequence matched other grape SSR-sequences in the GeneBank database. Genotyping was carried out in the parental lines and four offspring of two pseudo-test-cross populations, Cabernet Sauvignon x Seyval and Chardonnay x Bianca, and a further six other grape genotypes (V. vinifera Sultanina, Merlot, Syrah, Müller-Thurgau, Vitis Regent and V. riparia Gloire de Montpellier). A total of 108 microsatellites showed easily scorable alleles and 100 of them segregated according to a configuration suitable for mapping in either cross. A further 8 SSRs, although unsuitable for mapping in those crosses, showed polymorphism in the other genotypes tested. This set of markers was used, along with 75 microsatellites of other repeat-types, to fingerprint 46 offspring of the cross Chardonnay x Bianca. For each full-sib, individual heterozygosity and distance in repeat units between pairs of alleles at each locus (mean d²) were calculated as a tool for predicting highly outbred recombinant individuals. Six microsatellites with segregation ratios significantly distorted towards the lack of homozygous sibs were identified and mapped to linkage groups LG 3 and LG 5. Estimation of heterozygosity at genome-wide level and genotyping at loci for which homozygous sibs are discriminated against are discussed for marker-assisted background selection in outcrossing grapevines.     

Plant cells do not properly recognize animal gene polyadenylation signals

Summary We have introduced chimeric genes containing polyadenylation signals from a human gene and two animal virus genes into tobacco cells. We see, in all three cases, inefficient and aberrant utilization of the foreign polyadenylation signals. We find that a chimeric gene carrying the polyadenylation site of the human growth hormone gene is polyadenylated at three sites in the vicinity of the site that is polyadenylated in human cells. A chimeric gene containing the polyadenylation site from the adenovirus 5 E1A gene is polyadenylated at a site 11 bases downstream from that reported in animal cells. A gene carrying the polyadenylation site from the SV40 early region is polyadenylated some 80 bases upstream from the site that is polyadenylated in animal cells. In all three cases, related mRNAs ending at flanking authentic plant polyadenylation sites can be detected, indicating that the foreign polyadenylation signals are inefficiently utilized in tobacco cells.     

Cytoplasmic male sterility in barley

Summary The effect of the msm1 cytoplasm of barley (Hordeum vulgare L.) on kernel protein and lysine was studied using the near-isogenic, unrestored derivatives of seven barley varieties. With normal lysine varieties, Adorra, Bomi, CI 4362, and Hankkija's Eero, the msm1 cytoplasm produced an average of one percentage point more protein than did the normal cytoplasm of the same varieties. There was no difference between the two cytoplasms with respect to their effect on the lysine content. With high lysine varieties, Bomi Risø mutant 13, Bomi Risø mutant 1508, and CI 3947, msm1 produced almost one percentage point more protein but protein with a somewhat decreased lysine content.Induced partial spike fertility in normal Adorra was found to be associated with lysine in meal (r=–0.999), with protein in meal (r=–0.984), and with lysine in protein (r=0.941). Removal of the spikes on the secondary tillers affected both the protein and its lysine content. It is suggested that good spike fertility is an important pre-requisite when selecting high lysine and/or high protein segregants or mutants.     

Viability and activity in readily culturable bacteria: a review and discussion of the practical issues

In microbiology the terms viability and culturability are often equated. However, in recent years the apparently self-contradictory expression viable-but-nonculturable (VBNC) has been applied to cells with various and often poorly defined physiological attributes but which, nonetheless, could not be cultured by methods normally appropriate to the organism concerned. These attributes include apparent cell integrity, the possession of some form of measurable cellular activity and the apparent capacity to regain culturability. We review the evidence relating to putative VBNC cells and stress our view that most of the reports claiming a return to culturability have failed to exclude the regrowth of a limited number of cells which had never lost culturability. We argue that failure to differentiate clearly between use of the terms viability and culturability in an operational versus a conceptual sense is fuelling the current debate, and conclude with a number of proposals that are designed to help clarify the major issues involved. In particular, we suggest an alternative operational terminology that replaces VBNC with expressions that are internally consistent.     

Genetic control of heading date and spikelet number in common wheat (T. aestivum L.) line ‘Noa’

Summary The phenology and build-up of spikelet number under 10 h day-length were studied in five wheat lines: the multispikelet line Noa, the regular line Mara, the F₁ hybrid between them and monosomics 2D of Mara and of this hybrid (lacking the 2D chromosome of Mara). Noa had a longer spike development phase, a higher initial number of spikelet primordia and a slower rate of spikelet production than Mara. The F₁ hybrid was similar to Noa in its high initial number of spikelets and to Mara in its high rate of spikelet production. This hybrid had a shorter spikelet phase than both parents. Deletion of one dose of the Mara 2D chromosome from either Mara or the F₁ hybrid caused a reduction in the rate of spikelet production and an increase in the duration of the spikelet phase. These effects were due to the reduced dosage of the 2D chromosome. However, in the F₁ hybrid this deletion also caused an increase in the spike development phase — an indication that Noa carries on its 2D chromosome a recessive gene for late heading date which acts on the spike development phase. This gene of Noa is independent of the day-length sensitive gene ppd, and is different from Noas dominant gene for large initial number of spikelets.     

Nucellar callus of ‘Femminello’ lemon,selected for tolerance toPhoma tracheiphila toxin,shows enhanced release of chitinase and glucanase into the culture medium

Phoma tracheiphila is the causative agent of the disease mal secco. Citrus cultivars differ substantially in respect to their sensitivity to the pathogenP. tracheiphila and its toxin. Some cultivars (e.g., Femminello lemon) are inherently sensitive while others (e.g., Tarocco orange) are tolerant. Cell lines derived from nucellar tissue of Femminello, Tarocco and a cell line selected for tolerance to the fungal toxin (Femminello-S) were used to study host-pathogen interaction. Our results showed that calli or conditioned media of Tarocco and Femminello-S inhibited the size of co-cultivated fungal colonies when compared to Femminello. In addition, conditioned medium of Tarocco as well as FemminelloS, but not Femminello, promoted bursting of hyphal tips. A ten-fold increase in chitinase and glucanase enzymatic activity, as evaluated by radiometric assay and laminarin hydrolysis respectively, was detected in Femminello-S extracellular extracts as compared to Femminello. An increase in chitinase was also shown by immunoblot analysis. Our findings suggest a positive correlation between the presence of chitinase and glucanase in the conditioned media of the cultured cells and the tolerance of those cells toP. tracheiphila toxin.     

Effect of LiCl on compression and tension properties ofPophyra perforata tissue

The mechanical properties of various differentiated regions of thePorphyra perforata thallus and the effect of LiCl were studied by performing compression and tension tests. Among the various differentiated tissues, the holdfast area was high in its compressive modulus of elasticity and tensile modulus of elasticity, possibly related to its thick matrix. Vegetative non-dividing tissue and vegetative dividing tissue were the most flexible and strong, showing the highest percentage elongation at break and tensile strength. The patch area, which is a transition zone leading to sexually mature tissue, had moderate values of tensile properties. Meanwhile, sexually differentiated male and female tissues had the highest compressive modulus of elasticity and lowest tensile properties. Thes tisues tended to crumble easily. Treatments in LiCl, as used for DNA extraction, resulted in a decrease in both compressive modulus of elasticity (87%) and tensile modulus of elasticity (54%). After treatment of tissue for chromosome staining in a method using LiCl, there was a marked decrease in tensile modulus of elasticity (49%), while the compressive modulus of elasticity remained unchanged. Such mechanical changes verify the softening effect of LiCl on the seaweedP. perforata tissue.Author for correspondence     

Photoreactivation of the UV light effects on growth of Scots pine (Pinus sylvestris L.) seedlings

Summary Ultraviolet-B light (UV-B) and ultraviolet-A light (UV-A) at higher doses exert a strong inhibitory (toxic) effect on axis growth in Scots pine (Pinus sylvestris L.) seedlings. This effect is unrelated to control of growth rate by phytochrome. Rather, after a toxic UV dose growth of the pine seedling no longer responded to phytochrome. Both, the effect of UV-B as well as the inhibiting effect of UV-A could be photoreactivated by blue light (B). The action of UV-A was 2 fold: (i) it exerted a toxic effect which could be photoreactivated by B, and (ii) applied after UV-B it photoreactivated to some extent the toxic UV-B effect. Obviously, the UV-A range causes a toxic effect, and at the same time is capable of photoreactivating the toxic UV effect. At higher doses the toxic effect prevails.     

Estudio comparativo de la inmunoelectroforesis (IEF) y de la inmunoelectroosmoforesis-inmunodifusion (IEOF-ID) aplicadas al diagnostico de la paracoccidioidomicosis

Se diferencian 5 arcos de precipitatión en la técnica de inmunoelectroforesis (IEF) luego de enfrentar sueros de 16 pacientes de paracoccidioidomicosis a la paracoccidioidina. El arco l presente en todos los sueros es asimilado a arcos presuntamente específicos descritos previamente. En immunoelectroosmoforesis-inmunodifusión (IEOF-ID) se observaron bandas anódicas y bandas catódicas en todos los casos. La comparación de los resultados obtenidos con las 2 técnicas reveló siempre un mayor número de arcos en IEOF-ID lo que se debería a la aparición de los arcos de localizatión catódica 1 y 2 del inmunoelectroforegrama, tanto en el lado anódico como en el catódico de las láminas de IEOF-ID.

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Summary Immunoelectroporesis (IEF) and immunoelectroosmophoresis-immunodiffusion (IEOP-ID) (1) were comparatively used in the diagnosis of 16 patients with mycologically proved paracocidioidomycosis.In IEF, 5 different precipitin arcs were found and identified with arable numbers. Arc. 1, cathodic, present in all the patients, is assimilated to specific arcs previously described by other authors.In IEOP-ID both cathodic and anodic arcs were observed in all the sera.A high number of precipitin arcs were revealed by IEOP-ID technique in comparison to IEF in every case. This should be due to the presence of cathodic arcs 1 and 2 of the immunoelectrophoregram at both sides of the IEOP-ID preparate.

     

A linkage map with RFLP and isozyme markers for almond

Inheritance and linkage studies were conducted with seven isozyme genes and 120 RFLPs in the F₁ progeny of a cross between almond cultivars Ferragnes and Tuono. RFLPs were detected using 57 genomic and 43 cDNA almond clones. Eight of the cDNA probes corresponded to known genes (extensin, prunin (2), -tubulin, endopolygalacturonase, oleosin, actin depolymerizing factor and phosphoglyceromutase). Single-copy clones were found more frequently in the cDNA (65%) than in the genomic libraries (26%). Two maps were elaborated, one with the 93 loci heterozygous in Ferragnes and another with the 69 loci heterozygous in Tuono. Thirty-five loci were heterozygous in both parents and were used as bridges between both maps. Most of the segregations (91%) were of the 11 or 1111 types, and data were analyzed as if they derived from two backcross populations. Eight linkage groups covering 393 cM in Ferragnes and 394 in Tuono were found for each map. None of the loci examined in either map was found to be unlinked. Distorted segregation ratios were mainly concentrated in two linkage groups of the Ferragnes map.     

Comparative study on the chloroplast,mitochondrial and nuclear genome differentiation in two cultivated rice species,Oryza sativa and O. glaberrima,by RFLP analyses

Summary Restriction fragment length polymorphisms of chloroplast (ct), mitochondrial (mt) and nuclear DNA were investigated using eight cultivars of Oryza sativa and two cultivars of O. glaberrima. Relative variability in the nuclear and cytoplasmic genomes was estimated by a common measure, genetic distance. Based on the average genetic distances among ten cultivars for each genome, the evolutionary variabilities of the mitochondrial and nuclear genomes were found to be almost the same, whereas the variability of the chloroplast genome was less than half that of the other two genomes. Cluster analyses on ct and mt DNA variations revealed that chloroplast and mitochondrial genomes were conservative within a taxon and that their differentiations were well-paralleled with respect to each other. For nuclear DNA variation, an array of different degrees of differentiation was observed in O. sativa, in contrast with little variation in O. glaberrima. As a whole, differentiation between O. sativa and O. glaberrima was clearly observed in all three genomes. In O. sativa, no notable difference was found between the cultivars Japonica and Javanica, whereas a large differentiation was noticed between Japonica (including Javanica) and Indica. In all three genomes, the average genetic distances within Indica were much larger than those within Japonica (including Javanica), and almost similar between Japonica (including Javanica) and Indica. These facts indicate that differentiation in O. sativa was due mainly to Indica.     

Epistatic expression of quantitative trait loci (QTL) in soybean [Glycine max (L.) Merr.] determined by QTL association with RFLP alleles

Quantitative trait values for seed oil and protein content or for maturity were measured in recombinant inbred lines (RIL) of soybean derived from a cross between two soybean cultivars: Minsoy PI 27890 and Noir 1 PI 290136. Seed oil was found to be inversely correlated to protein content. By analyzing DNA from plants with extreme phenotypes, we were able to identify quantitative trait loci (QTL) for these traits as being linked to several restriction fragment length polymorphism (RFLP) loci, including R183 for oil and protein content and R79 for maturity. Cumulative distributions of trait values were graphed for those RIL with Minsoy alleles and for those with Noir 1 alleles. As already suggested by the alleles found associated with extreme phenotypes, the distributions were consistent with an independent and additive expression of the maturity QTL linked to R79. That is, the cumulative distributions for plants with Minsoy alleles and for plants with Noir 1 alleles were similar in shape, but the entire Noir 1 curve had been shifted to later maturity dates. In contrast, the trait distributions for a locus affecting oil and protein content linked to R183 were not compatible with an additive model. These results suggest that this approach can be used for rapid identification of QTLs with epistatic expression.     

Sources and inheritance of resistance to leaf curl virus in Lycopersicon

Summary One hundred and twenty-two varieties, lines and wild accessions of Lycopersicon were screened under three different regimes during the autumn/winter season of 1982–83 and 1983–84 for resistance to tomato leaf curl virus (TLCV). L. hirsutum f. glabratum (B6013) and L. hirsutum f. typicum (A1904) proved to be highly resistant to TLCV in all three environments. Various accessions of L. peruvianum were also highly resistant. L. pimpinellifolium (A1921) exhibited no TLCV symptoms within 90 days. Of the cultivated varieties, Acc 99 exhibited the minimim score for susceptibility; AC 142, Collection No. 2, Kalyanpur Angurlata and HS 101 had a low rating for virus incidence. The inheritance of resistance was studied in the interspecific crosses between a TLCV resistant line of L. pimpinellifolium (A1921) and five (HS 101, HS 102, HS 110, Pusa Ruby and Punjab Chhuhara) susceptible cultivars of L. esculentum. Parents, F₁, F₂ and backcross progenies were artificially inoculated with local strains of TLCV using vector the viruliferious whitefly, Bemisia tabaci (Genn.). Data indicated that the resistance of L. pimpinellifolium (A 1921) is monogenic and incompletely dominant over susceptibility.     

Ecological theories and Dutch nature conservation

This paper aims to achieve insight into various ecological theories in the Netherlands which have different, and sometimes opposing, views on the conservation of nature. Interviews, publications and archival research brought to light four separate theories: vitalistic/holistic, dynamic, cybernetic and chaos. Diversity is reached through stability according to vitalistic/holistic and cybernetic theories, but through change and instablility according to the dynamic and chaos theories. These two groups are working apart, and continue to have their own ideas. Prediction of the future is only possible with the vitalistic/holistic and cybernetic theories. Ecologists who adhere to these theories feel responsible and able in different ways to change ecological nature towards desirable end goals. The other two theories, dynamic and chaos, appear to be less activist.     

Water stress-induced oxidative damage and antioxidant responses in micropropagated banana plantlets

Oxidative injury and antioxidant responses were investigated in two banana genotypes (Musa AAA Berangan and Musa AA Mas) subjected to 40 % PEG-induced water stress. PEG treatment resulted in oxidative injury, as expressed in increased lipid peroxidation and reduced membrane stability index, in both cultivars; however, greater oxidative injury was detected in Mas. Under PEG treatment, catalase activity and glutathione reductase activity were enhanced in both cultivars, but were higher in Mas. Ascorbate peroxidase activity was enhanced in Berangan under water stress, but was unaffected in Mas. Meanwhile, superoxide dismutase activity was inhibited in both cultivars under water stress, but higher activity was detected in Berangan. Higher ascorbate peroxidase and superoxide dismutase activities were associated with greater protection against water stress-induced oxidative injury.     

Donor tissue and culture condition effects on mesophyll protoplasts of Medicago sativa

Mesophyll protoplasts were produced from clones of two cultivars of Medicago sativa, Rangelander and Regen S. Protoplasts from the Regen S clone generally gave rise to calli while those from the Rangelander clone would undergo direct embryogenesis. Effects of plant growth conditions, donor tissue pretreatment and protoplast culture conditions on mesophyll protoplast production and subsequent development patterns were investigated. The major factor determining whether or not mesophyll protoplasts would be produced from either of the clones was the pretreatment in water of shoots excised from the donor plants. Pretreatment in water containing growth regulators did not alter protoplast production or development in the Regen S clone. Pretreatment of the Rangelander clone shoots with abscisic acid or naphthaleneacetic acid was slightly beneficial to embryo production while pretreatment with benzylaminopurine was detrimental. Altered leaf morphology induced by growth condition changes did not affect mesophyll protoplast production or subsequent development patterns when shoots were pretreated in water. Culture of protoplasts in liquid droplets or solid agar medium increased low density protoplast survival and subsequent embryo production in the Rangelander clone.     

Mutagenic effectiveness and efficiency of EMS,DES and gamma-rays in rice

Summary Data on chlorophyll mutation frequency after treatment with EMS, DES and gamma-rays and sequential administration of gamma-rays and the two alkylating agents in three varieties of rice have been used to work out quantitatively the effectiveness and efficiency of each mutagen and combination treatment. For effectiveness, the order is EMS > DES and for efficiency it is EMS > DES > gamma-rays. In some sequential treatments (Gamma-rays + DES in IR8 and Basmati; DES + gamma-rays in IR8 and Jhona; Gamma-rays + EMS in IR8 and Basmati; and EMS + gamma-rays in IR8, Jhona and Basmati) mutation frequency is more than additive (synergistic) but these treatments are decisively less efficient because of their relatively high injurious effects in the M₁. generation. EMS induces more albinas than gamma-rays do. The mutational spectrum patterns induced by gamma-rays and DES are alike. In general, combination treatments tend to increase the frequency of albinas over other types of chlorophyll mutants.     

DNA markers tightly linked to a gall midge resistance gene (Gm2) are potentially useful for marker-aided selection in rice breeding

We have developed a polymerase chain reaction (PCR)-based assay that could effectively reduce the time period required to screen and select for Gall Midgeresistant rice lines under field conditions. The primers for the assay were designed on the basis of sequence information of two phenotype specific random amplified polymorphic DNA fragments which were found to be tightly linked to Gall Midge biotype-1 resistance gene (Gm2). The two RAPD fragments, F8₁₇₀₀ in the susceptible parent ARC6650 and F10₆₀₀ in the resistant parent Phalguna, were identified after screening 5450 loci using 520 random primers on genomic DNAs of ARC6650 and Phalguna. These primers, when used in a multiplexed PCR, amplified specifically a 1.7-kb and 0.6-kb fragment in the susceptible and resistant parents, respectively. When this assay was performed on genomic DNAs of 44 recombinant inbred lines derived from ARC6650 x Phalguna and 5 lines derived from other crosses where one of the parents was Phalguna, ARC6650 or their derivatives, the primers amplified a 1.7-kb fragment in all of the susceptible lines or a 0.6-kb fragment in all of the resistant ones. These markers can be of potential use in the marker-aided selection of Gall Midge biotype-1 resistant phenotypes. As screening for resistance can now be conducted independent of the availability of insects, the breeding of resistant varieties can be hastened.     

Differential susceptibility in some sorghum (Sorghum vulgar) genotypes to zinc deficiency in soil

Summary Nine genotypes of sorghum were screened for their relative susceptibility to zinc deficiency in soil. All the varieties were highly responsive to zinc application, but behaved differently under zinc stress conditions in respect of dry-matter production, zinc concentration and uptake. The varieties, JS-263, 29/1, JS-20 and Son 127×SL-245 and CSH-1 were relatively more susceptible to zinc deficiency than 3677×6352, Son 3×SL 209, CSH-3 and 2077×1151. re]19730102     

Frequency and spectrum of chlorophyll mutants induced in rice by chemical mutagens

Summary Dry seeds of rice varieties T(N)1, IR 8 and Sona, with stabilised moisture content and presoaked in distilled water, were treated with chemical mutagens MMS, dMS, dEMS and dES with the purpose of evaluating chlorophyll mutation frequency and spectrum. In the M₂ generation, mutants occurred in 24 lines of 'T(N)1; 94 lines of IR 8 but only in six of Sona. They include albino, viridis, xantha and other categories of which viridis was predominant. dES was found to be most effective of all mutagens used in all the three varieties and varietal differences were observed.