In vitro pollen germination of species and two interspecific hybrids from the genus Brassica

In vitro pollen germination of five species and two interspecific hybrids from the genus Brassica was tested in four media. Genetically fixed differences in the demands for optimal pollen germination among species were found. The experiments were designed to define optimal content of mineral salts, sugar, and PEG for every investigated species or hybrids. The differences found among species are discussed in relation to the evolutionary trend.     

Studies on the crossability relationships of some spinous Solanums

Summary Cytogenetic studies involving six species of spinous Solanums have been made using the parameters of crossability/fertility relationships and chromosome pairing in parental and derived hybrid genomes. These studies have made it possible to integrate hybrid reproductive performance with observed cytological phenomena and draw inferences on concerned intergenomic homeologies. It has been found that species' differences, as they exist, are ascribable to chromosome repatterning, in their divergence and maintenance as distinct taxa. In the light of these studies, the grouping of these species has to be reconsidered.     

Multivariate analysis applied to tomato hybrid production

Summary Twenty characters were measured on 60 tomato varieties cultivated in the open-air and in polyethylene plastic-house. Data were analyzed by means of principal components, factorial discriminant methods, Mahalanobis D² distances and principal coordinate techniques. Factorial discriminant and Mahalanobis D² distances methods, both of which require collecting data plant by plant, lead to similar conclusions as the principal components method that only requires taking data by plots. Characters that make up the principal components in both environments studied are the same, although the relative importance of each one of them varies within the principal components. By combining information supplied by multivariate analysis with the inheritance mode of characters, crossings among cultivars can be experimented with that will produce heterotic hybrids showing characters within previously established limits.     

Albumin extinction followed by de novo methylation of its gene in somatic hybrids of a rat hepatoma

We have previously identified an Msp I site at the 5′ end of the rat albumin gene whose undermethylation is necessary but not sufficient for stable albumin expression in rat hepatoma cells [1]. We have also shown that the extinction of albumin expression in somatic hybrids is not the result of methylation at this site, since for two different crosses, rapid extinction was found to occur in the absence of any de novo methylation of the previously active gene[2]. In the present study, we examine albumin expression and albumin gene methylation for independent hybrid clones isolated from crosses between albumin expressing rat hepatoma cells and cells of two different non-expressing lines. The cells from hybrid clones of both crosses are characterized by stable extinction of albumin expression. Moreover, we find that de novo methylation of the “extinguished” albumin gene can occur in somatic hybrids, but only some weeks after the gene has ceased to be expressed.     

Human β-glucuronidase: Assignment of the structural gene to chromosome 7 using somatic cell hybrids

-Glucuronidase (GUS) has become an important enzyme model for the genetic study of molecular disease, enzyme realization, and therapy, and for the biogenesis and function of the lysosome and lysosomal enzymes. The genetics of human -glucuronidase was investigated utilizing 188 primary man-mouse and man-Chinese hamster somatic cell hybrids segregating human chromosomes. Cell hybrids were derived from 16 different fusion experiments involving cells from ten different and unrelated individuals and six different rodent cell lines. The genetic relationship of GUS to 28 enzyme markers representing 19 linkage groups was determined, and chromosome studies on selected cell hybrids were performed. The evidence indicates that the -glucuronidase gene is assigned to chromosome 7 in man. Comparative linkage data in man and mouse indicate that the structural gene GUS is located in a region on chromosome 7 that has remained conserved during evolution. Involvement of other chromosomes whose genes may be important in the final expression of GUS was not observed. A tetrameric structure of human -glucuronidase was demonstrated by the formation of three heteropolymers migrating between the human and mouse molecular forms in chromosome 7 positive cell hybrids. Linkage of GUS to other lysosomal enzyme genes was investigated. -Hexosaminidase HEX _B) was assigned to chromosome 5; acid phosphatase₂ (ACP ₂) and esterase A₄ (ES-A ₄) were assigned to chromosome 11; HEX _A was not linked to GUS; and -galactosidase (-GAL) was localized on the X chromosome. These assignments are consistent with previous reports. Evidence was not obtained for a cluster of lysosomal enzyme structural genes. In demonstrating that GUS was not assigned to chromosome 9 utilizing an X/9 translocation segregating in cell hybrids, the gene coding for human adenylate kinase₁ was confirmed to be located on chromosome 9.Supported by NIH Grants HD 05196, GM 20454, and GM 06321, by NSF Grant BMS 73-07072, and by HEW Maternal and Child Health Service, Project 417.     

Relationship between species in the genus Rosa, section Pimpinellifoliae

The morphology of twelve species of Rosa is described and similarities between these species are assessed. Possible origins of the tetraploid species from diploid species are indicated on grounds of comparative morphology.
The wild origins of living and herbarium specimens are given in order to supplement published data on geographical distribution.
Meiosis in pollen mother cells, viability of pollen grains at anthesis and ability to set seed was studied in several F₁ hybrids: no indication of complete or even partial sterility was found. Reproductive isolation is therefore unlikely to be maintained by reduced fertility of interspecific hybrids.
Three species are reduced to synonymy with three other species, being retained as subspecific taxa. Two species are transferred from section Pimpinellifoliae to section Cinnamomeae.     

Isolation of somatic hybrids by cloning Nicotiana heterokaryons in nurse cultures

Fusion of Nicotiana knightiana Goodsp. and kanamycin resistant Nicotiana sylvestris Speg. et Com. protoplasts was induced by polyethylene glycol treatment. Heterokaryons were isolated by micropipette and transferred to nurse cultures of albino cells. Colonies originating from the heterokaryons could subsequently be distinguished by their green colour. The somatic hybrid nature of four such colonies was confirmed by isoenzyme pattern, kanamycin resistance and restored morphogenic potential. An additional kanamycin resistant line with characteristic Nicotiana knightiana isoenzymes was also found indicating that the drug resistance in the kanamycin resistant parent is under cytoplasmic control.Abbreviations ADH alcohol dehydrogenase - Nk Nicotiana knightiana     

Molecular characterisation of inter and intra-specific somatic hybrids of potato using randomly amplified polymorphic DNA (RAPD) markers

Summary Protoplast fusion allows the transfer of both mono- and polygenic traits between species that are sexually incompatible. This approach has particular relevance for potato, and somatic hybridisation has been used to introduce a range of disease resistance genes from sexually incompatible wild species into the cultivated potato gene pool. In addition, protoplast fusion allows the resynthesis of tetraploid genotypes from pre-selected diploid or dihaploid donor parents. A limiting factor for the efficient exploitation of this technology in potato breeding is the difficulty of unequivocally identifying nuclear hybrids (heterokaryons). In order to facilitate the identification of hybrids at an early stage following fusion, Randomly Amplified Polymorphic DNA markers (RAPDs) have been used to characterise molecularly both inter- and intra-specific somatic hybrids of potato. RAPD markers detect naturally occurring polymorphism in the donor genotypes and utilise short oligonucleotide primers of arbitrary nucleotide sequence in combination with the polymerase chain reaction (PCR). The exploitation of RAPDs in the characterisation of both somatic and sexual hybrids is discussed.     

The inheritance of salt exclusion in woody perennial fruit species

Citrus and grapevines are salt-sensitive perennial crops. Damage caused by salinity is due mostly to accumulation of excessive concentrations of salt (Na- and Cl ions) in shoot tissues. In both crops, however, some rootstock varieties can restrict the accumulation of salt in scion leaves and stems. Salt-excluding rootstocks, however, are often deficient with regard to other desirable characteristics and as such their use is limited. As a consequence, we have conducted a range of crosses within both crops to select new salt-excluding hybrids which may have potential as new rootstocks and also to investigate the inheritance of salt exclusion in these woody perennials.In citrus, both Cl-ion and Na-ion exclusion has been observed as a continuous character and progenies segregate widely for their ability to restrict the accumulation of these ions in shoot tissues. The ability to exclude Cl ions appears to be independent of the ability to exclude Na ions. Thus a good Cl-ion excluder is not necessarily a good Na-ion excluder and vice versa. It has been possible, however, to select new salt-excluding citrus hybrids which perform as well as and sometimes better than parent varieties when grafted with a common scion and grown in artificially salinised field plots.In grapevines, the research has concentrated on the ability for Cl-ion exclusion and depending on the Cl-ion-excluding parent chosen this is inherited as either a polygenic or monogenic trait. In crosses between Vitis champini (Cl-ion excluder) and Vitis vinifera (Cl-ion accumulator), the ability to restrict Cl-ion accumulation in shoot tissues segregates widely with some offspring transgressing the performance of either parent. In crosses and backcrosses involving V. berlandieri and V. vinifera, however, hybrids segregate as either Cl-ion excluders or accumulators suggesting the effect of a major dominant gene for Cl-ion exclusion from V. berlandieri. This was evident from both field and glasshouse experiments although possible modifying genes from V. vinifera may affect the expression of this gene under glasshouse conditions in some crosses.     

Advances in cereal protoplast research

Beginning in 1986, plants have been regenerated from protoplasts of all of the important cereal species, including wheat, rice, maize, and barley, and grasses such as sugarcane. In addition, somatic hybrids/cybrids as well as transgenic plants with introduced useful agronomic traits have been obtained in several instances. This rapid and impressive progress in the genetic manipulation of cereals has been made possible by two critical technical advances during the past decade: the establishment of embryogenic suspension cultures as a source of totipotent protoplasts and the direct delivery of DNA into protoplasts for genetic transformation.