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Summary: Sterols have long been associated with diverse fields, such as cancer treatment, drug development, and plant growth; however, their underlying mechanisms and functions remain enigmatic. Here, we unveil a critical role played by a GmNF‐YC9‐mediated CCAAT‐box transcription complex in modulating the steroid metabolism pathway within soybeans. Specifically, this complex directly activates squalene monooxygenase (GmSQE1), which is a rate‐limiting enzyme in steroid synthesis. Our findings demonstrate that overexpression of either GmNF‐YC9 or GmSQE1 significantly enhances soybean stress tolerance, while the inhibition of SQE weakens this tolerance. Field experiments conducted over two seasons further reveal increased yields per plant in both GmNF‐YC9 and GmSQE1 overexpressing plants under drought stress conditions. This enhanced stress tolerance is attributed to the reduction of abiotic stress‐induced cell oxidative damage. Transcriptome and metabolome analyses shed light on the upregulation of multiple sterol compounds, including fucosterol and soyasaponin II, in GmNF‐YC9 and GmSQE1 overexpressing soybean plants under stress conditions. Intriguingly, the application of soybean steroids, including fucosterol and soyasaponin II, significantly improves drought tolerance in soybean, wheat, foxtail millet, and maize. These findings underscore the pivotal role of soybean steroids in countering oxidative stress in plants and offer a new research strategy for enhancing crop stress tolerance and quality from gene regulation to chemical intervention. [ABSTRACT FROM AUTHOR]

Biotic stresses, like plant diseases and insect pests, and abiotic stresses, such as drought and salt, heavily threaten plant growth and influence crop yield and quality. Osmotic stress resulting from water deficiency in the soil is one of major hurdles. Nuclear Factor Y (NF-Y) is a heterotrimeric protein, consisting of NF-YA, NF-YB, and NF-YC in plant. NF-Y plays significant roles in the pathway responding to osmotic stresses in plants. We acquired a gene GmNF-YCa, a member of regulatory subunit NF-YC family in soybean (Glycine max L.), by screening a soybean cDNA library using yeast two-hybrid system. The full sequence of GmNF-YCa is 864 bp, encoding 287 amino acids and having a NF-YC motif, belonging to NF-YC subfamily. GmNF-YCa is a hydrophilic protein with a molecular weight of 31.6 kD and isoelectric point of 5.07, and containing a transmembrane domain and no any signal peptides. Sequence analysis showed that NF-YC subfamily was highly conserved among various species. Promoters of GmNF-YCa contained various abiotic stresses and light responsive elements, such as ARE, Box 4, GATA-motif, Box I, ACE, ABRE, and CAT-Box. According to tissue-specific analysis, GmNF-YCa had the highest expression level in germination stage. Quantitative real-time PCR suggested that GmNF-YCa was induced by sucrose stress and mannitol treatment. GmNF-YCa was transformated to Arabidopsis successfully by Agrobacterium-mediated method and the overexpressed Arabidopsis was prepared for the function characterization analysis. Overexpression of GmNF-YCa could improve tolerance of transgenic Arabidopsis to osmotic stress in the germination stage, and enhance root development with more lateral roots in sucrose and mannitol treatments. [ABSTRACT FROM AUTHOR]

Agrobacterium rhizogenes is a soil bacteria with extensive infectivity, which can infect almost all dicotyledonous plants and a few monocotyledonous plants to induce root nodules. This is caused by the root-inducing plasmid, which contains genes responsible for the autonomous growth of root nodules and crown gall base synthesis. Structurally, it is similar to the tumor-inducing plasmid in that it mainly contains the Vir region, the T-DNA region, and the functional region of crown gall base synthesis. Its T-DNA is integrated into the nuclear genome of the plant with the assistance of Vir genes, causing hairy root disease in the host plant and the formation of hairy roots. The roots produced by Agrobacterium rhizogenes -infested plants are characterized by a fast growth rate, high degree of differentiation, physiological, biochemical, and genetic stability, and ease of manipulation and control. In particular, the hairy root system is an efficient and rapid research tool for plants that have no affinity for transformation by Agrobacterium rhizogenes and low transformation efficiency. The establishment of germinating root culture system for the production of secondary metabolites in the original plants through the genetic transformation of natural plants mediated by root-inducing plasmid in Agrobacterium rhizogenes has become a new technology combining plant genetic engineering and cell engineering. It has been widely used in a variety of plants for different molecular purposes, such as pathological analysis, gene function verification, and secondary metabolite research. Chimeric plants obtained by induction of Agrobacterium rhizogenes that can be expressed instantaneously and contemporarily are more rapidly obtained, compared to tissue culture and stably inheritable transgenic strains. In general, transgenic plants can be obtained in approximately one month., Competing Interests: Competing interestsThere are no conflicts of interest or competing interests., (Copyright © 2023 The Authors; exclusive licensee Bio-protocol LLC.)

Objective: To evaluate the animal model of the multidrug resistant glioma cell line C6/adr for further in vivo studies., Methods: The rat glioma cells C6 and multidrug resistance cells C6/adr were cultured in vitro and implanted into the brain of S-D rats. After implantation, all these animals were examined continually with magnetic resonance imaging (MRI) and histological examination. The growth procedure of intracranial implanted glioma and the survival span of the animal model were evaluated. The statistical analysis was made between the survival data of the two cell lines., Results: The symptoms of intracranial hypertension did not occur until 4 weeks after inoculation. The MRI findings of the implanted glioma in the rat brain were much earlier than the abnormal behavior observed. Pathological results after inoculation demonstrated the MRI findings. The two cell lines had similar growth characteristics and no significant differences in survival times., Conclusion: These results suggest that by means of MRI and histology the growth procedure of the implanted glioma in vivo be successfully observed. All these data will proved to be a useful basis for study of glioma in vivo.