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Abstract: Plant biofactories are a promising platform for sustainable production of high-value compounds, among which are insect sex pheromones, a green alternative to conventional insecticides in agriculture. Recently, we have constructed transgenic Nicotiana benthamiana plants (“Sexy Plants”, SxP) that successfully produce a blend of moth (Lepidoptera) sex pheromone compounds (Z)-11-hexadecen-1-ol and (Z)-11-hexadecenyl acetate. However, efficient biosynthesis of sex pheromones resulted in growth and developmental penalty, diminishing the potential for commercial use of SxP in biomanufacturing. To gain insight into the underlying molecular responses, we analysed the whole-genome transcriptome and evaluated it in relation to growth and pheromone production in low- and high-producing transgenic plants of v1.0 and v1.2 SxP lines. In our study, high-producing SxPv1.2 plants accumulated the highest amounts of pheromones but still maintained better growth compared to v1.0 high producers. For an in-depth biological interpretation of the transcriptomic data, we have prepared a comprehensive functional N. benthamiana genome annotation as well as gene translations to Arabidopsis thaliana, enabling functional information transfer by using Arabidopsis knowledge networks. Differential gene expression analysis, contrasting pheromone producers to wild-type plants, revealed that while only a few genes were differentially regulated in low-producing plants, high-producing plants exhibited vast transcriptional reprogramming. They showed signs of stress-like response, manifested as downregulation of photosynthesis-related genes and significant differences in expression of hormonal signalling and secondary metabolism-related genes, the latter presumably leading to previously reported volatilome changes. Further network analyses confirmed stress-like response with activation of jasmonic acid and downregulation of gibberellic acid signalling, illuminating the possibility that the observed growth penalty was not solely a consequence of a higher metabolic burden imposed upon constitutive expression of a heterologous biosynthetic pathway, but rather the result of signalling pathway perturbation. Our work presents an example of comprehensive transcriptomic analyses of disadvantageous stress signalling in N. benthamiana biofactory that could be applied to other bioproduction systems.
Keywords: transcriptomics, plant biotechnology, jasmonic acid, growthstress tradeoffs, network analysis, growth penalty, insect sex pheromone
Published in DiRROS: 17.07.2024; Views: 419; Downloads: 240
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Abstract: In the Mediterranean Sea, the Strait of Messina (MS) is a very peculiar area, connecting highly different regions and representing a privileged observatory for an early comprehension and assessment of ecosystems shifts. It is hypothesized that the outbreaks observed near the coast of many sites in the Mediterranean Sea may be the result of transport of permanent populations of P. noctiluca in pelagic waters to the coast, caused by specific hydrodynamic conditions. By both visual observations and numerical experiments our objective is twofold: (A) to help clarify whether the basin of the Aeolian Islands Archipelago (AIA), in the Southern Tyrrhenian Sea (STS), may be the site from which large populations of P. noctiluca are transported to the MS, and (B) to evaluate whether the upwelling turbulent system of the MS can be an energetic opportunity for this species. It should offer a rich habitat without jeopardizing the overall survival of the population, that is subject to stranding due to strong currents. Although very different, the two involved ecosystems (AIA and MS ) are complementary for the success of Pelagia noctiluca life cycle. Outputs obtained by coupling the 3D hydrodynamic model (SHYFEM) with a Lagrangian particle tracking model support the hypothesis of a connectivity between these two ecosystems, particularly in the first half of the year, indicating the coastal areas around the AIA as potential optimal source location for Pelagia larval stages. We support the very attractive hypothesis that two connected systems exist, the former one favours Pelagia's reproduction and acts as a nursery and the latter favours its growth due to higher productivity. We speculate that the reproductive population of the AIA is not permanent, but is renewed every year by individuals who have fed and quickly grown in the MS and who are passively transported by downwelling along canyon "corridors".
Keywords: coastal systems, coastal water, Tyrrhenian Sea
Published in DiRROS: 17.07.2024; Views: 343; Downloads: 248
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Abstract: In this Collective Article on alien and cryptogenic diversity in the Mediterranean Sea we report a total of 19 species belonging to nine Phyla and coming from nine countries. Several of these records concern fish species, and of particular interest are the first records of: Terapon puta for Italian waters; Pteragopus trispilus from Malta; Plotosus lineatus from Cyprus; and the northernmost Mediterranean record of Lagocephalus sceleratus. The northernmost Mediterranean record was also reported for the sea urchin Diadema setosum. The portunid crab Thalamita poissonii was recorded for the first time in Libya. The copepod Pseudodiaptomus marinus was recorded for the first time in the Marmara Sea. The polychaete Branchiomma luctuosum was recorded for the first time from the Mediterranean coast of France. The alien anemone Diadumene lineata was recorded for the first time from Slovenia. The macroalgae Sargassum furcatum was recorded for the first time from Italy. The new Mediterranean records here reported help tracing abundance and distribution of alien and cryptic species in the Mediterranean Sea.
Keywords: alien diversity, cryptogenic diversity, new species, Mediterranean Sea
Published in DiRROS: 17.07.2024; Views: 356; Downloads: 226
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Abstract: MicroRNAs (miRNAs) are small noncoding RNAs, which modulate the abundance and spatiotemporal accumulation of target mRNAs at transcriptional and post-transcriptional levels and through that play important roles in several biological processes in plants. Here we show that in polyploid species, CRISPR/Cas9 system can be used for fine-tuning of miRNA expression, which can have broader range of applications compared to knock-out mutants. We established the complete pipeline for CRISPR-Cas9-mediated modulation of miRNA expression in potato. It consists of (1) design and assembly of dual sgRNA CRISPR/Cas9 constructs, (2) transient transfection of protoplasts following fast and efficient screening by high resolution melting analysis to select functional sgRNAs, and (3) stable transformation of potato explants with functional sgRNAs and selection of regenerated transgenic lines with desired mutations and desired miRNA abundance based on sequencing and RT-qPCR. We show that miRNA-editing using dual sgRNA approach results in different types of mutations among transgenic lines but also in different alleles of the same plant, which are target site-dependent. The most frequent were short deletions, but we also detected 1-nt insertions (T or G), deletions between two sgRNAs and larger deletions. miRNA abundance correlates with the frequency and type of introduced mutations, as more extensive mutations in more alleles result in lower miRNA abundance. Interestingly, some mutated loci can generate alternative miRNAs, now novel targets were however predicted for those. In all transgenic lines with Cas9 expression, we detected mutations, suggesting high efficiency of Cas9-editing. We confirmed the miRNA-editing efficiency of our optimised approach in two different potato genotypes and three different loci.
Keywords: CRISPR/Cas9, hypersensitive response (HR)-conferred resistance, immune signaling, live cell imaging, Solanum tuberosum (potato), spatiotemporal analysis, stromules, virus resistance
Published in DiRROS: 17.07.2024; Views: 394; Downloads: 249
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Abstract: Four obligate cave fish species have been recorded from Zagros in Iran: Garra typhlops (Bruun and Kaiser, 1944), G. lorestanensis (Mousavi-Sabet and Eagderi, 2016), G. tashanensis (Mousavi-Sabet, Vatandoust, Fatemi and Eagderi, 2016) and Eidinemacheilus smithi (Greenwood, 1976). So far, the only known locality of the latter has been the Loven cave. Here, we extend the known range of the Zagros blind loach (E. smithi) by reporting a new locality, the Tuveh spring, located 31 km south of Loven. We combine morphological evidence with the calculated K2P genetic divergences of 1.39% between Tuveh and Loven to confirm that these populations are conspecific. Our discovery of the second E. smithi population has implications for the conservation of this rare and vulnerable species.
Keywords: fishes, caves, evolution, Irano-Anatolian hotspot
Published in DiRROS: 17.07.2024; Views: 348; Downloads: 218
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