Adaptive evolution of sperm bindin tracks egg incompatibility in neotropical sea urchins of the genus Echinometra

Bindin is a gamete recognition protein known to control species-specificsperm-egg adhesion and membrane fusion in sea urchins. Previousanalyses have shown that diversifying selection on bindin aminoacid sequence is found when gametically incompatible speciesare compared, but not when species are compatible. The presentstudy analyzes bindin polymorphism and divergence in the threeclosely related species of Echinometra in Central America: E.lucunter and E. viridis from the Caribbean, and E. vanbruntifrom the eastern Pacific. The eggs of E. lucunter have evolveda strong block to fertilization by sperm of its neotropicalcongeners, whereas those of the other two species have not.As in the Indo-West Pacific (IWP) Echinometra, the neotropicalspecies show high intraspecific bindin polymorphism in the samegene regions as in the IWP species. Maximum likelihood analysisshows that many of the polymorphic codon sites are under mildpositive selection. Of the fixed amino acid replacements, mosthave accumulated along the bindin lineage of E. lucunter. Weanalyzed the data with maximum likelihood models of variationin positive selection across lineages and codon sites, and withmodels that consider sites and lineages simultaneously. Ourresults show that positive selection is concentrated along theE. lucunter bindin lineage, and that codon sites with aminoacid replacements fixed in this species show by far the highestsignal of positive selection. Lineage-specific positive selectionparalleling egg incompatibility provides support that adaptiveevolution of sperm proteins acts to maintain recognition ofbindin by changing egg receptors. Because both egg incompatibilityand bindin divergence are greater between allopatric speciesthan between sympatric species, the hypothesis of selectionagainst hybridization (reinforcement) cannot explain why adaptiveevolution has been confined to a single lineage in the AmericanEchinometra. Instead, processes acting to varying degrees withinspecies (e.g., sperm competition, sexual selection, and sexualconflict) are more promising explanations for lineage-specificpositive selection on bindin.     

PURIFICATION OF A SEX‐SPECIFIC LECTIN INVOLVED IN GAMETE BINDING OF AGLAOTHAMNION CALLOPHYLLIDICOLA (RHODOPHYTA)1

Egg and sperm binding and correct recognition is the first stage for successful fertilization. In red algae, spermatial attachment to female trichogynes is mediated by a specific binding between the lectin(s) distributed on the surface of trichogyne and the complementary carbohydrates on the spermatial surface. A female‐specific lectin was isolated from Aglaothamnion callophyllidicola by agarose‐bound fetuin affinity chromatography. Two proteins, 50 and 14 kDa, eluted from the fetuin column were separated using a native‐polyacrylamide gel electrophoresis method and subjected to a gamete binding assay. The 50 kDa protein, which blocked spermatial binding to female trichogynes, was used for further analysis. Internal amino acid sequence of the 50 kDa protein was analyzed using matrix‐assisted laser desorption/ionization‐mass spectrometry and degenerated primers were designed based on the information. A full‐length cDNA encoding the lectin was obtained using rapid amplification of cDNA ends polymerase chain reaction (PCR). The cDNA was 1552 bp in length and coded for a protein of 450 amino acids with a deduced molecular mass of 50.7 kDa, which agreed well with the protein data. Real‐time PCR analysis showed that this protein was up‐regulated about 10‐fold in female thalli. As the protein was novel and showed no significant homology to any known proteins, it was designated Rhodobindin.     

When genes move farther than offspring: gene flow by male gamete dispersal in the highly philopatric bat species Thyroptera tricolor

For species characterized by philopatry of both sexes, mate selection represents an important behaviour for inbreeding avoidance, yet the implications for gene flow are rarely quantified. Here, we present evidence of male gamete‐mediated gene flow resulting from extra‐group mating in Spix's disc‐winged bat, Thyroptera tricolor, a species which demonstrates all‐offspring philopatry. We used microsatellite and capture–recapture data to characterize social group structure and the distribution of mated pairs at two sites in southwestern Costa Rica over four breeding seasons. Relatedness and genetic spatial autocorrelation analyses indicated strong kinship within groups and over short distances (<50 m), resulting from matrilineal group structure and small roosting home ranges (~0.2 ha). Despite high relatedness among‐group members, observed inbreeding coefficients were low (F_IS = 0.010 and 0.037). Parentage analysis indicated mothers and offspring belonged to the same social group, while fathers belonged to different groups, separated by large distances (~500 m) when compared to roosting home ranges. Simulated random mating indicated mate choice was not based on intermediate levels of relatedness, and mated pairs were less related than adults within social groups on average. Isolation‐by‐distance (IBD) models of genetic neighbourhood area based on father–offspring distances provided direct estimates of mean gamete dispersal distances () > 10 roosting home range equivalents. Indirect estimates based on genetic distance provided even larger estimates of , indicating direct estimates were biased low. These results suggest extra‐group mating reduces the incidence of inbreeding in T. tricolor, and male gamete dispersal facilitates gene flow in lieu of natal dispersal of young.     

Voluntary DNA-based information exchange and contact services following donor conception: an analysis of service users’ needs

Medical science has enabled the creation of families through the use of donor conception but some lifelong policy and practice implications are only recently being recognized. Research and practice have shown that donor conception can, for some, carry substantial long-term consequences. In this paper we present findings from a questionnaire-based study that sought to shed light on donor-conceived adults’ and gamete donors’ views on service and support needs when searching for genetic relatives with the aid of DNA testing. The findings demonstrate the complexity and sensitivity of providing services in this newly emerging area of need. Such provision requires collaboration between very different disciplines and agencies (scientific and psychosocial), introduces the potential for blurring of lines of accountability and responsibility, and highlights the challenges of identifying appropriate funding streams. In addition, the findings demonstrate the opportunities and limitations afforded by the use of DNA in identifying unknown genetic relatives.     

Development of a novel recessive genetic male sterility system for hybrid seed production in maize and other cross‐pollinating crops

We have developed a novel hybridization platform that utilizes nuclear male sterility to produce hybrids in maize and other cross‐pollinating crops. A key component of this platform is a process termed Seed Production Technology (SPT). This process incorporates a transgenic SPT maintainer line capable of propagating nontransgenic nuclear male‐sterile lines for use as female parents in hybrid production. The maize SPT maintainer line is a homozygous recessive male sterile transformed with a SPT construct containing (i) a complementary wild‐type male fertility gene to restore fertility, (ii) an α‐amylase gene to disrupt pollination and (iii) a seed colour marker gene. The sporophytic wild‐type allele complements the recessive mutation, enabling the development of pollen grains, all of which carry the recessive allele but with only half carrying the SPT transgenes. Pollen grains with the SPT transgenes exhibit starch depletion resulting from expression of α‐amylase and are unable to germinate. Pollen grains that do not carry the SPT transgenes are nontransgenic and are able to fertilize homozygous mutant plants, resulting in nontransgenic male‐sterile progeny for use as female parents. Because transgenic SPT maintainer seeds express a red fluorescent protein, they can be detected and efficiently separated from seeds that do not contain the SPT transgenes by mechanical colour sorting. The SPT process has the potential to replace current approaches to pollen control in commercial maize hybrid seed production. It also has important applications for other cross‐pollinating crops where it can unlock the potential for greater hybrid productivity through expanding the parental germplasm pool.     

Characterization and biological properties of L-HGP, a glycoprotein from the amphibian oviduct with acrosome-stabilizing effects

BACKGROUND INFORMATION: The role of the jelly coat that surrounds the amphibian oocytes has been widely discussed, but is poorly understood. The presence of the jelly coat is essential for fertilization. However, the structure and function of the molecules that comprise the jelly coat have not been thoroughly documented. L-HGP (low-molecular-mass highly glycosylated protein) is a highly glycosylated protein that is present in the jelly coat of the toad, Bufo arenarum, oocytes and diffuses to the surrounding media. L-HGP, when purified from egg water, protects the sperm acrosome from breakdown induced by hypotonic solutions. RESULTS: L-HGP is an acidic glycoprotein, formed by two different subunits, linked by disulphide bonds. We raised polyclonal antibodies in rabbits against the deglycosylated protein. We determined that L-HGP is secreted along the oviduct, being hence present in all the jelly layers. The molecular mass of L-HGP is higher in the most cephalic region of the oviduct. The lower-M(r) L-HGP isoform, produced in the caudal regions of the oviduct, presents an acrosome-protecting property. L-HGP is produced by secretory cells in the oviduct and is deposited on the cilia at the oviduct lumen. CONCLUSIONS: Biochemical characterization of L-HGP has been carried out. It is synthesized by secretory cells in the oviduct and, when secreted, is deposited over the ciliated surface of the cells. The lower-M(r) isoform, secreted by the caudal region of the oviduct, protects acrosome integrity. This isoform diffuses into the medium. The role of the higher-M(r) L-HGP isoform in fertilization remains unknown.     

Multiple gene genealogies reveal asymmetrical hybridization and introgression among strongylocentrotid sea urchins

The evolution of incompatibilities between eggs and sperm is thought to play important roles in establishing and maintaining reproductive isolation among species of broadcast-spawning marine invertebrates. However, the effectiveness of gametic isolation in initiating the speciation process and/or in limiting the introgression of genes among species at later stages of divergence remains largely unknown. In the present study, we collected DNA sequence data from five loci in four species of Strongylocentrotus sea urchins ( S. droebachiensis , S. pallidus , S. purpuratus , and S. franciscanus ) to test whether the susceptibility of S. droebachiensis eggs to fertilization by heterospecific sperm results in gene flow between species. Despite the potential for introgression, a small but statistically significant signal of introgression was observed only between the youngest pair of sister taxa ( S. pallidus and S. droebachiensis ) that was strongly asymmetrical (from the former into the latter). No significant gene flow was observed for either S. purpuratus or S. franciscanus despite the ability of their sperm to readily fertilize the eggs of S. droebachiensis . Our results demonstrate that asymmetrical gamete compatibilities in strongylocentrotids can give rise to asymmetrical patterns of introgression but suggest that gamete traits alone cannot be responsible for maintaining species integrities. The genetic boundaries between strongylocentrotid urchin species in the northeast Pacific appear to be related to postzygotic isolating mechanisms that scale with divergence times and not intrinsic gametic incompatibilities per se .     

Determination of n-l-1 Gamete Transmission Rate of Trisomics and Location of Gene Controlling 2n Gamete Formation in Chinese Cabbage （Brassica rapa）

A set of trisomics of Chinese cabbage was used for determining the n-I-1 gamete transmission rate and locating the gene controlling 2n gamete formation on the corresponding chromosome. The results showed that the transmission rates of extra chromosomes in different trisomics varied from 0% to 15.38% by male gametes and from 0% to 17.39% by female gametes. Of the nine F2 populations derived from the hybridizations between each trisomic and Bp058 （2n gamete material）, only Tri- 4xBp058 showed that the segregation ratio of plants without 2n gamete formation to plants with 2n gamete formation was 10.38：1, which fitted the expected segregation ratio of the trisomics （AAa） based on the 7.37% of n＋l gamete transmission through female and 5.88% through male. In other populations the segregation ratios varied from 2.48：1 to 3.72：1, which fitted the expected 3：1 segregation ratio of the bisomics （Aa）. These results suggested that the gene controlling 2n gamete formation in Chinese cabbage Bp058 was located on chromosome 4. Further trisomic analysis based on the chromosome segregation and the incomplete stochastic chromatid segregation indicated that the gene locus was tightly linked to the centromere.     

Relationships between sperm morphometry and sperm motility in the Atlantic salmon

Relationships between spermatozoal design and swimming behaviour were investigated using the significant natural variance in sperm traits in Atlantic salmon Salmo salar. In vitro motility and fertilization experiments were conducted with 86 Atlantic salmon to measure sperm form and function under natural fertilization conditions. Spermatozoal traits of Atlantic salmon showed narrow variance within individuals but differed extensively between samples: mean sperm length varied from 32·3 to 39·5 μm, mean velocity ranged from 18 to 127 μm s⁻¹, and ejaculate longevity varied from 18 to 78 s. In addition to variation in sperm morphometry between fish, a negative relationship was also found between sperm head length and flagellum length. This natural variation in sperm form and function between males is counter-intuitive since measures are from a single Atlantic salmon population where all males are adapted to a common fertilization environment. No evidence was found that longer sperm, or sperm with longer flagella, achieved faster swimming velocities. Also no evidence was found for a trade-off between mean sperm velocity and ejaculate longevity. There were significant negative associations, however, between sperm total and flagellum length and ejaculate longevity, so that males with longer sperm had shorter-lived gametes. This finding has previously been reported in a study across fish species, supporting the theory that increased hydrostatic forces generated by longer flagella may trade against sperm cell longevity.     

Sexual reproduction: nuclear cycles and life-histories with particular reference to lower eukaryotes

Sexual reproduction is defined as the union of two haploid nuclei each derived from one of two different meioses. The implications of this definition for the interpretation of sexuality in coenocytes and homothallic haploids is discussed. The separation of the concept of the nuclear cycle from that of the life-history is advocated, and a new terminology for the nuclear cycle is proposed. The distinction between the terms life-cycle and life-history is reiterated.