Linkage disequilibrium at the APA insecticidal seed protein locus of common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Background  

An interesting seed protein family with a role in preventing insect herbivory is the multi-gene, APA family encoding the α-amylase inhibitor, phytohemagglutinin and arcelin proteins of common bean (Phaseolus vulgaris). Variability for this gene family exists and has been exploited to breed for insect resistance. For example, the arcelin locus has been successfully transferred from wild to cultivated common bean genotypes to provide resistance against the bruchid species Zabrotes subfasciatus although the process has been hampered by a lack of genetic tools for and understanding about the locus. In this study, we analyzed linkage disequilibrium (LD) between microsatellite markers at the APA locus and bruchid resistance in a germplasm survey of 105 resistant and susceptible genotypes and compared this with LD in other parts of the genome.     

Secretion of flagellin by the LEE-encoded type III secretion system of enteropathogenic Escherichia coli

Background  

Enteropathogenic Escherichia coli (EPEC) is an attaching and effacing (A/E) pathogen that possesses a type III secretion system (T3SS) encoded within the locus of enterocyte effacement (LEE). The LEE is essential for A/E lesion formation and directs the secretion and translocation of multiple LEE-encoded and non-LEE encoded effector proteins into the cytosol of infected cells. In this study we used proteomics to compare proteins exported to the culture supernatant by wild type EPEC E2348/69, a ΔespADB mutant and a ΔescF T3SS mutant.     

Estimation of loci involved in non-shattering of seeds in early rice domestication

Rice (Oryza sativa L.) is widely cultivated around the world and is known to be domesticated from its wild form, O. rufipogon. A loss of seed shattering is one of the most obvious phenotypic changes selected for during rice domestication. Previously, three seed-shattering loci, qSH1, sh4, and qSH3 were reported to be involved in non-shattering of seeds of Japonica-type cultivated rice, O. sativa cv. Nipponbare. In this study, we focused on non-shattering characteristics of O. sativa Indica cv. IR36 having functional allele at qSH1. We produced backcross recombinant inbred lines having chromosomal segments from IR36 in the genetic background of wild rice, O. rufipogon W630. Histological and quantitative trait loci analyses of abscission layer formation were conducted. In the analysis of quantitative trait loci, a strong peak was observed close to sh4. We, nevertheless, found that some lines showed complete abscission layer formation despite carrying the IR36 allele at sh4, implying that non-shattering of seeds of IR36 could be regulated by the combination of mutations at sh4 and other seed-shattering loci. We also genotyped qSH3, a recently identified seed-shattering locus. Lines that have the IR36 alleles at sh4 and qSH3 showed inhibition of abscission layer formation but the degree of seed shattering was different from that of IR36. On the basis of these results, we estimated that non-shattering of seeds in early rice domestication involved mutations in at least three loci, and these genetic materials produced in this study may help to identify novel seed-shattering loci.     

Genetic analysis of hybridization and introgression between wild mongoose and brown lemurs

Background  

Hybrid zones generally represent areas of secondary contact after speciation. The nature of the interaction between genes of individuals in a hybrid zone is of interest in the study of evolutionary processes. In this study, data from nuclear microsatellites and mitochondrial DNA sequences were used to genetically characterize hybridization between wild mongoose lemurs (Eulemur mongoz) and brown lemurs (E. fulvus) at Anjamena in west Madagascar.     

Comparative analysis of <Emphasis Type="Italic">Mycobacterium avium</Emphasis> subsp. <Emphasis Type="Italic">paratuberculosis</Emphasis> isolates from cattle,sheep and goats by short sequence repeat and pulsed-field gel electrophoresis typing

Background  

Mycobacterium avium subsp. paratuberculosis (Map) causes the chronic enteritis called paratuberculosis mainly in cattle, sheep and goats. Evidences that point out an association between Map and Crohn's Disease in humans are increasing. Strain differentiation among Map isolates has proved to be difficult and has limited the study of the molecular epidemiology of paratuberculosis. In order to asses the usefulness of the PCR based short sequence repeat (SSR) analysis of locus 1 and locus 8 in the epidemiological tracing of paratuberculosis strains we here compare for the first time the results of SSR and SnaBI-SpeI pulsed-field gel electrophoresis (PFGE) typing methods in a set of 268 Map isolates from different hosts (cattle, sheep, goats, bison, deer and wild boar).     

Quorum Sensing Contributes to Seed‐to‐Seedling Transmission of Acidovorax citrulli on Watermelon

Based on the observation that Acidovorax citrulli switches from saprobic to pathogenic growth for seed‐to‐seedling transmission of bacterial fruit blotch of cucurbits (BFB), we hypothesized that quorum sensing (QS) was involved in the regulation of this process. Using aacI (luxI homologue) and aacR (luxR homologue) mutants of AAC00‐1, we investigated the role of QS in watermelon seed colonization and seed‐to‐seedling transmission of BFB. aacR and aacI mutants of AAC00‐1 colonized germinating watermelon seed at wild‐type levels; however, BFB seed‐to‐seedling transmission was affected in a cell density‐dependent manner. There were no significant differences in BFB seedling transmission between watermelon seed infiltrated with approximately 1 × 10⁶ CFU of AAC00‐1, the aacR or aacI deletion mutants (95.2, 94.9 and 98.3% BFB incidence, respectively). In contrast, when seed inoculum was reduced to approximately 1 × 10³ CFU/seed, BFB seed‐to‐seedling transmission declined to 34.3% for the aacI mutant, which was significantly less than the wild type (78.6%). Interestingly, BFB seed‐to‐seedling transmission for the aacR mutant was not significantly different to the wild‐type strain. These data suggest that QS plays a role in regulation of genes involved in seed‐to‐seedling transmission of BFB.     

The origin and evolution of <Emphasis Type="Italic">ARGFX</Emphasis>homeobox loci in mammalian radiation

Background  

Many homeobox genes show remarkable conservation between divergent animal phyla. In contrast, the ARGFX (Arginine-fifty homeobox) homeobox locus was identified in the human genome but is not present in mouse or invertebrates. Here we ask when and how this locus originated and examine its pattern of molecular evolution.     

Invasin of Edwardsiella tarda is essential for its haemolytic activity,biofilm formation and virulence towards fish

Aims

The aim of this study was to investigate the role of invasin in a bacterial fish pathogen Edwardsiella tarda.

Methods and Results

In this study, an in‐frame deletion mutant of invasin (Δinv) in Edw. tarda H1 was constructed through double crossover allelic exchange to explore the function of invasin in virulence to fish. Meanwhile, an invasin overexpression strain (inv⁺) was obtained by electrotransformation of a low‐copy plasmid pACYC184 carrying the intact invasin into the Δinv mutant. Several virulence‐associated characters of the mutants and wild‐type strain were tested. Compared with the wild‐type H1, haemolytic activity and biofilm formation were decreased in Δinv, while increased significantly in inv⁺. In addition, the invasin overexpressing strain inv⁺ exhibited increased internalization into Epithelioma Papulosum Cyprini (EPC) cells. Moreover, in zebrafish model, Δinv showed decreased virulence compared with H1, while inv⁺ restored the virulence of wild type completely.

Conclusions

The results demonstrated that invasin of Edw. tarda plays essential roles in haemolytic activity, biofilm formation, adherence, internalization and pathogenicity of this bacterium.

Significance and Impact of the Study

This study revealed the role of invasin in Edw. tarda infection and provided useful information for further unveiling the pathogenesis of Edw. tarda.     

Mechanisms of haplotype divergence at the <Emphasis Type="Italic">RGA08</Emphasis> nucleotide-binding leucine-rich repeat gene locus in wild banana <Emphasis Type="Italic">(Musa balbisiana)</Emphasis>

Background  

Comparative sequence analysis of complex loci such as resistance gene analog clusters allows estimating the degree of sequence conservation and mechanisms of divergence at the intraspecies level. In banana (Musa sp.), two diploid wild species Musa acuminata (A genome) and Musa balbisiana (B genome) contribute to the polyploid genome of many cultivars. The M. balbisiana species is associated with vigour and tolerance to pests and disease and little is known on the genome structure and haplotype diversity within this species. Here, we compare two genomic sequences of 253 and 223 kb corresponding to two haplotypes of the RGA08 resistance gene analog locus in M. balbisiana "Pisang Klutuk Wulung" (PKW).     

Polymorphism at the apical membrane antigen 1 locus reflects the world population history of <Emphasis Type="Italic">Plasmodium vivax</Emphasis>

Background  

In malaria parasites (genus Plasmodium), ama-1 is a highly polymorphic locus encoding the Apical Membrane Protein-1, and there is evidence that the polymorphism at this locus is selectively maintained. We tested the hypothesis that polymorphism at the ama-1 locus reflects population history in Plasmodium vivax, which is believed to have originated in Southeast Asia and is widely geographically distributed. In particular, we tested for a signature of the introduction of P. vivax into the New World at the time of the European conquest and African slave trade and subsequent population expansion.     

Post-pruning shoot growth increases fruit abscission and reduces stem carbohydrates and yield in macadamia

Background and Aims

There is good evidence for deciduous trees that competition for carbohydrates from shoot growth accentuates early fruit abscission and reduces yield but the effect for evergreen trees is not well defined. Here, whole-tree tip-pruning at anthesis is used to examine the effect of post-pruning shoot development on fruit abscission in the evergreen subtropical tree macadamia (Macadamia integrifolia, M. integrifolia × tetraphylla). Partial-tree tip-pruning is also used to test the localization of the effect.

Methods

In the first experiment (2005/2006), all branches on trees were tip-pruned at anthesis, some trees were allowed to re-shoot (R treatment) and shoots were removed from others (NR treatment). Fruit set and stem total non-structural carbohydrates (TNSC) over time, and yield were measured. In the second experiment (2006/2007), upper branches of trees were tip-pruned at anthesis, some trees were allowed to re-shoot (R) and shoots were removed from others (NR). Fruit set and yield were measured separately for upper (pruned) and lower (unpruned) branches.

Key Results

In the first experiment, R trees set far fewer fruit and had lower yield than NR trees. TNSC fell and rose in all treatments but the decline in R trees occurred earlier than in NR trees and coincided with early shoot growth and the increase in fruit abscission relative to the other treatments. In the second experiment, fruit abscission on upper branches of R trees increased relative to the other treatments but there was little difference in fruit abscission between treatments on lower branches.

Conclusions

This study is the first to demonstrate an increase in fruit abscission in an evergreen tree in response to pruning. The effect appeared to be related to competition for carbohydrates between post-pruning shoot growth and fruit development and was local, with shoot growth on pruned branches having no effect on fruit abscission on unpruned branches.     

Differences in the BAL proteome after <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> infection in wild type and SP-A-/- mice

Background  

Surfactant protein-A (SP-A) has been shown to play a variety of roles related to lung host defense function. Mice lacking SP-A are more susceptible to infection than wild type C57BL/6 mice. We studied bronchoalveolar lavage (BAL) protein expression in wild type and SP-A-/- mice infected with Klebsiella pneumoniae by 2D-DIGE.     

Wild peas vary in their cross-compatibility with cultivated pea (Pisum sativum subsp. sativum L.) depending on alleles of a nuclear–cytoplasmic incompatibility locus

Key message

Divergent wild and endemic peas differ in hybrid sterility in reciprocal crosses with cultivated pea depending on alleles of a nuclear ‘speciation gene’ involved in nuclear–cytoplasmic compatibility.

Background

In hybrids between cultivated and wild peas, nuclear–cytoplasmic conflict frequently occurs. One of the nuclear genes involved, Scs1, was earlier mapped on Linkage Group III.

Results

In reciprocal crosses of seven divergent pea accessions with cultivated P. sativum, some alleles of Scs1 manifested incompatibility with an alien cytoplasm as a decrease in pollen fertility to about 50 % in the heterozygotes and lack of some genotypic classes among F2 segregants. Earlier, we defined monophyletic evolutionary lineages A, B, C and D of pea according to allelic state of three markers, from nuclear, plastid and mitochondrial genomes. All tested representatives of wild peas from the lineages A and C exhibited incompatibility due to Scs1 deleterious effects in crosses with testerlines of P. sativum subsp. sativum (the common cultivated pea) at least in one direction. A wild pea from the lineage B and a cultivated pea from the lineage D were compatible with the testerline in both directions. The tested accession of cultivated P. abyssinicum (lineage A) was partially compatible in both directions. The Scs1 alleles of some pea accessions even originating from the same geographic area were remarkably different in their compatibility with cultivated Pisum sativum cytoplasm.

Conclusion

Variability of a gene involved in reproductive isolation is of important evolutionary role and nominate Scs1 as a speciation gene.     

Molecular evolution of dimeric α-amylase inhibitor genes in wild emmer wheat and its ecological association

Background  

α-Amylase inhibitors are attractive candidates for the control of seed weevils, as these insects are highly dependent on starch as an energy source. In this study, we aimed to reveal the structure and diversity of dimeric α-amylase inhibitor genes in wild emmer wheat from Israel and to elucidate the relationship between the emmer wheat genes and ecological factors using single nucleotide polymorphism (SNP) markers. Another objective of this study was to find out whether there were any correlations between SNPs in functional protein-coding genes and the environment.     

Occurrence of Alternaria japonica on Seeds of Wild and Cultivated Rocket

In vitro evaluation was carried out on seed samples of wild and cultivated rocket cultivars, most frequently grown in Italy, and obtained from farms affected by the leaf spot caused by Alternaria japonica in Piedmont and Lombardy during the fall of 2010. Twelve seed samples were collected and assayed for the presence of A. japonica. The pathogen was isolated only from not disinfected seeds. Among the two seed samples of cultivated rocket (Eruca vesicaria), only one was infected by A. japonica at a level of one infected seed out of 800. Four out of ten samples of wild (Diplotaxis tenuifolia) rocket seeds were contaminated by A. japonica with the highest level of infection detected in a single sample of 3 out of 800. All tested isolates of A. japonica obtained from seeds were pathogenic on both wild and cultivated rocket.     

Diversity and selective sweep in the OsAMT1;1 genomic region of rice

Background  

Ammonium is one of the major forms in which nitrogen is available for plant growth. OsAMT1;1 is a high-affinity ammonium transporter in rice (Oryza sativa L.), responsible for ammonium uptake at low nitrogen concentration. The expression pattern of the gene has been reported. However, variations in its nucleotides and the evolutionary pathway of its descent from wild progenitors are yet to be elucidated. In this study, nucleotide diversity of the gene OsAMT1;1 and the diversity pattern of seven gene fragments spanning a genomic region approximately 150 kb long surrounding the gene were surveyed by sequencing a panel of 216 rice accessions including both cultivated rice and wild relatives.     

A mutation at the rb gene, lowering ADPGPPase activity, affects storage product metabolism of pea seed coats

Seed coat development was studied on two nearisogenic linesof peas (Pisum sativum L.): RbRb (wild type, round seed) andrbrb (wrinkled seed). A mutation at the rb locus modifies thedry seed shape and reduces the starch content of the embryo.This mutation is now known to affect the activity of ADPGlucosepyrophosphorylase, a key enzyme in the starch biosynthetic pathway.We have investigated the effects of the rb mutation on seedcoat development and found that the mutation reduces the growthrate and starch content in this organ. However, experimentson the kinetics of ¹⁴C-sucrose loading showed that starch synthesisfrom unloaded sucrose occurred in the seed coat for both mutantand wild-type lines. In addition, the sucrose concentrationwas increased and amino acid concentration decreased such thatthe nutritional balance of the embryos was affected. However,osmolality of the seed coat cells was not affected, suggestinga regulatory process which allows the maintenance of the importof assimilates in the seeds of either line. Key words: ADPGlucose pyrophosphorylase, seed coat, seed development, starch metabolism, wrinkled seed