Use of the SSLP-based method for detection of rare apomictic events in a sexual AtSERK1 transgenic Arabidopsis population

Here we present a screening method to evaluate the potential of genes to transfer aspects of apomixis into sexual crop plants. Based on the assumption that an apomictic progeny is an exact genetic replica of the mother plant we employed a set of single sequence length polymorphism (SSLP) markers to identify individuals displaying heterozygosity fixation in segregating sexual populations as an indication of rare apomictic events. Here we present the results of such a study using the Arabidopsis thaliana SOMATIC EMBRYOGENESIS RECEPTOR KINASE 1 (AtSERK1) gene expressed under the control of the AtLTP1 promoter in sexual Arabidopsis plants. In one of the three tested F2 transgenic populations expressing the AtLTP1::AtSERK1 construct we observed two plants with heterozygosity maintenance for the full set of SSLP markers indicating a possible clonal inheritance. However, as their offspring revealed a close to binomial segregation for a number of heterozygous loci, it was concluded that these two putative apomictic plants either lost their clonal ability in the next generation or resulted from incidental recombination events displaying the genotype of the parent. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.     

Description of a new species of Nitrosococcus

A new species of Nitrosococcus is described. It resembles Nitrosococcus oceanus in shape, size, and ultrastructure of the cells. However, the new species has a more pronounced salt requirement, corresponding to its natural habitats. Two strains were isolated from a salt lake in Saudi Arabia and a salt lagoon in the Mediterranean Sea, respectively. In contrast to N. oceanus, both isolates of the new species were unable to utilize urea as ammonia source. Both species also differed in gelelectrophoretic cell protein patterns. The name N. halophilus is proposed.     

Determining genetic origins of aberrant progeny from facultative apomictic Kentucky bluegrass using a combination of flow cytometry and silver-stained RAPD markers

Seeded plants that reproduce through facultative apomixis produce two types of progeny: (1) apomictic progeny genetically identical to the maternal genotype, and (2) aberrant progeny genetically different from the maternal genotype. Aberrant progeny have at least nine different genetic origins depending on gametic ploidy level and whether fertilization was self, cross, or absent. Multiple genetic origins of aberrant progeny complicate the results of basic and applied genetic studies. Determining the genetic origin of progeny plants using traditional techniques, such as cytology, embryology, and segregational studies, is technically difficult in Kentucky bluegrass. We have found that two relatively new techniques, flow cytometry and silver-stained RAPD (ssRAPD) markers, are powerful tools for rapidly determining the genetic origins of aberrant Kentucky bluegrass progeny. Our application of these techniques demonstrate that (1) flow cytometry accurately distinguishes progeny ploidy levels, and (2) ssRAPD markers distinguish progeny resulting from cross-fertilization. Therefore, a combination of flow cytometry and ss-RAPD data would be useful for most genetic studies of aberrant individuals. Moreover, ssRAPD s were found to be of value for measuring the loss of genetic markers from polyhaploids and quantifying the inheritance of parental genomes in polydiploid B_n (n+n) and polytriploid B_III (2n+n) hybrids. Quantifying shared ss-RAPD markers may also be useful for determining genetic relatedness between varieties and germplasm sources.     

Developmental expression of ASG- 1 during gametogenesis in apomictic guinea grass (Panicum maximum)

We have used Western blue-visualized in situ-hybridization (ISH) to monitor the expression of apomixis-specific gene-1 (ASG-1, GenBank accession number AB000809) during gametogenesis in obligate-sexual and facultative-apomictic (aposporic) genotypes of guinea grass (Panicum maximum). The in situ-analysis revealed that ASG-1 is not expressed in the ovule during early floral development in both, the facultative apomicts (A1 stage) and the obligate sexuals (S1 stage). With the appearance of the aposporous initial cell(s) in the ovule of the apomictic type (A2-1 stage), ASG-1 expression is strong and specific to this apomixis-specific cell. ASG-1 expression continued through different stages of aposporous embryo sac development (A2-2 stage), indicating that the gene may play a role in this developmental process. Regular embryo sacs in sexual types did not show hybridization signals (S2 stage). However, strong ASG-1 expression was detected in immature pollen grains and young embryos in both reproductive types, suggesting that ASG-1 may be an allele derived from the obligate-sexual wild type. Expression in pollen grains faded with maturation. In a heterologous system, using Paspalum notatum, a facultative-aposporic tropical grass (bahia grass), identical results were obtained. The results are discussed in view of the fact, that ASG-1 shows some homology to genes known to be seed- or embryo-specific or involved in processes related to cell growth.     

Chromosomal and embryological analyses in sexual x apomictic hybrids ofPanicum maximum Jacq.

Summary Cytological analyses in series of crosses between 7 sexual pistillate and 8 apomictic staminate parents of speciesPanicum maximum (Gramineae) are reported. Although these 15 progenitors were tetraploid (2n = 32), 2 dihaploids (2n = 16), 45 hexaploids (2n = 48) and 5 octoploids (2n = 64) were observed among 333 progeny plants. The role of unreduced gametes as the originators of polyploidy is discussed in relation to the so-called elements of apomixis. The 2 dihaploids appeared to be sexual while the hexaploids and octoploids were all apomictic. At the tetraploid level sexual and apomictic hybrids segregated in a ratio close to 11. These results were then compared to those already obtained from studies on other tropical grasses and indicate a simple genetic determinism for gametophytic apomixis.     

Mode of reproduction is detected by Parth1 and Sex1 SCAR markers in a wide range of facultative apomictic Kentucky bluegrass varieties

Gametophytic apomixis in Kentucky bluegrass (Poa pratensis L.) involves the parthenogenetic development of unreduced eggs from aposporic embryo sacs. Marker-assisted selection for the mode of reproduction in P. pratensis would avoid costly and time-consuming phenotypic progeny tests. We developed and tested two SCAR primer pairs that are associated with the mode of reproduction in P. pratensis. The SCAR primers identified the apomictic and sexual genotypes among progenies of sexual x apomictic crosses with very low bias. Furthermore, when tested on a wide range of Italian and exotic P. pratensis germplasm, they were able to unequivocally distinguish sexual from apomictic genotypes. This system should, therefore, allow new selection models to be set up in this species.     

Unusual microtubular cytoskeleton of apomictic embryo sac of Chondrilla juncea L.

Summary. The mature apomictic embryo sac of Chondrilla juncea is highly vacuolated and demonstrates a polarization similar to that of the amphimictic gametophyte. The microtubule cytoskeleton of this embryo sac is uncharacteristic and relatively weak. The microtubules are positioned along cell walls and resemble cortical microtubules of somatic cells. They do not form the parallel, brushlike structures observed around the filiform apparatus of synergids in the amphimictic embryo sac. In the apomictic embryo sac, the microtubules of both the egg cell and the central cell develop a cortical-like structure, which is entirely different from the radial arrangement observed around the nuclei in the amphimictic embryo sac. Correspondence and reprints: Department of Plant Cytology and Embryology, Jagiellonian University, Grodzka 52, 31-044 Kraków, Poland.     

Molecular markers shared by diverse apomictic Pennisetum species

Two molecular markers, a RAPD (randomly amplified polymorphic DNA) and a RFLP/STS (restriction fragment length polymorphism/sequence-tagged site), previously were found associated with apomictic reproductive behavior in a backcross population produced to transfer apomixis from Pennisetum squamulatum to pearl millet. The occurrence of these molecular markers in a range of 29 accessions of Pennisetum comprising 11 apomictic and 8 sexual species was investigated. Both markers were specific for apomictic species in Pennisetum. The RFLP/STS marker, UGT 197, was found to be associated with all taxa that displayed apomictic reproductive behavior except those in section Brevivalvula. Neither UGT197 nor the cloned RAPD fragment OPC-04₆₀₀ hybridized with any sexually reproducing representatives of the genus. The cloned C04₆₀₀ was associated with 3 of the 11 apomictic species, P. ciliare, P. massaicum, and P. squamulatum. UGT197 was more consistently associated with apomictic reproductive behavior than OPC04₆₀₀ or cloned C04₆₀₀, thus it could be inferred that UGT197 is more closely linked to the gene(s) for apomixis than the cloned C04₆₀₀. The successful use of these probes to survey other Pennisetum species indicates that apomixis is a trait that can be followed across species by using molecular means. This technique of surveying species within a genus will be useful in determining the relative importance of newly isolated markers and may facilitate the identification of the apomixis gene(s).     

Low intraspecific aggressiveness in two obligate plant-ant species

Little is known about the aggressiveness of plant-ants typically living in isolated trees nor about how that aggressiveness varies based on this isolation. Here, we examine intra- and interspecific aggressiveness between workers of two Allomerus species associated with two different myrmecophytes. In both cases, the level of intraspecific aggressiveness is very low whatever the distance separating the tested nests, while interspecific conflicts are always violent. Similar patterns of aggressiveness have been reported in various ant species, but the strictly arboreal life of Allomerus ants associated with the isolation of their adult colonies highlight different ecological conditions that might explain the lack of aggressiveness between conspecifics. Received 18 December 2007; revised 12 March 2008; accepted 13 March 2008.     

Description of two biovars in the Rhizobium galegae species: biovar orientalis and biovar officinalis

Twenty-six Rhizobium galegae strains, representing the center of origin of the host plants Galega orientalis and G. officinalis as well as other geographic regions, were used in a polyphasic analysis of the relationships of R. galegae strains. Phage typing, lipopolysaccharide (LPS) profiling, pulsed field gel electrophoresis (PFGE) profiling and rep-PCR (use of repetitive sequences as PCR primers for genomic fingerprinting) with REP and ERIC primers investigated nonsymbiotic properties, whereas plasmid profiling and hybridisation with a nif gene probe, and with nodB, nodD, nod box and an IS sequence from the symbiotic region as probes, were used to reveal the relationships of symbiotic genes. The results were used in pairwise calculations of distances between the strains, and the distances were visualised as a dendrogram. Indexes of association were compared for all tests pooled, and for chromosomal tests and symbiotic markers separately, to display the input of the different categories of tests on the grouping of the strains. Our study shows that symbiosis related genetic traits in R. galegae divide strains belonging to the species into two groups, which correspond to strains forming an effective symbioses with G. orientalis and G. officinalis respectively. We therefore propose that Rhizobium galegae strains forming an effective symbiosis with Galega orientalis are called R. galegae bv. orientalis and strains forming an effective symbiosis with Galega officinalis are called R. galegae bv. officinalis.     

An apomictic polyhaploid obtained from a pearl millet x Pennisetum squamulatum apomictic interspecific hybrid

Summary In a research program to transfer apomixis from Pennisetum squamulatum Fresen to pearl millet, P. americanum L. Leeke, a polyhaploid plant (2n=21) was discovered in the uniform open-pollinated progeny of an apomictic interspecific hybrid (2n = 41) between pearl millet and P. squamulatum. The polyhaploid was shorter, less vigorous and was smaller morphologically than its maternal parent. It probably originated by parthenogenetic development of a reduced gametophyte in the apomictic interspecific hybrid. The most common metaphase I chromosome association in the polyhaploid was 4 bivalents plus 13 univalents. Irregular chromosome distribution, tripolar spindles, bridges and fragments were observed at anaphase I and telophase I. The polyhaploid was male-sterile and partially female- fertile having multiple aposporous embryo sacs in 95% of the ovules. Seed set was 3% when open-pollinated and 33% when pollinated with pearl millet pollen. Low seed set was due to competition among multiple embryos developing in the same ovule. Seventeen progeny from seed produced under open-pollination on the polyhaploid each had 2n=21 chromosomes and were morphologically uniform and identical to the female parent. The expression of obligate apomixis in the polyhaploid conditioned by the P. squamulatum genome between the simplex and duplex condition indicates that apomictic reproduction is possible in nonpolyploid plants.     

Harnessing apomictic reproduction in grasses: what we have learned from Paspalum

Background

Apomixis is an alternative route of plant reproduction that produces individuals genetically identical to the mother plant through seeds. Apomixis is desirable in agriculture, because it guarantees the perpetuation of superior genotypes (i.e. heterotic hybrid seeds) by self-seeding without loss of hybrid vigour. The Paspalum genus, an archetypal model system for mining apomixis gene(s), is composed of about 370 species that have extremely diverse reproductive systems, including self-incompatibility, self-fertility, full sexual reproduction, and facultative or obligate apomixis. Barriers to interspecific hybridization are relaxed in this genus, allowing the production of new hybrids from many different parental combinations. Paspalum is also tolerant to various parental genome contributions to the endosperm, allowing analyses of how sexually reproducing crop species might escape from dosage effects in the endosperm.

Scope

In this article, the available literature characterizing apomixis in Paspalum spp. and its use in breeding is critically reviewed. In particular, a comparison is made across species of the structure and function of the genomic region controlling apomixis in order to identify a common core region shared by all apomictic Paspalum species and where apomixis genes are likely to be localized. Candidate genes are discussed, either as possible genetic determinants (including homologs to signal transduction and RNA methylation genes) or as downstream factors (such as cell-to-cell signalling and auxin response genes) depending, respectively, on their co-segregation with apomixis or less. Strategies to validate the role of candidate genes in apomictic process are also discussed, with special emphasis on plant transformation in natural apomictic species.     

雄性不育无籽刺梨花药发育过程中生理特性的初步研究

为探究无籽刺梨(Rosasterilis)雄性不育原因,采用1%I2-KI染色法观察花粉活性,并对无籽刺梨和正常可育刺梨(R.roburghii)花药不同发育时期的生理生化指标进行了研究。结果表明,无籽刺梨的败育花粉占95.5%,刺梨的正常花粉占99%。刺梨花药的可溶性蛋白质、可溶性糖和脯氨酸含量在各时期的总体变化趋势相似,可溶性淀粉含量呈上升趋势,而无籽刺梨花药的可溶性蛋白质、可溶性糖、淀粉和脯氨酸含量在各时期的变化无规律可循,且花粉成熟期这4种物质的含量均明显低于刺梨,即花粉成熟期缺少各营养物质的积累。在花药发育过程中,无籽刺梨的SOD活性均低于刺梨;MDA含量呈上升趋势,且上升幅度比刺梨大;MDA含量和POD活性均高于刺梨。因此,营养物质的匮乏和酶系统的紊乱可能是导致无籽刺梨雄性不育的原因。     

福建省刺齿虫兆属（弹尾纲：长角虫兆科）一新种描述

记述福建省刺齿属1新种:拟中华刺齿Homidia pseudosinensis sp.nov.。该新种的鉴定特征有胸部第2节后侧具近似M型的横条带,下唇毛序及腹部第4节毛序。本文提供了该物种的特征图及福建省刺齿属分种检索表。     

Spermatogenesis inDrosophila hydei: A genetic survey

Summary We constructed balancer-chromosomes for the large autosomes ofDrosophila hydei and screened more than 16000 chromosomes for male sterile mutations in order to dissect spermatogenesis genetically. 365 mutants on the X chromosome and the autosomes 2, 3, and 4 were recovered and analysed cytologically in squash preparations under phase-contrast optics. The majority of the mutations allows a rather advanced differentiation of the spermatozoa. At the light-microscopical level, it is possible to classify these mutations with respect to individualization, coiling or motility of the mutant spermatozoa. In contrast, a small number of mutants exhibits conspicuous, pleiotropic phenotypes. Gonial divisions, the shaping of the spermatocyte nucleus and male meiotic divisions are controlled by X chromosomal or autosomal genes which can mutate to male sterile alleles. A number of nonallelic 3^rd chromosome male sterile mutations interfere with the unfolding of the Y chromosomal lampbrush loops. Other autosomal male sterile mutations modify the morphology of these lampbrush loops. Another group of mutations inhibits the formation of the nebenkern while the development of the spermatid nucleus and the flagellum can proceed. Such male sterile mutations can decouple the development of nucleus, protein body, nebenkern, and flagellum of the spermatid. Thus, we can describe spermatogenesis inDrosophila as the coordinate execution of the individual developmental programs of the different components of the spermatozoon.     

Description of Enterobacter ludwigii sp. nov., a novel Enterobacter species of clinical relevance

A new species, Enterobacter ludwigii, is presented on the basis of the characteristics of 16 strains, which were isolated from clinical specimens. These bacteria form a distinct genetic cluster in phylogenetic analyses of the population structure of the Enterobacter cloacae complex. As determined by DNA-DNA cross-hybridization experiments in microplates, this genetic cluster can be delineated from the other species of the E. cloacae complex with deltaTm values equal to or above 5 degrees C with Enterobacter hormaechei being the closest relative. The bacteria are gram-negative, fermentative, motile rods with the general characteristics of the genus Enterobacter and the E. cloacae complex in particular. E. ludwigii can be differentiated from the other Enterobacter species by its growth on myo-inositol and 3-0-methyl-D-glucopyranose. The type strain is EN-119 (= DSM 16688T = CIP 108491T).     

Interspecific variation in the defensive responses of obligate plant-ants: experimental tests and consequences for herbivory

The aggressive behavior of ants that protect plants from herbivores in exchange for rewards such as shelter or food is thought to be an important form of biotic defense against herbivory, particularly in tropical systems. To date, however, no one has compared the defensive responses of different ant taxa associated with the same plant species, and attempted to relate these differences to longer-term efficacy of ant defense. We used experimental cues associated with herbivory—physical damage and extracts of chemical volatiles from leaf tissue—to compare the aggressive responses of two ant species obligately associated with the Amazonian myrmecophyte Tococa bullifera (Melastomataceae). We also conducted a colony removal experiment to quantify the level of resistance from herbivores provided to plants by each ant species. Our experiments demonstrate that some cues eliciting a strong response from one ant species elicited no response by the other. For cues that do elicit responses, the magnitude of these responses can vary interspecifically. These patterns were consistent with the level of resistance provided from herbivores to plants. The colony removal experiment showed that both ant species defend plants from herbivores: however, herbivory was higher on plants colonized by the less aggressive ant species. Our results add to the growing body of literature indicating defensive ant responses are stimulated by cues associated with herbivory. However, they also suggest the local and regional variation in the composition of potential partner taxa could influence the ecology and evolution of defensive mutualisms in ways that have previously remained unexplored.     

Sexual tetraploid and apomictic pentaploid populations ofHieracium pilosella (Compositae)

Five species are recognized inHieracium subgen.Pilosella sect.Pilosellina Fries. Four are diploid (2x, 2n = 18), one (H. pilosella L.) is highly variable morphologically and cytologically (from 2x to 10x), in its mode of reproduction (self-incompatibility, agamospermy, amphimixis, apo-amphimixis) and in its hybridization pattern. A part of this huge agamic complex was analysed by comparing sexual 4x and apomictic 5x plants (crossing and germination experiments, measurements of vegetative reproduction by stolons etc.). In the experimental garden apomictic 5x produced more stolons than the sexual 4x plants and the total length of the stolons per rosette was greater. However, in nature, the competitive potential of the sexual plants seems to be higher, presumably as a result of the higher mortality of ramets in 5x. Sexual 4x plants often grow in dense and grazed grass vegetation, whereas 5x apomicts often occur in dunes with patchy vegetation. Apomicts produce more capitula per rosette, and sexual rosettes form only about 60% of the number of viable achenes as compared to apomictic ones. Therefore, apomicts appear to be characterized by a greater colonizing ability than sexual plants. Apomictic plants produce equal numbers of viable achenes under conditions of both open pollination and isolation. Sexual plants do not form any viable achenes after isolation and produce a somewhat lower percentage of achenes after open pollination than do apomictics. 5xreproduce exclusively apomictically. Apo-amphimixis was never observed in pentaploids and only very rarely in tetraploids. Addition hybrids are very rare. The cross sexual 4x × apomictic 5x failed in 70% of the attempts, but the recombination of genomes carrying genes for apomixis is possible and results in apomictic 4x and sexual 5x, both with a reduced number of viable achenes. In nature sexual and apomictic plants may occur in close proximity. In such cases the germination rate of the achenes of 4x and 5x is lower; this may indicate that apomictic plants fertilize sexual plants in nature (unidirectional gene-flow). 5x plants form euploid gametes carrying two or three genomes. The results of the crossing experiments can be explained in terms ofNogler's theory of monogenic inheritance of apospory.Variation and evolution inHieracium subg.Pilosella sect.Pilosellina I.     

中国真开氏摇蚊属一新种及一新记录种记述（双翅目：摇蚊科）

记述真开氏摇蚊属Eukiefferiella Thienemann 1新种:盖真开氏摇蚊E. celata sp. nov.和1种中国新记录种: 翘叶真开氏遥蚊E. brehmi Gouin, 1943;提出兵库真开氏摇蚊E. togaeuquinta Sasa & Okazawa, 1992为翘叶真开氏摇蚊 E. brehmi Gouin,1943的新异名。