Study on Heat Sensitivity of Esterase Isozymes in Maise Inbreds

Crude extracts of 44 maize inbred seeds were treated under different temperatures, i. e. 50, 55, 60, 65 and 70℃, in order to determine their heat sensitivity of esterase isozymes. The results showed large discrepancy of heat sensitivity of anodic esterase isozymes between different or same relative mobility bands of isozyme, but similar inactivating temperature (65℃) of the cathodic ones for different inbreds. The heat sensitivity of isozymes for inbreds may be adopted in the identification of variation and purity of inbred and hybrid seeds of maize.     

Genetic and non-genetic factors affecting anther culture of Brussels sprouts (Brassica oleracea var. gemmifera)

Summary Eight inbred lines of Brussels sprouts and ten F₁ hybrids derived from them were tested for their response to anther culture. From 5–19 plants per genotype were tested, and each plant was tested on 3–6 separate occasions. Results from the inbred lines were broadly similar to those from the F₁ hybrids, despite the inbreds producing fewer buds and having a higher frequency of anther deformities. The maximum embryo yield from an inbred line was 215 embryos per 100 anthers, and from a hybrid was 275. From estimation of the variance components it was calculated that, for both inbreds and hybrids, about half the total variation was genetic whereas variation due to plants within genotypes and to occasions within plants were each about 13% of the total. The narrow sense heritability of responsiveness to anther culture (estimated by the proportion of variation between inbred lines which was genetic) was 0.48, and there was partial dominance for this character. In three cases the hybrid outyielded the better inbred, and this heterosis may well be due to dispersed dominant genes.     

RFLP analyses of early-maturing European maize germ plasm

Summary Thirty inbred lines representing a wide range of early-maturing European elite germ plasm of maize (Zea mays L.) were assayed for RFLPs using 203 clone-enzyme combinations (106 DNA clones with restriction enzymes EcoR1 and HindIII). The genetic materials comprised 14 flint, 12 dent, and 4 lines of miscellaneous origin. Objectives were to (1) characterize the genetic diversity for RFLPs in these materials, (2) compare the level of genetic diversity found within and between the flint and the dent heterotic groups, and (3) examine the usefulness of RFLPs for assigning inbreds to heterotic groups. All but two DNA clones yielded polymorphism with at least one restriction enzyme. A total of 82 and 121 clone-enzyme combinations gave single-banded and multiple-banded RFLP patterns, respectively, with an average of 3.9 and 7.7 RFLP patterns per clone-enzyme combination across all 30 inbreds, respectively. Genetic similarity (GS) between lines, estimated from RFLP data as Dice's similarity coefficient, showed considerable variation (0.32 to 0.58) among unrelated inbreds. The mean GS for line combinations of type flint x dent (0.41) was significantly smaller than for unrelated flint lines (0.46) and dent lines (0.46), but there was considerable variation in GS estimates of individual line combinations within each group. Cluster and principal coordinate analyses based on GS values resulted in separate groupings of flint and dent lines in accordance with phylogenetic information. Positioning of lines of miscellaneous origin was generally consistent with expectations based on known breeding behavior and pedigrees. Results from this study corroborated that RFLP data can be used for assigning inbreds to heterotic groups and revealing pedigree relationships among inbreds.     

Relationship of restriction fragment length polymorphisms to single-cross hybrid performance of maize

Summary Isozymes and restriction fragment length polymorphisms (RFLPs) have been proposed for use in varietal identification and selection for agronomic traits. Although the use of isozymes for these purposes has been well documented, evaluation of the efficacy of RFLP technology as applied to crop improvement is far from complete. This investigation was conducted to study the relationship between RFLP-derived genotypes and heterotic patterns of a group of maize (Zea mays L.) inbred lines. A total of 22 inbreds was crossed to four testers (B73, B76, Mo17, and Va26) in combinations that minimized crossing within heterotic groups. Forty-seven single-cross progeny were subsequently evaluated for several agronomic traits (including grain yield and moisture, ear height, and root lodging) over 2–4 consecutive years at two to four Iowa locations in a randomized complete-block design. The inbred lines were subjected to RFLP analysis, which involved 47 genomic clones and the restriction enzymes EcoRI and HindIII. Hybrid RFLP patterns were predicted from their inbred parents. Modified Roger's distances were computed to estimate genetic distance among the inbred lines. Principal component analysis facilitated ascertainment of relative dispersion of the inbreds based on the frequency of variants at specific RFLP loci. Evident associations of variants with genes affecting agronomic traits were identified by principal component regression analysis, in which adjusted hybrid means were regressed on the matrix of hybrid variants frequencies. The hybrid means were adjusted by removing environmental effects, using residuals as dependent variables in the regression analysis. Results from this study suggest that RFLP analysis may be of value in allocating maize inbreds to heterotic groups, but no relationship between RFLP-based genetic distance and hybrid performance was apparent. Principal component regression identified variants potentially linked to genes that control specific agronomic traits.Joint contribution: USDA-ARS and Journal Paper No. J-13590 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011, USA. Projects No. 2818 and 2778     

Variation in embryo size of inbred lines of carrot

Mean embryo lengths and coefficients of variation (CVs) of embryo length of inbred carrot lines differed between years but not between the two sets of inbreds tested nor between male-sterile and maintainer lines. Estimates of CVs were of similar magnitude to those previously obtained for experimental and commercial open-pollinated seed lots, but estimates of mean embryo length were lower. Thus greater genetic uniformity did not lead to lower CVs of embryo size of the inbreds.     

Effects of selection for resistance to Sesamia nonagrioides on maize yield,performance and stability under infestation with Sesamia nonagrioides and Ostrinia nubilalis in Spain

A maize synthetic population was improved for resistance to the Mediterranean corn borer (MCB, Sesamia nonagrioides) while maintaining yield. The objectives of this research were to investigate whether yield and yield stability of the maize synthetic population named EPS12 were affected by selection for MCB resistance; also to determine which genotypic and environmental covariates could explain the genotype (G), environment (E) and genotype × environment (GE) effects for yield under corn borer infestation. Plants from three cycles of selection and their testcrosses to three inbred testers (A639, B93 and EP42) were evaluated at two locations in 2 years, under MCB and European corn borer infestations. After selection EPS12 was a more stable genotype. Hybrids derived from crosses between B93 and inbreds obtained from the initial cycles of selection could be recommended for cultivation in northern Spain. The yield of crosses between cycles of selection and testers increased when there were fewer days with temperatures >25°C and higher mean maximum temperatures. Differences in yield among these genotypes were mostly explained by resistance to corn borer attack. In general, among EPS12-derived materials, genetic characteristics that contribute to increased grain yield were also responsible for increased abiotic stress tolerance.     

Mating speed and the interplay between female and male courtship responses inDrosophila melanogaster (Diptera: Drosophilidae)

The objective of this study was to compare measures of general activity and sexual behavior for various genotypes within a strain of Drosophila melanogaster, which had known differences in mating speed. Three inbred lines of D. melanogaster differed significantly in mating speed when tested in female-choice and in single-pair experiments. Analyses of locomotor activity and sexual activity of females and males revealed no significant differences between the inbred lines. An analysis of the interplay between female and male courtship behaviors enabled the examination of signal-response differences between the inbred lines. The inbred lines with intermediate and slow mean mating speed showed a decreased number of significant transitions between female and male behavioral responses. This decrease was more severe in the slow mating line. Further, the intermediate- and slow-mating females and males displayed courtship responses toward signals of the opposite sex that were different from those of the fastmating line. Models of the relationship between behavioral activity and mating speed in Drosophila are discussed and a different explanation for variation in mating speed among the three inbred lines is considered.     

Cytology of inbreds and f 1 hybrids of pearl millet

Summary The meiotic behaviour of chromosomes was studied in four inbred lines and their F ₁ hybrids of P. typhoides. The inbred lines showed a decrease in mean chiasma frequency when compared with the population plants, but differed from one another in their mean chiasma frequencies. Together with the decrease in mean chiasma frequency the inbreds showed variation in mean chiasma frequencies. The inbred lines showed a number of meiotic abnormalities such as extra chromosomes, extra fragments, desynapsis, persistent nucleoli and differential condensation of chromosomes. The F ₁ hybrids of these inbreds exhibited heterosis for chiasma frequency. All the F ₁'s had mean chiasma frequencies higher than the means of the respective participating parents. The F ₁'s, however, differed in the degree of heterosis exhibited. In the F ₁ hybrids, the variation in mean chiasma frequency, both between plants and between PMC's within plants, was significantly lower than that of the inbred lines. The effect of environment was studied in the inbred lines and their F ₁ hybrids. The mean chiasma frequencies of the inbred lines were significantly lower, and the variation in mean chiasma frequencies was greater, in the stress season. The mean chiasma frequencies of F ₁'s did not show any significant differences between the two seasons. The F ₁'s exhibited less variation in mean chiasma frequency than the inbred lines, showing that F ₁'s were developmentally more stable. The F ₁'s did not show any meiotic abnormalities in either season.     

A test of quantitative genetic theory using Drosophila- effects of inbreeding and rate of inbreeding on heritabilities and variance components

Inbreeding is expected to decrease the heritability within populations. However, results from empirical studies are inconclusive. In this study, we investigated the effects of three breeding treatments (fast and slow rate of inbreeding - inbred to the same absolute level - and a control) on heritability, phenotypic, genetic and environmental variances of sternopleural bristle number in Drosophila melanogaster. Heritability, and phenotypic, genetic and environmental variances were estimated in 10 replicate lines within each of the three treatments. Standard least squares regression models and Bayesian methods were used to analyse the data. Heritability and additive genetic variance within lines were higher in the control compared with both inbreeding treatments. Heritabilities and additive genetic variances within lines were higher in slow compared with fast inbred lines, indicating that slow inbred lines retain more evolutionary potential despite the same expected absolute level of inbreeding. The between line variance was larger with inbreeding and more than twice as large in the fast than in the slow inbred lines. The different pattern of redistribution of genetic variance within and between lines in the two inbred treatments cannot be explained invoking the standard model based on selective neutrality and additive gene action. Environmental variances were higher with inbreeding, and more so with fast inbreeding, indicating that inbreeding and the rate of inbreeding affect environmental sensitivity. The phenotypic variance decreased with inbreeding, but was not affected by the rate of inbreeding. No inbreeding depression for mean sternopleural bristle number was observed in this study. Considerable variance between lines in additive genetic variance within lines was observed, illustrating between line variation in evolutionary potential.     

Variation of DNA fingerprints among accessions within maize inbred lines and implications for identification of essentially derived varieties.

Genetic distances (GDs) based on molecular markers are important parameters for identifying essentially derived varieties (EDVs). In this context information about the variability of molecular markers within maize inbred lines is essential. Our objectives were to (1) determine the variation in the size of simple sequence repeat (SSR) fragments among different accessions of maize inbreds and doubled haploid (DH) lines, (2) attribute the observed variation to genetic and marker system-specific sources, and (3) investigate the effect of SSR fragment size differences within maize lines on the GD between maize lines and their consequences for the identification of essentially derived varieties. Two to five accessions from nine inbred lines and five DH lines were taken from different sources or drawn as independent samples from the same seed lot. Each accession was genotyped with 100 SSR markers that evenly covered the whole maize genome. In total, 437 SSR fragments were identified, with a mean of 4.4 alleles per locus. The average polymorphic information content (PIC) was 0.58. GD estimates between two accessions of the same genotype ranged from 0.00 to 0.12 with an average of 0.029 for inbred lines and 0.001 for DH lines. An average of 11.1 SSRs was polymorphic between accessions of the same inbred line due to non-amplification (8.1 SSRs), heterogeneity (4.0 SSRs) or unknown alleles (2.6 SSRs). In contrast to lab errors, heterogeneity contributed considerably to the observed variation for GD. In order to decrease the probability to be suited for infringing an EDV threshold by chance, we recommend to increase the level of homogeneity of inbred lines before applying for plant variety protection.     

The influence of morphological differences between sunflower inbred lines on their SCA effects for yield components

Genetic distances between seven oil sunflower inbred lines expressed in the variability of five morphological traits were analysed in terms of their effect on the specific combining ability (SCA) effects in these lines for yield component traits. The multivariate analysis of variance (MANOVA) and other related multidimensional methods were used to recognize the differences between genotypes with respect to morphological traits. Canonical analysis was applied for a graphical configuration of parental lines and diallele hybrids in the two-dimensional space. As a measure of the difference between parental lines with respect to the analysed morphological traits jointly, i.e. as a measure of the morphological distance between these forms, Mahalanobis' distance was used. Phenotypic differences between lines were estimated using Mahalanobis' distance for each pair of crossed lines calculated for all the analysed morphological traits. The effect of morphological differences between inbred lines on the mean SCA effect for individual yield components, measured by the linear regression, was significant for oil yield per plant and for the number of seeds per plant.     

Genetic diversity for restriction fragment length polymorphisms and heterosis for two diallel sets of maize inbreds

Summary Changes that may have occurred over the past 50 years of hybrid breeding in maize (Zea maize L.) with respect to heterosis for yield and heterozygosity at the molecular level are of interest to both maize breeders and quantitative geneticists. The objectives of this study were twofold: The first, to compare two diallels produced from six older maize inbreds released in the 1950's and earlier and six newer inbreds released during the 1970's with respect to (a) genetic variation for restriction fragment length polymorphisms (RFLPs) and (b) the size of heterosis and epistatic effects, and the second, to evaluate the usefulness of RFLP-based genetic distance measures in predicting heterosis and performance of single-cross hybrids. Five generations (parents, F₁; F₂, and backcrosses) from the 15 crosses in each diallel were evaluated for grain yield and yield components in four Iowa environments. Genetic effects were estimated from generation means by ordinary diallel analyses and by the Eberhart-Gardner model. Newer lines showed significantly greater yield for inbred generations than did older lines but smaller heterosis estimates. In most cases, estimates of additive x additive epistatic effects for yield and yield components were significantly positive for both groups of lines. RFLP analyses of inbred lines included two restriction enzymes and 82 genomic DNA clones distributed over the maize genome. Eighty-one clones revealed polymorphisms with at least one enzyme. In each set, about three different RFLP variants were typically found per RFLP locus. Genetic distances between inbred lines were estimated from RFLP data as Rogers' distance (RD), which was subdivided into general (GRD) and specific (SRD) Rogers' distances within each diallel. The mean and range of RDs were similar for the older and newer lines, suggesting that the level of heterozygosity at the molecular level had not changed. GRD explained about 50% of the variation among RD values in both sets. Cluster analyses, based on modified Rogers' distances, revealed associations among lines that were generally consistent with expectations based on known pedigree and on previous research. Correlations of RD and SRD with f₁ performance, specific combining ability, and heterosis for yield and yield components, were generally positive, but too small to be of predictive value. In agreement with previous studies, our results suggest that RFLPs can be used to investigate relationships among maize inbreds, but that they are of limited usefulness for predicting the heterotic performance of single crosses between unrelated lines.Joint contribution from Cereal and Soybean Research Unit, USDA, Agricultural Research Service and Journal Paper no. J-13929 of the Iowa Agric and Home Economics Exp Stn, Ames, IA 50011. Projects no. 2818 and 2778A.E.M. is presently at the Iowa State University on leave from University of Hohenheim, D-7000 Stuttgart 70, Federal Republic of Germany     

Relationship between maize stem structural characteristics and resistance to pink stem borer (Lepidoptera: Noctuidae) attack

The pink stem borer, Sesamia nonagrioides (Lefebvre), is one of the most important insect pests of maize (Zea mays L.) in northwestern Spain. The objectives of this work were to evaluate, at different times during the growth of maize, structural traits related to the entry point and tissues on which larvae feed and to determine the relationship between these structural traits and the stem borer resistance. Six inbred lines with different levels of stem resistance to S. nonagrioides were evaluated in several trials. Potential structural resistance factors included rind and pith puncture resistance (RPR and PPR), rind thickness, length of the meristematic area (LMA), and pith parenchyma interlumen thickness (PPIT). Surprisingly, the inbred lines that showed the strongest stalks, EP42 and EP47, were not stem resistant to pink stem borer attack, while the stem resistant inbreds A509, CM151, and PB130 were among the least resistant to rind puncture. There were no significant differences among resistant and susceptible inbreds for the rind thickness. However, the susceptible inbred EP42 had the softest internode pith, and the resistant inbred PB130 showed the hardest, as was expected. Susceptible inbred lines in general showed higher values for the LMA, while the PPIT was important for individual inbreds. The results suggest that the usefulness of these characters as estimators of pink stem borer resistance is limited to some genotypes. Besides, even among those genotypes, other mechanisms of resistance that do not involve stalk strength could be present. Among the traits considered, the LMA was the most promising as an indicator of resistance to pink stem borer, although further experimentation is necessary.     

Correction for non-linear relationships between root size and short term Pi uptake in genotype comparisons

Variation in phosphate (Pi) absorption rates, root length and root mass was investigated in 25 genotypes or inbred lines of Arabidopsis thaliana (L.) Heynh. All variables differed among lines (P<0.0001), and larger plants with larger roots and shoots had faster Pi uptake rates (influx) (P<0.0001). Within genotypes, influx was non-linearly related to root size. Increases in influx were not directly proportional to increases in root size; with a doubling in root length or biomass, there was only a 1.30 or 1.27 concomitant increase in influx, respectively. Because inbred lines are genetically homogeneous, this within-genotype pattern was random, and not genetically based.Influx has traditionally been standardized for variation in plant size by dividing by root length or biomass. This assumes a linear relationship between influx and root size. As uptake rate was non-linearly dependent on root size in A. thaliana, this resulted in a spurious negative correlation between influx per cm or mg root and root length or biomass. We present a general method that standardizes influx for variation in root size within genetically homogenous genotypes. This method determines and corrects for random non-linear relationships between influx and root size. In A. thaliana, influx per unit root length or biomass differed among lines (P<0.0001), but these differences did not correlate with root length or biomass. These data contradict the common interpretation in previous literature that species or cultivars with small roots have evolved greater influx per unit size of root.     

Factors affecting field-scale production of seed of F1 hybrid Brussels sprouts

The two main factors involved in field-scale production of seed of F₁ hybrid Brussels sprouts are pollen availability and honeybee behaviour. Pollen availability depends upon the extent of winter survival of the parent inbreds, their flowering times, plant size and on the number of flowers per plant. Plant losses varied between inbreds and between sites and seasons. Differences in the commencement of flowering time in pairs of inbreds varied from 7 to 21 days, and plant size affected flower number. In hybrid seed-production there was a direct relationship between the number of mature flowers on each inbred and the percentage of non-hybrid seed produced from that inbred. Bees were highly selective in their visits to inbreds, and a mean selfing-to crossing-movement ratio of 30:1 was observed. This behaviour, together with pollen availability, greatly influenced the production of ‘sibs’. Radioactive experiments showed that the amount of cross-pollen carried by a bee decreased by about 30% at each flower visited but radioactive pollen grains were detected on the tenth flower visited. Of a number of factors investigated as possibly influencing bee behaviour, differences in flower colour and plant height were associated with discrimination between inbred lines.     

A Temperature Effect on the Expression of Genotypic Differences in Flowering Induction in Antirrhinum majus

Three inbred lines of Antirrhinum majus and the F₁ generationsof crosses between them were scored for flowering time in ninegreenhouse and garden experiments. The inbreds fell into twogroups, the difference between which showed a marked interactionwith environments. The two lines (An 105, An 106) which weresimilar were earlier flowering than the third line (An 101)in those environments which produced generally early flowering,but were later than An 101 when the environmental conditionsled to late flowering. In all cases the F₁ generations wereat least as early as the earlier parent. Cabinet experiments identified temperature as the importantenvironmental factor when day length was kept constant at 16h. At 25°C, An 105 and An 106 were earlier than An 101 whileat 12°C, at which temperature the average flowering timewas much later, An 101 was earlier. The F₁ generations werelike An 105 and An 106 at 25°C but showed heterosis at 12°C. The differences in flowering time between genotypes were inall cases established by the time the first reproductive budsbecame visible (budding time). Thus a temperature differencewell outside the range of vernalizing temperatures is havinga striking effect on the phenotypic expression of genotypicdifferences determining the transition from vegetative to floweringdevelopment.     

Phenotypic plasticity of body size in a temperate population ofDrosophila melanogaster: When the temperature—size rule does not apply

A natural population ofDrosophila melanogaster in southern France was sampled in three different years and 10 isofemale lines were investigated from each sample. Two size-related traits, wing and thorax length, were measured and the wing/thorax ratio was also calculated. Phenotypic plasticity was analysed after development at seven different constant temperatures, ranging from 12‡C to 31‡C. The three year samples exhibited similar reaction norms, suggesting a stable genetic architecture in the natural population. The whole sample (30 lines) was used to determine precisely the shape of each reaction norm, using a derivative analysis. The practical conclusion was that polynomial adjustments could be used in all cases, but with different degrees: linear for the wing/thorax ratio, quadratic for thorax length, and cubic for wing length. Both wing and thorax length exhibited concave reaction norms, with a maximum within the viable thermal range. The temperatures of the maxima were, however, quite different, around 15‡C for the wing and 19.5‡C for the thorax. Assuming that thorax length is a better estimate of body size, it is not possible to state that increasing the temperature results in monotonically decreasing size (the temperature-size rule), although this is often seen to be the case for genetic variations in latitudinal clines. The variability of the traits was investigated at two levels—within and between lines—and expressed as a coefficient of variation. The within-line (environmental) variability revealed a regular, quadratic convex reaction norm for the three traits, with a minimum around 21‡C. This temperature of minimum variability may be considered as a physiological optimum, while extreme temperatures are stressful. The between-line (genetic) variability could also be adjusted to quadratic polynomials, but the curvature parameters were not significant. Our results show that the mean values of the traits and their variance are both plastic, but react in different ways along a temperature gradient. Extreme low or high temperatures decrease the size but increase the variability. These effects may be considered as a functional response to environmental stress.     

Recovery of maize (Zea mays L.) inbreds and hybrids from chilling stress of various duration: photosynthesis and antioxidant enzymes

The differences between two maize (Zea mays L.) inbred lines and their F1 hybrids in their response to chilling periods of various duration (1, 2, 3 or 4 weeks) and subsequent return to optimum temperatures were analysed by the measurement of the photosystem (PS) 1 and 2 activity, the photosynthetic pigments' content and the activity of antioxidant enzymes. The PS2 activity and the chlorophyll content decreased in plants subjected to 3 or 4 weeks of chilling, but not in those subjected to 1 or 2 weeks of chilling. This decrease was more pronounced in inbreds compared to their hybrids. The activity of superoxide dismutase did not much change with the increasing length of chilling period in the inbreds but decreased in the hybrids, the glutathione reductase activity increased in both types of genotypes but more in the inbred lines, while for ascorbate peroxidase and catalase the changes in parents-hybrids relationship did not show any specific trend. The PS1 activity and the carotenoids' content was not much affected.     

Genetical variation for enzyme activity in a population of Drosophila melanogaster. VII. Genotype-environment interaction for alcohol dehydrogenase (ADH) activity

Genotype-environment interaction was detected for ADH activity amongst a set of 18 highly inbred lines of Drosophila melanogaster which had been extracted from the laboratory population, "Texas". The genotype-environment interaction for ADH activity was not wholly associated with genotype-environment interaction for body weight or total protein level. Detailed analyses of the responses of the individual inbred lines in ADH activity in relation to the environmental index, ej and following the procedure of Jinks and Pooni (1979), showed substantial diversity in the form of response. Lines homozygous for the AdhF allele were more environmentally sensitive than AdhS/S lines. Amongst the 16 AdhS/S lines, models of linear, quadratic or two intersecting-straight-lines were used to illustrate the varied responses of genotypes to the environment. The heterogeneity in the response characteristics of the inbred lines was attributed to variations in the conditions of culture media normally present within populations and laboratories. Moreover the non-linear responses shown by some lines to the environment are consistent with a model of genotype-environment interaction for ADH activity mediated by varied genotype-specific sensitivities to different environmental factors.     

RFLP studies of genetic relationships among inbred lines of the cultivated sunflower,Helianthus annuus L.: evidence for distinct restorer and maintainer germplasm pools

One-hundred-and-eighty-one nuclear DNA probes were used to examine restriction-fragment length polymorphism in inbred lines of the cultivated sunflower (Helianthus annuus L.). The probes were from six libraries: two genomic libraries — one made with PstI and the other with HindIII, and four cDNA libraries — from etiolated plantlets, green leaves, ovaries, petals and anthers. Total DNA from 17 inbred lines representing an overview of the genetic stocks of sunflower, including restorer and maintainer lines of the classical cytoplasmic male sterility, was digested with four different restriction enzymes and probed in 331 probe-enzyme combinations. Of 181 clones analysed, 73 probes were found to be polymorphic. Genetic distances between inbreds were calculated from the resultant proportion of shared bands and submitted to principal component analysis and the UPGMA tree-making method. The RFLP analysis allowed a clear differentiation between restorer and maintainer lines of the cytoplasmic male sterility, together with a grouping of some of the genotypes from the same origin. The analysis of the accuracy of distance estimation as a function of the number of probe-enzyme combinations used, indicates that 40–50 combinations ensure a confidence level of near 95%. Considering the inbreds as representatives of the range of cultivated inbreds, estimates of gene diversity, as well as estimates of average gene diversity between and within the sets of restorer and maintainer lines, were calculated. Estimation of gene diversity showed that the available genetic variability in cultivated sunflower, based on allelic frequencies, is lower than that of other plants (H=0.20). Moreover, we show that the proportion of genetic variability due to the difference between maintainer and restorer lines (D_m) is about 2%.