Rab11-FIP3 binds dynein light intermediate chain 2 and its overexpression fragments the Golgi complex

The mechanochemical forces that move and position intracellular organelles and their intermediates in eukaryotic cells are provided by molecular motor proteins which include the cytoplasmic dynein-1 motor complex. Recently, we identified the Rab11 GTPase effector protein Rab11-FIP3 (henceforth, FIP3) as a novel binding-partner for dynein light intermediate chain 1 (DLIC-1, gene symbol DYNC1LI1), a subunit of cytoplasmic dynein-1. Here, we show that FIP3 also binds the dynein light intermediate chain 2 subunit (DLIC-2, gene symbol DYNC1LI2). We show that like DLIC-1, DLIC-2 binds the amino-terminal 435 amino acids of FIP3 and that FIP3 links Rab11a to DLIC-2. We also show that FIP3 recruits DLIC-2 onto membranes and that DLIC-2 is necessary for the accumulation of endocytosed-transferrin (Tfn) at the pericentrosomal endosomal-recycling compartment (ERC). Finally, we demonstrate that overexpression of FIP3 fragments the Golgi complex by sequestering cytoplasmic dynein-1. In conclusion, we have identified FIP3 as the first membrane-associated interacting-partner for DLIC-2 and propose that this interaction serves to control endosomal trafficking from sorting endosomes to the ERC.     

Trade-off between foliage and tuber resistance to Phthorimaea operculella in wild potatoes

Beet leafhopper, Circulifer tenellus (Baker) (Homoptera: Cicadellidae), is the only known North American vector of beet curly top virus (Geminiviridae), which causes major economic losses in a number of crops including sugar beet, tomato, beans, and peppers. Beet curly top virus is a phloem-limited, persistently transmitted, circulative geminivirus. The strain/species of curly top virus used in this study is the CFH strain, also referred to as beet severe curly top virus (BSCTV). The direct current (DC) electrical penetration graph technique was used to determine the specific stylet penetration behavior associated with inoculation of BSCTV. Viruliferous leafhoppers were allowed to feed on healthy 3–4-week-old sugar beet plants until specific electrical penetration graph waveforms were produced, at which point feeding was artificially terminated. A series of comparisons between leafhoppers that produced different combinations of waveforms clearly implicated waveform D1 as the only waveform correlated with inoculation of BSCTV. All successful inoculations contained waveform D1, and 56 out of 64 leafhoppers that produced waveform D1 successfully inoculated test plants. Eighty-five leafhoppers did not produce waveform D1 and none of these inoculated BSCTV. While the occurrence of waveform D1 appears to be necessary for BSCTV inoculation, there was no correlation between duration of waveform D1 and inoculation success rate. The correlation of waveform D1 and BSCTV inoculation found in this study implies that waveform D1 is associated with phloem salivation.     

Symbiont-mediated adaptation by planthoppers and leafhoppers to resistant rice varieties

For over 50 years, host plant resistance has been the principal focus of public research to reduce planthopper and leafhopper damage to rice in Asia. Several resistance genes have been identified from native varieties and wild rice species, and some of these have been incorporated into high-yielding rice varieties through conventional breeding. However, adaptation by hoppers to resistant rice has been phenomenally rapid, and hopper populations with virulence against several resistance genes are now widespread. Directional genetic selection for virulent hoppers seems unlikely given the rapid pace of adaptation reported from field and laboratory studies. Among the alternative explanations for rapid hopper adaptation are changes (genetic, epigenetic, or community structure) in endosymbiont communities that become advantageous for planthoppers and leafhoppers that feed on resistant rice varieties. This review examines the nature of these symbiont communities and their functions in planthoppers and leafhoppers—focusing on their likely roles in mediating adaptation to plant resistance. Evidence from a small number of experimental studies suggests that bacterial and eukaryotic (including yeast-like) symbionts can determine or mediate hopper virulence on rice plants and that symbiont functions could change over successive generations of selection on both resistant and susceptible plants. The review highlights the potential complexity of rice hopper–symbiont interactions and calls for a more careful choice of research materials and methods to help reduce this complexity. Finally, the consequences of symbiont-mediated virulence adaptation for future rice breeding programs are discussed.     

GRAB is a binding partner for the Rab11a and Rab11b GTPases

Co-ordination of Rab GTPase function has emerged as a crucial mechanism in the control of intracellular trafficking processes in eukaryotic cells. Here, we show that GRAB/Rab3IL1 [guanine nucleotide exchange factor for Rab3A; RAB3A interacting protein (rabin3)-like 1], a protein that has previously be shown to act as a GEF (guanine nucleotide exchange factor) for Rab3a, Rab8a and Rab8b, is also a binding partner for Rab11a and Rab11b, but not the closely related Rab25 GTPase. We demonstrate that exogenous expression of Rab11a and Rab11b shift GRAB’s distribution from the cytoplasm onto membranes. We find that the Rab11a/Rab11b-binding region of GRAB lies within its carboxy-terminus, a region distinct from its GEF domain and Rab3a-binding region. Finally, we describe a GRAB deletion mutant (GRAB_Δ223–228) that is deficient in Rab11-binding ability. These data identify GRAB as a dual Rab-binding protein that could potentially link Rab3 and Rab11 and/or Rab8 and Rab11-mediated intracellular trafficking processes.     

Uptake and metabolism of 6-benzylaminopurine in shoot cultures of

Shoots of Gerbera jamesonii Bolus, cultured in vitro were induced to multiply by the addition of 2.22 μ M 6-benzylaminopurine (BAP) to the medium. Shoots growing in the presence of [¹⁴C]-BAP were harvested every 4 days and sub-divided into petioles plus apices, laminae and basal callus. The 3- and 9-glucosides, 9-riboside and a novel compound, the 9-ribosylglucoside were characterised as metabolites of BAP by mass spectrometry. The 9-riboside was the principal metabolite formed initially in petioles plus apicies and callus but further metabolism varied depending on tissue type. After 24 days the major metabolite in the callus was the 9-ribosylglucoside, with only trace amounts of the other metabolites present. In contrast, both 3-glucoside and 9-ribosylglucoside accumulated in petioles plus apices whereas the 3-glucoside accumulated in laminae.     

Effect of Dietary Prebiotic (Mannan Oligosaccharide) Supplementation on the Caecal Bacterial Community Structure of Turkeys

The identification of specific bacterial species influenced by mannan oligosaccharide (MOS) supplementation may assist in the formulation of new and improved diets that promote intestinal health and improve bird performance, offering suitable alternatives to antimicrobials in feed for sustainable poultry production. This study has been conducted to evaluate the use of a MOS compound derived from the yeast cell wall of Saccharomyces cerevisiae on turkey performance, bacterial community structure and their phylogenetic associations. A 42-day turkey trial was carried out on birds fed control and MOS-supplemented diets. Bird performance data (weight gains, feed consumption and feed efficiency ratios) were collected, and caecal contents were extracted from randomly caught poults on days 28, 35 and 42 posthatch. Bird performance data showed no improvements as a result of dietary supplementation. Automated ribosomal intergenic spacer analysis (ARISA) revealed the bacterial community structure to be significantly altered on days 28 and 35 posthatch but not day 42 as a result of dietary supplementation. This technique was coupled with 16S rRNA gene sequence analysis to elucidate phylogenetic identities of bacteria. The dominant bacteria of the caecum on all days in both treatment groups were members of phylum Firmicutes, followed by the Bacteroidetes and Proteobacteria phyla, respectively. Statistical analysis of the 16S rRNA gene libraries showed that the composition of the MOS clone library differed significantly to the control on day 35 posthatch. It can be concluded that MOS alters the bacterial community structure in the turkey caecum.     

Transient association of MCM complex proteins with the nuclear matrix during initiation of mammalian DNA replication

The minichromosome maintenance complex (MCM2-7) is the putative DNA helicase in eukaryotes, and essential for DNA replication. By applying serial extractions to mammalian cells synchronized by release from quiescence, we reveal dynamic changes to the sub-nuclear compartmentalization of MCM2 as cells pass through late G1 and early S phase, identifying a brief window when MCM2 becomes transiently attached to the nuclear-matrix. The data distinguish 3 states that correspond to loose association with chromatin prior to DNA replication, transient highly stable binding to the nuclear-matrix coincident with initiation, and a post-initiation phase when MCM2 remains tightly associated with chromatin but not the nuclear-matrix. The data suggests that functional MCM complex loading takes place at the nuclear-matrix.     

Comparative Efficiency of the Fenwick Can and Schuiling Centrifuge in Extracting Nematode Cysts from Different Soil Types

The Fenwick can and Schuiling centrifuge are widely used to extract nematode cysts from soil samples. The comparative efficiencies of these two methods during cyst extraction have not been determined for different soil types under different cyst densities. Such information is vital for statutory laboratories that must choose a method for routine, high-throughput soil monitoring. In this study, samples of different soil types seeded with varying densities of potato cyst nematode (Globodera rostochiensis) cysts were processed using both methods. In one experiment, with 200 ml samples, recovery was similar between methods. In a second experiment with 500 ml samples, cyst recovery was higher using the Schuiling centrifuge. For each method and soil type, cyst extraction efficiency was similar across all densities tested. Extraction was efficient from pure sand (Fenwick 72%, Schuiling 84%) and naturally sandy soils (Fenwick 62%, Schuiling 73%), but was significantly less efficient from clay-soil (Fenwick 42%, Schuiling 44%) and peat-soil with high organic matter content (Fenwick 35%, Schuiling 33%). Residual moisture (<10% w/w) in samples prior to analyses reduced extraction efficiency, particularly for sand and sandy soils. For each soil type and method, there were significant linear relationships between the number of cysts extracted and the numbers of cysts in the samples. We discuss the advantages and disadvantages of each extraction method for cyst extraction in statutory soil laboratories.