Overexpression of Galactinol Synthase 1 From Solanum Commersonii (ScGolS1) Confers Freezing Tolerance In Transgenic Potato

Potato (Solanum tuberosum L.) is the fourth largest food crop in the world. Low temperature causes serious damage to potato plants every year, and freezing tolerance has become a hot spot in potato research. Galactinol synthase (GolS) is a key enzyme in the synthesis of raffinose family oligosaccharides (RFOs), and plays an important role in the response of plants to abiotic stress. In this study, the ScGolS1 gene from S. commersonii was cloned and introduced into the S. tuberosum cultivars ‘Atlantic’ and ‘Desiree’ via Agrobacterium-mediated transformation. Phenotyping assay showed that overexpression of the ScGolS1 could significantly improve freezing tolerance in transgenic potato plants. Further physiological and biochemical results showed that the relative conductivity, malondialdehyde (MDA) content, and 3,3'-Diaminobenzidine (DAB) staining of the transgenic lines decreased, and the plant survival rate increased compared with wild type (WT). Moreover, CBF1, CBF2, CBF3, CBF downstream cold responsive genes COR413, COR47 and ERF transcription factor genes ERF3, ERF4, ERF6 in the ethylene signaling pathway were all induced by freezing treatment, while higher levels were observed in ScGolS1 overexpression lines compared with WT. In addition, other genes such as MIPS, STS and RS genes from RFO metabolic pathway and some sugars content were altered in response to freezing treatment. This indicates that overexpression of the ScGolS1 gene induced both the regulation of the ethylene signaling pathway and the metabolism of raffinose series oligosaccharides, regulating the balance of sugar composition and improved anti-peroxidation capacity, and thereby improved freezing tolerance in potato. These results provide theoretical support and genetic resources for freezing tolerance breeding in potato.

Proteomic Analysis of Auricularia auricula-judae Under Freezing Treatment Revealed Proteins and Pathways Associated With Melanin Reduction

Auricularia auricula-judae is an edible nutrient-rich mushroom, which is a traditional medicinal resource in China. It is known that environment stimuli will affect the production of melanin by A. auricula-judae, but the mechanism of the effects of freezing treatment on melanin accumulation remains unknown. In the present study, the synthesis of melanin in A. auricula-judae was analyzed by physiological assays and a proteomics approach. Our findings showed that a longer freezing treatment causes a lighter color of A. auricula-judae fruiting bodies. The proteomic analysis showed that proteins involved in glycolysis/gluconeogenesis, tyrosine metabolism, ribosome, and arginine biosynthesis might contribute to the color differences in the A. auricula-judae after freezing treatment. This work will be expected to provide valuable information on the physiological and molecular mechanisms of freezing treatment on the color quality of A. auricula-judae.

Negligible risk of zoonotic anisakid nematodes in farmed fish from European mariculture, 2016 to 2018

Background The increasing demand for raw or undercooked fish products, supplied by both aquaculture and fisheries, raises concerns about the transmission risk to humans of zoonotic fish parasites. This has led to the current European Union (EU) Regulation No 1276/2011 amending Annex III of Regulation (EC) No 853/2004 and mandating a freezing treatment of such products. Zoonotic parasites, particularly anisakid larvae, have been well documented in wild fish. Data on their presence in European aquaculture products, however, are still scarce, except for Atlantic salmon (Salmo salar), where the zoonotic risk was assessed as negligible, exempting it from freezing treatment. Aim To evaluate the zoonotic Anisakidae parasite risk in European farmed marine fish other than Atlantic salmon. Methods From 2016 to 2018 an observational parasitological survey was undertaken on 6,549 farmed fish including 2,753 gilthead seabream (Sparus aurata), 2,761 European seabass (Dicentrarchus labrax) and 1,035 turbot (Scophthalmus maximus) from 14 farms in Italy, Spain and Greece. Furthermore, 200 rainbow trout (Oncorhynchus mykiss) sea-caged in Denmark, as well as 352 seabream and 290 seabass imported in Italy and Spain from other countries were examined. Fish were subjected to visual inspection and candling. Fresh visceral organs/fillet samples were artificially digested or UV pressed and visually examined for zoonotic anisakid larvae. Results No zoonotic parasites were found in any of the fish investigated. Conclusions The risk linked to zoonotic Anisakidae in the examined fish species from European mariculture appears negligible. This study laid the groundwork for considerations to amend the current EU regulation.

Effect of Barley Antifreeze Protein on Dough and Bread during Freezing and Freeze-Thaw Cycles

In order to verify the cryoprotective effect of an antifreeze protein (BaAFP-1) obtained from barley on bread dough, the effect of BaAFP-1 on the rheological properties, microstructure, fermentation, and baking performance including the proofing time and the specific volume of bread dough and bread crumb properties during freezing treatment and freeze-thaw cycles were analysed. BaAFP-1 reduced the rate of decrease in storage modulus and loss modulus values during freezing treatment and freeze-thaw cycles. It influenced the formation and the shape of ice formed during freezing and inhibited ice recrystallization during freeze-thaw. BaAFP-1 maintained gas production ability and gas retention properties, protected gluten network and the yeast cells from deterioration caused by ice formation and ice crystals recrystallisation in dough samples during freezing treatment and freeze-thaw treatment. It slow down the increase rate of hardness of bread crumb. The average area of pores in bread crumbs decreased significantly (p < 0.05) as the total number of pores increased (p < 0.05), and the addition of BaAFP-1 inhibited this deterioration. These results confirmed the cryoprotective activity of BaAFP-1 in bread dough during freezing treatment and freeze-thaw cycles.

Transcriptional reprogramming of the bud-mutation loquat YongLu emerging freezing resistant

Background: Freezing seriously affects loquat, an originally subtropical fruit. Here, a wide-spread cultivar loquat bud mutation ‘Yonglu’ (YL) was found to be more resistant to freezing than the parental ‘Ruantiaobaisha’ (RT), especially the fruits. The freezing resistance mechanism of YL was analyzed by physiological and transcriptomic methods. Results: After freezing treatment, only the fruits of YL showed a significant accumulation of proline (which increased by 1.5-fold). Interestingly, abscisic acid (ABA) accumulation in YL showed no difference after freezing treatment. Moreover, the stomatal density, area, and apertures of YL significantly decreased. All these results suggested that the proline content and stomatal closure contributed to the enhanced freezing tolerance. Further, the transcriptional profiles of freezing tissues were obtained and compared. Functional enrichment analysis showed that aside from basic nutrients and energy metabolisms, such as citrate cycle and oxidative phosphorylation, the fruits and flowers of YL have different systems to enhance freezing tolerance, suggesting a progressive freezing-resistant network. Signaling transduction of nearly all hormones, including ABA, was significantly differentially expressed. Several key genes, such as PP2C and SnRK2, which are crucial for transducing ABA signals, were significantly down- and upregulated, respectively. But the genes of neither ABA biosynthesis nor ABA transduction pathway significantly differentially expressed between two freezing varieties, suggesting they contribute limit in enhancing freezing tolerance of YL. Conclusion: Concludingly, the bud mutation YL loquat has stronger freezing tolerance than its parental RT due to the increasing proline and more effective regulation of stomatal closure. Furthermore, the bud mutation YL loquat reprogrammed its entire transcriptional profiles to enhance their abilities to adapt to freezing stress. However, ABA biosynthesis, ABA transduction pathway and CBF pathway, which are common reasons in cold response, were not the main reason for the increasing freezing tolerance of YL. There must be unknown regulation mechanisms for the bud mutation to freezing resistant.