Do sterile females affect the sexual performance of sterile males of Anastrepha ludens (Diptera: Tephritidae)?

The release of sterile males to suppress the Mexican fruit fly Anastrepha ludens (Loew) population was simulated in field cages to compare the sexual performance of sterile males with wild females in the presence and absence of sterile females. Using different release ratios of sterile and wild individuals, this study showed that the presence of sterile females negatively affected the sexual performance of sterile males in mating with wild females. We found that the sterility induced was proportional to the magnitude of the sterile/fertile ratio. These results indicate that the release of sterile males without sterile females can significantly improve the efficiency of the sterile insect technique for the control of A. ludens.     

A Dominant Gene for Male Sterility in Salvia miltiorrhiza Bunge

A natural male sterile mutant of Salvia miltiorrhiza (Labiatae, Sh-B) was found during field survey in 2002. Our objective was to analyze its genetic mechanism for producing F1 hybrid seeds and to develop a molecular marker linked to male sterile gene for selection of a hybrid parent line. The segregation ratios of male sterile plants to fertile plants in the progenies of both testcross and backcross were 1:1 in continuous experiments conducted in 2006–2009. The male sterile Sh-B was heterozygous (Msms). The male sterile plants could capture most pollen (2 granule/cm²·24 h) with row ratio (female : male 2 : 1) within 45-cm distance and harvest the largest amount of 6495 g hybrid seeds per hectare. We also developed DNA markers linked to the male sterile gene in a segregating population using bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) techniques. The segregating population was subjected to BSA-AFLP with 128 primer combinations. One out of fourteen AFLP markers (E11/M4208) was identified as tightly linked to the dominant male sterile gene with a recombination frequency of 6.85% and at a distance of 6.89 cM. This marker could be converted to PCR-based assay for large-scale selection of fertile plants in MAS (marker-assisted selection) at the seedling stage. Blastn analysis indicated that the male sterile gene sequence showed higher identity with nucleotides in Arabidopsis chromosome 1–5, and was more likely to encode S-adenosylmethionine-dependent methyltransferase, in which DNA methylation regulated the development of plant gametogenesis.     

Prefiltration enhances performance of sterile filtration for glycoconjugate vaccines

Recent studies have reported very low capacity during sterile filtration of glycoconjugate vaccines due to rapid fouling of the sterile filter. The objective of this study was to explore the potential for significantly increasing the capacity of the sterile filter through the use of an appropriate prefilter. Data were obtained using prefilters with different pore size and chemistry, with the sterile filtration performed at constant filtrate flux using 0.22 μm nominal pore size Durapore® polyvinylidene difluoride membranes. Prefiltration through 5 μm pore size Durapore® or Nylon prefilters nearly eliminated the fouling of the sterile filter, leading to more than a 100-fold reduction in the rate of pressure increase for the sterile filter. This dramatic improvement in sterile filter performance was due to the removal of large components (greater than 1 μm in size) as confirmed by dynamic light scattering. These results demonstrate the potential of using large pore size prefilters to significantly enhance the performance of the sterile filtration process for the production of important glycoconjugate vaccines.     

Male sterility and restored fertility in annual mercuries, relations with sex differentiation

The paper summarizes the researches conducted on male sterility in Mercurialis annua. Totally sterile individuals are very scarce in the dioecious species showing as the other Mercuries, unisexual flowers devoid of rudiments of the opposite sex. From one sterile male mutant, a ‘sterile series’ was conducted and genetics was studied. Sterile, semisterile, restored fertile male lines were constructed as well as female lines containing the inducer gene of male sterility, both fertility restorers and the sensitive cytoplasm. Morphology and ontogeny of these isogenic lines were presented. Male sterile anthers (empty) present a splitted tapetum and an abnormal meiotic end. Restored fertile male lines were normal. The relative abundance of auxin and cytokinins was studied. A specific cytokinin pathway measured as a background in fertile lines, the cis-oxidized pathway characterised the ‘sterile series’. Restoration of normal meiosis and tapetum appeared for the highest quantities of cis-zeatin (669 ng instead of 192 ng/100 g fresh weight in totally sterile). Auxin quantities were abundant compared with the normal males. Gene expression in the ‘sterile series’ was also compared with the fertile lines. t-RNAs specific for normal females were expressed in the male ‘sterile series’. Hybridization kinetics and in vitro translations pf poly(A)⁺RNAs demonstrate specific sequences for each line. Comparisons between identical organs (normal fertile male/restored fertile male or normal female/female of the ‘sterile series’) exhibited nearly 10% differences. The results suggest that for stamen development, a cascade of regulators probably exists: sex genes acting on the induction of stamen or pistil, then genes for sterility/restoration of fertility acting in anthers. Fertility-sterility regulators control the synthesis of a specific cytokinin pathway. The new hormonal signals are linked to several specific genes expressed in the floral morphology characterizing each line of the ‘sterile series’.     

Genetic mapping of sterile genes with epistasis in backcross designs

The mapping of sterile genes is an essential issue, which should be solved for the investigation of sterility mechanism in wide hybridization of plants. However, the methods formerly developed cannot address the problem of mapping sterile loci with epistasis. In this study, we developed a new method to map sterile genes with epistasis in wide hybridizations of plants using a backcross design. The maximum likelihood method was used to estimate the parameters of recombination fractions and effects of sterile genes, and the convergent results of these parameters were obtained using the expectation maximization (EM) algorithm. The application and efficiency of this method were tested and demonstrated by a set of simulated data and real data analysis. Results from the simulation experiments showed that the method works well for simultaneously estimating the positions and effects of sterile genes, as well as the epistasis between sterile genes. A real data set of a backcross (BC) population from an interspecific hybrid between cultivated rice, Oryza sativa, and its wild African relative, Oryza longistaminata, was analyzed using the new method. Five sterile genes were detected on the chromosomes of 1, 3, 6, 8 and 10, and significant epistatic effects were found among the four pairs of sterile genes.     

太空诱变玉米核不育突变体矮化的遗传及外施赤霉素分析

为了解太空诱变玉米核不育突变体矮化的遗传规律和原因,该研究以不育突变体为母本,自交系178、478为父本,对测交 F1、F2群体进行育性鉴定和株高分析,对 F2可育株进行基因型和株高分析,对姊妹交后代分离群体进行育性鉴定和株高、雄穗长度、节间数、节间长度分析,同时,还对姊妹交后代分离群体进行施赤霉素处理,调查育性和株高的变化。结果表明：178和478背景下的 F1表现出与测交母本一样的极显著差异;在178和478核背景下的 F2中,不育株株高极显著矮于可育株,两核背景下的不育株间株高差异不显著,而可育株间株高差异极显著;F2中纯合和杂合可育株的株高差异不显著;姊妹交后代分离群体中不育株株高、雄穗长度、节间数和节间长度极显著小于可育株;外施赤霉素的不育株在苗期表现出对赤霉素一定的敏感性,但株高最终未恢复正常高度。因此,得出该突变体矮化表现稳定,与不育性状并存,且不受细胞核背景的影响;核不育基因对植株株高的矮化无剂量效应;突变体的矮化与雄穗长度、节间数和节间长度有关;突变体不完全属于赤霉素不敏感型,其矮化并不是单一缺乏赤霉素而引起。该研究结果为认识太空诱变玉米核不育突变体矮化的遗传和生理机制提供了参考。     

Expression of female sterility in alfalfa (<Emphasis Type="Italic">Medicago sativa</Emphasis> L.)

Female sterility associated with the presence of callose in the nucellus at anthesis was studied in an F₁ progeny of two alfalfa plants displaying 5 and 81% ovule sterility. Transgressive segregation was observed and 100% sterile plants were obtained. Two of the sterile plants were used for cytological analyses on sectioned and stain-cleared whole ovules, in comparison to a 100% fertile full sib plant. The first sign of sterility was callose deposition in the nucellus cell walls surrounding the sporogenous cells of the young ovules. At the same stage, no trace of callose was present in ovule primordia of the fertile plant. Megaspore mother cells differentiated in both fertile and sterile ovules and meiosis was initiated, as indicated by chromatin patterning typical of a zygotene stage. However, meiosis was never completed in the sterile plants. In the control, callose was deposited around the meiocyte and as sects between the cells of the dyads and tetrads during meiosis, and disappeared after the completion of meiosis; an embryo sac developed and female fertility was normal. In the sterile ovules, some nucellus cells enlarged and callose accumulation continued forming thick deposits. At anthesis, the sterile ovules lacked an embryo sac and showed massive callose accumulation in the nucellus. Male fertility was normal in female-sterile plants, thus a female-specific arrest of sporogenesis appears to be the cause of sterility. Pistil development was aberrant in some sterile genotypes, even with arrested pistil growth in early flower buds.     

A Molecular Method to Discriminate between Mass-Reared Sterile and Wild Tsetse Flies during Eradication Programmes That Have a Sterile Insect Technique Component

Background

The Government of Senegal has embarked several years ago on a project that aims to eradicate Glossina palpalis gambiensis from the Niayes area. The removal of the animal trypanosomosis would allow the development more efficient livestock production systems. The project was implemented using an area-wide integrated pest management strategy including a sterile insect technique (SIT) component. The released sterile male flies originated from a colony from Burkina Faso.

Methodology/Principal Findings

Monitoring the efficacy of the sterile male releases requires the discrimination between wild and sterile male G. p. gambiensis that are sampled in monitoring traps. Before being released, sterile male flies were marked with a fluorescent dye powder. The marking was however not infallible with some sterile flies only slightly marked or some wild flies contaminated with a few dye particles in the monitoring traps. Trapped flies can also be damaged due to predation by ants, making it difficult to discriminate between wild and sterile males using a fluorescence camera and / or a fluorescence microscope. We developed a molecular technique based on the determination of cytochrome oxidase haplotypes of G. p. gambiensis to discriminate between wild and sterile males. DNA was isolated from the head of flies and a portion of the 5’ end of the mitochondrial gene cytochrome oxidase I was amplified to be finally sequenced. Our results indicated that all the sterile males from the Burkina Faso colony displayed the same haplotype and systematically differed from wild male flies trapped in Senegal and Burkina Faso. This allowed 100% discrimination between sterile and wild male G. p. gambiensis.

Conclusions/Significance

This tool might be useful for other tsetse control campaigns with a SIT component in the framework of the Pan-African Tsetse and Trypanosomosis Eradication Campaign (PATTEC) and, more generally, for other vector or insect pest control programs.     

A fertile revertant from petaloid cytoplasmic male-sterile carrot has a rearranged mitochondrial genome

 A spontaneously derived fertile plant was recovered from a petaloid cytoplasmic male-sterile (CMS) carrot inbred line. Genetic analysis indicated a single nuclear gene was responsible for the restoration to fertility. Within a family segregating for the nuclear restorer in combination with the sterility-inducing cytoplasm, fertile plants were recovered that could not restore fertility when crossed to sterile genotypes. Genetic analysis indicated cytoplasmic reversion for fertility, and Southern analysis, comparing mtDNA organization of the fertile revertant and its CMS progenitor, identified mitochondrial genome rearrangements. Hybridization of cosmids representing a 108-kb subgenomic circle of the sterile line to DNA of a fertile maintainer and fertile revertant lines indicated a similar mtDNA organization for these genotypes that was distinct from that of the sterile line. Six restriction fragments totalling 43.2 kb were common to the fertile maintainer and revertant and absent in the sterile; other restriction fragments totalling 38.2 kb were present only for the sterile line. Unique fragments of low stoichiometry, two for the fertile maintainer and three for the revertant, distinguished these lines. The reversion to fertility in the sterile line could have resulted from the amplification of a mitochondrial submolar genome highly homologous to that found in the fertile maintainer line. Received: 4 October 1997/Accepted: 12 December 1997     

Capture of Mediterranean fruit flies (Diptera: Tephritidae) in dry traps baited with a food-based attractant and Jackson traps baited with trimedlure during sterile male release in Guatemala

Captures of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), in Jackson traps baited with trimedlure were compared with captures in cylindrical open-bottom dry traps baited with a food-based synthetic attractant (ammonium acetate, putrescine, and trimethylamine). Tests were conducted in Guatemala during a sterile male release program in an area where wild flies were present in low numbers. More wild and sterile females were captured in food-based traps, and more wild and sterile males were captured in trimedlure traps. The food-based traps captured almost twice as many total (male plus female) wild flies as the trimedlure traps, but the difference was not significant. Females made up approximately 60% of the wild flies caught in the food-based attractant traps; the trimedlure traps caught no females. The ratio of capture of males in trimedlure traps to food-based traps was 6.5:1 for sterile and 1.7:1 for wild flies. Because fewer sterile males are captured in the food-based traps, there is a reduction in the labor-intensive process of examining flies for sterility. The results indicate that traps baited with food-based attractants could be used in place of the Jackson/trimedlure traps for C. capitata sterile release programs because they can monitor distributions of sterile releases and detect wild fly populations effectively; both critical components of fruit fly eradication programs by using the sterile insect technique.     

Bisexual Hybrid Sterility in DROSOPHILA MELANOGASTER

A new type of hybrid sterility was investigated in D. melanogaster . Matings between strain 27 males from Para Wirra, South Australia, and Canton-S females produce 70–80% fully sterile male and female progeny. Strain 27 males produce sterile progeny when crossed to females of other geographic origins, but produce fertile progeny when crossed to a second sympatric strain. The sterility is avoided by lower rearing temperatures. Heat shock and tetracycline produce no improvement in the fertility of the hybrids. Normal flies produce sterile progeny when injected with, or fed, homogenates of sterile flies. A combination of maternal and paternal factors may interact to produce sterile hybrids by inhibiting gonad development.     

The Sterile Insect Technique for Controlling Populations of Aedes albopictus (Diptera: Culicidae) on Reunion Island: Mating Vigour of Sterilized Males

Reunion Island suffers from high densities of the chikungunya and dengue vector Aedes albopictus. The sterile insect technique (SIT) offers a promising strategy for mosquito-borne diseases prevention and control. For such a strategy to be effective, sterile males need to be competitive enough to fulfil their intended function by reducing wild mosquito populations in natura. We studied the effect of irradiation on sexual maturation and mating success of males, and compared the sexual competitiveness of sterile versus wild males in the presence of wild females in semi-field conditions. For all untreated or sterile males, sexual maturation was completed within 13 to 20 h post-emergence and some males were able to inseminate females when 15 h old. In the absence of competition, untreated and sterile males were able to inseminate the same number of virgin females during 48 h, in small laboratory cages: an average of 93% of females was inseminated no matter the treatment, the age of males, and the sex ratio. Daily mating success of single sterile males followed the same pattern as for untreated ones, although they inseminated significantly fewer females after the ninth day. The competitiveness index of sterile males in semi-field conditions was only 0.14 when they were released at 1-day old, but improved to 0.53 when the release occurred after a 5-day period in laboratory conditions. In SIT simulation experiments, a 5∶1 sterile to wild male ratio allowed a two-fold reduction of the wild population’s fertility. This suggests that sterile males could be sufficiently competitive to mate with wild females within the framework of an SIT component as part of an AW-IPM programme for suppressing a wild population of Ae. albopictus in Reunion Island. It will be of interest to minimise the pre-release period in controlled conditions to ensure a good competitiveness without increasing mass rearing costs.     

Study of male sterility in Taiwania cryptomerioides Hayata (Taxodiaceae)

Summary. A study of male sterility over a period of three consecutive years on a conifer species endemic to Taiwan, Taiwania cryptomerioides Hayata (Taxodiaceae), was done for this article. With the aids of fluorescence and electron microscopic observations, the ontogenic processes in the fertile and sterile microsporangia are compared, using samples collected from Chitou Experimental Forest and Yeou-Shoei-Keng Clonal Orchard of the National Taiwan University, Nantou, Taiwan. The development of male strobili occurred from August to the end of March. Microsporogenesis starts with the formation of the archesporium and ends with the maturation of 2-celled pollen grains within the dehiscing microsporangium. Before meiosis, there was no significant difference in ultrastructure between the fertile and sterile microsporangia. Asynchronous pollen development with various tetrad forms may occur in the same microsporangium of either fertile or sterile strobili. However, a callose wall was observable in the fertile dyad and tetrad, but not in the sterile one. After dissolution of the callose wall, the fertile microspores were released into the locule, while some sterile microspores still retained as tetrads or dyads with intertwining of exine walls in the proximal faces. As a result, there was no well developed lamellated endexine and no granulate ectexine or intine in the sterile microspores. Eventually, the intracellular structures in sterile microspores were dramatically collapsed before anthesis. The present study shows that the abortion in pollen development is possibly attributed to the absence of the callose wall. The importance of this structure to the male sterility of T. cryptomerioides is discussed. Correspondence and reprints: Department of Life Science, National Taiwan University, 106 Taipei, Taiwan.     

Ectopic expression of <Emphasis Type="Italic">OsMADS1</Emphasis> caused dwarfism and spikelet alteration in rice

In rice, an E-class gene, OsMADS1, acts to specify the identities of the lemma and palea. In this study, the OsMADS1 gene with a CaMV35S promoter was transformed into a japonica cultivar, Zhonghua 11. All transgenic plants successfully showed similar phenotypes, including dwarfism, distorted panicles, decreased numbers of branches and spikelets, and elongated sterile lemma. Histological analysis showed that the elongated sterile lemma developed with silicified epidermal and sclerenchymal cells, which were lacking in the wild-type sterile lemma, suggesting that the elongated sterile lemma had assumed the identity of the lemma or palea. Some marker genes were subjected to a detailed analysis of the distribution of their expression among the lemma, palea and sterile lemma. DROOPING LEAF (DL) and OsMADS6 genes were only expressed in the normal lemma or palea, respectively. In the elongated sterile lemma, a high level of DL gene expression was detected, while no expression of OsMADS6 was found, implying that the sterile lemma transformed into the lemma but not the palea. These results provide clues to elucidate the mechanism of evolution from lemma to sterile lemma in rice. qPCR analysis also suggested that the ectopic expression of OsMADS1 induced abnormal brassinosteroid and gibberellin acid activation, and then resulted in developmental defects in the stem and panicle.     

Fusion-mediated combination of Ogura-type cytoplasmic male sterility with Brassica napus plastids using X-irradiated CMS protoplasts

X-irradiated protoplasts of a Brassica napus line carrying the Ogura Raphanus sativus male sterile cytoplasm were fused to protoplasts of male fertile B. napus cv. Olga. Plants were regenerated from six out of 34 randomly selected clones. In one clone, Bn(RS)26, a plant with male sterile flowers was obtained. Mitochondria of this plant are non-parental as revealed by DNA-DNA hybridization using a species specific probe. Its chloroplasts, however, derive from the fertile parent which results in loss of the sensitivity to low temperatures associated with R. sativus plastids in the male sterile parent. The novel cytoplasm of the Bn(RS)26 cybrid was transmitted through seed.Abbreviations CMS cytoplasmic male sterile - PEG polyethylene glycol - mtDNA mitochondrial DNA - cpDNA chloroplast DNA     

A field cage study of the optimal age for release of radio-sterilized Aedes albopictus mosquitoes in a sterile insect technique program

The use of the sterile insect technique (SIT) is being considered as an additional tool for the control of Aedes albopictus (Skuse) (Diptera: Culicidae), the vector of Chikungunya and Dengue viruses in Mauritius. The aim of this study was to assess the competitiveness value of sterile males of different age and under various release conditions. Three release ratios were tested with sterile males of either 1, 3, or 5 days old at release. The competition of sterile males against same age or a mixed age population of fertile males (which is more representative of the field situation) was also investigated. The participation in mating (observed through single female oviposition), the average-induced sterility, and the male competitiveness index indicated that 3-day-old sterile males have the best balance between survival and mating capacity, and should therefore be the favored age of release in the field. Reduction in the cage fertility was obtained at 5-to-1 release ratio; however, it is speculated that at least a 10-fold ratio of sterile-to-fertile males should be chosen to induce substantial sterility in the wild Ae. albopictus population in the SIT pilot release site in Mauritius. Interestingly, this study showed for the first time that the age of the fertile male population against which the sterile males compete is a very important parameter that can significantly affect the sexual performance of sterile males, leading to overestimation of their competitiveness values.     

A new method for hybrid seed production based on cytoplasmic male sterility combined with a lethal gene and a female sterile pollenizer in Capsicum annuum L.

Summary A new improved method for hybrid seed production was successfully tested. This method is based on using a cytoplasmic male sterile line possessing a lethal gene with action that can be easily inhibited and a female sterile pollenizer. The lethal gene ensures 100% purity of the F₁ crop. The female sterile pollenizer provides a permanent abundant flowering with excess of pollen grains that leads to increased hybrid seed production without additional labour expenses. The described scheme is applicable for other crops as well.     

Genetic diagnosis of cytoplasmic male sterile cybrid plants of rice

Twelve Japanese rice cultivars were converted to CMS by asymmetric protoplast fusion with MTC-5A, the cytoplasm of which was derived from an indica rice, Chinsurah Boro II. With the exception of the cybrids that had a nucleus from Hoshiyutaka, most of these cybrid plants were sterile. The unique sequence downstream from the mitochondrial atp6 of MTC-5A was specifically amplified in the sterile cybrid plants by PCR. All progenies of the cybrid plants carrying this unique sequence were sterile. On the other hand, in some of the sterile cybrid plants in which the unique sequence was not amplified by PCR, fertility was recovered in their progenies. Somaclonal mutation may have caused sterility in these cybrids. Only the cybrid plants that had the unique sequence detected by PCR were CMS. Thus, the CMS plants can be selected rapidly and easily by PCR, at an early stage of plant regeneration. Soon after transplanting the regenerated plants to a green house, fertile cybrids and sterile cybrids produced by somaclonal mutation can be removed. These findings also show that the unique region downstream from atp6 is tightly linked with the CMS phenotype.     

The selective florivory of Erioscelis emarginata matches its role as a pollinator of Philodendron

Flowers commonly face the dilemma of needing to be attractive to pollinators but unattractive to nectar robbers or florivores. When the pollinator has chewing mouthparts, there is also a considerable risk of the pollinator consuming the flowers. Scant field records indicate that the florivorous beetle Erioscelis emarginata (Mannerheim) (Coleoptera: Scarabaeidae: Cyclocephalini), which pollinates Philodendron spp., feeds selectively on the sterile staminate flowers. Here, the hypothesis was tested that E. emarginata prefers sterile staminate flowers of Philodendron bipinnatifidum Schott and Philodendron melinonii Brongn. ex Regel (Araceae) over fertile flowers. This study also examined whether such a preference exists regardless of the proportion of fertile vs. sterile flowers in each Philodendron species. Analysis of nutritional and defensive compounds plus scanning electron microscopy on each flower type were performed. The feeding preference of florivores for either P. bipinnatifidum or P. melinonii was also examined. In both species, sterile flowers were significantly more consumed than fertile ones. The sterile zone of P. bipinnatifidum was significantly larger than that of P. melinonii, and sterile staminate flowers of both plants were consumed at similar rates. Calcium oxalate was markedly low in sterile flowers, which also presented smaller papillae than the other flowers. This study presents evidence that the balance between pollination and florivory has most likely evolved through a strong feeding preference for less‐defended sterile flowers regardless of the size of the sterile zone.     

Seasonal variation of the capture and field distribution of sterile Anastrepha ludens in Tamaulipas,Mexico

Anastrepha ludens (Loew) (Diptera: Tephritidae) is one of the most important citrus pest in Mexico. The sterile insect technique (SIT) is used against pest populations of fruit flies for suppression, eradication, containment and prevention to reduce damages in fruit‐growing areas. In this study, we analyzed the seasonal variation of captures and field distribution of sterile A. ludens released in different seasons of the year in north‐eastern Mexico. Chilled releases were conducted by air at constant densities per ha on a citrus area for a period of 32 weeks that included the coldest and warmest seasons that is winter, spring and summer. Multilure traps baited with torula yeast pellets were used to capture sterile flies. Fly capture data were compared over the three seasons and correlated with climate. The lowest number of captures of the sterile insect occurred in the summer and the highest in winter and spring. High and low temperatures were negatively correlated with fly captures. Field distribution was also negatively correlated with high temperatures in summer, but no relationships were observed in winter and spring. No relationships were observed between rainfall with capture and field distribution of sterile flies. These results indicate that summer is a season involving agro‐ecological and environmental constraints for the capture and field distribution of sterile flies. This study may be useful for enhancing release strategies and optimizing economic resources in north‐eastern Mexico. Further research on the behaviour of sterile flies under stressful environments is suggested.