Bringing in the controversy : re-politicizing the de-politicized strategy of ethics committees

Human/animal relations are potentially controversial and biotechnologically produced animals and animal-like creatures – bio-objects such as transgenics, clones, cybrids and other hybrids – have often created lively political debate since they challenge established social and moral norms. Ethical issues regarding the human/animal relations in biotechnological developments have at times been widely debated in many European countries and beyond. However, the general trend is a move away from parliamentary and public debate towards institutionalized ethics and technified expert panels. We explore by using the conceptual lens of bio-objectification what effects such a move can be said to have. In the bio-objectification process, unstable bio-object becomes stabilized and receives a single “bio-identity” by closing the debate. However, we argue that there are other possible routes bio-objectification processes can take, routes that allow for more open-ended cases. By comparing our observations and analyses of deliberations in three different European countries we will explore how the bio-objectification process works in the context of animal ethics committees. From this comparison we found an interesting common feature: When animal biotechnology is discussed in the ethics committees, technical and pragmatic matters are often foregrounded. We noticed that there is a common silence around ethics and a striking consensus culture. The present paper, seeks to understand how the bio-objectification process works so as to silence complexity through consensus as well as to discuss how the ethical issues involved in animal biotechnology could become re-politicized, and thereby made more pluralistic, through an “ethos of controversies”

Simultaneous silencing of <i>GaPDS</i> and <i>GaANR</i> in a single plant with the VIGS system.

<p>A, The phenotypes of plants inoculated with pYL156 (i), pYL156:<i>PDS</i> + pYL156:<i>ANR</i> (ii), pYL156:<i>PDS-ANR</i> (iii), and pYL156:<i>PDS</i> (iv), and pYL156:<i>ANR</i> (v). B, Relative transcript levels of <i>PDS</i> and <i>ANR</i> in systemic leaves of plants infiltrated with pYL156:<i>PDS</i>, pYL156:<i>ANR</i>, and pYL156:<i>PDS</i> + pYL156:<i>ANR</i>, and pYL156:<i>PDS-ANR</i>. The CK value was set as 100%. C, Relative levels of TRV RNA2 in systemic leaves of plants infiltrated with pYL156:<i>PDS</i>, pYL156:<i>ANR</i>, and pYL156:<i>PDS</i> + pYL156:<i>ANR</i>, and pYL156:<i>PDS-ANR</i>. The CK value at 10 d post-inoculation (dpi) was set at 1. Error bars represent standard deviations (n = 3 biological replicates) in (B) and (C).</p

Optimal factors for Agrobacterium-mediated <i>GaPDS</i> VIGS in <i>G</i><i>. barbadense</i>.

<p>A, The photobleaching phenotype of cotton leaves triggered by <i>GaPDS</i> VIGS. The pYL156 vector was used as a vector control; WT, wild type. B–E, The percentage of plants showing photobleaching was affected by light intensity, photoperiod, seedling age, and OD value of Agrobacterium cultures. Means ± standard deviation labeled with different letters are significantly different at the 0.05 level.</p

Agrobacterium-mediated TRV VIGS of two marker genes, <i>GaPDS</i> and <i>GaCLA1</i>, in <i>G</i><i>. barbadense</i>.

<p>A, Phenotypes of plants inoculated with pYL156:<i>CLA1</i> or pYL156:<i>PDS</i> vectors. The pYL156 vector was used as a vector control. B, Three cotton cultivars exhibited the photobleaching phenotype triggered by <i>GaPDS</i> or <i>GaCLA1</i> gene silencing to differing extents. C, Relative transcript levels of <i>PDS</i> and <i>CLA1</i> in systemic leaves of plants infiltrated with pYL156:<i>PDS</i> or pYL156:<i>CLA1</i>. The CK value was set at 100%. D, Total chlorophyll content in photobleached leaves. Error bars represent standard deviations (n = 3 biological replicates) in (C) and (D).</p

Data_Sheet_1.docx

<p>The tomato resistance gene Tm-2² encodes a coiled coil-nucleotide binding site-leucine rich repeat type resistance protein and confers effective immune response against tobamoviruses by detecting the presence of viral movement proteins (MPs). In this study, we show that the Nicotiana benthamiana Heat shock protein 90-kD (Hsp90) interacts with Tm-2². Silencing of Hsp90 reduced Tm-2²-mediated resistance to Tobacco mosaic virus (TMV) and the steady-state levels of Tm-2² protein. Further, Hsp90 associates with SGT1 in yeast and in plant cells. These results suggest that Hsp90-SGT1 complex takes part in Tm-2²-mediated TMV resistance by functioning as chaperone to regulate Tm-2² stability.</p

TRV-induced silencing of the anthocyanidin and proanthocyanidin biosynthetic genes <i>ANS</i> and <i>ANR</i>.

<p>a–d, Plants infiltrated with the vector control (CK), pYL156:<i>ANS</i> and pYL156:<i>ANR</i> showed different phenotypes in systemic leaves (a–c) and stems (d). e–g, DMACA stained leaves. h, Relative transcript levels of <i>ANS</i> and <i>ANR</i> in systemic leaves of plants infiltrated with pYL156:<i>ANS</i> and pYL156:<i>ANR</i>. The CK value was set at 100%. Error bars represent standard deviations (n = 3 biological replicates). White arrows indicate pink leaf veins (c) and stem (d).</p

Plant Bax Inhibitor-1 interacts with ATG6 to regulate autophagy and programmed cell death

<p>Autophagy is an evolutionarily conserved catabolic process and is involved in the regulation of programmed cell death during the plant immune response. However, mechanisms regulating autophagy and cell death are incompletely understood. Here, we demonstrate that plant Bax inhibitor-1 (BI-1), a highly conserved cell death regulator, interacts with ATG6, a core autophagy-related protein. Silencing of <i>BI-1</i> reduced the autophagic activity induced by both <i>N</i> gene-mediated resistance to <i>Tobacco mosaic virus</i> (TMV) and methyl viologen (MV), and enhanced <i>N</i> gene-mediated cell death. In contrast, overexpression of plant BI-1 increased autophagic activity and surprisingly caused autophagy-dependent cell death. These results suggest that plant BI-1 has both prosurvival and prodeath effects in different physiological contexts and both depend on autophagic activity.</p

Type I J-Domain NbMIP1 Proteins Are Required for Both <i>Tobacco Mosaic Virus</i> Infection and Plant Innate Immunity

<div><p>Tm-2² is a coiled coil-nucleotide binding-leucine rich repeat resistance protein that confers durable extreme resistance against <i>Tomato mosaic virus</i> (ToMV) and <i>Tobacco mosaic virus</i> (TMV) by recognizing the viral movement protein (MP). Here we report that the <i>Nicotiana benthamiana</i> J-domain MIP1 proteins (NbMIP1s) associate with tobamovirus MP, Tm-2² and SGT1. Silencing of <i>NbMIP1s</i> reduced TMV movement and compromised <i>Tm-2²</i>-mediated resistance against TMV and ToMV. Furthermore, silencing of <i>NbMIP1s</i> reduced the steady-state protein levels of ToMV MP and Tm-2². Moreover, NbMIP1s are required for plant resistance induced by other <i>R</i> genes and the nonhost pathogen <i>Pseudomonas syringae pv. tomato</i> (<i>Pst</i>) DC3000. In addition, we found that SGT1 associates with Tm-2² and is required for <i>Tm-2²</i>-mediated resistance against TMV. These results suggest that NbMIP1s function as co-chaperones during virus infection and plant immunity.</p></div

NbMIP1s are required for plant immunity mediated by multiple <i>R</i> genes and general elicitors.

<p>(<b>A</b>) Silencing of <i>NbMIP1s</i> resulted in larger TMV-GFP infection foci in transgenic <i>N</i>-containing <i>N. benthamiana</i> plants (<i>NN</i>) plants. Photos were taken under normal or UV light at 7 dpi. (<b>B</b>) Silencing of <i>NbMIP1s</i> compromised <i>N</i>-mediated resistance to TMV-GFP in <i>NN</i> plants and TMV-GFP spread into the upper non-inoculated leaves at 14 dpi. (<b>C</b>) RT-PCR to confirm the presence of TMV-GFP RNA in the upper, non-inoculated leaves of <i>NbMIP1s</i>-silenced <i>NN</i> plants. (<b>D</b>) Silencing of <i>NbMIP1s</i> delayed HR mediated by several <i>R</i> genes and nonhost <i>Pseudomonas syringae pv. tomato</i> DC3000 (<i>Pst</i> DC3000). All R-Avr pairs were expressed in wild type plants. Transient co-expression of Pto with avrPto, or Cf9 with avr9 was performed using agroinfiltration and <i>Pst</i> DC3000 was inoculated in <i>NbMIP1s</i>-silenced and non-silenced control plants. The leaf images of <i>Pst</i> DC3000 were taken at 12 hpi, and the other images corresponding to R-Avr pairs were taken at 3 dpi. Scale bars represent 1 cm. (<b>E</b>) Silencing of <i>NbMIP1s</i> compromised nonhost resistance against <i>Pst</i> DC3000 and caused more bacterial growth. Bacterial growth was monitored at 0, 1, 2 and 3 dpi. Each data point represents the mean ± SEM of 3 replicate samples (** p<0.01, Student's <i>t</i>-test). All experiments were performed at least three times using three or more plants in each experiment.</p

<i>Tm-2²</i>-mediated resistance against TMV requires <i>NbMIP1s</i>.

<p>(<b>A</b>) Phenotype of <i>NbMIP1s</i>-silenced and TRV control <i>N. benthamiana</i> plants. <i>NbMIP1s</i>-silenced <i>N. benthamiana</i> plants developed dwarfed stems and crinkled leaves compared to TRV-infected control plants. (<b>B</b>) Leaves of <i>NbMIP1</i>-silenced and TRV control <i>N. benthamiana</i> plants. The leaf edge of <i>NbMIP1s</i>-silenced plants curled downward but TRV-infected control leaves looked normal (right side). Photos were taken at 14 days post agroinfiltration for VIGS. (<b>C</b>) Real time RT-PCR to confirm the suppression of <i>NbMIP1s</i>, and the <i>Actin</i> mRNA levels were used as internal controls. <i>NbMIP1</i> VIGS: silencing using pTRV2-<i>NbMIP1</i>. (<b>D</b>) Silencing of <i>NbMIP1s</i> caused the appearance of TMV-GFP infection foci and visible HR lesions in the inoculated leaves of <i>NbMIP1s</i>-silenced <i>Tm-2²</i>-containing TM#1 plants. (<b>E</b>) Silencing of <i>NbMIP1s</i> compromised <i>Tm-2²</i>-mediated resistance against TMV, and TMV-GFP spread from the inoculated leaves into the upper non-inoculated leaves of <i>NbMIP1s</i>-silenced TM#1 plants. TRV-infected TM#1 plants were used as negative controls. Photos were taken at 10 days post TMV-GFP infection (dpi). Scale bars represent 1 cm. (<b>F</b>) RT-PCR was performed to confirm the presence of TMV-GFP in systemic leaves of <i>NbMIP1s</i>-silenced TM#1 plants.</p