Table_1_Genome-wide identification of terpenoid synthase family genes in Gossypium hirsutum and functional dissection of its subfamily cadinene synthase A in gossypol synthesis.xlsx

Plant terpenoid synthase (TPS) family genes participate in metabolite synthesis, hormones, gossypol, etc. Here, we genome-widely identified TPS family genes in 12 land plant species. Four hundred and thirty TPS-related genes were divided into seven subfamilies. The TPS-c in Bryophytes was suggested to be the earliest subfamily, followed by the TPS-e/f and TPS-h presence in ferns. TPS-a, the largest number of genes, was derived from monocotyledonous and dicotyledonous plants. Collinearity analysis showed that 38 out of the 76 TPS genes in G. hirsutum were collinear within G. arboreum and G. raimondii. Twenty-one GhTPS-a genes belong to the cadinene synthase (GhCDN) subfamily and were divided into five groups, A, B, C, D, and E. The special cis-elements in the promoters of 12 GhCDN-A genes suggested that the JA and ethylene signaling pathways may be involved in their expression regulation. When 12 GhCDN-A genes were simultaneously silenced through virus-induced gene silencing, the glandular color of GhCDN-A-silenced plants was lighter than that of the control, supported by a gossypol content decrease based on HPLC testing, suggesting that GhCDN-A subgroup genes participate in gossypol synthesis. According to RNA-seq analysis, gossypol synthesis-related genes and disease-resistant genes in the glandular variety exhibited upregulated expression compared to the glandless variety, whereas hormone signaling-related genes were downregulated. All in all, these results revealed plant TPS gene evolution rules and dissected the TPS subfamily, GhCDN-A, function in gossypol synthesis in cotton.</p

Presentation_1_Genome-wide identification of terpenoid synthase family genes in Gossypium hirsutum and functional dissection of its subfamily cadinene synthase A in gossypol synthesis.pptx

Plant terpenoid synthase (TPS) family genes participate in metabolite synthesis, hormones, gossypol, etc. Here, we genome-widely identified TPS family genes in 12 land plant species. Four hundred and thirty TPS-related genes were divided into seven subfamilies. The TPS-c in Bryophytes was suggested to be the earliest subfamily, followed by the TPS-e/f and TPS-h presence in ferns. TPS-a, the largest number of genes, was derived from monocotyledonous and dicotyledonous plants. Collinearity analysis showed that 38 out of the 76 TPS genes in G. hirsutum were collinear within G. arboreum and G. raimondii. Twenty-one GhTPS-a genes belong to the cadinene synthase (GhCDN) subfamily and were divided into five groups, A, B, C, D, and E. The special cis-elements in the promoters of 12 GhCDN-A genes suggested that the JA and ethylene signaling pathways may be involved in their expression regulation. When 12 GhCDN-A genes were simultaneously silenced through virus-induced gene silencing, the glandular color of GhCDN-A-silenced plants was lighter than that of the control, supported by a gossypol content decrease based on HPLC testing, suggesting that GhCDN-A subgroup genes participate in gossypol synthesis. According to RNA-seq analysis, gossypol synthesis-related genes and disease-resistant genes in the glandular variety exhibited upregulated expression compared to the glandless variety, whereas hormone signaling-related genes were downregulated. All in all, these results revealed plant TPS gene evolution rules and dissected the TPS subfamily, GhCDN-A, function in gossypol synthesis in cotton.</p

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”

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

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

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

<i>AtWuschel</i> Promotes Formation of the Embryogenic Callus in <i>Gossypium hirsutum</i>

<div><p>Upland cotton (<i>Gossypium hirsutum</i>) is one of the most recalcitrant species for <i>in vitro</i> plant regeneration through somatic embryogenesis. Callus from only a few cultivars can produce embryogenic callus (EC), but the mechanism is not well elucidated. Here we screened a cultivar, CRI24, with high efficiency of EC produce. The expression of genes relevant to EC production was analyzed between the materials easy to or difficult to produce EC. Quantitative PCR showed that CRI24, which had a 100% EC differentiation rate, had the highest expression of the genes <i>GhLEC1</i>, <i>GhLEC2</i>, and <i>GhFUS3</i>. Three other cultivars, CRI12, CRI41, and Lu28 that formed few ECs expressed these genes only at low levels. Each of the genes involved in auxin transport (<i>GhPIN7</i>) and signaling (<i>GhSHY2</i>) was most highly expressed in CRI24, with low levels in the other three cultivars. WUSCHEL (WUS) is a homeodomain transcription factor that promotes the vegetative-to-embryogenic transition. We thus obtained the calli that ectopically expressed <i>Arabidopsis thaliana Wus</i> (<i>AtWus</i>) in <i>G. hirsutum</i> cultivar CRI12, with a consequent increase of 47.75% in EC differentiation rate compared with 0.61% for the control. Ectopic expression of <i>AtWus</i> in CRI12 resulted in upregulation of <i>GhPIN7</i>, <i>GhSHY2</i>, <i>GhLEC1</i>, <i>GhLEC2</i>, and <i>GhFUS3</i>. <i>AtWus</i> may therefore increase the differentiation potential of cotton callus by triggering the auxin transport and signaling pathways.</p></div

Analysis of gene expression in the non-transgenic calli of the four cultivars.

<p>Analysis of gene expression in the non-transgenic calli of the four cultivars.</p

<i>AtWus</i> expression in calli of CRI12.

<p><i>AtWus</i> expression in calli of CRI12.</p