Preparation and comparison of biological properties of recombinant carp (Cyprinus carpio) growth hormone and its Cys-123 to Ala mutant

Carp growth hormone (cGH) cDNA, in which Cys-123 was mutated to Ala, was prepared, transferred to the expression vector, expressed in Escherichia coli and the mutant was purified to homogeneity. The mutation only slightly improved yield of the monomeric fraction, indicating that Cys-123 is not involved in improper refolding. As compared to cGH, the mutant (cGH-C123A) exhibited lower binding affinity toward homologous liver receptors and lower bioactivity in a 3T3-F442A preadipocyte bioassay despite the fact that both hormones exhibited almost identical cross-reactivity with anti-cGH antibodies. These results, along with those of a structural comparison to hGH, suggest that Cys-123 is located in the hydrophobic core of the hormone, and is most likely affecting the conformation of the binding site. Dimeric forms of the hormone and its mutant were less active than their respective monomers. Homologous binding experiments using a carp liver microsomal fraction revealed a single receptor population with Kd = 0.77 nM and B_max = 241 fmol/mg microsomal protein.

---

Résumé Un ADN complémentaire (cDNA) de l'hormone de croissance de carpe (cGH), dans lequel l'acide aminé Cys-123 a été muté en Ala, a été préparé, inséré dans un vecteur d'expression, et exprimé dans Escherichia coli. Le mutant a ensuite été purifié jusqu'à homogénéité. La mutation améliore seulement faiblement la production de la fraction monomérique, indiquant que le Cys-123 n'est pas impliqué dans un repliement erroné. Comparé à la cGH, sa forme mutée (cGH-C123A) montre une plus faible affinité de liaison vis à vis de récepteurs hépatiques homologues, et une plus faible activité biologique dans un test réalisé sur des préadipocytes 3T3-F442A; cela en dépit du fait que les deux hormones présentent des réactions croisées presque identiques avec un anticorps anti-cGH polyclonale. Ces résultats, associés à une comparaison à la structure de l'hGH, suggèrent que le Cys-123 est localisé dans la partie hydrophobique de l'hormone, et affecte, le plus vraisemblablement, la conformation du site de liaison. Les formes dimériques de l'hormone et de sa forme mutée sont moins actives que leurs monomères respectifs. Les études de liaison homologue, réalisées avec des fractions microsomales de foie, révèlent une population unique de récepteurs de Kd = 0,77 nM et de B_max = 241 fmol/mg de proteine microsomale.

     

Milling Energy Polymorphism in Hordeum spontaneum Koch in Israel and its Potential Utilization in Breeding for Malting Quality

Variation for grain milling energy in Hordeum vulgare is associated with differences in grain size, shape and composition. Genetic polymorphisms within and between populations were found across the ecogeographical range of Hordeum spontaneum in Israel. Low milling energy estimates were positively correlated with and predictable by the aridity index. Milling energy in H. spontaneum, collected in Israel, ranged from the lowest values seen in making barley to twice the highest values seen in feed cultivars. It is concluded that H. spontaneum elite genotypes could be effectively identified by milling energy for use in the breeding of new cultivars.     

Genetic resources of wild cereals in Israel and vicinity. I. Phenotypic variation within and between populations of wild wheat,Triticum dicoccoides

Summary Populations of wild emmer wheat, Triticum dicoccoides, in Israel, originating from diverse habitats, and tested earlier for allozyme (Nevo et al., 1982) and disease resistance polymorphisms (Moseman et al., 1983a, 1983b; Nevo et al., 1984a, 1984b), were compared and contrasted for performance in agronomically important phenotypic traits. The traits compared involved 10 variables comparing germination, earliness, biomass and yield variables. The field experiments were conducted in 1980, 1981 and 1982 in two relatively standardized and contrasting environments: mesic (Mount Carmel, Haifa) and xeric (Acedat Farm, and Sede Boqer, in the northern Negev desert). The experimental design involved 12 population quadrangles at Avedat Farm in 1980, and rows of randomized genotypes of five populations in both Haifa and Sede Boqer in 1981 and 1982.The results indicate that the characters studied are partly genetically determined. Striking genetic variation was found between and, at least in some characters, also within populations in each site, whereas remarkable environmental variation including genetic-environmental interaction was found between the mesic and the xeric sites. We conclude that natural populations of wild emmer wheat in Israel vary not only in genetic polymorphisms of allozymes and disease resistance, but also in quantitative traits of agronomic importance. These traits are economically significant and should be conserved and utilized in wheat crop improvement.     

Genetic diversity in wild barley (Hordeum spontaneum C. Koch) in the Near East: a molecular analysis using Random Amplified Polymorphic DNA (RAPD) markers

We analyzed the genetic diversity in 88 genotypes from 20 populations of wild barley (Hordeum spontaneum C. Koch) from Israel, Turkey and Iran, by randomly amplified polymorphic DNA (RAPD). Twenty two of the 33 primers used yielded scorable products with 1–11 polymorphic bands. No duplicate patterns were found except for four haplotypes.When the total genetic diversity was estimated, 75% of the variation detected was partitioned within the 88 genotypes and 25% among the populations. When variation between countries was assessed, no substantial differences were found, because most of the variation detected (97%) was partioned within the 20 populations and the remainder among countries. The results of this limited survey indicate that the extensive genetic diversity is present in natural stands of wild barley throughout the Fertile Crescent.     

Genetic Resources for Salt Tolerance in the Wild Progenitors of Wheat (Triticum dicoccoides) and Barley (Hordeum spontaneum) in Israel

Salt tolerance was tested in the progenitors of cultivated cereals, wild barley (Hordeum spontaneum) and wild emmer wheat (Triticum dicoccoides) from Israel. Plants from five selected populations of H. spontaneum from the Mediterranean Coastal Plain and northern Negev desert, were grown on 250 and 350 mM of NaCl. Likewise, five populations of T. dicoccoides from the eastern Samaria steppes, Mt. Hermon and Mt. Carmel, were grown on 175 and 250 mM of NaCl. Here we report on superior genotypes of H. spontaneum, ripening at 350 mM NaCl (= 60 % sea water), and of T. dicoccoides ripening at 250 mM (— 40 % sea water). We are proceeding now with both genetical and physiological studies aimed at chromosomally-locating salt tolerant genes and unravelling the mechanism(s) of salt resistance in these wild cereals.     

Genetic resources of wild cereals in Israel and vicinity. II. Phenotypic variation within and between populations of wild barley,Hordeum spontaneum

Summary Populations of wild barley, Hordeum spontaneum, in Israel, originating from diverse habitats, and tested earlier for allozyme (Nevo et al., 1979a) and disease resistance polymorphisms (Moseman et al., 1984; Nevo et al., 1984a), were compared and contrasted for performance in agronomically important phenotypic traits. The traits compared involved 10 variables comparing germination, earliness, numerical and weight variables of biomass and yield. The field experiments were conducted in 1980, 1981 and 1982 in two relatively standardized and contrasting environments: mesic (Mount Carmel, Haifa) and xeric Avedat farm, and Sede Boqer (both in the northen Negev desert). The experimental design involved 26 population quadrangles at Avedat farm (1980), and rows of randomized plants of 11 populations in both Haifa and Sede Boqer (1981 and 1982).The results indicate that the characters studied are partly genetically determined. Striking genetic variation was found between and within populations in each site, whereas remarkable environmental variation including genetic environmental interaction was found between the mesic and the xeric sites, as well as between populations and years. We conclude that natural populations of wild barley in Israel vary not only in genetic polymorphisms of allozymes and disease resistance, but also in quantitative traits of agronomie importance. These traits are economically significant and should be conserved and utilized in barley crop improvement.     

Amino-Acid Resources in the Wild Progenitor of Wheats, Triticum dicoccoides, in Israel — Polymorphisms and Predictability by Ecology and Isozymes

Amino acid contents were measured in 109 genotypes from 22 populations of wild emmer wheat, Triticum dicoccoides, across its ecological range in Israel. Plants were grown outdoors on mesic Mr. Carmel, in a standardized environment, without irrigation and fertilization. T. dicoccoides was high in lysine and isoleucine contents as compared to six other species of wild wheats, but low in threonine and proline. Significant correlations were found among lysine and the other essential amino acids. Significant differences between Israeli populations of T. dicoccoides were found for isoleucine, methionine, leucine and threonine, but not for lysine, arginine, proline and glutamic acid. Significant differences between regions were only found in methionine. Ecological factors and allozyme markers appear to be good guidelines for predicting the ecogeographical location and allozymic constitution of specific elite amino acid genotypes, either singly or in combination. T. dicoccoides may be used in the future as a promising genetic resource for genetically improving the nutritional value of cultivated wheats.     

Resistance of wild emmer wheat to stem rust: Ecological,pathological and allozyme associations

Summary Seedling resistance to wheat stem rust was determined in populations of wild emmer wheat, Triticum dicoccoides, and characterized by means of ecological factors and allozyme genotypes. Reactions to wheat stem rust were studied in 102 single plant accessions of T. dicoccoides from ten populations by inoculation with Puccinia graminis tritici race 14, isolate GSR-739. Six populations displayed different degrees of response polymorphism with reactions ranging from high resistance to complete susceptibility, whereas four populations contained only susceptible plants. In some of the accession, unexplained intrasib variation in resistance and intraplant variation of infection-types were found. Resistance to stem rust was negatively correlated with two ecological factors, altitude and number of Sharav (hot-dry) days which are unfavorable to disease development. Variation in stem rust response was shown to exist in ecogeographic regions where climatic variables enhanced the development of the fungus, conceivably maintained by natural selection. Likewise, allozyme genotypes, single or in multiple loci combinations, appeared to be associated with resistance or susceptibility to rust. Such association need to be verified by genetic studies in order to become established as useful markers.     

Genetic variation in seine escapability of the common carp

During the years 1970, 1971 and 1972, 26 genetically diverse groups of common carp were tested in communal ponds, i.e., all the tested groups were co-stocked into the same ponds. These groups included inbred and crossbred domesticated European carp, one representative of the Chinese Big-Belly race, and several hybrids between the two races. The results led to the following conclusions concerning the genetics of seine escapability. (1) The Chinese race is much more adept at escaping seines than the European race. (2) The hybrids between the two races are intermediate with incomplete dominance of the higher escapability of the Chinese race. (3) Genetic variation was found within the European race with a strong dominance, or heterosis, of higher escapability. (4) Body weight as well as growth capacity were not associated with seine escapability. The absence of correlation between growth capacity and seine escapability contradicts earlier results that suggested a strong genetic correlation between the two characters.