Method for Efficient Regeneration of Solvent Mixture: Extractive Heterogeneous-Azeotropic Distillation

The fine chemical and pharmaceutical industries use large amounts of various organic solvents in their manufacturing processes. By reusing them, production costs can be significantly reduced. If we can regenerate waste solvent mixtures, we have the opportunity to reuse them in the production process or in other production processes. Our study illustrates an efficient regeneration process using the example of a four-component solvent mixture. Calculations were performed in a professional process simulator to demonstrate that the highly non-ideal Water-Ethyl Alcohol-Methyl Ethyl Ketone-Ethyl Acetate solvent mixture can be efficiently decomposed into azeotropic pairs and thus regenerated by the extractive heterogeneous-azeotropic distillation technique.

A highly efficient regeneration, genetic transformation system and induction of targeted mutations using CRISPR/Cas9 in Lycium ruthenicum

Background CRISPR/Cas9 is a rapidly developing genome editing technology in various biological systems due to its efficiency, portability, simplicity and versatility. This editing technology has been successfully applied in in several important plants of Solanaceae such as tomato, tobacco, potato, petunia and groundcherry. Wolfberry ranked the sixth among solanaceous crops of outstanding importance in China following potato, tomato, eggplant, pepper and tobacco. To date, there has been no report on CRISPR/Cas9 technology to improve Lycium ruthenicum due to the unknown genome sequencing and the lack of efficient regeneration and genetic transformation systems. Results In this study, we have established an efficientregeneration and genetic transformation system of Lycium ruthenicum. We have used this system to validate target sites for fw2.2, a major fruit weight quantitative trait locus first identified from tomato and accounted for 30% of the variation in fruit size. In our experiments, the editing efficiency was very high, with 95.45% of the transgenic lines containing mutations in the fw2.2 target site. We obtained transgenic wolfberry plants containing four homozygous mutations and nine biallelic mutations in the fw2.2 gene. Conclusions These results suggest that CRISPR-based gene editing is effective for the improvement of black wolfberry traits, and we expect this approach to be routinely applied to this important economic fruit.

Improving Adventitious Shoot Regeneration and Transient Agrobacterium-Mediated Transformation of Apricot (Prunus armeniaca L.) Hypocotyl Sections

The improvement of previously described protocols for the regeneration of shoots from ‘Canino’ mature seed hypocotyl slices has been accomplished. The effects of different factors such as the part of the hypocotyl used, vacuum-infiltration, 2,4-Dichlorophenoxyacetic acid pulse, vacuum-infiltration and sonication on regeneration and transient transformation were analyzed. When the three slices obtained from the hypocotyls were evaluated separately on regeneration medium, the highest percentages of regenerating explants were achieved in the part close to the epicotyl and in the central part. On the other hand, sonication of the explants for 30 s followed by vacuum-infiltration during Agrobacterium infection for 20 min allowed for an increase in the transformation events. The application of these modifications to the procedure increased the regeneration efficiencies, and transient transformation events and may reduce the frequency of failed experiments. An efficient regeneration/transformation protocol could facilitate its use as a biotechnological technique for apricot breeding.

The influence of fencing on seedling establishment during reforestation of oak stands: a comparison of artificial and natural regeneration techniques including costs

In temperate Europe, oak-dominated forests are widespread, supporting high biodiversity and providing important ecosystem services. Insufficient natural regeneration has, however, been a concern for over a century. The objective of this study was to gain insights into differences in regeneration success using artificial and natural regeneration techniques for reforestation of oak (Quercus robur L.) stands. We monitored seedlings following planting, direct seeding and natural regeneration over five years in a randomized block experiment in southern Sweden with fenced and non-fenced plots. Fencing had a strong positive effect on height growth, especially for planted seedlings that were taller than the other seedlings and more frequently browsed in non-fenced plots. In contrast, there was little effect of fencing on survival, establishment rate and recruitment rate of seedlings. Due to aboveground damage on seedlings from voles, protection of acorns did not improve establishment rate following direct seeding. Under current circumstances at the site with a sparse shelterwood of old oaks, we conclude that natural regeneration was the most cost-efficient regeneration method. It resulted in the most seedlings at the lowest cost. However, regeneration success was heavily influenced by interference from herbaceous vegetation. With a small additional investment in vegetation control, the results might have been improved for planting and direct seeding.