Jianling Zhang

In plants, hexokinase (HXK) is a kind of bifunctional enzyme involved in sugar metabolism and sugar signal transduction that plays important roles in plant growth and development and stress response. Some HXK genes without a phosphorylation function have been found in Arabidopsis, tobacco, etc., but these genes have not been identified in tomato. Therefore, further genome-wide systematic identification and characterization is necessary for tomato HXK genes. In this study, six HXK genes were identified from the tomato genome distributed across six different chromosomes, named SlHXK1-6. Gene structure analysis showed that the SlHXK genes contain the same number of introns and exons. Gene duplication and collinearity analysis revealed two pairs of tandem repeats among SlHXKs, and a higher collinearity between tomatoes and potatoes were found. Response elements associated with phytohormones, abiotic stresses, and growth and development were identified in the promoter sequences of SlHXKs. Quantitative real-time PCR (qRT-PCR) results further indicated the potential role of SlHXKs in tomato development and stress responses. The expression levels of most SlHXKs were significantly induced by abiotic stress, hormone, and sugar solution treatments. In particular, the expression of SlHXK1 was significantly induced by various treatments. Functional complementation experiments were performed using HXK-deficient yeast strain YSH7.4-3C (hxk1, hxk2, and glk1), and the results showed that SlHXK5 and SlHXK6 were unable to phosphorylate glucose and fructose in yeast. In conclusion, these results provide valuable foundations for further exploring the sugar metabolism and sugar signal transduction mechanisms of HXK and the functions of SlHXK genes in various abiotic stresses, and some SlHXKs may be key genes for enhancing plants’ tolerance to abiotic stresses.

The bHLH (basic helix–loop–helix) transcription factors function as crucial regulators in numerous biological processes including abiotic stress responses and plant development. According to our RNA-seq analysis of tomato seedlings under salt stress, we found that, although the bHLH gene family in tomato has been studied, there are still so many tomato bHLH genes that have not been identified and named, which will hinder the later study of SlbHLHs. In total, 195 SlbHLHs that were unevenly distributed onto 12 chromosomes were identified from the tomato genome and were classified into 27 subfamilies based on their molecular features. The collinearity between SlbHLHs and interrelated orthologs from 10 plants further revealed evolutionary insights into SlbHLHs. Cis-element investigations of SlbHLHs promotors further suggested the potential roles of SlbHLHs in tomato development and stress responses. A total of 30 SlbHLHs were defined as the differentially expressed genes in response to salt stress by RNA-seq. The expression profiles of selected SlbHLHs were varyingly and markedly induced by multiple abiotic stresses and hormone treatments. These results provide valuable information to further understand the significance and intricacy of the bHLH transcription factor family, and lay a foundation for further exploring functions and possible regulatory mechanisms of SlbHLH members in abiotic stress tolerance, which will be significant for the study of tomato stress resistance and agricultural productivity.

Introduction: Expansins (EXPs) are essential components of the plant cell wall that function as relaxation factors to directly promote turgor-driven expansion of the cell wall, thereby controlling plant growth and development and diverse environmental stress responses. EXPs genes have been identified and characterized in numerous plant species, but not in sweetpotato.Results and methods: In the present study, a total of 59 EXP genes unevenly distributed across 14 of 15 chromosomes were identified in the sweetpotato genome, and segmental and tandem duplications were found to make a dominant contribution to the diversity of functions of the IbEXP family. Phylogenetic analysis showed that IbEXP members could be clustered into four subfamilies based on the EXPs from Arabidopsis and rice, and the regularity of protein motif, domain, and gene structures was consistent with this subfamily classification. Collinearity analysis between IbEXP genes and related homologous sequences in nine plants provided further phylogenetic insights into the EXP gene family. Cis-element analysis further revealed the potential roles of IbEXP genes in sweetpotato development and stress responses. RNA-seq and qRT-PCR analysis of eight selected IbEXPs genes provided evidence of their specificity in different tissues and showed that their transcripts were variously induced or suppressed under different hormone treatments (abscisic acid, salicylic acid, jasmonic acid, and 1-aminocyclopropane-1-carboxylic acid) and abiotic stresses (low and high temperature).Discussion: These results provide a foundation for further comprehensive investigation of the functions of IbEXP genes and indicate that several members of this family have potential applications as regulators to control plant development and enhance stress resistance in plants.

MADS-box transcription factors act as the crucial regulators in plant organ differentiation. Crop yields are highly influenced by the flower number and fruit growth. However, flower identification is a very complex biological process, which involves many cascade regulations. The molecular mechanisms underlying the genetic regulation of flower identification in cultivated plants, such as tomato, are intricate and require further exploration. In this study, we investigated the vital function of a SEPALLATA (SEP) MADS-box gene, SlMBP21, in tomato sympodial inflorescence meristem (SIM) development for the conversion from SIMs to floral meristems (FMs). SlMBP21 transcripts were primarily accumulated in young inflorescence meristem, flowers, sepals, and abscission zones. The Ailsa Craig (AC⁺⁺) tomato plants with suppressed SlMBP21 mRNA levels using RNAi exhibited a large increase in flower number and fruit yields in addition to enlarged sepals and inhibited abscission zone development. Scanning electron microscopy (SEM) revealed that the maturation of inflorescence meristems (IMs) was repressed in SlMBP21-RNAi lines. RNA-seq and qRT-PCR analyses showed that numerous genes related to the flower development, plant hormone signal transduction, cell cycle, and cell proliferation et al. were dramatically changed in SlMBP21-RNAi lines. Yeast two-hybrid assay exhibited that SlMBP21 can respectively interact with SlCMB1, SFT, JOINTLESS, and MC, which play key roles in inflorescence meristems or FM development. In summary, our data demonstrate that SlMBP21 functions as a key regulator in SIM development and the conversion from SIMs to FMs, through interacting with other regulatory proteins to control the expression of related genes.

MADS-box transcription factors have crucial functions in numerous physiological and biochemical processes during plant growth and development. Previous studies have reported that two MADS-box genes, SlMBP21 and SlMADS1, play important regulatory roles in the sepal development of tomato, respectively. However, the functional relationships between these two genes are still unknown. In order to investigate this, we simultaneously studied these two genes in tomato. Phylogenetic analysis showed that they were classified into the same branch of the SEPALLATA (SEP) clade. qRT-PCR displayed that both SlMBP21 and SlMADS1 transcripts are preferentially accumulated in sepals, and are increased with flower development. During sepal development, SlMBP21 is increased but SlMADS1 is decreased. Using the RNAi, tomato plants with reduced SlMBP21 mRNA generated enlarged and fused sepals, while simultaneous inhibition of SlMBP21 and SlMADS1 led to larger (longer and wider) and fused sepals than that in SlMBP21-RNAi lines. qRT-PCR results exhibited that the transcripts of genes relating to sepal development, ethylene, auxin and cell expansion were dramatically changed in SlMBP21-RNAi sepals, especially in SlMBP21-SlMADS1-RNAi sepals. Yeast two-hybrid assay displayed that SlMBP21 can interact with SlMBP21, SlAP2a, TAGL1 and RIN, and SlMADS1 can interact with SlAP2a and RIN, respectively. In conclusion, SlMBP21 and SlMADS1 cooperatively regulate sepal development in tomato by impacting the expression or activities of other related regulators or via interactions with other regulatory proteins.

Expansins (EXPs) which are the essential components of plant cell wall function as relaxation factors to directly promote turgor-driven expansion of cell wall and control plant growth and development and diverse environmental stress responses. A lot of EXPs genes have been widely identified and characterized in numerous plant species, but not in sweetpotato. In the present study, a total of 59 EXPs genes that unevenly distributed onto 14 of 15 chromosomes were identified from sweetpotato genome, and segmental and tandem duplications were exhibited to act as dominant functions in IbEXPs diversity. Phylogenetic analysis exhibited that IbEXPs members were clustered into four subfamilies according to EXPs in Arabidopsis and rice and the regularity of motif, domain and gene structures was in accordance with the subfamily classification. The collinearity analysis between IbEXPs and related homologous sequences in 9 plants provided the phylogenetic insights of EXPs gene family. Cis-element analysis futhrer revealed the potential roles of IbEXPs in sweetpotato development and stress responses. Evidence from RNA-seq and qRT-PCR analysis of eight selected IbEXPs genes displayed their various tissue specificity in different tissues, and their transcripts were varyingly induced or suppressed under multiple hormone treatments (ABA, SA, JA and ACC) and abiotic stresses (low and high temperature). These results supply worthy foundations to further comprehensively investigate the exact functions of IbEXPs genes, and multiple members may function as promising regulators to control plant development and enhance the stress resistance of transgenic plants.

Background The bHLH (basic helix-loop-helix) transcription factors function as crucial regulators in numerous biological processes including abiotic stress responses and plant growth and development. Although SlbHLH gene family in tomato has been studied and the functions of some members have been explored, there are still many tomato SlbHLH genes that have not been identified and numbered, which will hinder the later study of SlbHLH genes. A genome-wide systematical identification and characterization is necessary for tomato SlbHLH genes. Results In the study, numerous genes were identified to be the potential and significant candidates that response to salt stress through RNA-seq analysis. 195 bHLH genes that unevenly distributed onto 12 chromosomes were identified from tomato genome. Phylogenetic relationships showed that SlbHLHs were classified into 26 subfamilies based on their conserved domain, motif compositions and gene structures. The collinearity between SlbHLHs and interrelated orthologs from ten plants further revealed evolutionary insights into SlbHLHs. Cis-elements investigations of SlbHLH gene promotors futhrer suggested the potential roles of SlbHLHs in tomato development and stress responses. The qRT-PCR analysis displayed that the expression of 12 selected SlbHLHs were varyingly and markedly induced by multiple abiotic stresses (salt, cold, heat and drought) and hormone treatments (SA, ABA, IAA, ACC and MeJA). Conclusions These results provide valuable foundations for further exploring possible regulatory mechanisms and functional characterizations of SlbHLHs, and numerous members may function as the potential regulators to enhance plant tolerance to abiotic stress.