Carotenoid Pigment Accumulation in Horticultural Plants

Environmental stresses, such as temperature, heavy metals, drought, cold, and microbial infections adversely damage various aspects of plant growth and development. Salinity and drought are among major hazardous factors, which adversity affects plant growth and productivity. Transcription factors, such as basic helix-loop-helix play critical roles in regulating plant physiological processes under abiotic stresses. In this study, we presented the characterization of a tomato SlbHLH22 gene under abiotic stresses such as drought and salinity. Plants overexpressing SlbHLH22 showed short height with small leaves and enhanced flavonoid accumulation. In wild type (WT) plant, the elevated levels of SlbHLH22 were detected under salt and D-mannitol stresses. Subcellular localization analysis revealed that SlbHLH22 protein was targeted to the nucleus in onion epidermal cells. Transactivation assay in yeast demonstrated that SlbHLH22 had transcriptional activation ability. The transgenic plants overexpressing SlbHLH22 displayed enhanced vigor and more tolerant to drought and salinity than WT. Overexpression of SlbHLH22 significantly peaked the activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) to minimize the impacts of reactive oxygen species such as H2O2, which was reduced significantly in transgenic plants along with Malondialdehyde (MDA). Moreover, the expression levels of ROS defense genes (SlPOD, SlCAT, SlSOD), ABA biosynthesis genes, proline biosynthesis, and flavonoids synthesis genes were also activated under salinity and drought. Taken together, our study implies that the overexpression of SlbHLH22 improved tomato plant stress resistance by improving ROS scavenging system, increasing osmotic potential and enhanced accumulation of secondary metabolites in tomato plants.

/pdf/dissecting-the-role-of-a-basic-helix-loop-helix-hc9u62bfl1.pdf

Dissecting the Role of a Basic Helix-Loop-Helix Transcription Factor, SlbHLH22 , Under Salt and Drought Stresses in Transgenic Solanum lycopersicum L.

The basic helix-loop-helix (bHLH) gene family is one of the largest transcription factor families in plants and is functionally characterized in diverse species. However, less is known about its functions in the economically important allopolyploid oil crop, Brassica napus. We identified 602 potential bHLHs in the B. napus genome (BnabHLHs) and categorized them into 35 subfamilies, including seven newly separated subfamilies, based on phylogeny, protein structure, and exon-intron organization analysis. The intron insertion patterns of this gene family were analyzed and a total of eight types were identified in the bHLH regions of BnabHLHs. Chromosome distribution and synteny analyses revealed that hybridization between Brassica rapa and Brassica oleracea was the main expansion mechanism for BnabHLHs. Expression analyses showed that BnabHLHs were widely in different plant tissues and formed seven main patterns, suggesting they may participate in various aspects of B. napus development. Furthermore, when roots were treated with five different hormones (IAA, auxin; GA3, gibberellin; 6-BA, cytokinin; ABA, abscisic acid and ACC, ethylene), the expression profiles of BnabHLHs changed significantly, with many showing increased expression. The induction of five candidate BnabHLHs was confirmed following the five hormone treatments via qRT-PCR. Up to 246 BnabHLHs from nine subfamilies were predicted to have potential roles relating to root development through the joint analysis of their expression profiles and homolog function. The 602 BnabHLHs identified from B. napus were classified into 35 subfamilies, and those members from the same subfamily generally had similar sequence motifs. Overall, we found that BnabHLHs may be widely involved in root development in B. napus. Moreover, this study provides important insights into the potential functions of the BnabHLHs super gene family and thus will be useful in future gene function research.

/pdf/genome-wide-survey-of-the-bhlh-super-gene-family-in-brassica-ae8c22oho7.pdf

Genome-wide survey of the bHLH super gene family in Brassica napus.

Proper flower development is essential for sexual reproductive success and the setting of fruits and seeds. The availability of a high quality genome sequence for pineapple makes it an excellent model for studying fruit and floral organ development. In this study, we sequenced 27 different pineapple floral samples and integrated nine published RNA-seq datasets to generate tissue- and stage-specific transcriptomic profiles. Pairwise comparisons and weighted gene co-expression network analysis successfully identified ovule-, stamen-, petal- and fruit-specific modules as well as hub genes involved in ovule, fruit and petal development. In situ hybridization confirmed the enriched expression of six genes in developing ovules and stamens. Mutant characterization and complementation analysis revealed the important role of the subtilase gene AcSBT1.8 in petal development. This work provides an important genomic resource for functional analysis of pineapple floral organ growth and fruit development and sheds light on molecular networks underlying pineapple reproductive organ growth.

/pdf/floral-transcriptomes-reveal-gene-networks-in-pineapple-d9rldp383r.pdf

Floral transcriptomes reveal gene networks in pineapple floral growth and fruit development

The transition from vegetative to reproductive growth is very important for early maturity in cotton. However, the genetic control of this highly dynamic and complex developmental process remains unclear. A high-resolution tissue- and stage-specific transcriptome profile was generated from six developmental stages using 72 samples of two early-maturing and two late-maturing cotton varieties. The results of histological analysis of paraffin sections showed that flower bud differentiation occurred at the third true leaf stage (3TLS) in early-maturing varieties, but at the fifth true leaf stage (5TLS) in late-maturing varieties. Using pairwise comparison and weighted gene co-expression network analysis, 5312 differentially expressed genes were obtained, which were divided into 10 gene co-expression modules. In the MElightcyan module, 46 candidate genes regulating cotton flower bud differentiation were identified and expressed at the flower bud differentiation stage. A novel key regulatory gene related to flower bud differentiation, GhCAL, was identified in the MElightcyan module. Anti-GhCAL transgenic cotton plants exhibited late flower bud differentiation and flowering time. GhCAL formed heterodimers with GhAP1-A04/GhAGL6-D09 and regulated the expression of GhAP1-A04 and GhAGL6-D09. GhAP1-A04- and GhAGL6-D09-silenced plants also showed significant late flowering. Finally, we propose a new flowering regulatory pathway mediated by GhCAL. This study elucidated the molecular mechanism of cotton flowering regulation and provides good genetic resources for cotton early-maturing breeding.

High-resolution temporal dynamic transcriptome landscape reveals a GhCAL-mediated flowering regulatory pathway in cotton (Gossypium hirsutum L.).

The overexpression of SlbHLH22 functioned in controlling flowering time, accelerated fruit ripening, and produced more ethylene-producing phenotypes in tomato. Flowering and fruit ripening are two complex transition processes regulated by various internal and external factors that ultimately lead to fruit maturation and final seed dispersal. The basic helix–loop–helix (bHLH) transcription factor is the largest TF gene family in plants that controls various biological and developmental aspects, but the actual roles of these genes have not been fully studied. Here, we performed a functional characterization of the bHLH gene SlbHLH22 in tomato. SlbHLH22 was fully expressed in tomato flowers, while a moderate expression level was also observed in fruits at different developmental stages. Overexpression of the SlbHLH22 gene revealed that it is highly involved in controlling flowering time, through the activation of the SlSFT or SlLFY genes, and promoting fruit ripening and improved carotenoid accumulation. The expression patterns of carotenoid-related genes (SlPYS1) were also upregulated in transgenic tomato fruits. In transgenic tomato fruit, we observed clear changes in colour from green to orange with enhanced expression of the SlbHLH22 gene. SlbHLH22 was upregulated under exogenous ACC, IAA, ABA, and ethephon. Overexpression of SlbHLH22 also promotes ethylene production. Moreover, ethylene biosynthesis and perception genes (SlACO3, SlACS1, SlACS2, SlACS4, SlACS1a, SlEIN1, SlEIN2, SlEIN3, SlEIN4, SlETR2, SlETR3, SlSAM3, and SlSAMS) were upregulated. Ripening-related genes (SlAP2a, SlCNR, SlNOR, SlMYB, and SlTAG) were consistent in their expression pattern in transgenic plants. Finally, our study provides evidence that tomato bHLH genes play an important role in flowering, fruit ripening, and development.

Overexpression of a basic helix-loop-helix transcription factor gene, SlbHLH22, promotes early flowering and accelerates fruit ripening in tomato (Solanum lycopersicum L.).

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Overexpression of a basic helix-loop-helix transcription factor gene, SlbHLH22, promotes early flowering and accelerates fruit ripening in tomato (Solanum lycopersicum L.).