Transcription Factor Gene Family Research Articles

Genome-wide analysis of the heat shock transcription factor gene family in Sorbus pohuashanensis (Hance) Hedl identifies potential candidates for resistance to abiotic stresses

Heat shock transcription factors (Hsfs) are essential regulators of plant responses to abiotic stresses, growth, and development. However, all the Hsf family members have not been identified in Sorbus pohuashanensis. Therefore, the aim of this study was to identify the Hsf family members in S. pohuashanensis and examine their expression under abiotic stress conditions through the integration of gene structure, phylogenetic relationships, chromosome location, and expression patterns. Bioinformatics-based methods, identified 33 Hsfs in S. pohuashanensis. Phylogenetic analysis of Hsfs from S. pohuashanensis and other species revealed that they were more closely related to apples and white pears, followed by Populus trichocarpa, and most distantly related to Arabidopsis. Moreover, the Hsfs were clustered into three major groups: A, B, and C. Gene structure and conserved motif analysis revealed a high degree of conservation among members of the same class. Collinearity analysis revealed that segmental duplication played an essential role in increasing the size of the SpHsfs gene family in S. pohuashanensis. Additionally, several cis-acting elements associated with growth and development, hormone response, and stress were found in the promoter region of SpHsfs genes. Furthermore, expression analysis in various tissues of S. pohuashanensis showed that the genes were closely associated with heat, drought, salt stress, growth, and developmental processes. Overall, these results provide valuable information on the evolutionary relationships of the Hsf gene family. These genes stand as strong functional candidates for further studies on the resistance of S. pohuashanensis to abiotic stresses.

Heat Stress Resistance Mechanisms of Two Cucumber Varieties from Different Regions.

High temperatures affect the yield and quality of vegetable crops. Unlike thermosensitive plants, thermotolerant plants have excellent systems for withstanding heat stress. This study evaluated various heat resistance indexes of the thermotolerant cucumber (TT) and thermosensitive cucumber (TS) plants at the seedling stage. The similarities and differences between the regulatory genes were assessed through transcriptome analysis to understand the mechanisms for heat stress resistance in cucumber. The TT plants exhibited enhanced leaf status, photosystem, root viability, and ROS scavenging under high temperature compared to the TS plants. Additionally, transcriptome analysis showed that the genes involved in photosynthesis, the chlorophyll metabolism, and defense responses were upregulated in TT plants but downregulated in TS plants. Zeatin riboside (ZR), brassinosteroid (BR), and jasmonic acid (JA) levels were higher in TT plants than in TS. The heat stress increased gibberellic acid (GA) and indoleacetic acid (IAA) levels in both plant lines; however, the level of GA was higher in TT. Correlation and interaction analyses revealed that heat cucumber heat resistance is regulated by a few transcription factor family genes and metabolic pathways. Our study revealed different phenotypic and physiological mechanisms of the heat response by the thermotolerant and thermosensitive cucumber plants. The plants were also shown to exhibit different expression profiles and metabolic pathways. The heat resistant pathways and genes of two cucumber varieties were also identified. These results enhance our understanding of the molecular mechanisms of cucumber response to high-temperature stress.

Biological Function and Stress Response Mechanism of MYB Transcription Factor Family Genes

Biological Function and Stress Response Mechanism of MYB Transcription Factor Family Genes

Heat shock transcription factor (Hsf) gene family in common bean (Phaseolus vulgaris): genome-wide identification, phylogeny, evolutionary expansion and expression analyses at the sprout stage under abiotic stress

BackgroundCommon bean (Phaseolus vulgaris) is an essential crop with high economic value. The growth of this plant is sensitive to environmental stress. Heat shock factor (Hsf) is a family of antiretroviral transcription factors that regulate plant defense system against biotic and abiotic stress. To date, few studies have identified and bio-analyzed Hsfs in common bean.ResultsIn this study, 30 Hsf transcription factors (PvHsf1–30) were identified from the PFAM database. The PvHsf1–30 belonged to 14 subfamilies with similar motifs, gene structure and cis-acting elements. The Hsf members in Arabidopsis, rice (Oryza sativa), maize (Zea mays) and common bean were classified into 14 subfamilies. Collinearity analysis showed that PvHsfs played a role in the regulation of responses to abiotic stress. The expression of PvHsfs varied across different tissues. Moreover, quantitative real-time PCR (qRT-PCR) revealed that most PvHsfs were differentially expressed under cold, heat, salt and heavy metal stress, indicating that PvHsfs might play different functions depending on the type of abiotic stress.ConclusionsIn this study, we identified 30 Hsf transcription factors and determined their location, motifs, gene structure, cis-elements, collinearity and expression patterns. It was found that PvHsfs regulates responses to abiotic stress in common bean. Thus, this study provides a basis for further analysis of the function of PvHsfs in the regulation of abiotic stress in common bean.

Evolution and functional diversification of R2R3-MYB transcription factors in plants.

R2R3-MYB genes (R2R3-MYBs) form one of the largest transcription factor gene families in the plant kingdom, with substantial structural and functional diversity. However, the evolutionary processes leading to this amazing functional diversity have not yet been clearly established. Recently developed genomic and classical molecular technologies have provided detailed insights into the evolutionary relationships and functions of plant R2R3-MYBs. Here, we review recent genome-level and functional analyses of plant R2R3-MYBs, with an emphasis on their evolution and functional diversification. In land plants, this gene family underwent a large expansion by whole genome duplications and small-scale duplications. Along with this population explosion, a series of functionally conserved or lineage-specific subfamilies/groups arose with roles in three major plant-specific biological processes: development and cell differentiation, specialized metabolism, and biotic and abiotic stresses. The rapid expansion and functional diversification of plant R2R3-MYBs are highly consistent with the increasing complexity of angiosperms. In particular, recently derived R2R3-MYBs with three highly homologous intron patterns (a, b, and c) are disproportionately related to specialized metabolism and have become the predominant subfamilies in land plant genomes. The evolution of plant R2R3-MYBs is an active area of research, and further studies are expected to improve our understanding of the evolution and functional diversification of this gene family.

Analysis of Transcriptome Sequencing and MYB Transcription Factor Family in <i>Rhododendron lapponicum</i>

MYB transcription factor is the largest family of transcription factors in plants, and they are involved in the regulation of plant growth and development, secondary metabolism, adversity stress and other biological processes. So far, there is no the study on the MYB transcription factor of  Rhododendron lapponicum . In this study, the  Rhododendron lapponicum  variety ‘Fuli Jinling’ transcriptome was generated by SMRT sequencing technology. A total of 15.37 Gb data was obtained, and 75 002 transcript sequences were obtained by removing redundancy. 71 155, 33 653 and 30 359 transcripts were assigned to the Nr, GO and COG databases, respectively. Based on the transcriptome sequencing data, 64 transcription factor gene families were identified, including 220 MYB genes. According to the structural characteristics, the MYB gene is divided into four categories, including 1R-MYB, R2R3-MYB, R1R2R3-MYB and 4R-MYB. The amino acid sequence of MYB transcription factor contains 20 conserved elements. Phylogenetic analysis showed that the MYB genes of  Rhododendron lapponicum  could be divided into 28 subclasses. In this study, we first used SMRT sequencing technology to generate the  Rhododendron lapponicum  transcriptome. In this study, single molecule real time (SMRT) technology was used to sequence the transcriptome of the  Rhododendron lapponicum  variety 'Fuli Jinling', and the obtained transcript sequences were functionally annotated and classified, and 220  MYB  genes for bioinformatics analysis, these related results have certain reference significance.

Neoadjuvant Chemotherapy Induces Genomic and Transcriptomic Changes in Ovarian Cancer.

The growing use of neoadjuvant chemotherapy to treat advanced stage high-grade serous ovarian cancer (HGSOC) creates an opportunity to better understand chemotherapy-induced mutational and gene expression changes. Here we performed a cohort study including 34 patients with advanced stage IIIC or IV HGSOC to assess changes in the tumor genome and transcriptome in women receiving neoadjuvant chemotherapy. RNA sequencing and panel DNA sequencing of 596 cancer-related genes was performed on paired formalin-fixed paraffin-embedded specimens collected before and after chemotherapy, and differentially expressed genes (DEG) and copy-number variations (CNV) in pre- and post-chemotherapy samples were identified. Following tissue and sequencing quality control, the final patient cohort consisted of 32 paired DNA and 20 paired RNA samples. Genomic analysis of paired samples did not reveal any recurrent chemotherapy-induced mutations. Gene expression analyses found that most DEGs were upregulated by chemotherapy, primarily in the chemotherapy-resistant specimens. AP-1 transcription factor family genes (FOS, FOSB, FRA-1) were particularly upregulated in chemotherapy-resistant samples. CNV analysis identified recurrent 11q23.1 amplification, which encompasses SIK2. In vitro, combined treatment with AP-1 or SIK2 inhibitors with carboplatin or paclitaxel demonstrated synergistic effects. These data suggest that AP-1 activity and SIK2 copy-number amplification are induced by chemotherapy and may represent mechanisms by which chemotherapy resistance evolves in HGSOC. AP-1 and SIK2 are druggable targets with available small molecule inhibitors and represent potential targets to circumvent chemotherapy resistance. SIGNIFICANCE: Genomic and transcriptomic analyses identify increased AP-1 activity and SIK2 copy-number amplifications in resistant ovarian cancer following neoadjuvant chemotherapy, uncovering synergistic effects of AP-1 and SIK2 inhibitors with chemotherapy.

Genome-Wide Identification and Functional Characterization of GATA Transcription Factor Gene Family in Alternaria alternata.

In the present study, we identified six GATA transcription factors (AaAreA, AaAreB, AaLreA, AaLreB, AaNsdD, and AaSreA) and characterized their functions in response to environmental stress and virulence in the tangerine pathotype of Alternaria alternata. The targeted gene knockout of each of the GATA-coding genes decreased the growth to varying degrees. The mutation of AaAreA, AaAreB, AaLreB, or AaNsdD decreased the conidiation. All the GATA transcription factors were found to be required for tolerance to cumyl hydroperoxide and tert-butyl-hydroperoxide (oxidants) and Congo red (a cell-wall-destructing agent). Pathogenicity assays assessed on detached citrus leaves revealed that mutations of AaAreA, AaLreA, AaLreB, or AaNsdD significantly decreased the fungal virulence. A comparative transcriptome analysis between the ∆AreA mutant and the wild-type strain revealed that the inactivation of AaAreA led to alterations in the expression of genes involved in a number of biological processes, including oxidoreductase activity, amino acid metabolism, and secondary metabolite biogenesis. Taken together, our findings revealed that GATA-coding genes play diverse roles in response to environmental stress and are important regulators involved in fungal development, conidiation, ROS detoxification, as well as pathogenesis. This study, for the first time, systemically underlines the critical role of GATA transcription factors in response to environmental stress and virulence in A. alternata.

Genome-wide identification and expression profile analysis of trihelix transcription factor family genes in response to abiotic stress in sorghum [Sorghum bicolor (L.) Moench

BackgroundTranscription factors, including trihelix transcription factors, play vital roles in various growth and developmental processes and in abiotic stress responses in plants. The trihelix gene has been systematically studied in some dicots and monocots, including Arabidopsis, tomato, chrysanthemum, soybean, wheat, corn, rice, and buckwheat. However, there are no related studies on sorghum.ResultsIn this study, a total of 40 sorghum trihelix (SbTH) genes were identified based on the sorghum genome, among which 34 were located in the nucleus, 5 in the chloroplast, 1 (SbTH38) in the cytoplasm, and 1 (SbTH23) in the extracellular membrane. Phylogenetic analysis of the SbTH genes and Arabidopsis and rice trihelix genes indicated that the genes were clustered into seven subfamilies: SIP1, GTγ, GT1, GT2, SH4, GTSb8, and orphan genes. The SbTH genes were located in nine chromosomes and none on chromosome 10. One pair of tandem duplication gene and seven pairs of segmental duplication genes were identified in the SbTH gene family. By qPCR, the expression of 14 SbTH members in different plant tissues and in plants exposed to six abiotic stresses at the seedling stage were quantified. Except for the leaves in which the genes were upregulated after only 2 h exposure to high temperature, the 12 SbTH genes were significantly upregulated in the stems of sorghum seedlings after 24 h under the other abiotic stress conditions. Among the selected genes, SbTH10/37/39 were significantly upregulated, whereas SbTH32 was significantly downregulated under different stress conditions.ConclusionsIn this study, we identified 40 trihelix genes in sorghum and found that gene duplication was the main force driving trihelix gene evolution in sorghum. The findings of our study serve as a basis for further investigation of the functions of SbTH genes and providing candidate genes for stress-resistant sorghum breeding programmes and increasing sorghum yield.

Post-Embryonic Lateral Organ Development and Adaxial-Abaxial Polarity Are Regulated by the Combined Effect of ENHANCER OF SHOOT REGENERATION 1 and WUSCHEL in Arabidopsis Shoots.

The development of above-ground lateral organs is initiated at the peripheral zone of the shoot apical meristem (SAM). The coordination of cell fate determination and the maintenance of stem cells are achieved through a complex regulatory network comprised of transcription factors. Two AP2/ERF transcription factor family genes, ESR1/DRN and ESR2/DRNL/SOB/BOL, regulate cotyledon and flower formation and de novo organogenesis in tissue culture. However, their roles in post-embryonic lateral organ development remain elusive. In this study, we analyzed the genetic interactions among SAM-related genes, WUS and STM, two ESR genes, and one of the HD-ZIP III members, REV, whose protein product interacts with ESR1 in planta. We found that esr1 mutations substantially enhanced the wus and stm phenotypes, which bear a striking resemblance to those of the wus rev and stm rev double mutants, respectively. Aberrant adaxial–abaxial polarity is observed in wus esr1 at relatively low penetrance. On the contrary, the esr2 mutation partially suppressed stm phenotypes in the later vegetative phase. Such complex genetic interactions appear to be attributed to the distinct expression pattern of two ESR genes because the ESR1 promoter-driving ESR2 is capable of rescuing phenotypes caused by the esr1 mutation. Our results pose the unique genetic relevance of ESR1 and the SAM-related gene interactions in the development of rosette leaves.

Genome-wide identification of the DNA-binding one zinc finger (Dof) transcription factor gene family and their potential functioning in nitrogen remobilization in tea plant (Camellia sinensis L.)

The DNA-binding one zinc finger (Dof) proteins are a type of transcription factor (TF) specific to plants that are involved in various aspects of their biological processes. Although the Dof TF has been identified in a variety of species, its systemic analysis in tea (Camellia sinensis L.) is lacking and its potential role in this plant's nitrogen remobilization remains unclear. Here, we carried out a genome-wide identification of Dofs in tea and identified 41 CsDofs at the chromosome level. Their genetic structure, conserved motif, and synteny relations were analyzed and visualized, using information extracted from a newly available genome database. Phylogenetic analysis of Dofs from seven different plant species revealed that Dof members could be clustered into six groups. Heatmap analysis showed that CsDofs have similar expression patterns among different species of genus Camellia, and tissue-specific transcription patterns among tea tissues. Expression patterns of CsDofs in mature leaves during the development of new shoots revealed that most of CsDofs peaked on March 24 when the new shoots developed into one bud and two leaves. Furthermore, the expression pattern of CsDof13/16/29/33/34/35/36/39 was significantly correlated with that of genes involved in N metabolism, such as GDH or GS, indicating these CsDofs might play roles in regulating the expression of genes participating in N metabolism of tea. Taken together, this study provides information on the genome-wide identification of the CsDofs’ family in tea, and identifies putative candidates possible involved in N remobilization in mature leaves during the development of new shoots in spring.

Genome-Wide Identification and Expression Profiling of the BZR Transcription Factor Gene Family in Nicotiana benthamiana.

Brassinazole-resistant (BZR) family genes encode plant-specific transcription factors (TFs), play essential roles in the regulation of plant growth and development, and have multiple stress-resistance functions. Nicotiana benthamiana is a model plant widely used in basic research. However, members of the BZR family in N. benthamiana have not been identified, and little is known about their function in abiotic stress. In this study, a total of 14 BZR members were identified in the N. benthamiana genome, which could be divided into four groups according to a phylogenetic tree. NbBZRs have similar exon-intron structures and conserved motifs, and may be regulated by cis-acting elements such as STRE, TCA, and ARE, etc. Organ-specific expression analysis showed that NbBZR members have different and diverse expression patterns in different tissues, and most of the members are expressed in roots, stems, and leaves. The analysis of the expression patterns in response to different abiotic stresses showed that all the tested NbBZR members showed a significant down-regulation after drought treatment. Many NbBZR genes also responded in various ways to cold, heat and salt stress treatments. The results imply that NbBZRs have multiple functions related to stress resistance.

Transporters and transcription factors gene families involved in improving nitrogen use efficiency (NUE) and assimilation in rice (Oryza sativa L.).

Nitrogen (N) as a macronutrient is an important determinant of plant growth. The excessive usage of chemical fertilizers is increasing environmental pollution; hence, the improvement of crop's nitrogen use efficiency (NUE) is imperative for sustainable agriculture. N uptake, transportation, assimilation, and remobilization are four important determinants of plant NUE. Oryza sativa L. (rice) is a staple food for approximately half of the human population, around the globe and improvement in rice yield is pivotal for rice breeders. The N transporters, enzymes indulged in N assimilation, and several transcription factors affect the rice NUE and subsequent yield. Although, a couple of improvements have been made regarding rice NUE, the knowledge about regulatory mechanisms operating NUE is scarce. The current review provides a precise knowledge of how rice plants detect soil N and how this detection is translated into the language of responses that regulate the growth. Additionally, the transcription factors that control N-associated genes in rice are discussed in detail. This mechanistic insight will help the researchers to improve rice yield with minimized use of chemical fertilizers.

Genome-Wide Comparative Analysis of R2R3 MYB Gene Family in Populus and Salix and Identification of Male Flower Bud Development-Related Genes.

The MYB transcription factor (TF) family is one of the largest plant transcription factor gene family playing vital roles in plant growth and development, including defense, cell differentiation, secondary metabolism, and responses to biotic and abiotic stresses. As a model tree species of woody plants, in recent years, the identification and functional prediction of certain MYB family members in the poplar genome have been reported. However, to date, the characterization of the gene family in the genome of the poplar’s sister species willow has not been done, nor are the differences and similarities between the poplar and willow genomes understood. In this study, we conducted the first genome-wide investigation of the R2R3 MYB subfamily in the willow, identifying 216 R2R3 MYB gene members, and combined with the poplar R2R3 MYB genes, performed the first comparative analysis of R2R3 MYB genes between the poplar and willow. We identified 81 and 86 pairs of R2R3 MYB paralogs in the poplar and willow, respectively. There were 17 pairs of tandem repeat genes in the willow, indicating active duplication of willow R2R3 MYB genes. A further 166 pairs of poplar and willow orthologs were identified by collinear and synonymous analysis. The findings support the duplication of R2R3 MYB genes in the ancestral species, with most of the R2R3 MYB genes being retained during the evolutionary process. The phylogenetic trees of the R2R3 MYB genes of 10 different species were drawn. The functions of the poplar and willow R2R3 MYB genes were predicted using reported functional groupings and clustering by OrthoFinder. Identified 5 subgroups in general expanded in woody species, three subgroups were predicted to be related to lignin synthesis, and we further speculate that the other two subgroups also play a role in wood formation. We analyzed the expression patterns of the GAMYB gene of subgroup 18 (S18) related to pollen development in the male flower buds of poplar and willow at different developmental stages by qRT-PCR. The results showed that the GAMYB gene was specifically expressed in the male flower bud from pollen formation to maturity, and that the expression first increased and then decreased. Both the specificity of tissue expression specificity and conservation indicated that GAMYB played an important role in pollen development in both poplar and willow and was an ideal candidate gene for the analysis of male flower development-related functions of the two species.

OMICS approaches towards understanding plant's responses to counterattack heavy metal stress: An insight into molecular mechanisms of plant defense

Under constant climatic change and the existing environment, plants are exposed to several adverse factors such as extreme temperatures, high salt, heavy metals (HMs), drought, and pathogens. Prolonged exposure to such adverse conditions results in phytotoxicity and overall retardation of growth and development. Excess of HMs is one of the significant factors that affects plant growth significantly. Plants develop several defense mechanisms at the cellular and physiological levels to survive HMs stress. However, these internal defense mechanisms are not enough to overcome excessive HMs stress. Plants generate various secondary messengers to activate cell signaling, which results in the activation of several transcriptional responses associated with plant defense. Different transcription factors (TFs) gene families such as WRKY, my elob lastosis (MYB), basic leucine zipper domain (bZIP), basic helix-loop-helix (bHLH), APETALA2/ethylene-responsive element binding protein (AP2/EREBP), Cys2-His2 (C2H2), receptor genes/proteins, and kinases, like mitogen-activated protein kinase (MAPK), play an essential role in the regulation of stress using hormone-mediated pathways under HMs stress. Additionally, phytohormones and their signaling network also contribute to regulating cellular functions at the molecular level to combat HMs stress. Several mechanisms have been explored to understand the defense mechanism of plants under HMs stress. However, many more advanced molecular approaches are required to explore these defense mechanisms. Herein, we have described OMICS’ approaches to improve the plants' internal defense system to counteract HMs stress.

Genome-wide analysis of the MADS-box gene family in Rhododendron hainanense Merr. and expression analysis under heat and waterlogging stresses

The MADS-box transcription factor gene family plays an important role in the growth and development of plants as well as in biotic and abiotic stress. We conducted a genome-wide identification of the members of the MADS-box family of Rhododendron hainanense to determine the molecular evolutionary characteristics of the MADS-box gene family and the expression profile of this gene family under different abiotic stress conditions. A total of 55 MADS-box gene family members with complete domains were identified, which were unevenly distributed on 13 chromosomes. The construction of a phylogenetic tree showed that the 55 MADS-box proteins were divided into five categories and distributed on 8 chromosomes unevenly. Conserved motif analysis shows that motif 1 and motif 2 are highly conserved in most RhMADS proteins. Proteins belonging to the same category show similar gene structures. Analysis of gene replication events suggested that there were one pair of tandem repeats and eight pairs of fragment repeat events. In addition, analysis of cis-acting elements and protein interaction networks indicated that some genes are involved in abiotic stress processes in plants. The transcriptome data were used to analyze the expression of the members of the MADS-box family under different temperatures and waterlogging period treatments, which were then verified by qRT-PCR. The expressions of RhMADS24, RhMADS25, RhMADS39 and RhMADS44 were significantly regulated under temperature and waterlogging stresses. The identified MADS-box candidate genes of R. hainanense play an important role in resisting abiotic stress and lay the foundation for future applications in breeding and gardening.

RecA gene genetic diversity and its regulatory element analysis: The case of Vibrio cholerae

In conventional systems, indicator organisms are used to monitor drinking water quality which is unspecific in detecting pathogens. On the contrary, molecular techniques play vital role in these aspects by providing precise genetic information about particular pathogen of interest via proper biomarkers. The aim of this study was to undertake genetic relatedness and promoter region analysis for V. cholerae strains based on recA gene sequences which were retrieved from NCBI. Sequences homologous were ensured using BLASTn from RefSeq sequence in the NCBI using V. cholerae serotype O1 strain N16961 as a reference. Accordingly, considering only complete and coding sequences with query coverage of greater than 90% and E-value of less than 5%, a total of 109 sequences were recovered from NCBI. The genetic diversity analysis was carried out using MEGA X. The analysis revealed more than 99% sequence similarity among V. cholerae strains. From promoter regions analyses, five motifs shared by all (100%) of the promoter input sequences were identified. Using TOMTOM web application, motif_1was compared to a known prokaryotes' transcription factor database to search for its similarity with known regulatory motifs and was shown to serve as abundant binding site for helix-turn-helix (HTH) transcription factor gene family to regulate expression of V. cholerae recA genes. Furthermore, our analysis revealed that the V. cholerae recA gene promoter regions are CpG islands rich. In conclusion, all V. cholerae strains shared a very high degree of recA gene sequence similarity apart from other vibrios, which indicates that these non-virulent strains may be the remnant of pathogenic strains with the loss of toxin encoding genes.

Molecular Manipulation of the MiR396/GRF Expression Module Alters the Salt Stress Response of Arabidopsis thaliana

We previously demonstrated that microRNA396 (miR396) abundance is altered in 15-day-old Arabidopsis thaliana (Arabidopsis) whole seedlings following their exposure to a 7-day salt stress treatment regime. We, therefore, used a molecular modification approach to generate two new Arabidopsis transformant populations with reduced (MIM396 plants) and elevated (MIR396 plants) miR396 abundance. The exposure of 8-day-old wild-type Arabidopsis whole seedlings and a representative plant line of the MIM396 and MIR396 transformant populations to a 7-day salt stress treatment regime revealed unique phenotypic and physiological responses to the imposed stress by unmodified wild-type Arabidopsis plants and the MIM396 and MIR396 transformat lines. A quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) approach was, therefore, applied to demonstrate that the plant line specific responses to salt stress likely stemmed from the unique molecular profile of each of the GROWTH REGULATING FACTOR (GRF) transcription factor gene family members which form posttranscriptional targets of miR396-directed expression regulation. RT-qPCR additionally revealed that, in 15-day-old Arabidopsis whole seedlings, the three previously identified putative target genes of miR396 belonging to the NEUTRAL/ALKALINE NONLYSOSOMAL CERAMIDASE-LIKE (NCER) gene family, including NCER1, NCER2, and NCER3, do not form targets of miR396-directed expression regulation at the posttranscriptional level. Taken together, the phenotypic and molecular analyses presented here demonstrate that alteration of the miR396/GRF expression module is central to the molecular response of Arabidopsis to salt stress.

Genome-Wide Identification of ARF Transcription Factor Gene Family and Their Expression Analysis in Sweet Potato.

Auxin response factors (ARFs) are a family of transcription factors that play an important role of auxin regulation through their binding with auxin response elements. ARF genes are represented by a large multigene family in plants; however, to our knowledge, the ARF gene family has not been well studied and characterized in sweet potatoes. In this study, a total of 25 ARF genes were identified in Ipomea trifida. The identified ItrARF genes’ conserved motifs, chromosomal locations, phylogenetic relationships, and their protein characteristics were systemically investigated using different bioinformatics tools. The expression patterns of ItfARF genes were analyzed within the storage roots and normal roots at an early stage of development. ItfARF16b and ItfARF16c were both highly expressed in the storage root, with minimal to no expression in the normal root. ItfARF6a and ItfARF10a exhibited higher expression in the normal root but not in the storage root. Subsequently, ItfARF1a, ItfARF2b, ItfARF3a, ItfARF6b, ItfARF8a, ItfARF8b, and ItfARF10b were expressed in both root types with moderate to high expression for each. All ten of these ARF genes and their prominent expression signify their importance within the development of each respective root type. This study provides comprehensive information regarding the ARF family in sweet potatoes, which will be useful for future research to discover further functional verification of these ItfARF genes.

BSCI-05. ABL2-HSF1-E2F signaling axis promotes lung adenocarcinoma brain metastasis

Brain metastases are the most common intracranial tumors in adults and are associated with increased patient morbidity and mortality. Limited therapeutic options are currently available for the treatment of brain metastasis. We have identified an actionable signaling pathway utilized by metastatic tumor cells whereby the transcriptional regulator Heat Shock Factor 1 (HSF1) drives a transcriptional program, divergent from its canonical role as the master regulator of the heat shock response, leading to enhanced expression of a subset of E2F transcription factor family gene targets. We showed that HSF1 is required for survival and outgrowth by metastatic lung cancer cells in the brain parenchyma. Unexpectedly, we identified the ABL2 tyrosine kinase as an upstream regulator of HSF1 protein expression, and showed that the Src-homology 3 (SH3) domain of ABL2 directly interacts with HSF1 protein at a non-canonical, proline-independent SH3 interaction motif. Importantly, knockdown of ABL2 impairs expression of HSF1 protein and HSF1-E2F transcriptional gene targets. Notably, we found that pharmacologic inhibition of the ABL kinases using selective ABL allosteric inhibitors, but not ATP-competitive inhibitors, ablates the physical interaction between ABL2 and HSF1, leading to markedly decreased expression of HSF1, E2F1 and E2F8 proteins in brain-metastatic lung cancer cells, and depletion of HSF1-E2F transcriptional targets. These findings highlight potential differences affecting intra- and inter-molecular protein-protein interactions induced by allosteric versus ATP-competitive kinase inhibitors that have important therapeutic implications. Importantly, the targetable nature of the ABL2-HSF1-E2F signaling network identifies ABL allosteric inhibitors as a potentially effective therapy for the treatment of metastatic lung cancers characterized by high expression of HSF1.