The GRAS transcription factor CsTL regulates tendril formation in cucumber.

Cucumber (Cucumis sativus, Cs) tendrils are slender vegetative organs that typically require manual removal to ensure orderly growth during greenhouse cultivation. Here, we identified cucumber tendril-less (tl), a Tnt1 retrotransposon-induced insertion mutant lacking tendrils. Map-based cloning identified the mutated gene, CsaV3_3G003590, which we designated as CsTL, which is homologous to Arabidopsis thaliana LATERAL SUPPRESSOR (AtLAS). Knocking out CsTL repressed tendril formation but did not affect branch initiation, whereas overexpression of CsTL resulted in the formation of two or more tendrils in one leaf axil. Although expression of two cucumber genes regulating tendril formation, Tendril (CsTEN) and Unusual Floral Organs (CsUFO), was significantly decreased in CsTL knockout lines, these two genes were not direct downstream targets of CsTL. Instead, CsTL physically interacted with CsTEN, an interaction that further enhanced CsTEN-mediated expression of CsUFO. In Arabidopsis, the CsTL homolog AtLAS acts upstream of REVOLUTA (REV) to regulate branch initiation. Knocking out cucumber CsREV inhibited branch formation without affecting tendril initiation. Furthermore, genomic regions containing CsTL and AtLAS were not syntenic between the cucumber and Arabidopsis genomes, whereas REV orthologs were found on a shared syntenic block. Our results revealed not only that cucumber CsTL possesses a divergent function in promoting tendril formation but also that CsREV retains its conserved function in shoot branching.

GRAS family member LATERAL SUPPRESSOR regulates the initiation and morphogenesis of watermelon lateral organs.

The lateral organs of watermelon (Citrullus lanatus), including lobed leaves, branches, flowers, and tendrils, together determine plant architecture and yield. However, the genetic controls underlying lateral organ initiation and morphogenesis remain unclear. Here, we found that knocking out the homologous gene of shoot branching regulator LATERAL SUPPRESSOR in watermelon (ClLs) repressed the initiation of branches, flowers, and tendrils and led to developing round leaves, indicating that ClLs undergoes functional expansion compared with its homologs in Arabidopsis (Arabidopsis thaliana), rice (Oryza sativa), and tomato (Solanum lycopersicum). Using ClLs as the bait to screen against the cDNA library of watermelon, we identified several ClLs-interacting candidate proteins, including TENDRIL (ClTEN), PINOID (ClPID), and APETALA1 (ClAP1). Protein-protein interaction assays further demonstrated that ClLs could directly interact with ClTEN, ClPID, and ClAP1. The mRNA in situ hybridization assay revealed that the transcriptional patterns of ClLs overlapped with those of ClTEN, ClPID, and ClAP1 in the axillary meristems and leaf primordia. Mutants of ClTEN, ClPID, and ClAP1 generated by the CRISPR/Cas9 gene editing system lacked tendrils, developed round leaves, and displayed floral diapause, respectively, and all these phenotypes could be observed in ClLs knockout lines. Our findings indicate that ClLs acts as lateral organ identity protein by forming complexes with ClTEN, ClPID, and ClAP1, providing several gene targets for transforming the architecture of watermelon.

Cucumber NUCLEAR FACTOR-YC2/-YC9 target translocon component CsTIC21 in chloroplast photomorphogenesis.

Light signals promote photomorphogenesis and photosynthesis, allowing plants to establish photoautotrophic growth. Chloroplasts are organelles responsible for photosynthesis in which light energy is converted into chemical energy and stored as organic matter. However, how light regulates chloroplast photomorphogenesis remains unclear. Here, we isolated a cucumber (Cucumis sativus L.) mutant albino seedling (as) from an ethyl methane sulfonate mutagenesis library with an albino phenotype. Map-based cloning revealed that the mutation occurred in a component of cucumber translocon at the inner membrane of chloroplasts (CsTIC21). Subsequently, virus-induced gene silencing and CRISPR/Cas9 analyses confirmed the association between the mutant gene and the as phenotype. Loss-of-function of CsTIC21 induces malformation of chloroplast formation, leading to albinism and death in cucumber. Notably, CsTIC21 transcription was very low in etiolated seedlings grown in the dark and was upregulated by light, with expression patterns similar to those of Nuclear factor-YC (NF-YC) genes. Here, 7 cucumber NF-YC family genes (CsNF-YC) were identified, among which the expression of 4 genes (CsNF-YC1, -YC2, -YC9, and -YC13) responded to light. Gene silencing of all CsNF-YC genes in cucumber indicated that CsNF-YC2, -YC9, -YC11-1, and -YC11-2 induced distinct etiolated growth and decreased chlorophyll content. Interaction studies verified that CsNF-YC2 and CsNF-YC9 target the CsTIC21 promoter directly and promote gene transcription. These findings provide mechanistic insights on the role of the NF-YCs-TIC21 module in chloroplast photomorphogenesis promoted by light in cucumber.

A YABBY gene CRABS CLAW a (CRCa) negatively regulates flower and fruit sizes in tomato.

CRABS CLAW (CRC) is a YABBY transcription factor that plays a pivotal role in carpel development and flower meristem determinacy. Here, we characterized a CRC homolog SlCRCa and elucidated its specific roles in tomato (Solanum lycopersicum). SlCRCa is highly expressed in the petals and stamens, and is responsive to gibberellin (GA) treatment. Overexpression of SlCRCa in tomato reduces the sizes of petals, stamens, and fruits, while the inverse phenotypes are induced by knockdown of SlCRCa. Furthermore, histological investigation suggests that the smaller or larger fruits in SlCRCa-overexpressing or SlCRCa-RNAi plants are mainly determined by the decreases or increases in cell layers and cell sizes in pericarp, respectively. Through transcriptome and qRT-PCR analyses, we speculate that SlCRCa inhibits cell division by regulating the transcription of cell division-related genes, and also suppresses cell expansion by modulating the expansin genes and GA pathway in tomato fruits. Besides, SlCRCa is involved in the feedback regulation of GA biosynthesis. Our findings reveal that SlCRCa negatively regulates fruit size by affecting cell division and cell expansion, and it is also an inhibitor of floral organ sizes in tomato.

Identification and characterization of the BEL1-like genes reveal their potential roles in plant growth and abiotic stress response in tomato.

BEL1-like (BELL) transcription factors, belonging to three-amino acid-loop-extension (TALE) superfamily, are ubiquitous in plants. BELLs regulate a wide range of plant biological processes, but the understanding of the BELL family in tomato (Solanum lycopersicum) remains fragmentary. In this study, a total of 14 members of the SlBELL family were identified in tomato. SlBELL proteins contained the conserved BELL and SKY domains that served as typical structures of the BELL family. Syntenic analysis indicated that the BELL orthologs between tomato and other dicots had close evolutionary relationships. Furthermore, the promoters of SlBELLs contained numerous cis-elements related to plant growth, development, and stress response. The SlBELL genes exhibited different tissue-specific expression profiles and responded to cold, heat, and drought stresses, implying their potential functions in regulating multiple aspects of plant growth, as well as in response to abiotic stresses. Through the interaction network prediction, we found that most SlBELL proteins displayed probable interactions with the KNOTTED1-like (KNOX) proteins, another kind of transcription factor in the TALE superfamily. These findings laid foundations for further dissection of the functions of SlBELL genes in tomato, as well as for exploration of the evolutionary relationships of BELL homologs among different plant species.

Identification and characterization of the CONSTANS (CO)/CONSTANS-like (COL) genes related to photoperiodic signaling and flowering in tomato.

CO is an important regulator of photoperiodic response and flowering. However, the biological functions of CO and COL genes in tomato (Solanum lycopersicum) remain elusive. Here we identified 13 members in CO/COL family from the tomato genome. They were divided into three groups, and each group had specific characteristics in gene structures and protein domains. The SlCO/SlCOL genes showed different tissue-specific expression patterns and circadian rhythms, indicating their functional diversity in tomato. Moreover, among 13 members, the expression of SlCOL, SlCOL4a, and SlCOL4b was negatively correlated with flowering time variation in ten tomato lines. Through interaction network prediction, we found three FLOWERING LOCUS T (FT) orthologs, SINGLE FLOWER TRUSS (SFT), FT-like (FTL), and FT-like 1 (FTL1), which functioned as candidate interactors of SlCOL, SlCOL4a, and SlCOL4b. Further expression analyses suggested that SFT coincided with the three SlCOL genes in ten tomato lines with varied flowering time. These findings implied that SlCOL, SlCOL4a, and SlCOL4b are potential flowering inducers in tomato, and SFT may act as their downstream target. Thus, our study built a foundation for understanding the precise roles of SlCO/SlCOL family in plant growth and development of tomato, especially in flowering.

The GAMYB-like gene SlMYB33 mediates flowering and pollen development in tomato.

GAMYBs are positive GA signaling factors that exhibit essential functions in reproductive development, particularly in anther and pollen development. However, there is no direct evidence of the regulation of any GAMYB in these biological processes in tomato (Solanum lycopersicum). Here, we identified a tomato GAMYB-like gene, SlMYB33, and characterized its specific roles. SlMYB33 is predominately expressed in the stamens and pistils. During flower development, high mRNA abundance of SlMYB33 is detected in both male and female organs, such as microspore mother cells, anthers, pollen grains, and ovules. Silencing of SlMYB33 leads to delayed flowering, aberrant pollen viability, and poor fertility in tomato. Histological analyses indicate that SlMYB33 exerts its function in pollen development in the mature stage. Further transcriptomic analyses imply that the knockdown of SlMYB33 significantly inhibits the expression of genes related to flowering in shoot apices, and alters the transcription of genes controlling sugar metabolism in anthers. Taken together, our study suggests that SlMYB33 regulates tomato flowering and pollen maturity, probably by modulating the expression of genes responsible for flowering and sugar metabolism, respectively.

Genome-wide analysis of Jatropha curcas MADS-box gene family and functional characterization of the JcMADS40 gene in transgenic rice.

BACKGROUND: Physic nut (Jatropha curcas), an inedible oilseed plant, is among the most promising alternative energy sources because of its high oil content, rapid growth and extensive adaptability. Proteins encoded by MADS-box family genes are important transcription factors participated in regulating plant growth, seed development and responses to abiotic stress. However, there has been no in-depth research on the MADS-box genes and their roles in physic nut. RESULTS: In our study, 63 MADS-box genes (JcMADSs) were identified in the physic nut genome, and classed into five groups (MIKCC, Mα, Mß, Mγ, MIKC*) according to phylogenetic comparison with Arabidopsis homologs. Expression profile analysis based on RNA-seq suggested that many JcMADS genes had the strongest expression in seeds, and seven of them responded in leaves to at least one abiotic stressor (drought and/or salinity) at one or more time points. Transient expression analysis and a transactivation assay indicated that JcMADS40 is a nucleus-localized transcriptional activator. Plants overexpressing JcMADS40 did not show altered plant growth, but the overexpressing plants did exhibit reductions in grain size, grain length, grain width, 1000-seed weight and yield per plant. Further data on the reduced grain size in JcMADS40-overexpressing plants supported the putative role of JcMADS genes in seed development. CONCLUSIONS: This study will be useful in order to further understand the process of MADS-box genes involved in regulating growth and development in addition to their functions in abiotic stress resistance, and will eventually provide a theoretical basis for the functional investigation and the exploitation of candidate genes for the molecular improvement of physic nut.

Trichoderma harzianum mitigates salt stress in cucumber via multiple responses.

Trichoderma harzianum T-soybean plays an important role in controlling soybean root rot disease. However, the mechanism by which it improves plant tolerance to salt stress is not clear. In this study, we investigated the possible mechanism of T-soybean in mitigating the damage caused by salt stress in Cucumis sativus L plants. Our results suggest that T-soybean improved salt tolerance of cucumber seedlings by affecting the antioxidant enzymes including peroxidase (POD) (EC 1.11.1.6), polyphenol oxidase (PPO) (EC 1.14.18.1), phenylalanine ammonia-lyase (PAL) (EC 4.3.1.5), catalase (CAT) (EC 1.11.1.6), superoxide dismutase (SOD) (EC 1.15.1.1), ascorbate peroxidase (APX) (EC 1.11.1.11), and glutathione reductase (GR) (EC 1.6.4.2), by increasing the levels of proline, soluble sugars, soluble protein, ascorbic acid (AsA) and chlorophyll as well as improving root activity. Treatment with T-soybean improved the ratio of glutathione (GSH)/oxidized glutathione (GSSG) and AsA/dehydroascorbate (DHA), and up-regulated the expression of CsAPX and CsGR genes involved in the AsA-GSH cycle. In addition, treatment with T-soybean increased the K+ content and K+/Na+ ratio while decreased the Na+ concentration and ethylene level. In summary, the improved salt tolerance of cucumber plants may be due to multiple mechanisms of T-soybean, such as the increase in reactive oxygen species (ROS) scavenging, as well as maintaining osmotic balance and metabolic homeostasis under salt stress.

The complete mitochondrial genome of the Border Collie dog.

Border Collie dog is one of the famous breed of dog. In the present work we report the complete mitochondrial genome sequence of Border Collie dog for the first time. The total length of the mitogenome was 16,730 bp with the base composition of 31.6% for A, 28.7% for T, 25.5% for C, and 14.2% for G and an A-T (60.3%)-rich feature was detected. It harbored 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes and one non-coding control region (D-loop region). The arrangement of all genes was identical to the typical mitochondrial genomes of dogs.

The complete mitochondrial genome of the Chinese rhesus macaques, Macaca mulatta lasiota.

In this study, the complete mitochondrial genome sequence of the Chinese rhesus macaques Macaca mulatta lasiota has been reported for the first time. The total length of the mitogenome was 16,561 bp. It contained the typical structure, including 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and 1 control region. The overall composition of the mitogenome was A (31.7%), G (12.8%), C (30.4%) and T (25.1%). The M. mulatta lasiota mitogenome had 21 tRNA genes folded into a typical clover-leaf secondary structure except for tRNA(Ser).

[Effects of enhanced UV-B radiation on leaf anthraquinones content and cell ultrastructure of Aloe vera L].

By using transmission electron microscopy and high performance liquid chromatography, this paper studied the effects of enhanced UV-B radiation on the leaf anthraquinones content and cell ultrastructure of Aloe vera L. After treated with enhanced UV-B radiation 6 hours per day for 20 days, the total anthraquinone content, barbaloin content, and aloe-emondin content in A. vera leaves increased by 31.8%, 11.3%, and 22.0%, respectively, chloroplast envelope membrane was slightly damaged, but the structure of other organelles had no significant change. It was suggested that UV-B radiation could promote the accumulation of anthraquinone in A. vera leaves, but had less effects on the leaf cell ultrastructure.

Effect of lanthanum ions (La3+) on ferritin-regulated antioxidant process under PEG stress.

The physiological effects of lanthanum(III) ions on the ferritin-regulated antioxidant process were studied in wheat (Triticum aestivum L.) seedlings under polyethylene glycol (PEG) stress. Treatment with 0.1 mM La3+ resulted in increased levels of chlorophyll, carotenoid, proline, ascorbate, and reduced glutathione. The activities of superoxide dismutase, catalase, ascorbate peroxidase, monodehydroascorbate reductase, dehydroascorbate reductase, glutathione reductase, and peroxidase were also increased after La3+ treatment. Treatment with La3+ seems to enhance the capacity of the reactive oxygen species scavenging system, affect the Fe2+ and Fe3+ electron-transfer process in ferritin, and restrain the formation of hydroxyl radical (OH.), alleviating the oxidative damage induced by PEG stress.

Molecular cloning and characterization of a novel MAP kinase gene in Chorispora bungeana.

Chorispora bungeana Fisch. and C.A. Mey (Chorispora bungeana) is a rare alpine subnival plant species that is highly capable of resisting freezing environment. Since it is a stress-tolerant plant, we investigated the participation of mitogen-activated protein kinases (MAPKs) as possible mediators of abiotic stresses. We have isolated from Chorispora bungeana a new MAPK cDNA CbMAPK3 which encodes a 369 amino-acid protein with moderate to high nucleotide sequence similarity to previously reported plant MAPK genes. CbMAPK3 contains all 11 of the MAPK conserved subdomains and the phosphorylation motif TEY. The transcripts of CbMAPK3 were detected and no tissue-specific expression were observed in both roots and leaves, The transcripts of CbMAPK3 accumulated highly and rapidly when Chorispora bungeana treated with cold (4 and -4 degrees C), ABA and salinity stress. These results indicate that the CbMAPK3 may play an important role in response to environmental stresses.