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Studies on Genetic Male-Sterile Soybeans : III. The Initiation of Monocarpic Senescence.

Soybean (Glycine max [L.] Merr.) germplasm, isogenic except for loci controlling male-sterility (ms(1)) and nodulation (rj(1)) was utilized to investigate the effects of reproductive tissue development and nitrogen source on the initiation of monocarpic senescence. The experimental genotypes (Ms(1)Rj(1), Ms(1)rj(1), ms(1)Rj(1), and ms(1)rj(1), were selected from a cross between N69-2774 and N59-5259, and were inbred to the F(5) generation. Green-house-grown plants were collected during the period of flowering (77 days after transplanting) until maturity (147 days after transplanting). Leaf tissues from the respective genotypes were analyzed at the various harvest dates for RNA, phenolic, and chlorophyll concentrations; acid protease activity; polypeptide banding patterns of chloroplast thylakoids; and chloroplastic ultrastructure.Regardless of nitrogen source, total chlorophyll concentrations declined between 77 and 119 days after transplanting, resulting in a 40% loss of chlorophyll per square centimeter in all genotypes. Leaf chlorophyll levels continued to decline at a constant rate in male-fertile genotypes, but remained at a constant level (26 micrograms chlorophyll per square centimeter) in male-sterile genotypes, for the remainder of the study. With increased leaf age, a gradual disruption of thylakoid structures was observed, particularly in chloroplasts from the male-fertile genotypes. Chloroplasts from the male-sterile genotypes appeared to lose starch grains but increased their number of chloroplastic lipid bodies with leaf aging. These data suggest that monocarpic senescence in soybeans was initiated at or before flowering. Although reproductive tissue development probably augmented the process, the response attributed to seed formation was not apparent until the mid-pod fill stage (119 days after transplanting). All genotypes had similar changes in other cellular components that are recognized as indicators of plant senescence regardless of whether the plants produced seed.

Journal Article↗

Studies on Genetic Male-Sterile Soybeans : IV. Effect of Male Sterility and Source of Nitrogen Nutrition on Accumulation, Partitioning, and Transport of Nitrogen.

Soybean (Glycine max [L.] Merr.) germplasm, isogenic except for loci controlling male sterility (ms(1)) and nodulation (rj(1)), was used to investigate the effects of reproductive tissue development and source of nitrogen nutrition on accumulation, transport, and partitioning of nitrogen in a greenhouse experiment. Nodulated plants were supplied nitrogen-free nutrient solution, and nonnodulated plants were supplied nutrient solution containing 20 millimolar KNO(3). Plants were sampled from flowering until maturity (77 to 147 days after transplanting).Accumulation rates of nitrogen in whole plants during reproductive growth were not significantly different among the four plant types. Nitrogen accumulation in the sterile, nonnodulated plants, however, ceased 2 weeks earlier than in fertile, nonnodulated or fertile and sterile, nodulated plants. This early cessation in nitrogen accumulation resulted in sterile, nonnodulated plants accumulating significantly less whole plant nitrogen by 133 days after transplanting (DAT) than fertile, nonnodulated plants. Thus, changing the site of nitrogen assimilation from nodules (N(2)-fixing plants) to roots and leaves (NO(3)-fed plants) resulted in similar whole-plant nitrogen accumulation rates in fertile and sterile plants, despite the absence of seed in the latter.Leaflet and stem plus petiole tissues of both types of sterile plants had significantly higher nitrogen concentrations after 119 DAT than both types of fertile plants. Significantly higher concentrations and exudation rates of nonureide, reduced-nitrogen in xylem sap of sterile than of fertile plants after 105 DAT were observed. These latter results indicated possible cycling of nonureide, reduced-nitrogen from the downward phloem translocation stream to the upward xylem translocation stream in roots of sterile plants. Collectively, these results suggest a lack of sinks for nitrogen utilization in the shoots of sterile plants. Hence, comparison of nitrogen accumulation rates for sterile and fertile plants does not provide a definitive test of the hypothesis that reproductive tissue development limits photosynthate availability for support of N(2) fixation and nitrate assimilation in determinate soybeans.Nitrogen assimilation during reproductive growth met a larger proportion of the reproductive-tissue nitrogen requirement of nitrate-dependent plants (73%) than of N(2)-fixing plants (63%). Hence, vegetative-tissue nitrogen mobilization to reproductive tissue was a more prominent process in N(2)-fixing than in nitrate-dependent plants. N(2)-fixing plants partitioned nitrogen to reproductive tissue more efficiently than nitrate-dependent plants as the reproductive tissues of the former and latter contained 65 and 55%, respectively, of the whole-plant nitrogen at the time that nitrogen accumulation in reproductive parts had ceased (133 DAT).

Journal Article↗

Studies on genetic male-sterile soybeans : v. Effects of male-sterility on the function and glycerolipid composition of chloroplast thylakoids.

Soybean (Glycine max L. Merr.) germplasm, isogenic except for loci controlling male sterility (ms(1)), was utilized to study the effects of reproductive development on certain aspects of photosynthesis. Plants were sampled at various times between flowering (77 days after transplanting) and maturity (147 days after transplanting). During that period photosynthetic rates declined more rapidly in the male-sterile genotypes than male-fertile genotypes; and after 105 days, the sterile genotypes maintained low but relatively constant carbon exchange rates. The decline of leaf photosynthesis in the male-sterile genotype occurred concomitantly with an inhibition of the photosynthetic electron transport chain associated with photosystem II. Changes in photosystem I activities, cytochrome f levels, and chlorophyll a/b ratios per se were not responsible for the decline in whole leaf photosynthesis. These conditions were independent of the source of nitrogen nutrition. Lipid analyses of the thylakoids revealed that a loss of phosphatidylglycerol was highly correlated with the inhibition of photosystem II activity. These results suggested a relation between the decline in leaf carbon exchange and the decline in photosynthetic electron transport activity.

Journal Article↗

Anticonvulsant activity of progesterone and neurosteroids in progesterone receptor knockout mice.

Many of the biological actions of progesterone are mediated through the progesterone receptor (PR), a nuclear transcription factor. Progesterone is well recognized to protect against seizures in animal models. Although this activity has been attributed to the progesterone metabolite allopregnanolone, a GABAA receptor-modulating neurosteroid with anticonvulsant properties, PRs could also play a role. Here, we used PR knockout (PRKO(-/-)) mice bearing a targeted deletion of the PR gene that eliminates both isoforms of the PR to investigate the contribution of the PR to the anticonvulsant activity of progesterone. The protective activity of progesterone was examined in female and male homozygous PRKO mice and isogenic wild-type controls in the pentylenetetrazol (PTZ), maximal electroshock, and amygdala-kindling seizure models. In all three models, the anticonvulsant potency of progesterone was undiminished in PRKO mice compared with control mice. On the contrary, there was a substantial increase in the anticonvulsant potency of progesterone in the PTZ and kindling models. The antiseizure activity of progesterone in PRKO mice was reversed by pretreatment with finasteride, a 5alpha-reductase inhibitor that blocks the metabolism of progesterone to allopregnanolone. Unlike progesterone, the neurosteroids allopregnanolone and allotetrahydrodeoxycorticosterone exhibited comparable anticonvulsant potency in PRKO and wild-type mice. The basis for the heightened progesterone responsiveness of PRKO mice is not attributable to pharmacokinetic factors, because the plasma allopregnanolone levels achieved after progesterone administration were not greater in the PRKO mice. These studies provide strong evidence that the PR is not required for the antiseizure effects of progesterone, which mainly occurs through its conversion to the neurosteroid allopregnanolone.

Animals↗

Vacuolar degradation of rat liver CYP2B1 in Saccharomyces cerevisiae: further validation of the yeast model and structural implications for the degradation of mammalian endoplasmic reticulum P450 proteins.

Mammalian hepatic cytochromes P450 (P450s) are endoplasmic reticulum (ER)-anchored hemoproteins with highly variable half-lives. CYP3A4, the dominant human liver drug-metabolizing enzyme, and its rat liver orthologs undergo ubiquitin (Ub)-dependent 26S proteasomal degradation after suicide inactivation or after heterologous expression in Saccharomyces cerevisiae. In contrast, rat liver CYP2C11 is degraded by the vacuolar "lysosomal" pathway when similarly expressed in yeast. The structural determinants that commit P450s to proteasomal or lysosomal degradation are unknown. To further validate S. cerevisiae as a model for exploring mammalian P450 turnover, the degradation of phenobarbital-inducible liver CYP2B1, an enzyme reportedly degraded via the rat hepatic autophagic-lysosomal pathway, was examined in a yeast strain (pep4delta) deficient in vacuolar degradation and its isogenic wild-type control (PEP4). Although CYP2B1 was equivalently expressed in both strains during early logarithmic growth, its degradation was retarded in pep4delta strain, remaining at a level 5-fold higher than that in PEP4 yeast when monitored at the stationary phase. No comparable CYP2B1 stabilization was detected in yeast genetically deficient in the ER Ub-conjugating enzyme Ubc6p or Ubc7p or defective in 19S proteasomal subunit Hrd2p. Thus, as in the rat liver, CYP2B1 is a target of vacuolar/lysosomal rather than proteasomal degradation in yeast, thereby further validating this model for mammalian P450 turnover. It is intriguing that a chimeric protein, CYP2B1-3A4CT, with the CYP3A4 C-terminal heptapeptide grafted onto the CYP2B1 C terminus, was proteasomally degraded after similar expression. Such diversion of CYP2B1 from its predominantly vacuolar degradation suggests that the CYP3A4 heptapeptide could either actively signal its proteasomal degradation or block its vacuolar proteolysis.

Amino Acid Sequence↗

Expression of the inlAB operon by Listeria monocytogenes is not required for entry into hepatic cells in vivo.

Listeria monocytogenes injected intravenously into mice is taken up in the liver, where hepatocytes serve as the principal site of intracellular replication. The factors effecting entry of L. monocytogenes into hepatic cells remain to be determined. Others have shown that the protein products of the inlAB (internalin) operon are required for maximum entry of L. monocytogenes into a number of cell lines in vitro. Likewise, we report here that expression of the inlAB operon was required for maximum uptake of L. monocytogenes by primary cultures of mouse hepatocytes. Uptake of an inlAB mutant strain of L. monocytogenes was approximately 10-fold less than that of the isogenic wild-type control. In contrast, inlAB expression was not a factor in (i) clearance of L. monocytogenes injected intravenously into mice and taken up in the liver, (ii) the distribution of L. monocytogenes among hepatocytes and nonparenchymal cells in the liver, or (iii) internalization of L. monocytogenes by hepatic cells in vivo. These latter findings suggest that infection of hepatic cells by L. monocytogenes in vivo does not require the protein products of the inlAB operon.

Animals↗

Internalin B promotes the replication of Listeria monocytogenes in mouse hepatocytes.

The uptake of Listeria monocytogenes by a variety of cell types in vitro is facilitated by the protein products of the inlAB (internalin) operon expressed by the organism. In the case of mouse hepatocytes, the extent to which inlAB expression influenced the uptake of Listeria in vitro was markedly dependent upon the ratio of bacteria to cells. At a ratio of 100:1, greater than 40-fold fewer transposon-induced inl4B mutant listeriae entered hepatocytes compared to the isogenic wild-type control; the difference was only fourfold, however, in cultures inoculated at a 1:1 ratio. Similarly, the uptake of in-frame inlB or inlAB deletion mutants differed only fourfold from the uptake of wild-type or inlA mutant Listeria at a 1:1 multiplicity of infection. Mutations affecting inlB or inlAB, on the other hand, resulted in a marked decrease in the capacity of Listeria to proliferate within mouse hepatocytes in vivo and in vitro. Electron micrographs of Listeria-infected hepatocytes demonstrated the impaired capacity of inlB mutants to escape from endocytic vacuoles and to enter the cytoplasm where proliferation occurs. These findings indicate that the protein product of inlB exerts a significant effect on the intracellular replication of Listeria.

Animals↗

The Ras-specific exchange factors mouse Sos1 (mSos1) and mSos2 are regulated differently: mSos2 contains ubiquitination signals absent in mSos1.

We have compared aspects of the mouse sos1 (msos1) and msos2 genes, which encode widely expressed, closely related Ras-specific exchange factors. Although an msos1 plasmid did not induce phenotypic changes in NIH 3T3 cells, addition of a 15-codon myristoylation signal to its 5' end enabled the resulting plasmid, myr-sos1, to induce approximately one-half as many foci of transformed cells as a v-H-ras control. By contrast, an isogenic myr-sos2 plasmid, which was made by fusing the first 102 codons from myr-sos1 at homologous sequences to an intact msos2 cDNA, did not induce focal transformation directly, although it could form foci in cooperation with c-H-ras. Pulse-chase experiments indicated that the half-life of Sos1 in NIH 3T3 cells was greater than 18 h, while that of Sos2 was less than 3 h. While in vitro-translated Sos1 was stable in a rabbit reticulocyte lysate, Sos2 was degraded in the lysate, as were each of two reciprocal chimeric Sos1-Sos2 proteins, albeit at a slower rate. In the lysate, Sos2 and the two chimeric proteins could be stabilized by ATPgammaS. Unlike Sos1, Sos2 was specifically immunoprecipitated by antiubiquitin antibodies. In a myristoylated version, the chimeric gene encoding Sos2 at its C terminus made a stable protein in NIH 3T3 cells and induced focal transformation almost as efficiently as myr-msos1, while the myristoylated protein encoded by the other chimera was unstable and defective in the transformation assay. We conclude that mSos2 is much less stable than mSos1 and is degraded by a ubiquitin-dependent process. A second mSos2 degradation signal, mapped to the C terminus in the reticulocyte lysate, does not seem to function under the growth conditions of the NIH 3T3 cells.

3T3 Cells↗

Prolonged improvement of total pancreatic allograft function by previous intrathymic bone marrow cell injection in rats.

Donor-specific induction of tolerance was previously achieved in the diabetic rat by intrathymic injection of pancreatic islets. It allowed a secondary islet graft in any site without immunosuppression. Since total pancreatic graft in man is metabolically more proficient than islet graft, we attempted tolerance induction for total vascularized pancreas transplantation in diabetic BN recipient rats by an intrathymic bone marrow cell (BMC) injection from Lewis donor rats, associated to an antilymphocyte antibody (ALS) administration. Control groups consisted of isogenic grafts, allogenic grafts without tolerance induction and allogenic grafts with ALS alone. In all grafted groups, mean blood glucose and plasma insulin were normalised within 24 h. Graft rejection (clinically suggested by diabetes recurrence and later confirmed by histology) appeared at 18 +/- 2 postoperative days in the absence of intrathymic BMC injection and at 36 +/- 8 days in the group with BMC injection (p < 0.05). Intrathymic bone marrow graft was successful in delaying rejection in our study.

Animals↗

Toll-like receptors in normal and cystic fibrosis airway epithelial cells.

Toll-like receptors (TLRs) mediate cellular responses to diverse microbial ligands. The distribution and function of TLRs in airway cells were studied to identify which are available to signal the presence of inhaled pathogens and to establish if differences in TLR expression are associated with the increased proinflammatory responses seen in cystic fibrosis (CF). Isogenic, polarized CF and control bronchial epithelial cell lines, human airway cells in primary culture, and cftr null and wild-type mice were compared. TLRs 1-10, MD2, and MyD88 were expressed in CF and normal cells. Only TLR2 transcription was modestly increased in CF as compared with normal epithelial cells following bacterial stimulation. TLR2 was predominantly at the apical surface of airway cells and was mobilized to cell surface in response to bacteria. TLR4 was present in a more basolateral distribution in airway cells, but appeared to have a limited role in epithelial responses. Lipopolysaccharide failed to activate nuclear factor-kappaB in these cells, and TLR2 dominant negative but not TLR4 dominant negative mutants inhibited activation by both Gram-negative and Gram-positive bacteria. Increased availability of TLR2 at the apical surfaces of CF epithelial cells is consistent with the increased proinflammatory responses seen in CF airways and suggests a selective participation of TLRs in the airway mucosa.

Animals↗

Effects of brown midrib 3 mutation in corn silage on dry matter intake and productivity of high yielding dairy cows.

The effects of enhanced in vitro neutral detergent fiber (NDF) digestibility of corn silage on dry matter intake (DMI) and milk yield were evaluated using 32 Holstein cows in a crossover design with 28-d periods. At the beginning of the experiment, cows were 89 d in milk and yielded 45.6 kg/d of milk. Experimental diets contained either brown midrib (bm3) corn silage or isogenic normal corn silage (control) at 44.6% of DM. The NDF digestibility estimated by 30-h in vitro fermentation was higher for bm3 corn silage by 9.7 units. Contents of NDF and lignin were lower for bm3 corn silage by 1.8 and 0.8 units, respectively. Diets were formulated to contain 19% crude protein and 31% NDF and to have a forage to concentrate ratio of 56:44. Daily DMI, milk yield (3.5% fat-corrected milk), and solids-corrected milk were 2.1, 2.6, and 2.7 kg higher, respectively, for cows fed bm3 corn silage. The milk protein and lactose contents were greater for bm3 treatment, but milk fat content was not. Individual milk yield responses of the cows to bm3 treatment were positively related to pretrial milk yield, and DMI response tended to be positively related to pretrial milk yield. Enhanced in vitro NDF digestibility was associated with higher energy intake, which resulted in increased milk yield.

Animals↗

Establishment of an isogenic human colon tumor model for NQO1 gene expression: application to investigate the role of DT-diaphorase in bioreductive drug activation in vitro and in vivo.

Many tumors overexpress the NQO1 gene, which encodes DT-diaphorase (NADPH:quinone oxidoreductase; EC 1.6.99.2). This obligate two-electron reductase deactivates toxins and activates bioreductive anticancer drugs. We describe the establishment of an isogenic human tumor cell model for DT-diaphorase expression. An expression vector was used in which the human elongation factor 1alpha promoter produces a bicistronic message containing the genes for human NQO1 and puromycin resistance. This was transfected into the human colon BE tumor line, which has a disabling point mutation in NQO1. Two clones, BE2 and BE5, were selected that were shown by immunoblotting and enzyme activity to stably express high levels of DT-diaphorase. Drug response was determined using 96-h exposures compared with the BE vector control. Functional validation of the isogenic model was provided by the much greater sensitivity of the NQO1-transfected cells to the known DT-diaphorase substrates and bioreductive agents streptonigrin (113- to 132-fold) and indoloquinone EO9 (17- to 25-fold) and the inhibition of this potentiation by the DT-diaphorase inhibitor dicoumarol. A lower degree of potentiation was seen with the clinically used agent mitomycin C (6- to 7-fold) and the EO9 analogs, EO7 and EO2, that are poorer substrates for DT-diaphorase (5- to 8-fold and 2- to 3-fold potentiation, respectively), and there was no potentiation or protection with menadione and tirapazamine. Exposure time-dependent potentiation was seen with the diaziquone analogs methyl-diaziquone and RH1 [2, 5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone], the latter being an agent in preclinical development. In contrast to the in vitro potentiation, there was no difference in the response to mitomycin C when BE2 and BE vector control were treated as tumor xenografts in vivo. This isogenic model should be valuable for mechanistic studies and bioreductive drug development.

Animals↗

Isogene specific oligo arrays reveal multifaceted changes in gene expression during grape berry (Vitis vinifera L.) development.

The transition from a green, hard, and acidic pericarp to a sweet, soft, coloured, and sugar-rich ripe fruit occurs in many unrelated fruit species. High throughput identification of differentially expressed genes in grape berry has been achieved by the use of 50-mers oligoarrays bearing a set of 3,200 Unigenes from Vitis vinifera to compare berry transcriptome at nine developmental stages. Analysis of transcript profiles revealed that most activations were triggered simultaneously with softening, occurring within only 24 h for an individual berry, just before any change in colouration or water, sugar, and acid content can be detected. Although most dramatically induced genes belong to unknown functional categories, numerous changes occur in the expression of isogenes involved in primary and secondary metabolism during ripening. Focusing on isogenes potentially significant in development regulation (hormonal control of transcription factor) revealed a possible role for several hormones (cytokinin, gibberellin, or jasmonic acid). Transcription factor analysis revealed the induction of RAP2 and WRKY genes at véraison, suggesting increasing biotic and abiotic stress conditions during ripening. This observation was strengthened by an increased expression of multiple transcripts involved in sugar metabolism and also described as induced in other plant organs during stress conditions. This approach permitted the identification of new isogenes as possible control points: a glutathione S-transferase exhibits the same expression profile as anthocyanin accumulation and a new putative sugar transporter is induced in parallel with sugar import.

Anthocyanins↗

The case for isogenic strains in toxicological screening.

The fundamental principle of the controlled experiment is that treated and control groups should be identical, with minimal within-group variability. Toxicologists recognise this and control age, body weight, disease and the physical environment of the test animals. However most toxicological screening in done with genetically variable outbred stocks, even though isogenic inbred strains, F1 hybrids or identical siblings are usually available. The result is poor experiments with the inevitable genetic differences between groups resulting in increased false positive and negative results, and no indication that the response is under genetic control. It is also illogical to treat genetic variation differently from other types of variation. The argument that it is essential to use outbred animals to model outbred man is illogical. If bacteria can be used to model man (as in the Ames test), so can inbred animals. The uncontrolled variation present in an outbred stock can not be used efficiently to increase the range of phenotypes tested because it also introduces "noise" which obscures experimental effects. The use of two or more isogenic strains gives a much more efficient experimental design with low "noise" and an indication of whether the response is under genetic control. Inbred and F1 hybrid strains (but not identical siblings) have the added advantage of an immortal genotype which outlives any individual animal. Such immortal genotypes may be studied in detail to gather background information. Toxicologists should treat genetics like every other variable and control it, using several isogenic strains in cases where testing needs to be done on more than one genotype.

Animals↗

Acquired and R-gene-mediated resistance against the potato aphid in tomato.

We examined the effects of three forms of host plant resistance in tomato, Lycopersicon esculentum, on the potato aphid, Macrosiphum euphorbiae. Mi-1.2, a resistance gene (R-gene) in tomato that deters aphid feeding, reduced the population growth of both potato aphid isolates tested, although it appeared to have a greater impact on isolate WU11 than on isolate WU12. The results suggest that there may be quantitative differences in virulence between these two aphid isolates. We also examined two distinct forms of acquired resistance in tomato, jasmonic acid (JA)-dependent and salicylic acid (SA)-dependent induced defenses. Exogenous foliar application of JA triggered expression of a JA-inducible proteinase inhibitor in tomato cultivars with and without Mi-1.2, although the effects of treatment on aphid performance differed between these cultivars. JA-treatment reduced aphid population growth on a susceptible tomato cultivar that lacks Mi-1.2, but did not significantly enhance or inhibit aphid control on a near-isogenic resistant tomato cultivar that carries this gene. Foliar application of an SA analog, benzothiadiazole (BTH), was used to induce SA-dependent defenses. BTH treatment reduced the population growth of both aphid isolates on a susceptible tomato cultivar, and also enhanced aphid control on a resistant cultivar. The results indicate that both SA- and JA-dependent acquired resistance in tomato have a direct negative effect on a phloem-feeding insect. Furthermore, this study demonstrates that acquired resistance and R-gene-mediated resistance can interact for enhanced suppression of insect herbivores.

Animals↗

Control of mouse U1 snRNA gene expression during in vitro differentiation of mouse embryonic stem cells.

Early in mouse development, two classes of U1 RNAs, mU1a and mU1b, are synthesized, but as development proceeds, transcription of the embryo-specific mU1b genes is selectively down-regulated to a barely detectable level. We show here that during in vitro differentiation of mouse embryonic stem (ES) cells, both exogenously introduced and endogenous U1b genes are subject to normal developmental regulation. Thus, ES cells represent a convenient isogenic system for studying the control of expression of developmentally regulated snRNA genes. Using this system, we have identified a region in the proximal 5'flanking region, located outside the PSE element, that is responsible for differential transcription of the mU1a and mU1b genes in both developing cells and transiently transfected NIH 3T3 cells.

3T3 Cells↗

Two-component systems of Helicobacter pylori contribute to virulence in a mouse infection model.

Helicobacter pylori encodes three histidine kinases and five response regulators belonging to the family of two-component regulatory systems which are involved in transcriptional control. Here we demonstrate that isogenic mutants of H. pylori P76 with deletions of the response regulator open reading frame (ORF) HP1365 and ORFs HP244, HP165, and HP1364 encoding histidine kinases are unable to colonize the stomachs of BALB/c mice, suggesting an essential role of these systems in the regulation of important virulence properties of H. pylori. Furthermore, we demonstrate that the genes under the control of the P(HP1408) and P(HP119) promoters which are regulated by the two-component system HP166-HP165 are not essential for single mutant colonization of mice but are required under competitive colonization conditions.

Animals↗

[Comparative contribution of different genetic factors in induction of MGE transpositions under isogenization].

Localization patterns of mobile genetic element 412 in polytene chromosomes of larvae from the control (riC) line, the balancer line, the F1 and F2 generations of the isogenization scheme, and 10 final isogenic lines were obtained and compared. The contributions of the recombination transfer of mobile genetic element copies from the balancer line, the outbreeding of control and balancer lines, and the inbreeding of isogenized lines to the rate of transposition were determined and estimated. These constituted < 0.187, < 0.30, and > 0.207 events per initial mobile genetic element copy per isogenized haploid genome per isogenization, respectively. During consecutive steps of isogenization (F1-F2-isogenic lines), the total transposition rate decreased: 2.09, 1.78, and 0.69. This was explained in terms of the existence of large selective and random losses in the variability of mobile genetic elements within the sites of their patterns during isogenization. The existence of a recombination transfer does not change the main conclusions and estimates regarding isogenization-induced transpositions.

Animals↗