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Sperling, A. L.

Publications and source records attributed to Sperling, A. L..

3 recordsLinked to original sources

Expansion of chromosome F heterochromatin in parthenogenetic Drosophila mercatorum

The dramatic transition to parthenogenetic reproduction is often accompanied by similarly dramatic changes in genome organization. Genomic changes may precede or succeed the onset of parthenogenesis and may directly or indirectly contribute to the success of this reproductive transition and fitness of parthenogenetic animals. To gain greater understanding of genomic changes accompanying the transition to parthenogenetic reproduction, we have characterized genomic differences between a sexually reproducing and parthenogenetic strain of Drosophila mercatorum. This revealed three large (2.6-9 Mbp) inversions present on Muller element B and a 30-fold length expansion of heterochromatin on Muller element F. Comparisons of these genomic changes and parthenogenetic ability in 18 D. mercatorum strains collected in South America, North America, Hawaii, and Africa indicates no clear correlation of these genomic changes with geographical origin. However, these changes are minimal in strains collected in Brazil suggesting this is the origin for the ancestral strain and that both chromosome inversions and increased heterochromatin have likely preceded the transition to parthenogenetic reproduction. Parthenogenetic ability correlates most strongly with expansion of chromosomal element F. We speculate that this increased heterochromatic environment of genes on element F influences gene expression to either enhance parthenogenesis directly or redirect the activity of another factor contributing to it, in a manner analogous to the known influence of heterochromatin on gene activity in position effect variegation.

evolutionary biology↗

The SUbventral-Gland master Regulator (SUGR) of nematode virulence

All pathogens must tailor their gene expression to their environment. Therefore, targeting host:parasite biology that regulates these changes in gene expression could open up routes to pathogen control. Here, we show that in the plant-parasitic nematode Heterodera schachtii, host signals (termed effectostimulins) within plant roots activate the master regulator sugr1. SUGR1, then, directly binds effector promoters, and orchestrates their production. Effector production, in turn, facilitates host entry, releasing more effectostimulins. These data show that gene expression during the very earliest stages of parasitism is defined by a feed forward loop for host entry. Importantly, we demonstrate that blocking SUGR1 blocks parasitism, underlining the SUGR1 signalling cascade as a valuable target for crop protection. Given that nematodes also parasitise humans and other animals, the potential impact is broad: disrupting effector production could, in principle, be applied to any pathogen that secrets effectors. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=146 SRC="FIGDIR/small/576598v1_ufig1.gif" ALT="Figure 1"> View larger version (32K): org.highwire.dtl.DTLVardef@cebedaorg.highwire.dtl.DTLVardef@153fe87org.highwire.dtl.DTLVardef@16b6957org.highwire.dtl.DTLVardef@d0e920_HPS_FORMAT_FIGEXP M_FIG C_FIG

pathology↗

Whole mount multiplexed visualization of DNA, mRNA, and protein in plant-parasitic nematodes

BackgroundPlant-parasitic nematodes compromise the agriculture of a wide variety of the most common crops worldwide. Obtaining information on the fundamental biology of these organisms and how they infect the plant has been restricted by the ability to visualize intact nematodes using small molecule stains, antibodies, or in situ hybridization. Consequently, there is limited information available about the internal composition of the nematodes or the biology of the effector molecules they use to reprogram their host plant. ResultsWe present the Sperling prep -a whole mount method for nematode preparation that enables staining with small molecules, antibodies, or in situ hybridization chain reaction. This method does not require specialized apparatus and utilizes typical laboratory equipment and materials. By dissociating the strong cuticle and interior muscle layers, we enabled entry of the small molecule stains into the tissue. After permeabilization, small molecule stains can be used to visualize the nuclei with the DNA stain DAPI and the internal structures of the digestive tract and longitudinal musculature with the filamentous actin stain phalloidin. The permeabilization even allows entry of larger antibodies, albeit with lower efficiency. Finally, this method works exceptionally well with in situ HCR. Using this method, we have visualized effector transcripts specific to the dorsal gland and the subventral grand of the sugar beet cyst nematode, Heterodera schachtii, multiplexed in the same nematode. ConclusionWe were able to visualize the internal structures of the nematode as well as key effector transcripts that are used during plant infection and parasitism. Therefore, this method provides an important toolkit for studying the biology of plant-parasitic nematodes.

plant biology↗