Molecular identification and the features of genetic diversity in interspecific hybrids of Amur sturgeon ( Acipenser schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus , and A. ruthenus × A. schrenckii ) based on variability of multilocus RAPD markers

The method of polymerase chain reaction with random primers (RAPD PCR) was used to identify the progeny of the crosses between three sturgeon species, Amur sturgeon (Acipenser schrenckii Brandt, 1869), Siberian sturgeon (A. baerii Brandt, 1869), and sterlet (A. ruthenus Linnaeus, 1758). Using ten primers, genetic variation in 70 yearlings, produced in seven individual crosses: Acipenser schrenckii × A. schrenckii, A. baerii × A. baerii, A. ruthenus × A. ruthenus, A. schrenckii × A. baerii, A. baerii × A. schrenckii, A. schrenckii × A. ruthenus, and A. ruthenus × A. schrenckii was described and evaluated. It was demonstrated that the samples composed of hybrids from individual crosses were more variable than the samples of parental species. On the other hand, pooled samples of hybrids from two cross directions were genetically less variable than the pooled samples of their parents. The three main features of the hybrid RAPD profiles identified included: (1) preservation of marker DNA fragments of both parents in one genome; (2) presence of specific DNA fragments, absent from both parents; and (3) dependence of the frequency of some DNA fragments from the cross direction. Multidimensional scaling clearly distinguishes in the space of three coordinates the individuals of original species and the hybrid progeny with differentiation in the groups of direct and backcross hybrids. Analysis of relationships (UPGMA and NJ) pointed to substantial differentiation between the species, as well as between the species and hybrid progeny. Close genetic relationships between direct and backcross hybrids were demonstrated. Multilocus RAPD markers in association with statistical methods are considered to be the useful tool for discrimination of interspecific hybrids of sturgeon. Possible reasons for the differences in the hybrid RAPD profiles are discussed. Original Russian Text ? K.V. Rozhkovan, G.N. Chelomina, E.I. Rachek, 2008, published in Genetika, 2008, Vol. 44, No. 11, pp. 1453–1460.     

Comparative study (AFLP and morphology) of three species of Prosopis of the Section Algarobia: P. juliflora, P. pallida, and P. limensis. Evidence for resolution of the “P. pallida–P. juliflora complex”

The problems of delimitation of species of Prosopis originate from the few morphological discontinuities which exist among some of them; some, however, originated as a result of wide distribution of germplasm without proper knowledge of the species, in particular, much material catalogued as P. juliflora, but being of other species, was distributed for reforestation projects worldwide. This work tests the morphological results obtained for P. pallida and P. limensis of the Peruvian–Ecuadorian coast and for P. juliflora of the Caribbean Basin of Colombia and Venezuela utilizing a study of AFLPs and a study of the morphology of plantlets developed in a conventional garden study. The phenogram obtained for the AFLPs demonstrates each of the three species to be a well differentiated cluster and the molecular variance between them is significantly greater than the variance within each species. Study of the plantlets also indicates statistically significant differences for four morphological characters between P. juliflora and the other two species (P. pallida and P. limensis). These results, in addition to the morphological differentiation evident between adult plants of P. pallida and P. limensis and the clear separation of these two species from P. juliflora, corroborate the genetic identity of the three taxa analyzed.     

Introgression of the low-gossypol seed & high-gossypol plant trait in upland cotton: Analysis of [(Gossypium hirsutum?×?G. raimondii)2?×?G. sturtianum] trispecific hybrid and selected derivatives using mapped SSRs

In order to select genotypes of Gossypium hirsutum genetically balanced and expressing the low-gossypol seed & high-gossypol plant trait introgressed from the Australian wild diploid species G. sturtianum, the [(G. hirsutum × G. raimondii)2 × G. sturtianum] triple hybrid was backcrossed to G. hirsutum and autopollinated to produce backcross and selfed progenies. Two hundred and six mapped SSR markers of G. hirsutum were used to monitor the introgression of SSR alleles specific to G. sturtianum and G. raimondii in the selected progenies. A high level of heterozygosity, varying from 25 to 100%, was observed for all G. sturtianum-specific SSR markers conserved in the most advanced progenies. These results indicate the existence of segregation distortion factors that are associated with the genes controlling the researched trait. This study represents a starting point to map the genes involved in the expression of the trait and better understand its genetic determinism.     

Androgenic response of genotypes selected from Capsicum annuum L.?×?C. chinense Jacq. hybrids

In research on androhaploids in the progeny of interspecific hybrids within the Capsicum genus, three genetically stable lines of F₇ generation, selected from the C. annuum L. × C. chinense Jacq. hybrid, were used. In the first line, only callus tissue was formed as a reaction to the conditions of culture. Cytometric analysis of this tissue revealed the presence of cells with DNA content in nuclei at the level of 1C to 16C. The tissue was mixoploid and non-embryogenic. Anthers of the other line did not respond. In the third one, nine embryos were obtained, and they developed into plants. By means of cytometric analysis, the 1C DNA level was found in eight of them and these were androgenic plants. The origin of one of the diploid plant was not established due to the homozygous character of the donor plants. The experiment results confirm the already known diversity of genotype reaction to the conditions of culture. It moreover point to the possibility of selection of the forms with an androgenic potency from interspecific hybrids.     

Transformation of androgenic-derived Festulolium plants (Lolium?perenne L.?×?Festuca?pratensis Huds.) by Agrobacterium?tumefaciens

Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. × Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium “Bx351” were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and β-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling. An erratum to this article can be found at     

Production of fertile somatic hybrids of Gossypium hirsutum?+?G. bickii and G. hirsutum?+?G. stockii via protoplast fusion

Fertile somatic hybrids were obtained via symmetric electrofusion of protoplasts from two combinations of tetraploid cotton (G. hirsutum cv. Coker 201, AD genome) and diploid wild cottons G. bickii (G genome) and G. stockii (E genome), respectively. Observation by morphological, flow cytometric analysis, chromosome counting and RAPD analysis of the tested hybrids of Coker 201 + G. bickii and Coker 201 + G. stockii confirmed the regenerated plants as hybrid status. Cytological investigation of the metaphase root-tip cells revealed there were 78 chromosomes in the hybrids. Flow cytometric analysis showed the tested plants had a relative DNA contents close to the total DNA contents of the two parents. RAPD analysis revealed the hybrids contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the hybrids was intermediate between the two fusion partners. The hybrid plants were successfully transferred to the soil, and they bloomed and set bolls. It is sure that the new hexaploids developed by cell fusion would contribute to cotton breeding through backcrossing with the elite genotypes of G. hirsutum.     

Phylogeny and expression profiling of CAD and CAD-like genes in hybrid Populus (P. deltoides × P. nigra): evidence from herbivore damage for subfunctionalization and functional divergence

Background  

Cinnamyl Alcohol Dehydrogenase (CAD) proteins function in lignin biosynthesis and play a critical role in wood development and plant defense against stresses. Previous phylogenetic studies did not include genes from seedless plants and did not reflect the deep evolutionary history of this gene family. We reanalyzed the phylogeny of CAD and CAD-like genes using a representative dataset including lycophyte and bryophyte sequences. Many CAD/CAD-like genes do not seem to be associated with wood development under normal growth conditions. To gain insight into the functional evolution of CAD/CAD-like genes, we analyzed their expression in Populus plant tissues in response to feeding damage by gypsy moth larvae (Lymantria dispar L.). Expression of CAD/CAD-like genes in Populus tissues (xylem, leaves, and barks) was analyzed in herbivore-treated and non-treated plants by real time quantitative RT-PCR.     

A ThCAP gene from Tamarix hispida confers cold tolerance in transgenic Populus (P. davidiana?×?P. bolleana)

The ThCAP gene, which encodes a cold acclimation protein, was isolated from a Tamarix hispida NaCl-stress root cDNA library; its expression patterns were then assayed by qRT-PCR in different T. hispida tissues treated with low temperature (4°C), salt (400 mM NaCl), drought (20% PEG6000) and exogenous abscisic acid (100 μM). Induction of ThCAP gene was not only responsive to different stress conditions but was also organ specific. When transgenic Populus (P. davidiana × P. bolleana) plants were generated, expressing ThCAP under regulation of the cauliflower mosaic virus CaMV 35S promoter, they had a greater resistance to low temperature than non-transgenic seedlings, suggesting that ThCAP might play an important role in cold tolerance.     

Obtaining and characterization of “Holospora curviuscula” and Holospora obtusa, bacterial symbionts of the macronuclei of Paramecium bursaria and Paramecium caudatum

The symbionts of the macronuclei of Paramecium bursaria and P. caudatum, “Holospora curviuscula” 02AZ16 and H. obtusa 88Ti, respectively, were obtained and investigated. The 16S rDNA nucleotide sequences of “Holospora curviuscula” were obtained for the first time. The differences in 16S rDNA (3.4%) suggest their classification within the genus Holospora. Molecular phylogenetic analysis of the symbionts revealed that these intranuclear symbionts of the ciliates belonged to the order Rickettsiales, forming within a compact cluster of related species.     

A comparative description and distribution of the flying fishes—Cypselurus poecilopterus, C. simus, and C. callopterus, sorted out into the species group of spotwing species of the subgenus Poecilocypselurus

The three species of flying fishes—Indo-Pacific Cypselurus poecilopterus, Central Pacific C. simus, and Eastern Pacific C. callopterus form a particular species group within the subgenus Poecilocypselurus. These species rather weakly differ in the body height, number of predorsal scales and number of gill rakers. The sculls of these three species are very much alike; there are just very small differences in the proportions of some bones and the number of skull lateral line canals and pores. It could be imagined that C. poecilopterus has appeared in the Indo-Malayan Archipelago area and thence extended in all directions (up to the coasts of the Eastern Africa, Northern and Eastern Australia, New Guinea and adjacent islands, the western islands of Polynesia and the waters of Japan). C. callopterus inhabiting warm waters of the Eastern Pacific and C. simus whose area is situated in the central part of the Pacific Ocean are seemingly derived from this species.     

Analysis of storage proteins (prolamines, puroindolines and Waxy) in common wheat lines Triticum aestivum L. × (Triticum timopheevii Zhuk. × Triticum tauschii) with complex resistance to fungal infections

Storage proteins (prolamines, puroindolines, and Waxy) were studied in common wheat introgression lines obtained with the use of the Saratovskaya 29 (S29) cultivar line and synthetic hexaploid wheat (Triticum timopheevii Zhuk. × T. tauschii) (Sintetik, Sin.) displaying complex resistance to fungal infections. Comparative analysis of storage proteins in the introgression lines of common wheat Triticum aestivum L. and in the parental forms revealed the only line (BC5) having a substitution at the Gli-B2 locus from Sintetik. Hybrid lines subjected to nine backcrosses with the recurrent parental form S29 and selections for resistance to pathogens can be considered as nearly isogenic for the selected trait and retaining the allelic composition of (1) prolamines responsible for the bread-making qualitiy, (2) puroindolines associated with grain texture, and (3) Waxy proteins responsible for nutritive qualities. These lines are valuable as donors of immunity in breeding programs without the loss of the quality of flour and grain as compared to the S29 line and are also important in searching for genes determining resistance to leaf and stem rust and to powdery mildew. The amphiploid has a number of characters (silent Glu-A1 locus and Ha genotype) that can negatively affect the quality of flour and grain and thus should be taken into account when choosing this donor.     

Prevalence of Escherichia coli, Salmonella sp. and Campylobacter sp. in the intestinal flora of farm-reared, restocked and wild red-legged partridges (Alectoris rufa): is restocking using farm-reared birds a risk?

For hunting purposes, several millions of red-legged partridges (Alectoris rufa) are released each year in Spain, and these releases have the potential to introduce new parasites and disease into wild populations. We studied the prevalence of Escherichia coli, Campylobacter sp. and Salmonella sp. in the intestinal flora of red-legged partridges from three different husbandry groups: farm-reared, restocked and natural populations. Prevalence of E. coli was significantly higher in farm-reared (45%, p = 0.01) and restocked partridges (60%, p < 0.001) than in wild ones (6%, p > 0.05). The prevalence of Campylobacter sp. (23%, 100 out of 444) did not differ significantly between these three husbandry groups, and Salmonella sp. was only detected in a group of partridge chicks on one of the farms studied (0.9%, 5 out of 544). These results suggest that farm-reared and restocked partridges can act as carriers of these three enteropathogens and highlight a potential risk of transmission to natural populations via the releases of farm-reared partridges. However, future investigations are needed regarding the relation of the isolated bacteria with zoonotic strains and dissemination of antibiotic resistant microorganisms, especially E. coli, and to better evaluate the effect that these three enteropathogens have on partridge health and on the success of restocking with farm-reared birds.     

Genetic evidence for the recognition of two fiddler crabs, Uca iranica and U.?albimana (Crustacea: Brachyura: Ocypodidae), from the northwestern Indian Ocean, with notes on the U.?lactea species-complex

The status of two poorly known fiddler crabs, Uca iranica Pretzmann, 1971, from the Persian Gulf, and U. albimana (Kossmann, 1877), from the Red Sea, was studied using two mitochondrial genes: the large subunit (16S) ribosomal (r)RNA and cytochrome oxidase subunit I (COI). A molecular phylogeny shows both U. iranica and U. albimana to be members of a monophyletic U. lactea species-complex containing six taxa, with three highly supported internal clades. Uca iranica and U. albimana are the closest genetically, but are different enough to be considered valid species (16S rRNA nucleotide divergence > 7.0%, and COI > 11.9%), and form a highly supported “western” clade with U. annulipes (in line with the original morphological concept). A West Pacific “eastern” clade includes U. lactea in the north and the more widely ranging U. perplexa. An Australian endemic species, U. mjoebergi, forms a third monotypic clade.     

A wild Marsh Warbler?×?Sedge Warbler hybrid (Acrocephalus palustris?×?A. schoenobaenus) in Norway documented with molecular markers

We present photographic and molecular evidence of a wild Marsh Warbler × Sedge Warbler (Acrocephalus palustris × A. schoenobaenus) hybrid that occurred over three breeding seasons (2007–2009) near Trondheim, Central Norway. The bird had the appearance of a Marsh Warbler but with some typical Sedge Warbler plumage traits. DNA analyses of a few plucked body feathers, using the COI barcode region (mtDNA) and conserved microsatellite loci, confirmed that the bird was a hybrid, with a Marsh Warbler mother and a Sedge Warbler father.     

In?vitro evaluation of the effect of the nematophagous fungi Duddingtonia flagrans , Monacrosporium sinense and Pochonia chlamydosporia on Schistosoma mansoni eggs

The in vitro effect of four isolates of the nematophagous fungi Duddingtonia flagrans (AC 001), Monacrosporium sinense (SF 53) and Pochonia chlamydosporia (VC 1 and VC 4) on eggs of Schistosoma mansoni was examined. One thousand S. mansoni eggs were plated on 2% water–agar with the grown isolates and control without fungus. After 7, 14 and 21 days, the eggs were removed and classified according to the following parameters: type 1, lytic effect without morphological damage to eggshell; type 2, lytic effect with morphological alteration of embryo and eggshell; and type 3, lytic effect with morphological alteration of embryo and eggshell, besides hyphal penetration and internal egg colonization. Significant differences (P < 0.01) were found among the studied fungal isolates for ovicidal activity, confirming type 3 effect for the isolates VC 1 and VC 4, which characterizes the ovicidal activity of a fungus. Type 3 effect was only found for P. chlamydosporia (VC 1 and VC 4) with 26.6 and 17.2%, 25.6 and 22.6%, 27.4 and 23.9% in the 7, 14 and 21 days respectively (P < 0.01). P. chlamydosporia can thus be a potential biological control agent for S. mansoni eggs.     

Comparative cytogenetic analysis of the “Sylvaemus” group of Apodemus (Rodentia, Muridae): A. sylvaticus from Sicily and A. flavicollis from the central Apennines

Chromosome mapping of three common markers, ie major and 5S rRNA genes (rDNA) and telomeric repeats, and conventional chromosome bandings were applied to two sibling species, Apodemus sylvaticus Linnaeus, 1758 and A. flavicollis Melchior, 1834, to further investigate intra- and interspecific karyological differentiation in the genus Apodemus. A slight variation of the rDNA-patterns was detected between the two Apodemus species. In both of them, the major NORs were located on autosome pairs 8, 11, 12 and 22, while the two other rDNA sites detected on chromosomes 7 and 21 were variable in, respectively, A. sylvaticus and A. flavicollis. Several tiny rDNA sites were present on the sex chromosomes in both species, but their incidence was lower in A. flavicollis. Single 5S rDNA chromosomal sites were conserved on chromosome pair 20. No interstitial sites of telomeric repeats were present in either species. In the Sicilian population of A. sylvaticus, the constitutive heterochromatin pattern corresponded to the “sylvaticus-E1” cytotype, while A. flavicollis had a species-specific pattern restricted to centromeres of all chromosomes. The results are discussed in relation to cytogenetic data available for the genus, with emphasis on the Sylvaemus group/subgenus.     

Screening of IR50?×?Rathu Heenati F7 RILs and Identification of SSR Markers Linked to Brown Planthopper (Nilaparvata lugens St?l) Resistance in Rice (Oryza sativa L.)

Brown planthopper (Nilaparvata lugens St?l) is one of the major insect pests of rice. A Sri Lankan indica rice cultivar Rathu Heenati was found to be resistant to all biotypes of the brown planthopper. In the present study, a total of 268 F₇ RILs of IR50 and Rathu Heenati were phenotyped for their level of resistance against BPH by the standard seedbox screening test (SSST) in the greenhouse. A total of 53 SSR primers mapped on the chromosome 3 were used to screen the polymorphism between the parents IR50 and Rathu Heenati, out of which eleven were found to be polymorphic between IR50 and Rathu Heenati. The eleven primers that have shown polymorphism between the IR50 and Rathu Heenati parents were genotyped in a set of five resistant RILs and five susceptible RILs along with the parents for co-segregation analysis. Among the eleven primers, two primers namely RM3180 (18.22 Mb) and RM2453 (20.19 Mb) showed complete co-segregation with resistance. The identification of SSR markers linked with BPH resistant could be used for the maker assisted selection (MAS) program in rice breeding and to map the resistant genes on rice chromosomes for further gene cloning.