Inheritance of gusA and neo genes in transgenic rice

Inheritance of foreign genes neo and gusA in rice (Oryza sativa L. cv. IR54 and Radon) has been investigated in three different primary (T₀) transformants and their progeny plants. T₀ plants were obtained by co-transforming protoplasts from two different rice suspension cultures with the neomycin phosphotransferase II gene [neo or aph (3) II] and the -glucuronidase gene (uidA or gusA) residing on separate chimeric plasmid constructs. The suspension cultures were derived from callus of immature embryos of indica variety IR54 and japonica variety Radon. One transgenic line of Radon (AR2) contained neo driven by the CaMV 35S promoter and gusA driven by the rice actin promoter. A second Radon line (R3) contained neo driven by the CaMV 35S promoter and gusA driven by a promoter of the rice tungro bacilliform virus. The third transgenic line, IR54-1, contained neo driven by the CaMV 35S promoter and gusA driven by the CaMV 35S.Inheritance of the transgenes in progeny of the transgenic rice was investigated by Southern blot analysis and enzyme assays. Southern blot analysis of genomic DNA showed that, regardless of copy numbers of the transgenes in the plant genome and the fact that the two transgenes resided on two different plasmids before transformation, the introduced gusA and neo genes were stably transmitted from one generation to another and co-inherited together in transgenic rice progeny plants derived from self-pollination. Analysis of GUS and NPT II activities in T₁ to T₂ plants provided evidence that inheritance of the gusA and neo genes was in a Mendelian fashion in one plant line (AR2), and in an irregular fashion in the two other plant lines (R3 and IR54-1). Homozygous progeny plants expressing the gusA and neo genes were obtained in the T₂ generation of AR2, but the homozygous state was not found in the other two lines of transgenic rice.     

Genetic markers for Atlantic salmon (Salmo salar L.): single locus inheritance and joint segregation analyses of minisatellite (VNTR) DNA loci

Relatively few genetic markers are available for detailed studies of Atlantic salmon. The detection of 12 distinct minisatellite DNA loci in this species (by 10 Atlantic salmon and brown trout derived probes) and subsequent inheritance analyses in two half-sib families are reported here. Disomic Mendelian inheritance was confirmed at all loci. Only a single aberrant progeny genotype (at Ssa -A60) was observed among 138 progeny screened. None of the loci was sex-linked. The tight linkage association Str -A22/1 with Str -A22/2, previously reported for brown trout, was found to be conserved in the Atlantic salmon genome. An additional male-specific linkage group, Ssa -A34 with Str -A9/2, was also noted. These highly polymorphic loci should find widespread use as chromosomal, individual, familial and, probably, population markers.     

Sex-related spatial kin structure in a spring population of grey-sided voles Clethrionomys rufocanus as revealed by mitochondrial and microsatellite DNA analyses

Polymerase chain reaction-directed mitochondria (mt) and microsatellite DNA analyses were performed to examine the kin structure in a spring population of grey-sided voles Clethrionomys rufocanus in Hokkaido, Japan. The spatial distribution of 81 voles in a trapping grid (about 1 ha) was estimated by using the catch-mark-release method. DNA samples were extracted from the toes clipped for individual identification. Maternal lineages of voles were unequivocally determined by the mtDNA haplotypes, as identified by nucleotide sequencing of the control region. Relatedness between individuals was estimated based on the genotype and allele frequencies at several microsatellite loci. Although the distribution of voles was uniform within the grid, neighbouring females were frequently from the same maternal lineage. Relatedness values between females correlated negatively with geographical distances. Combination of the two molecular markers revealed four clusters of closely related, matrilineal females in the population, whereas no such cluster was apparent in males. The present study first demonstrated a sex-related spatial kin structure in a natural population of the grey-sided vole.     

Development of a pollen-mediated transformation method forNicotiana glutinosa

The development is described of a new procedure to genetically transform plant species using the male gametophyte as a natural transformation vector. Our system avoids the need for complicated regeneration procedures thus making it broadly applicable. Naked plasmid DNA encoding kanamycin resistance and GUS activity was introduced by particle gun bombardment into mature pollen grains ofNicotiana glutinosa. Bombarded pollen was used for pollinations and the resulting seeds were selected for kanamycin resistance. Two different kanamycin-resistant plants, designated VIP A and VIP B, were obtained in two independent experiments. In VIP A, TR2-driven GUS activity was observed in vascular bundles, trichomes and in a small number of pollen grains. DNA gel blot analysis indicated that the introduced DNA was integrated independently into the genome of VIP A and VIP B. It was shown that male and female gametophyte development and seed set were highly aberrant in both VIP A and VIP B and that the offspring of self- and cross-pollinations did not contain the transgenes. This might be caused by a recombination event during the integration of the naked DNA resulting in a deletion of part of the target chromosome. After meiosis such a deletion is lethal for the gametes. Our observation that the transgenes were detected in DNA isolated from sporophytic tissues but not in DNA from VIP A and VIP B pollen grains is in line with this explanation. Future experiments designed to increase the frequency of transformation and to transfer the transgenes to the offspring are discussed.     

The significance of specific distributions and functions in models of quantitative inheritance

The main purpose of this paper is to attract attention to the fact that a mathematical model of quantitative inheritance can lead to qualitatively different results, if distributions and functions which in the model represent biological processes are assumed to take analytically different (even though qualitatively similar) forms. A simple model of the effect of environmental variation on the post-selection genotypic variance in a population under phenotypic stabilizing selection is considered. It is demonstrated that this model leads to qualitatively different conclusions depending on whether the phenotypic fitness function is assumed in a Gaussian or in a quadratic form.     

On the eigenvalue distribution of genetic and phenotypic dispersion matrices: Evidence for a nonrandom organization of quantitative character variation

A quantitative genetic model of random pleiotropy is introduced as reference model for detecting the kind and degree of organization in quantitative genetic variation. In this model the genetic dispersion matrix takes the form of G = BB ^T, where B is a general, real, Gaussian random matrix. The eigenvalue density of the corresponding ensemble of random matrices (_G) is considered. The first two moments are derived for variance-covariance matrices G as well as for correlation matrices R, and an approximate expression of the density function is given. The eigenvalue distribution of all empirical correlation matrices deviates from that of a random pleiotropy model by a very large leading eigenvalue associated with a size factor. However the frequency-distribution of the remaining eigenvalues shows only minor deviations in mammalian skeletal data. A prevalence of intermediate eigenvalues in insect data may be caused by the inclusion of many functionally unrelated characters. Hence two kinds of deviations from random organization have been found: a mammal like and an insect like organization. It is concluded that functionally related characters are on the average more tightly correlated than by chance (= mammal like organization), while functionally unrelated characters appear to be less correlated than by random pleiotropy (insect like organization).     

Esterases of laboratory-reared and field-collected cotton boll weevils,Anthonomus grandis Boh.: Polymorphism of adult esterases and formal genetics of esterase II

The esterases of the cotton boll weevil were separated by polyacrylamide gel electrophoresis into four major regions. These were named Est I–IV in order of migration from anode to origin. Polymorphism was observed in all regions. The Est II region was shown to consist of no more than two bands (fast and slow). The inheritance of the fast and slow bands of Est II was demonstrated to be controlled by codominant autosomal alleles. Analysis of the gene frequency of the Est II region showed that one field population was consistent with the Hardy-Weinberg law (P=0.995), while a second field population was not at equilibrium (P<0.001).This work was supported in part by a Faculty Research Grant from Memphis State University.     

地方性克汀病遗传方式的分析

本文通过1794人、176家系,340户的调查,检出地方性克汀病患者64例,应用分离分析和多基因阈值理论,证实本症属多基因遗传,其加权平均遗传率和标准误为78.51±7.53.     

Comparison of techniques for determining the effect of aluminium on the growth of,and the inheritance of aluminium tolerance in wheat

The effect of Al on the growth of plants derived from the F3 generation of a cross between Al tolerant (Waalt) and Al sensitive (Warigal) wheat cultivars, grown in low ionic strength nutrient solutions, were assessed by a number of methods viz; root length and haematoxylin stain after 3 days exposure to Al and plant top and root yields, and root length and visual assessment for Al damage after 4 weeks growth.Of these methods haematoxylin stain (3 days) and visual assessment at 4 weeks identified the same plants as being sensitive or tolerant to Al and clearly segregated the 2 populations. Consequently these 2 methods were used as standard techniques to determine the ability of the other methods to distinguish between tolerant and sensitive plants.The ratio of plant top: root yields clearly segregated the 2 populations. The 2 populations could not be clearly distinguished based on plant top or root yields, or on root length either after 3 days or 4 weeks exposure to Al.Within the population of tolerant plants, root length was significantly correlated with root weight (r²=0.86) and top weight (r²=0.71). None of these relationships were significant for the population of sensitive plants.These techniques were applied in a number of separate experiments on the F2 and F3 populations from a Waalt × Warigal cross. The results indicate that Al tolerance in wheat is inherited by a single gene and that this gene has incomplete dominance.     

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.