Addressing Reproducibility in Cryopreservation,and Considerations Necessary for Commercialization and Community Development in Support of Genetic Resources of Aquatic Species

For the past six decades a repeated cycle of developing new cryopreservation protocols or simply reinventing them to counteract a lack of reproducibility has led to hundreds of published studies that have offered little to the establishment of a genetic resources community for aquatic species. This has hampered repository development and inhibited industrial application. Most protocols were developed without standardized approaches, leading to irreproducible studies and questionable or meaningless comparisons. Thus cryopreservation of germplasm in aquatic species would greatly benefit from strategies to facilitate reproducibility. Our objectives were to: (1) identify major sources of irreproducibility across research, small‐scale, repository, and commercial‐scale development levels; (2) provide recommendations to address reproducibility challenges; and (3) offer suggestions on how researchers can directly influence commercial development and application of cryopreservation research. Sources of irreproducibility include lack of standardized procedural approaches, lack of standardized terminology, and lack of reporting guidelines. To address these challenges, we propose implementation of standard operating procedures, support of stock centers and internet content for development of training programs, and strengthening of the role of scientific journals and reviewers in reducing the frequency of irreproducible outcomes. Reproducibility is the foundation for quality management programs and product reliability, and therefore, standardization is necessary to assure efficient transition to commercial‐scale application and repository development. Progress can only be possible through community‐based approaches focused on coalescence and consensus of disparate groups involved in aquatic species cryopreservation and management of genetic resources.     

Anthocyanins of <Emphasis Type="Italic">Pithecellobium dulce</Emphasis> (Roxb.) Benth. Fruit Associated with High Antioxidant and α-Glucosidase Inhibitory Activities

Red arils of Pithecellobium dulce fruit, commonly known as guamuchil, show high antioxidant (AOx) and α-glucosidase inhibitory (IαG) activities, which have been mainly associated with the content of unknown anthocyanins. In this study, the AOx (i.e., DPPH and ABTS as Trolox equivalents, μmol TE/g) and IαG (as half-maximal inhibitory concentration, IC₅₀, mg/mL) activities of the anthocyanin-rich fraction (ARF) obtained from red arils were contrasted with those of the methanol extract (ME), and the main ARF anthocyanins were characterized by HPLC-DAD-ESI-MS, GC-MS and ¹H-NMR. The AOx and IαG values of the ARF (DPPH?=?597.8; ABTS?=?884.01; IαG?=?0.06) were better than those of the ME (DPPH?=?41.5; ABTS?=?142.3; IαG?=?17.5); remarkably, the ARF IαG value was about 42 times lower than that of acarbose. The main anthocyanins in ARF were pelargonidin 3-O-glucoside and cyanidin 3-O-glucoside. Thus, the consumption of red P. dulce arils could provide health benefits for prevention/treatment of chronic degenerative diseases such as diabetes.     

Physical, Chemical, and Antioxidant Activity Characterization of Pitaya (Stenocereus pruinosus) Fruits

Fruits with red and orange flesh of the columnar cactus pitaya (Stenocereus pruinosus) were studied to evaluate physical characteristics, total soluble solids, betalains and soluble phenols content, and antioxidant activity. Fruits had, in average, weight of 179.0 g, 9.8 °Brix, 9.4 % carbohydrates, 1.25 % protein, 0.11 % ethereal extract, 0.60 % crude fiber, and 0.62 % ash. Also, fruits resulted rich in Fe (22.8–27.8 mg/kg). Hue angle and contents of betacyanins, betaxanthins (μg/g dry sample), and total soluble phenols (mg GAE/g fresh sample) were 19.8°, 2860.0, 3210.0, and 1.6 in the red material, and 28.9°, 470.0, 2670.0, and 1.2, respectively, in the orange fruit. The antioxidant capacity was higher in the red material, since the ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) technique reported 1090.6 and 735.4 μmol of Trolox equivalents/g fresh flesh in red and orange fruits, respectively, while the oxygen radical absorbance capacity (ORAC) assay reported 7.84 and 5.16 μmol of Trolox equivalents/g fresh flesh, respectively. The chromatographic profile showed five betalains in red fruits, but only four of them were observed in those orange fleshed.     

Climate change and amphibian diversity patterns in Mexico

The aim of this article is to characterize at fine scale alpha and beta diversity patterns for Mexican amphibians and analyze how these patterns might change under a moderate climate-change scenario, highlighting the overall consequences for amphibian diversity at the country level. We used a geo-referenced database gathered from museums worldwide as a basis for climatic envelope models, based on the working assumptions that we can accurately model the ecological niche, niche conservatism, and that the future climate scenario is realistic. We generated six sets of models with different parameters (threshold of presence and dispersal capability) in the modelling processes. We simulated the base line (2000) and future scenarios for Mexican amphibian diversity (2020, 2050, 2080), using climate data layers constructed for Mexico. Using moving-window analyses of different sizes (9, 25, 100, 225 and 400 km²) we calculated beta diversity with R.H. Whittaker’s formula β = γ/α_mean. We analyzed the changes of alpha- and beta-diversity spatial patterns during these periods, and performed an analysis of sensitivity for the beta-diversity patterns. We found that setting low dispersal capability generates high levels of species extinctions, but the overall geographic pattern of beta diversity remained stable. Zones of high beta diversity resulted that were associated with topographic formations, while the values of beta diversity initially increased, and then declined over time. Extinctions (complete loss of range within country boundaries) were particularly intense during the period 2020–2050. The results implied that heterogeneous zones associated with mountain ranges will remain particularly important for amphibian diversity and thus constitute areas for continued conservation prioritization in the face of climate change.     

Resistance to <Emphasis Type="Italic">Cucumber green mottle mosaic virus</Emphasis> in <Emphasis Type="Italic">Cucumis sativus</Emphasis>

Cucumber green mottle mosaic virus (CGMMV) is a severe threat for cucumber production worldwide. At present, there are no cultivars available in the market which show an effective resistance or tolerance to CGMMV infection, only wild Cucumis species were reported as resistant. Germplasm accessions of Cucumis sativus, as well as C. anguria and C. metuliferus, were mechanically infected with the European and Asian strains of CGMMV and screened for resistance, by scoring symptom severity, and conventional RT-PCR. The viral loads of both CGMMV strains were determined in a selected number of genotypes using quantitative RT-PCR. Severe symptoms were found following inoculation in C. metuliferus and in 44 C. sativus accessions, including C. sativus var. hardwickii. Ten C. sativus accessions, including C. sativus var. sikkimensis, showed intermediate symptoms and only 2 C. sativus accessions showed mild symptoms. C. anguria was resistant to both strains of CGMMV because no symptoms were expressed and the virus was not detected in systemic leaves. High amounts of virus were found in plants showing severe symptoms, whereas low viral amounts found in those with mild symptoms. In addition, the viral amounts detected in plants which showed intermediate symptoms at 23 and 33 dpi, were significantly higher in plants inoculated with the Asian CGMMV strain than those with the European strain. This difference was statistically significant. Also, the amounts of virus detected over time in plants did not change significantly. Finally, the two newly identified partially resistant C. sativus accessions may well be candidates for breeding programs and reduce the losses produced by CGMMV with resistant commercial cultivars.     

Study of the genetic variability and similarity among and within Arachis villosulicarpa, A. pietrarellii and A. hypogaea through isoenzyme analysis

Arachis villosulicarpa is a perennial species cultivated for its soft and tasty seeds by indigenous inhabitants of the Mato Grosso State, Brazil. Besides A. hypogaea, this species is considered as the only species of Arachis which represents a valuable food source for human consumption. Due to the lack of knowledge concerning the genetic diversity of A. villosulicarpa, this study was conducted to evaluate the genetic variability of the accessions from the Germplasm Collection of CENARGEN/EMBRAPA (Brasília, DF, Brazil) and Instituto Agrono^mico (IAC, Campinas, SP, Brazil). In addition, the genetic similarity between A. villosulicarpa, the related wild species A. pietrarellii, and the cultivated peanut A. hypogaea cv. Tatu was evaluated. From the entire sample analyzed of A. villosulicarpa, the accession from Instituto Agrono^mico showed the highest indices of diversity for both enzymatic systems analyzed, pointing this accession as a promising source of genetic variabil ity that must be preserved in the Germplasm Bank. A high level of genetic similarity was observed between A. pietrarellii and A. villosulicarpa, supporting previous suggestions that A. pietrarellii could be the ancestral progenitor species of A. villosulicarpa or that both species originated from a common ancestor.     

Radionuclides and stable elements in the sediments of the Yesa Reservoir,Central Spanish Pyrenees

Purpose  

The sediments accumulated in the Yesa Reservoir (Central Spanish Pyrenees) have greatly decreased its water storage capacity and are a major threat to the sustainability of water resources in the region. This study examines the contents of radionuclides and stable elements in the reservoir sediments and relates their variations with the sediment composition and local sedimentary dynamics, particularly flood frequency and intensity, which are responsible for changes in the main supply and distribution of radionuclides in the basin.