GA-related Genes Research Articles

Prognosis and metabolism with a Golgi apparatus-related genes-based formula in breast cancer.

The Golgi apparatus (GA), an organelle that processes, sorts, and transports proteins synthesized by the endoplasmic reticulum, is also involved in many cellular processes associated with cancer, such as angiogenesis, the innate immune response, and tumor invasion and migration. We aimed to construct a breast cancer (BC) prognosis prediction model based on GA-related genetic information to evaluate the prognosis of patients with BC more accurately than existing models and to stratify patients for clinical therapy. In this study, The Cancer Genome Atlas-breast invasive carcinoma was used as the training cohort, and the Molecular Taxonomy of Breast Cancer International Consortium cohort was used as the validation cohort. Using bioinformatics methods, we constructed a GA-related gene risk score (GRS). The GRS was used to divide BC patients into a high-GRS group and a low-GRS group, and functional analysis, survival analysis, mutation analysis, immune landscape analysis, and metabolic analysis were performed to compare the 2 groups. Finally, a nomogram was constructed for clinical application. The genes in the GRS model were mainly related to the glucose metabolism pathway, and the main mutations in the 2 groups of patients were mutations in TP53 and CHD1. The mutation rate in the high-GRS group was greater than that in the low-GRS group. The high GRS group had higher tumor immune activity glycolysis; the pentose phosphate pathway tended to be the dominant metabolic pathways in this group, while fatty acid oxidation and glutamine catabolism tended to be dominant in the low-GRS group. GA-related genes were used to construct a prediction model for BC patients and had high accuracy in predicting prognosis. The mutations associated with the GRS are mainly TP53 and CDH1. Interestingly, the GRS is correlated with glucose metabolism in terms of gene expression and functional enrichment. In summary, the role of GRS-related genes in glucose metabolism is worthy of further study.

Characterization of the ddt1 Mutant in Rice and Its Impact on Plant Height Reduction and Water Use Efficiency.

Rice (Oryza sativa L.), a fundamental global staple, nourishes over half of the world's population. The identification of the ddt1 mutant in rice through EMS mutagenesis of the indica cultivar Shuhui527 revealed a dwarf phenotype, characterized by reduced plant height, smaller grain size, and decreased grain weight. Detailed phenotypic analysis and map-based cloning pinpointed the mutation to a single-base transversion in the LOC_Os03g04680 gene, encoding a cytochrome P450 enzyme, which results in a premature termination of the protein. Functional complementation tests confirmed LOC_Os03g04680 as the DDT1 gene responsible for the observed phenotype. We further demonstrated that the ddt1 mutation leads to significant alterations in gibberellic acid (GA) metabolism and signal transduction, evidenced by the differential expression of key GA-related genes such as OsGA20OX2, OsGA20OX3, and SLR1. The mutant also displayed enhanced drought tolerance, as indicated by higher survival rates, reduced water loss, and rapid stomatal closure under drought conditions. This increased drought resistance was linked to the mutant's improved antioxidant capacity, with elevated activities of antioxidant enzymes and higher expression levels of related genes. Our findings suggest that DDT1 plays a crucial role in regulating both plant height and drought stress responses. The potential for using gene editing of DDT1 to mitigate the dwarf phenotype while retaining improved drought resistance offers promising avenues for rice improvement.

Molecular characterization of Golgi apparatus-related genes indicates prognosis and immune infiltration in osteosarcoma.

The Golgi apparatus (GA) is crucial for protein synthesis and modification, and regulates various cellular processes. Dysregulation of GA can lead to pathological conditions like neoplastic growth. GA-related genes (GARGs) mutations are commonly found in cancer, contributing to tumor metastasis. However, the expression and prognostic significance of GARGs in osteosarcoma are yet to be understood. Gene expression and clinical data of osteosarcoma patients were obtained from the TARGET and GEO databases. A consensus clustering analysis identified distinct molecular subtypes based on GARGs. Discrepancies in biological processes and immunological features among the subtypes were explored using GSVA, ssGSEA, and Metascape analysis. A GARGs signature was constructed using Cox regression. The prognostic value of the GARGs signature in osteosarcoma was evaluated using Kaplan-Meier curves and a nomogram. Two GARG subtypes were identified, with Cluster A showing better prognosis, immunogenicity, and immune cell infiltration than Cluster B. A novel risk model of 3 GARGs was established using the TARGET dataset and validated with independent datasets. High-risk patients had poorer overall survival, and the GARGs signature independently predicted osteosarcoma prognosis. Combining risk scores and clinical characteristics in a nomogram improved prediction performance. Additionally, we discovered Stanniocalcin-2 (STC2) as a significant prognostic gene highly expressed in osteosarcoma and potential disease biomarker. Our study revealed that patients with osteosarcoma can be divided into two GARGs subgroups. Furthermore, we have developed a GARGs prognostic signature that can accurately forecast the prognosis of osteosarcoma patients.

Diversity of Gibberellin 2-oxidase genes in the barley genome offers opportunities for genetic improvement

IntroductionGibberellin (GA) is a vital phytohormone in regulating plant growth and development. During the “Green Revolution”, modification of GA-related genes created semi-dwarfing phenotype in cereal crops but adversely affected grain weight. Gibberellin 2-oxidases (GA2oxs) in barley act as key catabolic enzymes in deactivating GA, but their functions are still less known. ObjectivesThis study investigates the physiological function of two HvGA2ox genes in barley and identifies novel semi-dwarf alleles with minimum impacts on other agronomic traits. MethodsVirus-induced gene silencing and CRISPR/Cas9 technology were used to manipulate gene expression of HvGA2ox9 and HvGA2ox8a in barley and RNA-seq was conducted to compare the transcriptome between wild type and mutants. Also, field trials in multiple environments were performed to detect the functional haplotypes. ResultsThere were ten GA2oxs that distinctly expressed in shoot, tiller, inflorescence, grain, embryo and root. Knockdown of HvGA2ox9 did not affect plant height, while ga2ox8a mutants generated by CRISPR/Cas9 increased plant height and significantly altered seed width and weight due to the increased bioactive GA4 level. RNA-seq analysis revealed that genes involved in starch and sucrose metabolism were significantly decreased in the inflorescence of ga2ox8a mutants. Furthermore, haplotype analysis revealed one naturally occurring HvGA2ox8a haplotype was associated with decreased plant height, early flowering and wider and heavier seed. ConclusionOur results demonstrate the potential of manipulating GA2ox genes to fine tune GA signalling and biofunctions in desired plant tissues and open a promising avenue for minimising the trade-off effects of Green Revolution semi-dwarfing genes on grain size and weight. The knowledge will promote the development of next generation barley cultivars with better adaptation to a changing climate.

Genome-wide association studies and expression analysis of GA-related genes for exploring foot rot resistance in rice germplasm

Genome-wide association studies and expression analysis of GA-related genes for exploring foot rot resistance in rice germplasm

Salinomycin disturbs Golgi apparatus function and specifically affects cells in epithelial-to-mesenchymal transition.

Epithelial-to-mesenchymal transition (EMT) gives rise to cells with properties similar to cancer stem cells (CSCs). Targeting the EMT program to selectively eliminate CSCs is a promising way to improve cancer therapy. Salinomycin (Sal), a K+/H+ ionophore, was identified as highly selective towards CSC-like cells, but its mechanism of action and selectivity remains elusive. Here we show that Sal, similarly to monensin and nigericin, disturbs the function of the Golgi apparatus (GA). Sal alters the expression of GA-related genes and leads to marked changes in GA morphology, particularly in cells that underwent EMT. Moreover, GA disturbing agents severely affect post-translational modifications of proteins, including protein processing, glycosylation, and secretion. We discover that the alterations induced by GA disturbing agents specifically affect the viability of EMT cells. Collectively, our work reveals a novel vulnerability related to the EMT, suggesting an important role for the GA in the EMT, while targeting the GA represents a novel therapeutic approach against CSCs.

Transcriptome-Based Construction of the Gibberellin Metabolism and Signaling Pathways in Eucalyptus grandis × E. urophylla, and Functional Characterization of GA20ox and GA2ox in Regulating Plant Development and Abiotic Stress Adaptations.

Gibberellins (GAs) are the key regulators controlling plant growth, wood production and the stress responses in perennial woody plants. The role of GA in regulating the above-mentioned processes in Eucalyptus remain largely unclear. There is still a lack of systematic identification and functional characterization of GA-related genes in Eucalyptus. In this study, a total of 59,948 expressed genes were identified from the major vegetative tissues of the E. grandis × E. urophylla using transcriptome sequencing. Then, the key gene families in each step of GA biosynthesis, degradation and signaling were investigated and compared with those of Arabidopsis, rice, and Populus. The expression profile generated using Real-time quantitative PCR showed that most of these genes exhibited diverse expression patterns in different vegetative organs and in response to abiotic stresses. Furthermore, we selectively overexpressed EguGA20ox1, EguGA20ox2 and EguGA2ox1 in both Arabidopsis and Eucalyptus via Agrobacterium tumefaciens or A. rhizogenes-mediated transformation. Though both Arabidopsis EguGA20ox1- and EguGA20ox2-overexpressing (OE) lines exhibited better vegetative growth performance, they were more sensitive to abiotic stress, unlike EguGA2ox1-OE plants, which exhibited enhanced stress resistance. Moreover, overexpression of EguGA20ox in Eucalyptus roots caused significantly accelerated hairy root initiation and elongation and improved root xylem differentiation. Our study provided a comprehensive and systematic study of the genes of the GA metabolism and signaling and identified the role of GA20ox and GA2ox in regulating plant growth, stress tolerance, and xylem development in Eucalyptus; this could benefit molecular breeding for obtaining high-yield and stress-resistant Eucalyptus cultivars.

Brcd1 Is Associated with Plant Height through the Gibberellin Pathway in Brassica rapa L.

In crops such as Brassica rapa, the agronomic trait plant height affects the leaf light absorption rate, benefits substance accumulation, and increases production by increasing the biological yield of the population. The mechanism of plant height was adequately studied in Brassica napus and Arabidopsis, while Brassica rapa had been rarely reported. Herein, we studied a B. rapa dwarf mutant Brcd1YS. Compared with its wild-type, Yellow sarson, Brcd1YS showed shorter hypocotyls and slow growth, with inhibited root elongation and reduced numbers of lateral roots. Chlorophyll content determination and pollen viability detection indicated that the mutant’s chlorophyll content was higher than that of the wild-type; however, its pollen was inactive. Cytological identification showed that the number of cells in Brcd1YS leaves was significantly reduced and were arranged irregularly compared with those of the wild-type. Bulked Segregant RNA sequencing combined with conventional linkage mapped the dwarf mutation Brcd1YS to Chromosome A06, at position 21,621,766 to 24,683,923 bp. Application of exogenous gibberellic acid (GA) partially rescued the dwarf phenotype of Brcd1YS. GA-related genes BraA06g034110.3C and BraA06g033010.3C were identified as the most promising candidate genes. These results make a valuable contribution to our understanding of the mechanism of plant height determination in B. rapa, paving the way for further breeding of dwarf B. rapa.

Cucumber Strigolactone Receptor CsDAD2 and GA3 Interact to Regulate Shoot Branching in Arabidopsis thaliana L.

Previous studies identified that strigolactones (SLs) and gibberellins (GAs) interacted when controlling branching in plant shoots, but the underlying mechanism remains unknown. qRT-PCR analysis suggested that the SL receptor gene CsDAD2 was significantly upregulated in the leaves, stems, and nodes of cucumber after treatment with 50 mg/L of GA3. Furthermore, the CsDAD2 gene was cloned and introduced into wild-type Arabidopsis plants via Agrobacterium-mediated transformation. For the CsDAD2-OE lines, the endogenous content of GA3 was subsequently higher at the seedling stage, with the number of primary cauline branches also significantly increased at the maturity stage compared with WT. Additionally, GA-related genes were up-regulated in the first inter-nodes and the third nodes of the CsDAD2-OE lines, thus indicating that GA was metabolically active in these tissues. The expression of the branch inhibitor gene AtBRC1 decreased at the seedling stage as well as at the maturity stage of the CsDAD2-OE lines. These findings suggest that CsDAD2 might have important functions in the interactions between GAs and SLs as it can promote the accumulation of GAs in plant nodes and suppress the expression of BRC1, hence increasing primary cauline branching.

Manipulating GA-Related Genes for Cereal Crop Improvement.

The global population is projected to experience a rapid increase in the future, which poses a challenge to global food sustainability. The "Green Revolution" beginning in the 1960s allowed grain yield to reach two billion tons in 2000 due to the introduction of semi-dwarfing genes in cereal crops. Semi-dwarfing genes reduce the gibberellin (GA) signal, leading to short plant stature, which improves the lodging resistance and harvest index under modern fertilization practices. Here, we reviewed the literature on the function of GA in plant growth and development, and the role of GA-related genes in controlling key agronomic traits that contribute to grain yield in cereal crops. We showed that: (1) GA is a significant phytohormone in regulating plant development and reproduction; (2) GA metabolism and GA signalling pathways are two key components in GA-regulated plant growth; (3) GA interacts with other phytohormones manipulating plant development and reproduction; and (4) targeting GA signalling pathways is an effective genetic solution to improve agronomic traits in cereal crops. We suggest that the modification of GA-related genes and the identification of novel alleles without a negative impact on yield and adaptation are significant in cereal crop breeding for plant architecture improvement. We observed that an increasing number of GA-related genes and their mutants have been functionally validated, but only a limited number of GA-related genes have been genetically modified through conventional breeding tools and are widely used in crop breeding successfully. New genome editing technologies, such as the CRISPR/Cas9 system, hold the promise of validating the effectiveness of GA-related genes in crop development and opening a new venue for efficient and accelerated crop breeding.

Transcriptome analysis and identification of abscisic acid and gibberellin-related genes during seed development of alfalfa (Medicago sativa L.)

BackgroundAlfalfa (Medicago sativa) is a widely cultivated plant. Unlike many crops, the main goal of breeding alfalfa is to increase its aboveground biomass rather than the biomass of its seeds. However, the low yield of alfalfa seeds limits alfalfa production. Many studies have explored the factors affecting seed development, in which phytohormones, especially ABA and GAs, play an important role in seed development.ResultsHere, we performed a transcriptome analysis of alfalfa seeds at five development stages. A total of 16,899 differentially expressed genes (DEGs) were identified and classified into 10 clusters, and the enriched Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were determined. The contents of ABA, GA1, GA3, GA4 and GA7 in alfalfa seeds at five development stages were determined. In addition, 14 ABA-related DEGs and 20 GA-related DEGs were identified and analysed. These DEGs are involved in plant hormone pathways and play an important role in seed development. Moreover, morphological and physiological analyses revealed the dynamic changes during the development of alfalfa seeds.ConclusionOverall, our study is the first to analyse the transcriptome across various stages of seed development in alfalfa. The results of our study could be used to improve alfalfa seed yield. The key ABA and GA related-genes are potential targets for improving alfalfa seed yield via genetic engineering in the future.

Transcriptome Analysis Reveals the Role of GA3 in Regulating the Asynchronism of Floral Bud Differentiation and Development in Heterodichogamous Cyclocarya paliurus (Batal.) Iljinskaja.

Cyclocarya paliurus is an important medical plant owing to the diverse bioactive compounds in its leaves. However, the heterodichogamy with female and male functions segregation within protandry (PA) or protogyny (PG) may greatly affect seed quality and its plantations for medicinal use. To speculate on the factor playing the dominant role in regulating heterodichogamy in C. paliurus, based on phenotypic observations, our study performed a multi comparison transcriptome analysis on female and male buds (PG and PA types) using RNA-seq. For the female and male bud comparisons, a total of 6753 differentially expressed genes (DEGs) were detected. In addition, functional analysis revealed that these DEGs were significantly enriched in floral development, hormone, and GA-related pathways. As the dominant hormones responsible for floral differentiation and development, gibberellins (GAs) in floral buds from PG and PA types were quantified using HPLC-MS. Among the tested GAs, GA3 positively regulated the physiological differentiation (S0) and germination (S2) of floral buds. The dynamic changes of GA3 content and floral morphological features were consistent with the expression levels of GA-related genes. Divergences of GA3 contents at S0 triggered the asynchronism of physiological differentiation between male and female buds of intramorphs (PA-M vs. PA-F and PG-F vs. PG-M). A significant difference in GA3 content enlarged this asynchronism at S2. Thus, we speculate that GA3 plays the dominant role in the formation of heterodichogamy in C. paliurus. Meanwhile, the expression patterns of GA-related DEGs, including CPS, KO, GA20ox, GA2OX, GID1, and DELLA genes, which play central roles in regulating flower development, coincided with heterodichogamous characteristics. These results support our speculations well, which should be further confirmed.

2-Hydroxymelatonin Promotes Seed Germination by Increasing Reactive Oxygen Species Production and Gibberellin Synthesis in Arabidopsis thaliana.

It was recently reported that 2-hydroxymelatonin (2-OHM) is responsible for inducing reactive oxygen species (ROS) in plants. ROS are crucial molecules that promote germination through interaction with hormones such as gibberellic acid (GA). In this study, to confirm the pro-oxidant role of 2-OHM, we investigated its effect on seed germination in Arabidopsis thaliana (L.) Heynh. Columbia-0. We found that 2-OHM treatment stimulated seed germination by 90% and 330% in non-dormant and dormant seeds, respectively, whereas melatonin marginally increased germination (~13%) in both seed types compared to untreated control seeds. The germination promotion effects of exogenous 2-OHM treatment were due to increased ROS production followed by the induction of GA synthesis and expression of responsive genes. Accordingly, melatonin 2-hydroxylase (M2H), the gene responsible for 2-OHM synthesis, was strictly expressed only during the germination process. Further molecular genetic analyses using m2h knockout mutant and M2H overexpression clearly supported an increase in ROS triggered by 2-OHM, followed by increased expression of GA-related genes, which shortened the time to germination. Notably, 2-OHM application to m2h knockout mutant seeds fully recovered germination to levels comparable to that of the wild type, whereas melatonin treatment failed to increase germination. Together, these results indicate that 2-OHM is a pivotal molecule that triggers increased ROS production during seed germination, thereby enhancing germination via the GA pathway in Arabidopsis thaliana.

Phosphate Suppression of Arbuscular Mycorrhizal Symbiosis Involves Gibberellic Acid Signaling.

Most land plants entertain a mutualistic symbiosis known as arbuscular mycorrhiza with fungi (Glomeromycota) that provide them with essential mineral nutrients, in particular phosphate (Pi), and protect them from biotic and abiotic stress. Arbuscular mycorrhizal (AM) symbiosis increases plant productivity and biodiversity and is therefore relevant for both natural plant communities and crop production. However, AM fungal populations suffer from intense farming practices in agricultural soils, in particular Pi fertilization. The dilemma between natural fertilization from AM symbiosis and chemical fertilization has raised major concern and emphasizes the need to better understand the mechanisms by which Pi suppresses AM symbiosis. Here, we test the hypothesis that Pi may interfere with AM symbiosis via the phytohormone gibberellic acid (GA) in the Solanaceous model systems Petunia hybrida and Nicotiana tabacum. Indeed, we find that GA is inhibitory to AM symbiosis and that Pi may cause GA levels to increase in mycorrhizal roots. Consistent with a role of endogenous GA as an inhibitor of AM development, GA-defective N. tabacum lines expressing a GA-metabolizing enzyme (GA methyltransferase—GAMT) are colonized more quickly by the AM fungus Rhizoglomus irregulare, and exogenous Pi is less effective in inhibiting AM colonization in these lines. Systematic gene expression analysis of GA-related genes reveals a complex picture, in which GA degradation by GA2 oxidase plays a prominent role. These findings reveal potential targets for crop breeding that could reduce Pi suppression of AM symbiosis, thereby reconciling the advantages of Pi fertilization with the diverse benefits of AM symbiosis.

Phenotypic Characterization and Differential Gene Expression Analysis Reveal That Dwarf Mutant dwf Dwarfism Is Associated with Gibberellin in Eggplant

Dwarfism is a desirable trait in eggplant breeding, as it confers higher lodging resistance and allows simplified management and harvest. However, a few dwarf mutants have been reported, and the molecular mechanism underlying dwarfism in eggplant is completely unknown. Here, we report a dwarf mutant (dwf) isolated from an ethyl methyl sulfonate (EMS)-induced mutant library. The hypocotyl length, plant height, and length of internode cells of dwf were significantly decreased compared to those of the wild-type parent ‘14-345’ (WT). Differential gene expression analysis revealed that GA-related genes, including GA2ox and DELLA, were up-regulated whereas the gibberellin (GA3) content decreased in dwf. Moreover, exogenous GA3 treatment significantly increased the relative growth rate of dwf compared to WT, further indicating the important roles of GA in regulating the dwarf phenotype of dwf. Collectively, our findings shed light on GA-mediated dwarfism in dwf plants and offer a good germplasm that could be used for eggplant dwarfism breeding in the future.

Melatonin promotes seed germination under salt stress by regulating ABA and GA3 in cotton (Gossypium hirsutum L.)

Although previous studies have found that melatonin can promote seed germination, the phytohormone regulation mechanism by which exogenous melatonin mediates salt tolerance during cotton seed germination is still largely unknown. The effects of melatonin on germination traits and physiological parameters of GXM9 cotton seeds (Gossypium hirsutum L.) under three salt stress treatments (CK, germination of seeds pretreated with water alone; S, germination of seeds pretreated in 150 mM NaCl under salt stress; SM, germination of seeds pretreated in 20 μM melatonin under 150 mM NaCl solution) in the laboratory was investigated. The results showed that salt stress (150 mM) inhibited cotton seed germination and endogenous melatonin accumulation, and pretreatment with 20 μM exogenous melatonin enhanced the cotton germination rate and hypocotyl length as well as the content of endogenous melatonin during seed germination. This suggests that exogenous melatonin promotes seed germination from a morphological perspective. The contents of starch, α-amylase (EC3.3.1.1), β-galactosidase (EC3.2.1.23), abscisic acid (ABA), and gibberellin (GA) were determined simultaneously. The results showed that the α-amylase and β-galactosidase contents in the cotton seeds decreased by 56.97% and 20.18%, respectively, under salt stress compared with the control, while the starch content increased by 11.53% compared with the control at day 7. The ABA content increased by 25.18% and GA content decreased by 27.99% under salt stress compared with the control at 24 h. When exogenous melatonin was applied to the cotton seeds, the content of α-amylase and β-galactosidase increased by 121.77% and 32.76%, respectively, whereas the starch contents decreased by 13.55% compared with the S treatment at day 7. Similarly, the ABA content increased by 12.20% and the GA content increased by 4.77% at 24 h. To elucidate the molecular mechanism by which melatonin promotes seed germination under salt stress, the effects of ABA- and GA-related genes on plant hormone signal transduction were analyzed by quantitative real-time PCR and RNA sequencing. The results indicated that melatonin regulated the expression of ABA and GA genes in the plant signal transduction pathway, induced embryo root development and seed germination, and alleviated dormancy. The expression of the ABA signaling gene GhABF2 was up-regulated and GhDPBF2 was down-regulated, and the expression of GA signaling genes (e.g., GhGID1C and GhGID1B) was up-regulated by melatonin. In conclusion, melatonin enhances salt tolerance in cotton seeds by regulating ABA and GA and by mediating the expression of hormone-related genes in plant hormone signal transduction. This should help us to explore the regulatory mechanisms of cotton resistance and provide a foundation for the cultivation of new varieties.

Maize transcription factor ZmEREB20 enhanced salt tolerance in transgenic Arabidopsis

Soil salinity severely limits agricultural crop production worldwide. As one of the biggest plant specific transcription factor families, AP2/ERF members have been extensively studied to regulate plant growth, development and stress responses. However, the role of AP2/ERF family in maize salt tolerance remains largely unknown. In this study, we identified a maize AP2-ERF family member ZmEREB20 as a positive salinity responsive gene. Overexpression of ZmEREB20in Arabidopsis enhanced ABA sensitivity and resulted in delayed seed germination under salt stress through regulating ABA and GA related genes. ZmEREB20 overexpression lines also showed higher survival rates with elevated ROS scavenging toward high salinity. Furthermore, root hair growth inhibition by salt stress was markedly rescued in ZmEREB20 overexpression lines. Auxin transport inhibitor TIBA drastically enhanced root hair growth in ZmEREB20 overexpression Arabidopsis under salt stress, together with the increased expression of auxin-related genes, ion transporter genes and root hair growth genes by RNA-seq analysis. ZmEREB20 positively regulated salt tolerance through the molecular mechanism associated with hormone signaling, ROS scavenging and root hair plasticity, proving the potential target for crop breeding to improve salt resistance.

Overexpression of the Brassica rapa SRS7 gene in pot-type chrysanthemum [Chrysanthemum morifolium Ramat] reduces plant height

Retardation of plant growth is important in horticultural plant production and is generally achieved through the application of various chemical plant growth regulators (PGRs). Chrysanthemum plants grown in pots (pot-mums), are typically treated with PGRs to produce a compact form; however, these compounds can be hazardous to the environment and human health. In an effort to develop a method to produce compact chrysanthemum plants without using PGRs, we introduced the BrSRS7 gene from Brassica rapa L. ssp. pekinensis into chrysanthemum with its expression controlled by the promoter of the chrysanthemum actin gene (CmActin). As a first step, we evaluated the regeneration capacities of seven pot-mum cultivars and identified the Peace Copper (PC) cultivar as having the best regeneration efficiency. After transformation of the BrSRS7 gene into PC followed by selection and 5 rounds of vegetative subcultivation, 11 transgenic lines were identified for analysis. Internode lengths were reduced in all lines compared to control plants, but plant heights of some lines were similar to those of control plants. Four transgenic lines were selected for a sixth round of subcultivation and for further study. We determined the relative expression levels of gibberellin- and auxin-related genes in the four transgenic lines. Of the auxin-related genes, ARF2 and YUCCA10 showed higher expression in the transgenic lines than in the control. The expression levels of GA-related genes decreased overall in P-33, which showed the greatest decrease in height. Subjecting the transgenic lines to a dwarfing treatment with the PGR B9 showed that the plants could be reduced to actual marketable size with only a single dwarfing treatment. Our results demonstrate the possibility of using the BrSRS7 gene to produce compact chrysanthemum plants with reduced or no use of PGRs.

Generation and Transcriptome Profiling of Slr1-d7 and Slr1-d8 Mutant Lines with a New Semi-Dominant Dwarf Allele of SLR1 Using the CRISPR/Cas9 System in Rice

The rice SLR1 gene encodes the DELLA protein (protein with DELLA amino acid motif), and a loss-of-function mutation is dwarfed by inhibiting plant growth. We generate slr1-d mutants with a semi-dominant dwarf phenotype to target mutations of the DELLA/TVHYNP domain using CRISPR/Cas9 genome editing in rice. Sixteen genetic edited lines out of 31 transgenic plants were generated. Deep sequencing results showed that the mutants had six different mutation types at the target site of the TVHYNP domain of the SLR1 gene. The homo-edited plants selected individuals without DNA (T-DNA) transcribed by segregation in the T1 generation. The slr1-d7 and slr1-d8 plants caused a gibberellin (GA)-insensitive dwarf phenotype with shrunken leaves and shortened internodes. A genome-wide gene expression analysis by RNA-seq indicated that the expression levels of two GA-related genes, GA20OX2 (Gibberellin oxidase) and GA3OX2, were increased in the edited mutant plants, suggesting that GA20OX2 acts as a convert of GA12 signaling. These mutant plants are required by altering GA responses, at least partially by a defect in the phytohormone signaling system process and prevented cell elongation. The new mutants, namely, the slr1-d7 and slr1-d8 lines, are valuable semi-dominant dwarf alleles with potential application value for molecule breeding using the CRISPR/Cas9 system in rice.

Co-expression of glutamatergic and autism-related genes in the hippocampus of male mice with disturbances of social behavior

В настоящее время существует представление о вовлеченности глутаматергической системы (ГГ) в механизмы развития аутизма. В предыдущих исследованиях нами было показано, что негативный социальный опыт, приобретенный в ежедневных межсамцовых конфронтациях, приводит к нарушениям в социальном поведении: снижению коммуникативности, нарушению социализации, появлению стереотипных форм поведения, которые могут рассматриваться как симптомы аутистического спектра. В связи с этим целью нашей работы было изучение с помощью транскриптомного анализа изменений экспрессии генов, кодирующих белки, вовлеченные в функционирование глутаматергической системы, и генов, связанных с патологией аутизма (ГА), в гиппокампе. В эксперименте использовали животных с нарушениями социального поведения, вызванными повторным опытом социальных побед или поражений в ежедневных агонистических взаимодействиях. Для формирования групп животных с контрастными типами поведения использовали модель сенсорного контакта (хронического социального стресса). Полученные образцы мозга были секвенированы в ЗАО «Геноаналитика» (http://genoanalytica.ru/, Россия, Москва). Транскриптомный анализ показал, что у агрессивных животных снижается экспрессия генов Shank3, Auts2, Ctnnd2, Nrxn2, для которых показано участие в развитии аутизма, а также глутаматергического гена Grm4. В то же время у животных с негативным социальным опытом экспрессия ГА Shank2, Nlgn2, Ptcdh10, Reln, Arx возрастает. При этом ГГ (Grik3, Grm2, Grm4, Slc17a7, Slc1a4, Slc25a22), за исключением гена Grin2a, повышают свою экспрессию. Корреляционный анализ выявил статистически значимую взаимосвязь из- мененной экспрессии ГГ и ГА. Полученные результаты, с одной стороны, могут служить подтверждением участия ГГ в патофизиологии развития симптомов аутистического спектра, с другой – свидетельствовать о коэкспрессии ГГ и ГА в гиппокампе, развивающейся под влиянием социальной среды. Так как большинство ГА, изменивших экспрессию в настоящем исследовании, являются генами, связанными с клеточным скелетом и внеклеточным матриксом, в частности участвующими в формировании синапсов, а ГГ, изменившие свою экспрессию, – генами, кодирующими субъединицы рецепторов, то можно предположить, что вовлечение ГГ в патофизиологию аутизма происходит на уровне рецепторов.