Russian wheat aphid causes greater reduction in phloem transport capacity of barley leaves than bird cherry-oat aphid

The effects of feeding by the Russian wheat aphid (RWA), Diuraphis noxia Mordvilko and the Bird cherry-oat aphid (BCA), Rhopalosiphum padi L on the transport capacity of barley Hordeum vulgare L leaves were investigated and compared with a view to relating these effects to the visible symptoms shown by the respective infested plants. RWAcauses extensive chlorosis and necrosis on an infested plant whereas BCA causes no observable symptoms. Our results using the xenobiotic, phloem mobile fluorophore, 5, 6 carboxyfluorescein diacetate (5, 6-CFDA) revealed striking differences in damage to the transport of assimilates through the phloem by these two aphids. The result clearly suggests that short-term feeding by RWA causes a reduction in transport of assimilates and a more severe reduction or perhaps even permanent cessation of transport during long-term feeding. In contrast, feeding by BCA does not lead to a marked decrease in transport during short-term feeding period, however, a reduction in the transport was recorded during long-term feeding activities. These results perhaps suggest that damage to transport capacities of the barley leaves appears to be partly responsible for the observed symptoms in RWA-infested plants and the lack of them during BCAinfestations, symptoms such as reduction or cessation in transport of assimilates to growing tissues may lead to such observable symptoms

Russian wheat aphid causes greater reduction in phloem transport capacity of barley leaves than bird cherry-oat aphid

The effects of feeding by the Russian wheat aphid (RWA), Diuraphis noxia Mordvilko and the Bird cherry-oat aphid (BCA), Rhopalosiphum padi L on the transport capacity of barley Hordeum vulgare L leaves were investigated and compared with a view to relating these effects to the visible symptoms shown by the respective infested plants. RWAcauses extensive chlorosis and necrosis on an infested plant whereas BCA causes no observable symptoms. Our results using the xenobiotic, phloem mobile fluorophore, 5, 6 carboxyfluorescein diacetate (5, 6-CFDA) revealed striking differences in damage to the transport of assimilates through the phloem by these two aphids. The result clearly suggests that short-term feeding by RWA causes a reduction in transport of assimilates and a more severe reduction or perhaps even permanent cessation of transport during long-term feeding. In contrast, feeding by BCA does not lead to a marked decrease in transport during short-term feeding period, however, a reduction in the transport was recorded during long-term feeding activities. These results perhaps suggest that damage to transport capacities of the barley leaves appears to be partly responsible for the observed symptoms in RWA-infested plants and the lack of them during BCAinfestations, symptoms such as reduction or cessation in transport of assimilates to growing tissues may lead to such observable symptoms

Stronger induction of callose deposition in barley by Russian wheat aphid than bird cherry-oat aphid is not associated with differences in callose synthase or ≤-1,3-glucanase expression

The effects of infestation by the bird cherry-oat aphid (BCA), (Rhopalosiphum padi L) and the Russian wheat aphid (RWA) (Diuraphis noxia Mordvilko) on callose deposition and gene expression related to callose accumulation were investigated in barley (Hordeum vulgare L. cv. Clipper). The BCA, which gives no visible symptoms, induced very limited callose deposition, even after 14 days of infestation. In contrast, RWA, which causes chlorosis, white and yellow streaking and leaf rolling, induced callose accumulation already after 24h in longitudinal leaf veins. The deposition was pronounced after 72 h, progressing during 7 and 14 days of infestation. In RWA-infested source leaves, callose was also induced in longitudinal veins basipetal to the aphid-infested tissue, whereas in sink leaves, more callose deposition was found above the feeding sites. Nine putative callose synthase genes were identified in a data base search, of which eight were expressed in the leaves, but with similar level of expression in control and aphid-infested tissue. Four out of 12 examined β-1,3-glucanases were expressed in the leaves, and three of them were up-regulated in aphid-infested tissue. They were all more strongly induced by RWA than BCA. The results suggest that callose accumulation may be partly responsible for the symptoms resulting from RWA feeding and that a callose-inducing signal may be transported in the phloem. Furthermore it is concluded that the absence of callose deposition in BCA-infested leaves is not due to a stronger induction of callose-degrading β-1,3-glucanases in this tissue, as compared to RWA-infested leaves

Overexpression of the aphid-induced serine protease inhibitor <i>CI2c </i>gene in barley affects the generalist green peach aphid, not the specialist bird cherry-oat aphid

<div><p>Aphids are serious pests in crop plants. In an effort to identify plant genes controlling resistance against aphids, we have here studied a protease inhibitor, CI2c in barley (<i>Hordeum vulgare</i> L.). The <i>CI2c</i> gene was earlier shown to be upregulated by herbivory of the bird cherry-oat aphid <i>(Rhopalosiphum padi</i> L.<i>)</i> in barley genotypes with moderate resistance against this aphid, but not in susceptible lines. We hypothesized that CI2c contributes to the resistance. To test this idea, cDNA encoding <i>CI2c</i> was overexpressed in barley and bioassays were carried out with <i>R</i>. <i>padi</i>. For comparison, tests were carried out with the green peach aphid (<i>Myzus persicae</i> Sulzer), for which barley is a poor host. The performance of <i>R</i>. <i>padi</i> was not different on the <i>CI2c</i>-overexpressing lines in comparison to controls in test monitoring behavior and fecundity. <i>M</i>. <i>persicae</i> preference was affected as shown in the choice test, this species moved away from control plants, but remained on the <i>CI2c</i>-overexpressing lines. <i>R</i>. <i>padi</i>-induced responses related to defense were repressed in the overexpressing lines as compared to in control plants or the moderately resistant genotypes. A putative susceptibility gene, coding for a β-1,3-glucanase was more strongly induced by aphids in one of the <i>CI2c</i>-overexpressing lines. The results indicate that the CI2c inhibitor in overexpressing lines affects aphid-induced responses by suppressing defense. This is of little consequence to the specialist <i>R</i>.<i>padi</i>, but causes lower non-host resistance towards the generalist <i>M</i>. <i>persicae</i> in barley.</p></div

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals &lt;1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

The constitutive expression and induction of three β-1,3-glucanases by bird cherry-oat aphid in relation to aphid resistance in 15 barley breeding lines

In this study, 15 closely related barley genotypes were analyzed for the abundance of three β-1,3-glucanase transcripts immediately before and during infestation by the bird cherry-oat aphid (Rhopalosiphum padi L.). The barley lines are doubled haploid lines in backcross (BC) generations BC1 and BC2 from a cross between cultivar Lina and a wild barley accession. Previously, they have been characterized as susceptible (S) or resistant (R) to R. padi based on their ability to support nymphal growth. Here we also tested whether resistance was manifested as reduced aphid settling on the plants. Indeed, aphid numbers were lower on R than on S lines in all cases where there were significant differences between R and S lines. The choice of β-1,3-glucanase sequences is based on earlier results comparing two S and two R genotypes, suggesting that at least two of the three studied sequences are susceptibility factors. The comparisons of transcript abundance in plants with aphids showed for two of the β-1,3-glucanase sequences that there were several cases where an S genotype had significantly higher abundance than an R genotype, and in no case did an R line have significantly higher abundance than an S line. Thus, there was some support for the idea that β-1,3-glucanase sequences are susceptibility factors in the interaction between barley and R. padi