Regulation of seed oil accumulation by lncRNAs in Brassica napus.

BACKGROUND: Studies have indicated that long non-coding RNAs (lncRNAs) play important regulatory roles in many biological processes. However, the regulation of seed oil biosynthesis by lncRNAs remains largely unknown. RESULTS: We comprehensively identified and characterized the lncRNAs from seeds in three developing stages in two accessions of Brassica napus (B. napus), ZS11 (high oil content) and WH5557 (low oil content). Finally, 8094 expressed lncRNAs were identified. LncRNAs MSTRG.22563 and MSTRG.86004 were predicted to be related to seed oil accumulation. Experimental results show that the seed oil content is decreased by 3.1-3.9% in MSTRG.22563 overexpression plants, while increased about 2% in MSTRG.86004, compared to WT. Further study showed that most genes related to lipid metabolism had much lower expression, and the content of some metabolites in the processes of respiration and TCA (tricarboxylic acid) cycle was reduced in MSTRG.22563 transgenic seeds. The expression of genes involved in fatty acid synthesis and seed embryonic development (e.g., LEC1) was increased, but genes related to TAG assembly was decreased in MSTRG.86004 transgenic seeds. CONCLUSION: Our results suggest that MSTRG.22563 might impact seed oil content by affecting the respiration and TCA cycle, while MSTRG.86004 plays a role in prolonging the seed developmental time to increase seed oil accumulation.

An improved whole-cell biotransformation system for (S)-equol production.

(S)-equol, the most active metabolite of the soybean isoflavones in vivo, has exhibited various biological activities and clinical benefits. Existing studies on the heterologous biosynthesis of (S)-equol via the engineered E. coli constructed have been significantly progressed. In the present study, the engineered E. coli was further improved to be more suitable for (S)-equol production. The four enzymes involved in the biosynthesis of (S)-equol and another GDH for NADPH regeneration were combined to construct the recombinant E. coli BL21(DE3). The optimal conditions for (S)-equol production were explored, respectively. The yield of equol reached 98.05% with 1 mM substrate daidzein and 4% (wt/vol) glucose. Even when the substrate concentration increased to 1.5 mM, (S)-equol could maintain a high yield of 90.25%. Based on the 100 ml one-pot reaction system, (S)-equol was produced with 223.6 mg/L in 1.5 h. The study presented a more suitable engineered E. coli for the production of (S)-equol.

Fatty acid exporter 1 enhances seed oil content in Brassica napus.

Fatty acid exporter 1 (FAX1) is an initial transporter for fatty acid (FA), in charge of transporting FA from the inside of the plastid to the outside. Brassica napus (B. napus) has nineteen members in the FAX family, of which there are six FAX1 homologous genes. Here, we generated the BnaFAX1 CRISPR mutants (BnaA09.FAX1 and BnaC08.FAX1 were both edited) and overexpression (OE) plants of BnaA09.FAX1 in B. napus. The results showed that the FA content was increased by 0.6-0.9% in OE plant leaves, and the seed oil content was increased by 1.4-1.7% in OE lines, compared to WT. Meanwhile, the content of triacylglycerol, diacylglycerol, and phosphatidylcholine was significantly increased in OE seeds. Moreover, seedling biomass and plant height of OE plants were increased compared to WT plants. However, the traits above had no significant difference between the mutants and WT. These results suggest that BnaA09.FAX1 plays a role in improving seed oil accumulation and plant growth, while the function of BnaFAX1 may be compensated by other homologous genes of BnaFAX1 and other BnaFAX genes in the mutants. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-022-01346-0.

Controlling the localization of nanoparticles in assemblies of amphiphilic diblock copolymers.

We performed a dissipative particle dynamics (DPD) approach to study the self-assembly of AB diblock copolymer tethered nanoparticles (P) in dilute solutions. Different morphological aggregates, including spherical micelles, vesicles, disk-like micelles and rod-like micelles, were found by varying the interaction between block copolymers and nanoparticles. Most importantly, the nanoparticles can selectively localize in the different domains within the aggregates. When the repulsive interaction between block copolymers and nanoparticles aPA = aPB = 25, the nanoparticles are evenly distributed within the spherical micelles. While aPA or aPB increases, the nanoparticles gradually aggregate and separate from copolymers and then localize in the central portion of vesicular wall or disk-like and rod-like micelles. The degree of stretching of the tethered copolymer chains gradually grows with the increase of aPA or aPB, while the degree of stretching of solvophobic block B decreases when the morphologies change from spherical to disk-like micelles and further to rod-like micelles. This work illustrates that tuning the miscibility of copolymers and nanoparticles could be used to project the selective localization of nanoparticles within the aggregates self-assembled by diblock copolymer tethered nanoparticles in dilute solutions.

Conformation-dependent translocation of a star polymer through a nanochannel.

The translocation process of star polymers through a nanochannel is investigated by dissipative particle dynamics simulations. The translocation process is strongly influenced by the star arm arrangement as the polymer enters the channel, and a scaling relation between the translocation time [Formula: see text] and the total number of beads N tot is obtained. Qualitative agreements are found with predictions of the nucleation and growth model for linear block co-polymer translocation. In the intermediate stage where the center of the star polymer is at the channel entrance, the translocation time is found to have power law-dependence on the number of arms outside the channel and very weakly dependent on the number of arms in the channel. Increasing the total number of star arms also increases the star translocation time.

Formation and structural characteristics of thermosensitive multiblock copolymer vesicles.

The spontaneous vesicle formation of ABABA-type amphiphilic multiblock copolymers bearing thermosensitive hydrophilic A-block in a selective solvent is studied using dissipative particle dynamics (DPD) approach. The formation process of vesicle through nucleation and growth pathway is observed by varying the temperature. The simulation results show that spherical micelle takes shape at high temperature. As temperature decreases, vesicles with small aqueous cavity appear and the cavity expands as well as the membrane thickness decreases with the temperature further decreasing. This finding is in agreement with the experimental observation. Furthermore, by continuously varying the temperature and the length of the hydrophobic block, a phase diagram is constructed, which can indicate the thermodynamically stable region for vesicles. The morphological phase diagram shows that vesicles can form in a larger parameter scope. The relationship between the hydrophilic and hydrophobic block length versus the aqueous cavity size and vesicle size are revealed. Simulation results demonstrate that the copolymers with shorter hydrophobic blocks length or the higher hydrophilicity are more likely to form vesicles with larger aqueous cavity size and vesicle size as well as thinner wall thickness. However, the increase in A-block length results to form vesicles with smaller aqueous cavity size and larger vesicle size.

A new ab initio intermolecular potential energy surface and predicted rotational spectra of the Ne-H2S complex.

We report a new three-dimensional ab initio intermolecular potential energy surface for the Ne-H(2)S complex with H(2)S monomer fixed at its experimental average structure. Using the supermolecular approach, the intermolecular potential energies were evaluated at CCSD(T) (coupled cluster with single and double and perturbative triple excitations) level with large basis sets including bond functions. The full counterpoise procedure was employed to correct the basis set superposition error. The planar T-shaped global minimum is located at the intermolecular distance of 3.51 Å with a well depth of 71.57 cm(-1). An additional planar local minimum was found to be separated from the global minimum with an energy barrier of 23.11 cm(-1). In addition, two first-order and one second-order saddle points were also located. The combined radial discrete variable representation/angular finite basis representation method and the Lanczos algorithm were employed to evaluate the rovibrational energy levels for eight isotopic species of the Ne-H(2)S complexes. The rotational transition frequencies for the eight isotopomers were also determined for the ground and first vibrational excited states, which are all in very good agreement with the available experimental values.