Assessing genetic identity of sporophytic offspring of the brown alga Undaria pinnatifida derived from mono-crossing of gametophyte clones by use of amplified fragment length polymorphism and microsatellite markers

The haploid stage of gametophytes of the subtidal brown alga Undaria pinnatifida can be vegetatively propagated under favorable conditions. This unique characteristic makes it possible to establish independent gametophyte cell lines that are zoospore-derived. Sporophytic offspring can be generated through hybridizing the male and female gametophytes, which are derived from different cell lines. Accumulated experiences in this and other species in Laminariales demonstrated the applicability of this novel way to breed desired strains for open-sea cultivation. Sporophytic offspring originated from mono-crossing of male and female gametophyte clones were shown to have similar morphological characteristics under identical ambient conditions. However, there has been no report to relate this similarity on molecular levels. In this report, amplified fragment length polymorphism (AFLP) and microsatellite markers were used to analyze the genetic identity of sporophytic offspring of U. pinnatifida originated from two mono-crossing lines (M1 and M2), two self-breeding lines (S1 and S2) and one wild population (W). Totally 318 AFLP loci were revealed by use of 11 primer sets, of which 4.7%, 0.3%, 17.9%, 16.4% and 36.5% were polymorphic in M1, M2, S1, S2 and W, respectively. The pairwise genetic identity among the individuals of the same line was assessed. It was shown that offspring from mono-crossing lines had a higher degree of identity (95.6-100%) than self-breeding lines (87.7-98.4%) and the wild population (81.5-92.1%). Analysis by use of six microsatellite loci also revealed a higher genetic identity among individuals of the mono-crossing line, further confirming the results of AFLP analysis. Results from this investigation support, on molecular levels, the novel way to produce and maintain strains in U. pinnatifida by use of different gametophyte cell lines.

Characterization of two membrane-associated protease genes obtained from screening out-membrane protein genes of Flavobacterium columnare G(4)

In order to identify genes encoding the outer membrane proteins (OMPs) of the myxobacter Flavobacterium columnare G(4), the expression library of the bacterium was screened by using rabbit antisera developed against its OMPs. Positive colonies of Escherichia coli M15 containing fragments encoding the bacterial OMPs were selected for cloning the relevant genes by genomic walking methods. Two genes encoding a membrane-associated zinc metalloprotease and prolyl oligopeptidase are reported in this paper. The membrane-associated zinc metalloprotease gene (map) is 1800 bp in length, coding for 449 amino acids (aa). Despite the presence of a conserved motif HEXXH for all metalloproteases, the special HEXXH similar to 32 aa similar to E motif of the F. columnare G(4) Map and its low level of identity with other reported zinc-containing metalloproteases may imply that the membrane-associated zinc metalloprotease of F. columnare G(4) represents a new family of zincins. The gene encoding prolyl oligopeptidase (Pop), a serine proteinase, is 2352 bp in length, coding for 649 aa. Sequence homology analysis revealed that the Pop is also novel as it has <50% identity with other reported prolyl oligopeptidase family proteins. The present study represents the first to employ anti-fish bacterial OMP sera to screen genes of membrane-associated proteases of fish pathogenic bacteria, and to provide necessary information for the examination of the role of the two genes in the infection and pathogenesis of F. columnare.

Molecular cloning and response to laminarin stimulation of arginine kinase in haemolymph in Chinese shrimp, Fenneropenaeus chinensis

Arginine kinase (AK) was previously reported as a phosphagen-ATP phosphotransferase found in invertebrates. In this study, an 1184 bp cDNA was cloned and sequenced. It contained an open reading frame of 1068 bp that coded for 356 deduced amino acids of AK in Fenneropenaeus chinensis. The calculated molecular mass of AK is 40129.73 Da and pI is 5.92. The predicted protein showed a high level of identity to known AK in invertebrates and creatine kinase from vertebrates, which belong to a conserved family of ATP:guanidino phospho-transferases. In addition, AK protein in plasma of F. chinensis was identified using two-dimensional electrophoresis (2DE) and electrospray ionization mass spectrometry (ESI-MS) according to the calculated molecular mass and pI. AK was significantly decreased in the plasma of F. chinensis at 45 min and recovered at 3 It after laminarin injection as confirmed by 2DE and ESI-MS. The results showed that AK was one of the most significantly changed proteins on two-dimensional gel in the plasma proteins of F. chinensis at 45 min and 3 It after simulation. (c) 2005 Elsevier Ltd. All rights reserved.

面孔身份与面部表情识别中的区分度效应

Whether facial identity and facial expression was processed independently has long been a controversy. Studies at levels of experimental, neuropsychological, functional imaging and cell-recording all failed to consistently support either independent or interdependent processing. Present study proposed that familiarity and discriminability of facial identity and expression was important variable in mediating the relation between facial identity and facial expression recognition. Effects of familiarity on recognition of facial identity and expression had been examined (e.g. Ganel & Goshen-Gottstein, 2004) but the role of the discriminability in recognition of facial identity and expression has not yet been carefully examined. To examine the role of discriminability of facial identity and expression, 8 experiments were conducted with Garner’s speeded classification task in recognition of identity and expression of unfamiliar faces. The discriminability of facial identity and expression was manipulated, and the measurements of Garner interference and facilitation indicated that: 1. The discriminability of facial identity and expression mediate the relation between facial identity and expression recognition. Four possible discriminability combinations between identity and expression predicted 4 interference patterns between them. Low discriminability accounted for the interference either in facial identity judgment or in facial expression judgment task. 2. The measurements of eye movements indicated that either in facial identity or in facial expression recognition low discriminability led to a narrowly-distributed eye fixation pattern while high discriminability led to a widely-distributed eye fixation pattern. 3. By combining the morphing technique with the Garner paradigm, study 2 successfully demonstrated the linar relation between discriminability and Garner facilitation effects, confirmed the discriminability effects in the measurements of Garner facilitation effects.. 4. By providing the varying information of facial expression, study 2 revealed that varying information improved the discriminability of facial expression, and then enhanced the recognition of facial expression. All the results indicated that the discriminability of facial identity and expression could mediate the independent or interdependent processing between them, and the discriminability effects on recognition of identity and expression of unfamiliar faces was identified. The results from interference effects and facilitation effects both indicated that the dimensional relation between facial identity and expression was separable but not asymmetric claimed by previous studies（Schweinberger et al, 1998, 1999）. Absolutedly independent or interdependent processing between facial identity and expression recognition were both impossible, discriminability of identity and expression mediated the relation between them. The discriminability effects revealed in present study could explain the conflicts between existing findings well.