Promoting plant resilience against stress by engineering root microenvironment with Streptomyces inoculants.

Plant growth is directly influenced by biotic and abiotic stress factors resulting from environmental changes. Plant growth-promoting rhizobacteria (PGPR) have become a crucial area of research aimed at addressing these challenges. However, a knowledge gap exists regarding how PGPR impacts the microenvironments surrounding plant roots. The purpose of this study is to elucidate the effects of two distinct PGPR strains, Streptomyces griseorubiginosus BTU6 (known for its resistance to smut disease) and S. chartreusis WZS021, on sugarcane roots. Additionally, we compare the resultant modifications in the physicochemical characteristics of the rhizospheric soil and root architecture. The results reveal that following the inoculation of S. chartreusis WZS021, there was a significant increase in the active chemicals associated with nitrogen metabolism in sugarcane roots. This enhancement led to a substantial enrichment of nitrogen-cycling microbes like Pseudomonas and Gemmatimona. This finding supports earlier research indicating that S. chartreusis WZS021 enhances sugarcane's capacity to utilize nitrogen effectively. Furthermore, after treatment with S. chartreusis, Aspergillus became the predominant strain among endophytic fungi, resulting in alterations to their community structure that conferred drought resistance. In contrast, the relative abundance of Xanthomonas in the root environment decreased following inoculation with S. griseorubiginosus. Instead, Gemmatimona became more prevalent, creating a favorable environment for plants to bolster their resistance against disease. Notably, inoculations with S. chartreusis WZS021 and S. griseorubiginosus BTU6 led to substantial changes in the chemical composition, enzymatic activity, and microbial community composition in the soil surrounding sugarcane roots. However, there were distinct differences in the specific alterations induced by each strain. These findings enhance plant resilience to stress by shedding light on PGPR-mediated modifications in root microenvironments.

The complete chloroplast genome of Piper sarmentosum Roxburgh, 1820 (Piperaceae).

Piper sarmentosum Roxb. (Piperaceae) is a traditional medicinal herb native to Southeast Asia. The complete genome of P. sarmentosum was sequenced and characterized in this study with the aim of providing genomic resources for the evolution and molecular breeding of P. sarmentosum. It has a typical quadripartite structure, with a large single-copy (LSC) region of 88,979 bp, a small single-copy (SSC) region of 18,274 bp, and two copies of 27,068 bp inverted-repeat regions (IRa and IRb). A total of 130 genes were annotated, comprising 85 protein-coding genes (PCGs), 8 ribosomal RNA (rRNA) genes, and 37 transfer RNA (tRNA) genes. The phylogenetic tree showed that P. sarmentosum in the current study is closely related to Piper longum.

miR-222 enhances radiosensitivity of cancer cells by inhibiting the expression of CD47.

Radiotherapy is one of the most common and effective treatments for localized cancer. However, radiotherapy kills tumor cells while causing damage to surrounding normal cells. Enhancing the radiation sensitivity of tumor cells and reducing the radiation damage to normal cells is a difficult problem. Here, we find that the expression of a human microRNA (miRNA), hsa-miR-222, is upregulated in response to ionizing radiation. TargetScan analysis shows that the 3' UTR of CD47 is potentially targeted by miR-222. This prediction was validated by luciferase reporter and mutation assays. It was demonstrated that miR-222 negatively regulates CD47 expression at mRNA and protein levels, and overexpression of the miR-222 enhances cancer cell radiosensitivity by the CD47-pERK pathway in cancer cells. Our findings enrich the complex relationship between miRNA and CD47 in irradiation stress and shed light on the potential of miRNAs both for direct cancer therapeutics and as tools to sensitize tumor cells to radiotherapy.

The Solanum lycopersicum auxin response factor SlARF2 participates in regulating lateral root formation and flower organ senescence.

ARF2 as apleiotropic developmental regulator has been reported in Arabidopsis thaliana and tomato (Solanum lycopersicum). The present study showed SlARF2 transcripts in all tomato plant tissues but with higher accumulation in flowers. During bud-anthesis stages, SlARF2 transcripts showed a dynamic expression pattern in sepal, stamen, ovary and petal. Hormone treatment analysis suggested that SlARF2 transcript accumulation was positively regulated by auxin and gibberellic acid, and negatively regulated by ethylene in tomato seedlings. Phenotypes and molecular analyses of SlARF2-upregulated transgenic tomato indicated that SlARF2 regulated tomato lateral root formation and flower organ senescence may be partially mediated by regulating the gene expression of auxin and ethylene response factors. The data enlarges the functional characterization of SlARF2 in tomato, and broadens our understanding of auxin signaling in regulating plant growth and development.

Methylation of promoter of RBL1 enhances the radioresistance of three dimensional cultured carcinoma cells.

Three dimensional (3D) culture in vitro is a new cell culture model that more closely mimics the physiology features of the in vivo environment and is being used widely in the field of medical and biological research. It has been demonstrated that cancer cells cultured in 3D matrices are more radioresistant compared with cells in monolayer (2D). However, the mechanisms causing this difference remain largely unclear. Here we found that the cell cycle distribution and expression of cell cycle regulation genes in 3D A549 cells are different from the 2D. The higher levels of the promotor methylation of cell cycle regulation genes such as RBL1 were observed in 3D A549 cells compared with cells in 2D. The treatments of irradiation or 5-Aza-CdR activated the demethylation of RBL1 promotor and resulted in the increased expression of RBL1 only in 3D A549 cells. Inhibition of RBL1 enhanced the radioresistance and decreased the G2/M phase arrest induced by irradiation in 2D A549 and MCF7 cells. Overexpression of RBL1 sensitized 3D cultured A549 and MCF7 cells to irradiation. Taken together, to our knowledge, it is the first time to revealthat the low expression of RBL1 due to itself promotor methylation in 3D cells enhances the radioresistance. Our finding sheds a new light on understanding the features of the 3D cultured cell model and its application in basic research into cancer radiotherapy and medcine development.

SlTIR1 is involved in crosstalk of phytohormones, regulates auxin-induced root growth and stimulates stenospermocarpic fruit formation in tomato.

TIR1 and its homologs act as auxin receptors and play important roles in plant growth and development in Arabidopsis thaliana. An auxin receptor homolog Solanum lycopersicum TIR1 (SlTIR1) has been isolated from tomato cultivar Micro-Tom, and SlTIR1 over-expression results in parthenocarpic fruit formation. In this study, the promoter driving the ß-glucuronidase (GUS) expression vector was constructed and stably transformed into Micro-Tom seedlings. The SlTIR1 promoter driving GUS expression accumulated predominantly in the leaf and vasculature in transgenic seedlings. Promoter analysis identified an auxin-response element (AuxRE) and two gibberellic acid (GA)-response elements in the SlTIR1 promoter. Quantitative PCR showed that SlTIR1 transcript level was down-regulated by naphthaleneacetic acid, ethephon and abscisic acid and up-regulated by GA. Furthermore, because of the lack of ability to form reproductive seeds in SlTIR1 over-expressing Micro-Tom, this limits further exploration of potential roles of SlTIR1 in auxin signaling. Here, an antisense vector and an over-expression vector of the SlTIR1 gene were stably transformed into Micro-Tom and Ailsa Craig tomato, respectively. Phenotypes and physiological analyses indicated that SlTIR1 regulated primary root growth and auxin-associated lateral root formation in Micro-Tom. Meanwhile, SlTIR1 also stimulated abnormal seed development, so-called stenospermocarpy, in Ailsa Craig. Transcript accumulations of auxin-signaling genes determined by quantitative PCR were consistent with the idea that SlTIR1 regulated plant growth and development, partially mediated by controlling the mRNA levels of auxin-signaling genes. Our work demonstrates that SlTIR1 regulated auxin-induced root growth and stimulated stenospermocarpic fruit formation. SlTIR1 may be a key mediator of the crosstalk among auxin and other hormones to co-regulate plant growth and development.

Radiation induces premature chromatid separation via the miR-142-3p/Bod1 pathway in carcinoma cells.

Radiation-induced genomic instability plays a vital role in carcinogenesis. Bod1 is required for proper chromosome biorientation, and Bod1 depletion increases premature chromatid separation. MiR-142-3p influences cell cycle progression and inhibits proliferation and invasion in cervical carcinoma cells. We found that radiation induced premature chromatid separation and altered miR-142-3p and Bod1 expression in 786-O and A549 cells. Overexpression of miR-142-3p increased premature chromatid separation and G2/M cell cycle arrest in 786-O cells by suppressing Bod1 expression. We also found that either overexpression of miR-142-3p or knockdown of Bod1 sensitized 786-O and A549 cells to X-ray radiation. Overexpression of Bod1 inhibited radiation- and miR-142-3p-induced premature chromatid separation and increased resistance to radiation in 786-O and A549 cells. Taken together, these results suggest that radiation alters miR-142-3p and Bod1 expression in carcinoma cells, and thus contributes to early stages of radiation-induced genomic instability. Combining ionizing radiation with epigenetic regulation may help improve cancer therapies.

Ectopic expression a tomato KNOX Gene Tkn4 affects the formation and the differentiation of meristems and vasculature.

The KNOTTED-LIKE HOMEODOMAIN genes are involved in maintenance of the shoot apical meristem which produces the whole above-ground body of vascular plants. In this report, a tomato homolog gene, named as Tkn4 (a nucleus targeted transcription factor) was identified and characterized. By performing RT-PCR, the transcript level of Tkn4 was separately found in stem, root, stamen, stigma, fruit and sepal but hardly visible in the leaf. Besides, Tkn4 was induced by a series of plant hormones. Overexpression of Tkn4 gene in tomato resulted in dwarf phenotype and strongly repressed the formation of shoot apical meristem, lateral meristem and cambiums in transgenic lines. The transgenic lines had wrinkled leaves and anatomic analysis showed that there was no obvious palisade tissues in the leaves and the layer of cells changed in vascular tissue (xylem and phloem). To explore the regulation network of Tkn4, RNA-sequencing was performed in overexpression lines and wild type plants, by which many genes related to the synthesis and the signal transduction of cytokinin, auxin, gibberellin, ethylene, abscisic acid, and tracheary element differentiation or extracellular matrix synthesis were significantly regulated. Taken together, our results demonstrate that Tkn4 plays important roles in regulating the biosynthesis and signal transduction of diverse plant hormones, and the formation and differentiation of meristems and vasculature in tomato.

Reprogramming mediated radio-resistance of 3D-grown cancer cells.

In vitro 3D growth of tumors is a new cell culture model that more closely mimics the features of the in vivo environment and is being used increasingly in the field of biological and medical research. It has been demonstrated that cancer cells cultured in 3D matrices are more radio-resistant compared with cells in monolayers. However, the mechanisms causing this difference remain unclear. Here we show that cancer cells cultured in a 3D microenvironment demonstrated an increase in cells with stem cell properties. This was confirmed by the finding that cells in 3D cultures upregulated the gene and protein expression of the stem cell reprogramming factors such as OCT4, SOX2, NANOG, LIN28 and miR-302a, compared with cells in monolayers. Moreover, the expression of ß-catenin, a regulating molecule of reprogramming factors, also increased in 3D-grown cancer cells. These findings suggest that cancer cells were reprogrammed to become stem cell-like cancer cells in a 3D growth culture microenvironment. Since cancer stem cell-like cells demonstrate an increased radio-resistance and chemo-resistance, our results offer a new perspective as to why. Our findings shed new light on understanding the features of the 3D growth cell model and its application in basic research into clinical radiotherapy and medicine.

A feedback regulation between miR-145 and DNA methyltransferase 3b in prostate cancer cell and their responses to irradiation.

It is believed that epigenetic modification plays roles in cancer initiation and progression. Both microRNA and DNA methyltransferase are epigenetic regulation factors. It was found that miR-145 upregulates while DNMT3b downregulates in PC3 cells. Presence of any negative correlationship and their response to irradiation were investigated in the current study. We found that miR-145 downregulated DNMT3b expression by directly targeting the 3'-UTR of DNMT3b mRNA and knockdown of DNMT3b increased expression of miR-145 via CpG island promoter hypomethylation, suggesting that there is a crucial crosstalk between miR-145 and DNMT3b via a double-negative feedback loop. Responses of the miR-145 and DNMT3b to irradiation are a negative correlation. We also found that either overexpression of miR-145 or knockdown of DNMT3b sensitized prostate cancer cells to X-ray radiation. Our findings enrich the complex relationships between miRNA and DNMTs in carcinogenesis and irradiation stress. It also sheds light on the potential combination of ionizing radiation and epigenetic regulation in prostate cancer therapy.

The tomato SlIAA15 is involved in trichome formation and axillary shoot development.

The Aux/IAA genes encode a large family of short-lived proteins known to regulate auxin signalling in plants. Functional characterization of SlIAA15, a member of the tomato (Solanum lycopersicum) Aux/IAA family, shows that the encoded protein acts as a strong repressor of auxin-dependent transcription. The physiological significance of SlIAA15 was addressed by a reverse genetics approach, revealing that SlIAA15 plays multiple roles in plant developmental processes. The SlIAA15 down-regulated lines display lower trichome number, reduced apical dominance with associated modified pattern of axillary shoot development, increased lateral root formation and decreased fruit set. Moreover, the leaves of SlIAA15-inhibited plants are dark green and thick, with larger pavement cells, longer palisade cells and larger intercellular space of spongy mesophyll cells. The SlIAA15-suppressed plants exhibit a strong reduction in type I, V and VI trichome formation, suggesting that auxin-dependent transcriptional regulation is required for trichome initiation. Concomitant with reduced trichome formation, the expression of some R2R3 MYB genes, putatively involved in the control of trichome differentiation, is altered. These phenotypes uncover novel and specialized roles for Aux/IAAs in plant developmental processes, clearly indicating that members of the Aux/IAA gene family in tomato perform both overlapping and specific functions.

Functional identification of a novel F-box/FBA gene in tomato.

In plants and animals, the SCF-type ubiquitin protein ligases play an important role in many different physiological processes by regulating protein stability such as S-RNase-based self-compatibility, flower development, hormone responses and meiosis. This study identified an SlFbf gene in tomato that encodes 381 amino acid residues containing a typical F-box motif and an FBA_1 motif associated proteasome pathway; the transcripts of SlFbf was detected in all the tissues (root, stem, leaf, sepal, petal, stamen, pistil, green fruit, breaker fruit and red fruit), with the highest in stamen specifically during flowering stage; SlFbf responded to gibberellins, abscisic acid and light. Suppressed SlFbf leads to bigger pollen and less seeds showing that SlFbf might have an effect on fertilization through regulating stamen development. These findings provide more information about the functions of Fbf gene family.

The auxin receptor homologue in Solanum lycopersicum stimulates tomato fruit set and leaf morphogenesis.

TIR1 and its homologues act as auxin receptors and play a crucial role in auxin-mediated plant development. While the functions of auxin receptor genes have been widely studied in Arabidopsis thaliana, there has been no report on the consequences of TIR1 overexpression in plants that regulate fruit development. Here a putative tomato auxin receptor gene, homologous to Arabidopsis AtTIR1, is reported. This gene, designated as Solanum lycopersicum TIR1 (SlTIR1), was found to be expressed in all the parts of floral buds and flowers at anthesis stages. From bud to anthesis, SlTIR1 expression increases slightly in sepal tissue and decreases dramatically in stamen. From anthesis to post-anthesis when fruit set is expected to occur, the expression of SlTIR1 declines in the ovary and sepal. Overexpression of SlTIR1 results in a pleiotropic phenotype including parthenocarpic fruit formation and leaf morphology. Furthermore, SlTIR1 overexpression altered transcript levels of a number of auxin-responsive genes. The present data demonstrate that the tomato SlTIR1 gene plays an important role at the stages of flower-to-fruit transition and leaf formation.