Homozygous-viable pericentric inversions for genetic control of Lucilia Cuprina

Summary The isolation of homozygous-viable pericentric inversions for inclusion in field-female killing (FK) systems in Lucilia cuprina is described. From 7,236 irradiated chromosomes screened, 16 pericentric inversions were isolated. Four of these were viable as homozygotes. One of these, In (3LR) 14, possesses the properties required for inclusion in FK systems (tight linkage of one inversion break-point to the white-eye gene and substantial genetic exchange within the inversion in heterozygous females).     

Computer simulation of genetic control. Comparison of sterile males and field-female killing systems

Summary A computer program, GENCON, designed to simulate genetic control using field-female killing systems, is described. These systems incorporate sex-linked translocations and conditional lethal mutations. Genetic death in field populations is caused by semisterility of the translocation and by homozygosis of the mutations in females and non-translocation males of field origin. Simulations using the program compare the effectiveness, in populations regulated by density, of genetic control using this type of system with control using sterile-male release. At high release rates, sterile males cause more rapid suppression and earlier eradication than sex-linked translocation strains. However, if releases are interrupted before eradication, the rate of recovery of density-dependent populations is more rapid following sterile-male release than following suppression with translocation strains. In such populations, the cumulative population suppression (number of individuals killed) is greater with translocation-strain release than with sterile-male release. At low release rates, sex-linked translocation strains can be much more effective at suppressing and eradicating density-dependent populations than sterile males. In continental Australia, eradication of the sheep blowfly Lucilia cuprina is probably not practicable. A suppression campaign using sex-linked translocation strains could yield a higher benefit to cost ratio than one using sterile males.     

Chromosomal inversions and genetic control revisited: the use of inversions in sexing systems for higher Diptera

Summary Genetic sexing systems based on sex-linked translocations and deleterious mutations are subject to breakdown from genetic recombination in males. Including inversions in these strains may provide a solution to this problem, by ensuring selective elimination of recombinant products. Inversions could be used either in coupling to or in repulsion to the translocation. The latter system, requiring homozygous-viable inversions, would be more difficult to construct, but would offer several advantages not available with coupled translocation/inversion systems. A system proposed for the blowfly Lucilia cuprina is outlined, which combines homozygous-viable pericentric inversions in repulsion to existing sex-linked translocations. This system should both stabilize the genetic sexing system and increase the suppressive potential of such strains.     

Genetic control of Lucilia cuprina: analysis of field trial data using simulation techniques

Summary An analytical version of the genetic control simulation program GENCON has been used to further analyze the data obtained during field trials of genetic control of the sheep blowfly, Lucilia cuprina, in 1976–79. In the simulations, population trends from a nonrelease area were used as an estimate of the rates of increase that would have occurred in the target population if there had been no releases. Genetic data from the target area (frequencies of matings by released males) were used to predict the frequencies of descendants of released males, the resulting genetic death, and the effects of this on population trends. In simulations that assumed no migration and full survival and competitiveness of all field-reared descendants of released males (translocation-bearing males and males and females heterozygous for deleterious mutations), neither the predicted genetic changes nor the predicted population trends agreed well with the observed data. Further simulations suggested that reduced survival or competitiveness of field-reared descendants did not account for this disagreement, but that immigration of wild flies into the test areas was probably a major contributor to the failure to achieve suppression. However, immigration alone was not sufficient to explain all the differences between observed and expected results. Other plausible contributors to this failure were: (1) lower survival of translocation males due to the effects of a dieldrin resistance allele carried on the translocation, and (2) increased survival of immature stages of L. cuprina at low population densities.     

Distribution of genetic diversity in relation to chromosomal inversions in the malaria mosquito Anopheles gambiae

The epidemiology of malaria in Africa is complicated by the fact that its principal vector, the mosquito Anopheles gambiae, constitutes a complex of six sibling species. Each species is characterized by a unique array of paracentric inversions, as deduced by karyotypic analysis. In addition, most of the species carry a number of polymorphic inversions. In order to develop an understanding of the evolutionary histories of different parts of the genome, we compared the genetic variation of areas inside and outside inversions in two distinct inversion karyotypes of A. gambiae. Thirty-five cDNA clones were mapped on the five arms of the A. gambiae chromosomes with divisional probes. Sixteen of these clones, localized both inside and outside inversions of chromosome 2, were used as probes in order to determine the nucleotide diversity of different parts of the genome in the two inversion karyotypes. We observed that the sequence diversity inside the inversion is more than threefold lower than in areas outside the inversion and that the degree of divergence increases gradually at loci at increasing distance from the inversion. To interpret the data we present a selectionist and a stochastic model, both of which point to a relatively recent origin of the studied inversion and may suggest differences between the evolutionary history of inversions in Anopheles and Drosophila species.Correspondence to: K.D. Mathiopoulos     

An RFLP-based genetic map of pearl millet (Pennisetum glaucum)

Analysis of a sample of diverse pearl millet genotypes with 200 genomic DNA probes revealed this crop species to be extremely polymorphic. Among these genotypes, 85% of probes detected polymorphism using only two restriction enzymes, with an average pair-wise polymorphism between all of the probe-enzyme combinations of 56%. Two crosses were employed to construct an RFLP-based genetic map. In an intervarietal F₂ population, derived from a single F₁ plant, 181 loci were placed on a linkage map. The total length of this map, which comprised seven linkage groups, was 303 cM and the average map distance between loci was about 2 cM, although a few intervals in excess of 10 cM were present at the ends of a few linkage groups. Very few clones, including those which hybridized to more than one copy, detected more than one locus in the pearl millet genome. The analysis was complicated initially because 83 of the 181 loci mapped to a single linkage group. Analysis of a second cross identified a probable translocation breakpoint in the middle of this large linkage group.     

Structure and evolution of the genomes ofsorghum bicolor andZea mays

Cloned maize genes and random maize genomic fragments were used to construct a genetic map of sorghum and to compare the structure of the maize and sorghum genomes. Most (266/280) of the maize DNA fragments hybridized to sorghum DNA and 145 of them detected polymorphisms. The segregation of 111 markers was analyzed in 55 F₂ progeny. A genetic map was generated with 96 loci arranged in 15 linkage groups spanning 709 map units. Comparative genetic mapping of sorghum and maize is complicated by the fact that many loci are duplicated, often making the identification of orthologous sequences ambiguous. Relative map positions of probes which detect only a single locus in both species indicated that multiple rearrangements have occurred since their divergence, but that many chromosomal segments have conserved synteny. Some sorghum linkage groups were found to be composed of sequences that detect loci on two different maize chromosomes. The two maize chromosomes to which these loci mapped were generally those which commonly share duplicated sequences. Evolutionary models and implications are discussed.     

Structural differences in pericentric inversions. Application to a model of risk of recombinants

Summary The potential chromosomal imbalance in offspring of pericentric inversion heterozygotes can be evaluated by measuring (% of haploid autosomal length, % HAL) the chromosomal segments distal to the breakpoints in the inversion. These distal segments were measured in presently reported and published cases of pericentric inversions, divided into two ascertainment groups: (I) those ascertained through recombinant offspring and (II) those ascertained through balanced heterozygotes. The distal segments in group II inversions were significantly larger than those of group I, i.e., the potentially larger chromosomal imbalances were not observed in full-term offspring. These results are discussed in relation to the model of risk of abnormal offspring in the progeny of heterozygotes for structural rearrangements (the chromosome imbalance size-viability model). The mean distal segment sizes for group I and group II pericentric inversions were respectively not significantly different from the mean interchange segment size for a sample of reciprocal translocations divided into the same two ascertainment groups. It was concluded that the restrictions on the size (% HAL) of chromosomal imbalance in offspring surviving until term are similar whether this imbalance arises from reciprocal translocations or pericentric inversions.     

Development of a genetic sexing strain of Anopheles arabiensis for KwaZulu‐Natal,South Africa

An efficient sexing system is important for the release of sterile males for any control programme using the sterile insect technique. This study describes the development and characterization of a new genetic sexing strain from South Africa (GMK), needed for the planned implementation of such a programme in northern KwaZulu‐Natal Province. The base colony used was a locally modified laboratory strain of Anopheles arabiensis containing a sex‐linked gene conferring dieldrin resistance to male mosquitoes. Female A. arabiensis mosquitoes from northern KwaZulu‐Natal were mated with these males and backcrossed to introduce the dieldrin resistance gene to the Y chromosome. The resulting strain therefore had an overall genotype representing the local population but with the Y chromosome containing the dieldrin resistance gene. Life‐history characteristics, stability of the sex‐linked resistance marker, and reduction in dieldrin waste were investigated. The strain showed semi‐sterility exhibited by low egg hatch rates, faster development in the immature stages and longer adult survivorship compared with the parental strains. While the GMK strain carrying the dieldrin‐resistant gene was successfully established, the stability of the gene is limited, requiring periodic purification. Dieldrin waste can be limited by treating many more eggs than currently recommended.     

Practical genetic control strategies for industrial bioprocesses

Optimization of metabolism to maximize production of bio-based chemicals must consistently balance cellular resources for biocatalyst growth and desired compound synthesis. This mini-review discusses synthetic biology strategies for dynamically controlling expression of genes to enable dual-phase fermentations in which growth and production are separated into dedicated phases. Emphasis is placed on practical examples which can be reliably scaled to commercial production with the current state of technology. Recent case studies are presented, and recommendations are provided for environmental signals and genetic control circuits.     

Blood genetic systems in four Amazonian tribes.

Data on 31 genetic systems were obtained for 421 individuals belonging to the Arara, Araweté, Mundurucu, and Jamamadi tribes of northern Brazil. The Jamamadi depart farthest, and the Mundurucu least, from South American Indian averages. These data are analyzed together with those of 24 other Amazonian groups. Genetic distances and corresponding dendrograms indicate a cluster of 14 related tribes living north of the Amazon river. These genetic results show only a modest correlation with linguistic and geographic relationships among these groups.     

The use of bridging systems to increase genetic variability in compound chromosome strains for genetic control of Lucilia cuprina (Wiedemann)

Summary Genetic variability in the non-compound portion of the genomes of compound-chromosome (CC) strains intended for genetic control can be increased by the use of bridging strains which can be crossed to both CC and normal strains. Two bridging systems are described for chromosome-5 CC strains of Lucilia cuprina. The first system relies on the established viability and fertility of males trisomic for chromosome 5R. Males carrying the (5L.YL)23 half-translocation, a C(5R), and a normal chromosome 5 were crossed successfully to a CC strain and a normal strain. The second system uses a pair of reciprocal whole-arm 4;5 translocations to generate gametes disomic for 5R and nullosomic for 5L, which in combination with C(5L)-bearing gametes form viable near-euploid offspring with only small duplications and deficiencies. These offspring (C(5L); (4L.5R)357; (4R.5R)194; (4L.4R)) were crossed successfully with both CC and T(4;5)357/ + individuals. The latter were in turn crossed successfully with normal strains. The T(Y;5)23 system allows replacement of the non-CC genome with wild material more rapidly than the T(4;5)357/T(4;5)194 system, but unlike the latter does not allow replacement of the Y chromosome in the CC strain. The double translocation system is currently being used in L. cuprina.     

Cytogenetic analysis of three genetic sexing strains of Ceratitits capitata

Summary Polytene chromosomes of three genetic sexing strains of Ceratitis capitata were analyzed. The genetic sexing mechanism is based on a pupal color dimorphism (white-brown) and is the result of a reciprocal translocation between the Y chromosome and the autosome bearing the w locus (white pupal case). The analyzed polytene chromosomes were derived from two different pupal tissues, the orbital bristle and fat body cells. The Y chromosome is visible in both tissues, while the autosomes present a different banding pattern. Based on these features, the autosome breakpoints in the three Y; autosome translocations were mapped, and the homology of the translocated autosome in both tissues was established. In addition, the location of the break-points was compared to the stability of these three strains.     

Modeling resistance to genetic control of insects

The sterile insect technique is an area-wide pest control method that reduces pest populations by releasing mass-reared sterile insects which compete for mates with wild insects. Modern molecular tools have created possibilities for improving and extending the sterile insect technique. As with any new insect control method, questions arise about potential resistance. Genetic RIDL^®1 (Release of Insects carrying a Dominant Lethal) technology is a proposed modification of the technique, releasing insects that are homozygous for a repressible dominant lethal genetic construct rather than being sterilized by irradiation. Hypothetical resistance to the lethal mechanism is a potential threat to RIDL strategies' effectiveness. Using population genetic and population dynamic models, we assess the circumstances under which monogenic biochemically based resistance could have a significant impact on the effectiveness of releases for population control. We assume that released insects would be homozygous susceptible to the lethal genetic construct and therefore releases would have a built-in element of resistance dilution. We find that this effect could prevent or limit the spread of resistance to RIDL constructs; the outcomes are subject to competing selective forces deriving from the fitness properties of resistance and the release ratio. Resistance that is spreading and capable of having a significant detrimental impact on population reduction is identifiable, signaling in advance a need for mitigating action.     

害虫遗传防治的研究历史与现状

作为防治或根除重大害虫最为有效的手段之一,害虫遗传防治在世界范围内被广泛采用并取得了巨大成功。本文综述了不育昆虫技术、雌性致死系统和昆虫显性致死技术等经典害虫遗传防治策略的发展历史、技术特点和应用情况。近年来,许多新的分子生物手段被不断提出并整合到害虫遗传防治策略中,包括归巢核酸内切酶基因、锌指核酸酶、转录激活因子样效应因子核酸酶、CRISPR/Cas9系统、Medea元件、Killer-Rescue系统、Wolbachia-细胞质不亲和性系统等。基于这些新的工具手段,许多国家已经在不同程度上启动了下一代害虫遗传防治项目。而我国在该领域的研究相对薄弱,需要在借鉴国外成功经验的同时,进一步加强害虫遗传防治的基础和应用研究,从而实现本地有害生物的可持续治理和外来入侵生物的有效狙击,确保我国未来的粮食和生态安全。     

颗粒体病毒对小菜蛾自然种群的控制作用模拟

利用作用因子组配的生命表和种群系统控制理论,模拟了小菜蛾颗粒体病毒不同浓度和不同施用次数组合对菜心地小菜蛾自然种群的控制作用.结果表明,在蔬菜生态系统天敌作用得到逐渐恢复的前提下,如果不采用任何防治措施,春季菜心地小菜蛾自然种群经第一代的繁殖,第二代卵和幼虫数量为第一代的41倍;在小菜蛾第一代低龄幼虫期,喷施0.25 LE/L病毒2次,小菜蛾第二代卵和幼虫数量比对照显著下降,但下代小菜蛾种群增长仍保持在1以上;而喷施0.5 LE/L病毒2次,1 LE/L病毒1次两种组合处理,小菜蛾第二代种群增长倍数仅为0.13,显著降低田间小菜蛾种群数量,达到控制小菜蛾为害的目的.     

Direct evidence for suppression of recombination within two pericentric inversions in humans: a new sperm-FISH technique.

Crossover within a pericentric inversion produces reciprocal recombinant chromosomes that are duplicated/deficient for all chromatin distal to the breakpoints. In view of this fact, a new technique is presented for estimating the frequency of recombination within pericentric inversions. YAC probes were selected from within the q- and p-arm flanking regions of two human inversions, and two-color FISH analysis was performed on sperm from heterozygous inversion carriers. A total of 6,006 sperm were analyzed for chromosome 1 inversion (p31q12), and 3,168 were analyzed for chromosome 8 inversion (p23q22). Both inversions displayed suppression of crossing-over, although the amount of suppression differed between the two inversions. The recombination frequency of 13.1% recorded for chromosome 8 inversion was similar to the frequency of 11.4% previously estimated by the human/hamster-fusion method. For chromosome 1 inversion, the recombination frequency of 0. 4% reported here was below the limits of detection of the fusion technique. The simplicity of the FISH technique and the ease of scoring facilitate analysis of a sample-population size much larger than previously had been possible.     

Assessment of quality control parameters during mass rearing of a pupal colour genetic sexing strain of the Mediterranean fruit fly

Quality control charts were employed to monitor the quality of several parameters during mass rearing of the T : Y(wp ⁺)30C strain of Ceratitis capitata (Wiedemann), in which male and female flies emerge from brown and white puparia, respectively. Egg hatch and pupal recovery in this genetic sexing strain were below that reported for a wild-type Sohag strain, whereas pupal size, adult emergence and flight ability were higher. The overall recovery from eggs to adult flies capable of flight was similar in the two strains but egg production in T:Y(wp ⁺)30C was reduced to about 56% of that of Sohag. The quality control charts showed an acceptable level of variation within sets of data but the level between data sets was occasionally out of control. This indicated the presence of uncontrolled conditions during the rearing process, possibly arising from variations in the quality of the larval diet. It is concluded that the quality of T : Y(wp ⁺)30C renders this strain suitable for mass rearing in sterile insect technique (SIT) programmes requiring the elimination of the female flies prior to the release of sterilized males. The next step should involve further quality assessments both under field cage and under field release conditions. The importance of optimal and standardized dietary conditions is stressed in relation to the quality of mass reared medflies.

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Résumé Des normes de qualité ont été utilisées pour relever la valuer de plusieurs paramètres lors de l'élevage industriel de la souche T : Y(wp ⁺)30C de C. capitata Wied. dont les mâles et les femelles émergent respectivement de pupes brunes et blanches. Les taux d'éclosion et d'obtention de pupes sont plus faibles avec cette souche qu'avec la souche souvage Sohag; par contre, la taille des pupes, le taux d'émergence des adultes et l'aptitude au vol étaient plus élevés. La fréquence totale des adultes capables de voler par rapport au nombre initial d'ufs a été de 0,297 contre environ 0,311 pour Sohag. Cependant, la production d'ufs a été réduite à environ 56% de celle de Sohag. Les tableaux de contrôle de la qualité ont révélé un niveau de variation acceptable à l'interieur rieur des lots de données, mais la variabilité entre les lots excédait les normes admissibles. Ceci implique des paramètres incontrôlés pendant l'élevage de masse, probablement liés à des variations de la qualité de l'aliment. On peut en conclure que la qualité de la souche T : Y(wp ⁺)30C la rend utilisable pour un élevage de masse dans des programmes d'utilisation d'insectes stériles (SIT) exigeant l'élimination des femelles avant le lâcher de mâles stériles. La prochaine étape impliquera de nouvelles épreuves de qualité dans des cages en champ et dans les conditions de libération dans la nature. L'importance des conditions optimales d'une alimentation standardisée est soulignée par la qualité des mouches obtenues en élevage de masse.

     

遗传标记和免疫学方法在实验动物质量控制中的应用

生命科学发展迅速,实验动物级别逐渐提高,在保证生物制品实验结果可靠性前提下,完善动物由低级别向高级别过渡显得十分必要,最终目的达到实验动物标准化和动物实验规范化。对实验动物质量控制中的遗传标记和免疫学方法进行了综述。