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[This corrects the article DOI: 10.1371/journal.pone.0181962.].

High transformation efficiency is a prerequisite for study of gene function and molecular breeding. -mediated transformation is a preferred method in many plants. However, the transformation efficiency in soybean is still low. The objective of this study is to optimize -mediated transformation in soybean by improving the infection efficiency of and regeneration efficiency of explants. Firstly, four factors affecting infection efficiency were investigated by estimation of the rate of GUS transient expression in soybean cotyledonary explants, including concentrations, soybean explants, suspension medium, and co-cultivation time. The results showed that an infection efficiency of over 96% was achieved by collecting the at a concentration of OD = 0.6, then using an suspension medium containing 154.2 mg/L dithiothreitol to infect the half-seed cotyledonary explants (from mature seeds imbibed for 1 day), and co-cultured them for 5 days. The infection efficiencies for soybean varieties Jack Purple and Tianlong 1 were higher than the other six varieties. Secondly, the rates of shoot elongation were compared among six different concentration combinations of gibberellic acid (GA ) and indole-3-acetic acid (IAA). The shoot elongation rate of 34 and 26% was achieved when using the combination of 1.0 mg/L GA and 0.1 mg/L IAA for Jack Purple and Tianlong 1, respectively. This rate was higher than the other five concentration combinations of GA and IAA, with an 18 and 11% increase over the original laboratory protocol (a combination of 0.5 mg/L GA and 0.1 mg/L IAA), respectively. The transformation efficiency was 7 and 10% for Jack Purple and Tianlong 1 at this optimized hormone concentration combination, respectively, which was 2 and 6% higher than the original protocol, respectively. Finally, GUS histochemical staining, PCR, herbicide (glufosinate) painting, and QuickStix Kit for Liberty Link ( ) were used to verify the positive transgenic plants, and absolute quantification PCR confirmed the exogenous gene existed as one to three copies in the soybean genome. This study provides an improved protocol for -mediated transformation in soybean and a useful reference to improve the transformation efficiency in other plant species.

Breast and prostate cancer research to date has largely been predicated on the use of cell lines in vitro or in vivo. These limitations have led to the development of more clinically relevant models, such as organoids or murine xenografts that utilize patient‐derived material; however, issues related to low take rate, long duration of establishment, and the associated costs constrain use of these models. This study demonstrates that ex vivo culture of freshly resected breast and prostate tumor specimens obtained from surgery, termed patient‐derived explants (PDEs), provides a high‐throughput and cost‐effective model that retains the native tissue architecture, microenvironment, cell viability, and key oncogenic drivers. The PDE model provides a unique approach for direct evaluation of drug responses on an individual patient's tumor, which is amenable to analysis using contemporary genomic technologies. The ability to rapidly evaluate drug efficacy in patient‐derived material has high potential to facilitate implementation of personalized medicine approaches. This study describes an innovative preclinical model that has significant potential to accelerate translational cancer research outcomes. Breast and prostate tumors were cultured as patient‐derived explants (PDE), in a way that sustains the native tissue architecture, microenvironment, cell viability, and steroid receptor signaling. PDEs were responsive to hormones, therapeutic agents, and shRNA while in culture, allowing direct comparison of treatments within an individual's tumor using high‐throughput molecular approaches, including cistrome profiling.

Tetrastigma hemsleyanum is a perennial evergreen vine of the Vitaceae. The entire herb is used in traditional Chinese medicine as a broad-spectrum plant-based antibiotic, so it has high economic and social value. Wild T. hemsleyanum resources are scarce, so it has been declared an endangered and rare medicinal plant. Seed yield is low and vegetative propagation by cuttings results in limited plant production, so development of the T. hemsleyanum industry requires optimized propagation protocols and the development of new biotechnologies to proliferate this plant in commercial quantities. In this study, shoot organogenesis was successfully induced from leaves and petioles. Two plant growth regulators, 6-benzyladenine (BA) and thidiazuron, induced callus and adventitious shoots, but the ideal adventitious shoot induction medium was Murashige and Skoog (MS) medium containing 1.0 mg L−1 BA and 0.1 mg L−1 α-naphthaleneacetic acid (NAA). This resulted in a shoot proliferation coefficient (SPC) of 6.73 within 30 d at a light intensity of 100 µmol m−2 s−1. When light intensity was increased from 50 to 200 µmol m−2 s−1, SPC (7.35), chlorophyll a (Chl a), Chl b, and total Chl (a + b) content increased. On MS medium containing 0.1–2.0 mg L−1 NAA or indole-3-butyric acid, 100% of adventitious shoots formed adventitious roots. Plantlets showed no obvious morphological variation, and their survival exceeded 98% on a substrate of peat and river sand (v:v = 2:1). This study’s protocols allow for the mass production of adventitious shoots for conservation purposes, and potentially for the commercial propagation of T. hemsleyanum.

Despite many improvements in ovarian cancer diagnosis and treatment, until now, conventional chemotherapy and new biological drugs have not been shown to cure the disease, and the overall prognosis remains poor. Over 90% of ovarian malignancies are categorized as epithelial ovarian cancers (EOC), a collection of different types of neoplasms with distinctive disease biology, response to chemotherapy, and outcome. Advances in our understanding of the histopathology and molecular features of EOC subtypes, as well as the cellular origins of these cancers, have given a boost to the development of clinically relevant experimental models. The overall goal of this review is to provide a comprehensive description of the available preclinical investigational approaches aimed at better characterizing disease development and progression and at identifying new therapeutic strategies. Systems discussed comprise monolayer (2D) and three-dimensional (3D) cultures of established and primary cancer cell lines, organoids and patient-derived explants, animal models, including carcinogen-induced, syngeneic, genetically engineered mouse, xenografts, patient-derived xenografts (PDX), humanized PDX, and the zebrafish and the laying hen models. Recent advances in tumour-on-a-chip platforms are also detailed. The critical analysis of strengths and weaknesses of each experimental model will aid in identifying opportunities to optimize their translational value.

Physical function declines in old age, portending disability, increased health expenditures, and mortality. Cellular senescence, leading to tissue dysfunction, may contribute to these consequences of aging, but whether senescence can directly drive age-related pathology and be therapeutically targeted is still unclear. Here we demonstrate that transplanting relatively small numbers of senescent cells into young mice is sufficient to cause persistent physical dysfunction, as well as to spread cellular senescence to host tissues. Transplanting even fewer senescent cells had the same effect in older recipients and was accompanied by reduced survival, indicating the potency of senescent cells in shortening health- and lifespan. The senolytic cocktail, dasatinib plus quercetin, which causes selective elimination of senescent cells, decreased the number of naturally occurring senescent cells and their secretion of frailty-related proinflammatory cytokines in explants of human adipose tissue. Moreover, intermittent oral administration of senolytics to both senescent cell–transplanted young mice and naturally aged mice alleviated physical dysfunction and increased post-treatment survival by 36% while reducing mortality hazard to 65%. Our study provides proof-of-concept evidence that senescent cells can cause physical dysfunction and decreased survival even in young mice, while senolytics can enhance remaining health- and lifespan in old mice. Transfer of senescent cells into naive, young mice can induce physical dysfunction, and a senolytic can reverse this dysfunction and potently increase lifespan in aged mice.

The genus Opuntia includes more than 300 species, with the most economically important being Opuntia ficus-indica. In this study, the in vitro propagation potential of the prickly pear cactus (O. ficus-indica (L.) Mill.) was evaluated. For shoot initiation (6 weeks, MS medium, 25 ± 1 °C, 24 h dark), three cladode explant types based on areole number (none, one, several), five BA (2, 3, 4, 5 and 6 mg L-1) and four IAA concentrations (0, 1, 1.5 and 2 mg L-1) were tested. The results showed 100% shoot induction by cladodes with several areoles under 6 mg L-1 BA + IAA (1-2 mg L-1), compared to cladodes with one (63.33-66.67%) or none areole (26.66-30%). For multiplication of in vitro shoots derived from areole stimulation, four cytokinins [6-benzyladenine (BA), kinetin (Kin), thidiazuron (TDZ), forchlorfenuron (CPPU)] alone and combined with two auxins [indole-3-acetic acid (IAA), 2,4-dichlorophenoxy acetic acid (2,4-D)] were tested (8 weeks, Murashige-Skoog medium, 25 °C, 16h photoperiod). These results showed significantly higher shoot number (5.10) and length (6.19 cm) under 5 mg L-1 CPPU and 5 mg L-1 CPPU + 0.5 mg L-1 2,4-D, respectively. For in vitro rooting, three auxins [indole-3-butyric acid (IBA), indole-3-acetic acid (IAA), α-naphthaleneacetic acid (NAA)] were applied (5 weeks, MS medium, 25 °C, 16-h photoperiod) and the results showed higher root number (19.06) with 0.5 mg L-1 IBA. For acclimatization, rooted microshoots (ex vitro) placed in a peat: vermiculite substrate and non-rooted microshoots (ex vivo) to the same substrate but supplemented with different concentrations (0, 0.8, 1.2, 1.6, 2 g alveoli-1) of a controlled release fertilizer (CRF, 15%N-9%P2O5-12%K2O). After 10 weeks in the greenhouse (24 ± 5 °C/ 18 ± 5 °C), 100% rooting (ex vivo) and survival (ex vitro, ex vivo) were recorded, nonetheless 1.2 g alveoli-1 CRF gave higher ex vivo root number (23.05).

Plant gene editing is typically performed by delivering reagents such as Cas9 and single guide RNAs to explants in culture. Edited cells are then induced to differentiate into whole plants by exposure to various hormones. The creation of edited plants through tissue culture is often inefficient, time-consuming, works for only limited species and genotypes, and causes unintended changes to the genome and epigenome. Here we report two methods to generate gene-edited dicotyledonous plants through de novo meristem induction. Developmental regulators and gene-editing reagents are delivered to somatic cells of whole plants. This induces meristems that produce shoots with targeted DNA modifications, and gene edits are transmitted to the next generation. The de novo induction of gene-edited meristems sidesteps the need for tissue culture and promises to overcome a bottleneck in plant gene editing.Methods to induce edited somatic plant cells to form meristems circumvent tissue culture and enable genome editing of a wider set of plant species.

This study was conducted to develop a protocol for in vitro shoot multiplication and callus induction of various mung bean varieties to obtain enhanced phytochemical content with the help of elicitors. For shoot multiplication, two types of explants (shoot tips and nodal tips) of three varieties of mung bean (Mung NCM-13, MgAT-7, and MgAT-4) were used. Both types of explants from in vitro and in vivo sources were cultured on the MS medium supplemented with different concentrations (0.25–3.0 mg/L, increment of 0.5 mg/L) and combinations of BAP and IBA as independent treatments. For callus induction, leaf explants ( in vitro source) were cultured on MS medium supplemented with 2,4-D (1–3 mg/L) alone or in combination with BAP or NAA (0.5 and 1.0 mg/L). For the enhanced production of phenolics and glycosides, calli were cultured on MS media supplemented with zinc oxide (0.5 mg/L) and copper oxide nanoparticles (0.5 mg/L) as nano-elicitors. Results showed that in vitro explants responded better in terms of shoot length, number of shoots, and number of leaves per explant when compared to in vivo explants. Moreover, shoot tips were better than nodal explants to in vitro culturing parameters. All three varieties showed the optimized results in the MS medium supplemented with 1 mg/L BAP, while roots were produced only in cultures fortified with 1 mg/L IBA. The leaf explants of in vitro and soil-grown plantlets showed a maximum callogenic response of 90 and 80%, respectively, on MS medium supplemented with 2,4-D (3 mg/ml). Maximum phenolic content (101.4 μg of gallic acid equivalent/g) and glycoside content (34 mg of amygdalin equivalent/g of plant material) was observed in the calli cultured on MS medium supplemented with 3 mg/L of 2,4-D. Furthermore, the addition of zinc oxide (0.5 mg/L) and copper oxide (0.5 mg/L) nanoparticles to the callus culture medium significantly enhanced the phenolic content of Mung NCM-13 (26%), MgAT-7 (25.6%), and MgAT-4 (22.7%). Glycosidic content was also found to be increased in Mung NCM-13 (50%), MgAT-7 (37.5%), and MgAT-4 (25%) varieties when compared to the control. It is suggested that elicitation of in vitro cultures of mung beans with nanoparticles could be an effective strategy for the enhanced production of secondary metabolites.