Characterization and Potential Action Mode Divergences of Homologous ACO1 Genes during the Organ Development and Ripening Process between Non-Climacteric Grape and Climacteric Peach.

Ethylene is one crucial phytohormone modulating plants' organ development and ripening process, especially in fruits, but its action modes and discrepancies in non-climacteric grape and climacteric peach in these processes remain elusive. This work is focused on the action mode divergences of ethylene during the modulation of the organ development and ripening process in climacteric/non-climacteric plants. We characterized the key enzyme genes in the ethylene synthesis pathway, VvACO1 and PpACO1, and uncovered that their sequence structures are highly conserved, although their promoters exhibit important divergences in the numbers and types of the cis-elements responsive to hormones, implying various responses to hormone signals. Subsequently, we found the two have similar expression modes in vegetative organ development but inverse patterns in reproductive ones, especially in fruits. Then, VvACO1 and PpACO1 were further validated in promoting fruit ripening functions through their transient over-expression/RNAi-expression in tomatoes, of which the former possesses a weaker role than the latter in the fruit ripening process. Our findings illuminated the divergence in the action patterns and function traits of the key VvACO1/PpACO1 genes in the tissue development of climacteric/non-climacteric plants, and they have implications for further gaining insight into the interaction mechanism of ethylene signaling during the modulation of the organ development and ripening process in climacteric/non-climacteric plants.

Experimental study on particle movement and erosion behavior of the elbow in liquid-solid flow.

Recent investigations into the erosion of elbow junctions predominantly focus on identifying and predicting peak erosion points. Notably, these studies rely heavily on computational fluid dynamics methods, a valid approach but limited by its lack of empirical physical data. Additionally, the majority of these studies concentrate on the extrados, or outer curve of the elbow, neglecting the intrados or the inner curve. To provide a more comprehensive understanding of particle movements and the micro-mechanics of erosion on the elbow intrados, this study utilizes advanced observational technologies. High-speed camera technology, coupled with scanning electron microscopy, is employed to capture and record particle motion and micro-erosion patterns. The erosion rate is then estimated via the weight-loss method. The findings suggest that in low-speed liquid-solid flows (2.5 m/s), particles released from the intrados side of the elbow inlet exhibit a significant trajectory deviation from the centreline at an elbow angle of 60° from the inlet. Particles released from the extrados deviate towards the intrados side at an elbow angle of 30°. Secondary flow contributes to particle acceleration, unexpected trajectory deviation within the elbow, and an upward inclination in erosion on the intrados. The presence of partially overlapping scratches and cracks suggests that continuous ploughing and material fracturing are significant contributors to the micro-mechanics of erosion. When comparing the intrados and extrados, the extrados exhibits longer and shallower scratches, indicating a smaller impact angle. This research provides a more comprehensive understanding of particle trajectories and erosion patterns within elbow junctions during liquid-solid flows, offering new insights into the mechanisms underpinning these processes.

Comparative transcriptome analysis provides insight into regulation pathways and temporal and spatial expression characteristics of grapevine (Vitis vinifera) dormant buds in different nodes.

BACKGROUND: Bud dormancy is a strategic mechanism plants developed as an adaptation to unfavorable environments. The grapevine (Vitis vinifera) is one of the most ancient fruit vine species and vines are planted all over the world due to their great economic benefits. To better understand the molecular mechanisms underlying bud dormancy between adjacent months, the transcriptomes of 'Rosario Bianco' grape buds of 6 months and three nodes were analyzed using RNA-sequencing technology and pair-wise comparison. From November to April of the following year, pairwise comparisons were conducted between adjacent months. RESULTS: A total of 11,647 differentially expressed genes (DEGs) were obtained from five comparisons. According to the results of cluster analysis of the DEG profiles and the climatic status of the sampling period, the 6 months were divided into three key processes (November to January, January to March, and March to April). Pair-wise comparisons of DEG profiles of adjacent months and three main dormancy processes showed that the whole grapevine bud dormancy period was mainly regulated by the antioxidant system, secondary metabolism, cell cycle and division, cell wall metabolism, and carbohydrates metabolism. Additionally, several DEGs, such as VvGA2OX6 and VvSS3, showed temporally and spatially differential expression patterns, which normalized to a similar trend during or before April. CONCLUSION: Considering these results, the molecular mechanisms underlying bud dormancy in the grapevine can be hypothesized, which lays the foundation for further research.

Comparative study of DAM, Dof, and WRKY gene families in fourteen species and their expression in Vitis vinifera.

Bud dormancy is one of the most important defensive mechanisms through which plants resist cold stress during harsh winter weather. DAM, Dof, and WRKY have been reported to be involved in many biological processes, including bud dormancy. In the present study, grapevine (Vitis vinifera) and other thirteen plants (six woody plants and seven herbaceous plants) were analyzed for the quantity, sequence structure, and evolution patterns of their DAM, Dof, and WRKY gene family members. Moreover, the expression of VvDAM, VvDof, and VvWRKY genes was also investigated. Thus, 51 DAM, 1,205 WRKY, and 489 Dof genes were isolated from selected genomes, while 5 DAM, 114 WRKY, and 50 Dof duplicate gene pairs were identified in 10 genomes. Moreover, WGD and segmental duplication events were associated with the majority of the expansions of Dof and WRKY gene families. The VvDAM, VvDof, and VvWRKY genes significantly differentially expressed throughout bud dormancy outnumbered those significantly differentially expressed throughout fruit development or under abiotic stresses. Interestingly, multiple stress responsive genes were identified, such as VvDAM (VIT_00s0313g00070), two VvDof genes (VIT_18s0001g11310 and VIT_02s0025g02250), and two VvWRKY genes (VIT_07s0031g01710 and VIT_11s0052g00450). These data provide candidate genes for molecular biology research investigating bud dormancy and responses to abiotic stresses (namely salt, drought, copper, and waterlogging).

Characterization of VvSPL18 and Its Expression in Response to Exogenous Hormones during Grape Berry Development and Ripening.

The sequence and structure of grape SBP-box-like18 (VvSPL18) were identified and characterized to explore its regulatory roles during grape berry development and ripening. Homologous conservation across diverse plant species was observed, and its potential function and modulated roles in grapes were investigated. The results showed that VvSPL18 has an ORF sequence of 1,137 bp, encodes 378 amino acids, and is located on chromosome 14 of grapevine. VvSPL18 has the closest relationship with its homolog in soybeans. The promoter of VvSPL18 contains cis-elements responsive to gibberellins (GA) and salicylic acid (SA), indicating that this gene might respond to these hormones involved in the modulation of grape berry. VvSPL18 is mainly distributed in the nucleus. Expression profiles showed that VvSPL18 is highly expressed only at the veraison stage of the grape berry and is slightly expressed in other phases. RNA-seq data also revealed that VvSPL18 might participate in the modulation of grape berry development and ripening. Treatment with diverse hormones demonstrated that abscisic acid (ABA) had almost no effect on its expression, whereas naphthalene acetic acid (NAA) significantly upregulated its expression at the veraison stage. We also found that VvSPL18 has a GA-responsive cis-element but no NAA-responsive cis-element. GA could promote the expression of VvSPL18 with a peak at an earlier stage than NAA, suggesting that VvSPL18 responds faster to GA than to NAA. This result indicates that VvSPL18 might modulate berry development at this phase through an ABA-independent pathway, and it might directly respond to GA, but indirectly to NAA. Our findings provide insights into the functions of VvSPL18 in mediating grape berry development and ripening.

Identification of copper (Cu) stress-responsive grapevine microRNAs and their target genes by high-throughput sequencing.

MicroRNAs (miRNAs) are a class of single-stranded non-coding small RNAs (sRNAs) that are 20-24 nucleotides (nt) in length. Extensive studies have indicated that miRNAs play important roles in plant growth, development and stress responses. With more copper (Cu) and copper containing compounds used as bactericides and fungicides in plants, Cu stress has become one of the serious environmental problems that affect plant growth and development. In order to uncover the hidden response mechanisms of Cu stress, two small RNA libraries were constructed from Cu-treated and water-treated (Control) leaves of 'Summer Black' grapevine. Following high-throughput sequencing and filtering, a total of 158 known and 98 putative novel miRNAs were identified in the two libraries. Among these, 100 known and 47 novel miRNAs were identified as differentially expressed under Cu stress. Subsequently, the expression patterns of nine Cu-responsive miRNAs were validated by quantitative real-time PCR (qRT-PCR). There existed some consistency in expression levels of Cu-responsive miRNAs between high throughput sequencing and qRT-PCR assays. The targets prediction of miRNAs indicates that miRNA may regulate some transcription factors, including AP2, SBP, NAC, MYB and ARF during Cu stress. The target genes for two known and two novel miRNAs showed specific cleavage sites at the 10th and/or 11th nucleotide from the 5'-end of the miRNA corresponding to their miRNA complementary sequences. The findings will lay the foundation for exploring the role of the regulation of miRNAs in response to Cu stress and provide valuable gene information for breeding some Cu-tolerant grapevine cultivars.

Transcriptome analysis and physiological responses of the potato plantlets in vitro under red, blue, and white light conditions.

Light is an important factor for plant development and has serious effects on the growth, production and quality of potatoes. However, the physical and molecular mechanisms by which potato plantlets cope with different light qualities are not understood. In this study, the potato "Zhuanxinwu", which is a germplasm potato resource with a high anthocyanin content, was used for physiological and transcriptome profiling analyses to uncover the different mechanisms that occur in response to blue, red and white light conditions, with the white light condition serving as the control. Multiple growth indexes, protective enzyme activity and metabolite accumulation were measured. The results indicated that white light promoted a shift in biomass allocation away from tubers to leaves to enhance dry leaf matter and reduce tuber fresh/dry weight relative to the effects of blue or red light. The leaf area and anthocyanin content values were greater for plants grown in blue light than those grown in white or red light, suggesting that combinations of different spectra were more conducive to regulating potato growth. A total of 2220 differentially expressed genes (DEGs) were found among the three samples, and the DEGs in the three comparison sets were analyzed. A total of 1180 and 984 DEGs were identified in the red light (Red) and blue light (Blue) conditions compared to the control condition, respectively, and 359 DEGs overlapped between the two comparison sets (Blue_vs_White and Red_vs_White). Interestingly, the 24 most common overlapped DEGs were involved in photosynthesis, respiration, and reactive oxygen species (ROS) scavenging. Of these DEGs, four genes involved in photosynthesis and two genes involved in pigment synthesis were highly expressed, implying that some genes could be implemented to cope with different light spectra by regulating the expression of DEGs involved in the corresponding metabolic pathways. In conclusion, our study characterizes physiological responses of potato to different light qualities and identifies potential pathways and candidate genes involved in these responses, thus providing a basis for further research on artificial light regulation of potato plant growth.

Genetic diversity and relatedness among ornamental purslane (Portulaca L.) accessions unraveled by SRAP markers.

Ornamental purslanes (Portulaca L.) are a popular annual bedding and container plant for landscaping. Little information is available concerning the genetic characterization of ornamental purslane resources thus far. The purpose of this study was to investigate the genetic diversity and relationships present in a collection of ornamental purslanes from Portulaca umbraticola and P. grandiflora cultivated in China, using sequence-related amplified polymorphism (SRAP) markers. The genotyping showed that 16 SRAP primer combinations totally produced 261 informative fragments and averaged 16.31 per primer combination. The major allele frequency and Nei's gene diversity was calculated at 0.78 and 0.31 across the loci, indicative of a moderate low diversity. Both unweighted pair group method with arithmetic average (UPGMA) clustering and a Bayesian-based approach apparently assigned the whole accessions into two sub-groups: P. umbraticola and P. grandiflora, well concordant with the botanical classification and flower type. The findings provide a brandnew understanding of genetic diversity and population structure present in ornamental purslane, and benefit a sound design of breeding programs in future.

Abscisic acid, sucrose, and auxin coordinately regulate berry ripening process of the Fujiminori grape.

The aim of this study was to examine the effect of abscisic acid (ABA), sucrose, and auxin on grape fruit development and to assess the mechanism of these three factors on the grape fruit ripening process. Different concentrations of ABA, sucrose, and auxin were used to treat the grape fruit, and the ripening-related indices, such as physiological and molecular level parameters, were analyzed. The activity of BG protein activity was analyzed during the fruit development. Sucrose, ABA, and auxin influenced the grape fruit sugar accumulation in different ways, as well as the volatile compounds, anthocyanin content, and fruit firmness. ABA and sucrose induced, but auxin blocked, the ripening-related gene expression levels, such as softening genes PE, PG, PL, and CELL, anthocyanin genes DFR, CHI, F3H, GST, CHS, and UFGT, and aroma genes Ecar, QR, and EGS. ABA, sucrose, and glucose induced the fruit dry weight accumulation, and auxin mainly enhanced fruit dry weight through seed weight accumulation. In the early development of grape, starch was the main energy storage; in the later, it was glucose and fructose. Sucrose metabolism pathway-related gene expression levels were significant for glucose and fructose accumulation. BG protein activity was important in the regulation of grape ABA content levels. ABA plays a core role in the grape fruit development; sucrose functions in fruit development through two pathways: one was ABA dependent, the other ABA independent. Auxin blocked ABA accumulation to regulate the fruit development process.

Fertilization of Grapevine Based on Gene Expression.

The application of genetic information in agricultural production is an important issue, which is highly worthy of attention. Gene expression data can accurately reflect the growth and metabolic status of plants, with which we can predict and monitor the nutritional requirements of plants and then derive accurate fertilization strategies. In this study, to verify the feasibility and workability of gene information-based fertilization strategies and to figure out the specific nutritional requirements of grapevine ( L.) at various developmental stages, the expression levels of 13 N-P-K uptake and metabolism genes and their responses to fertilization during the flowering and berry development stages were validated by using quantitative polymerase chain reaction (PCR). The results showed that in the particular stages where N-P-K uptake and metabolism genes were highly expressed, these genes also showed more positive responses to fertilization and the grape quality was more dramatically improved. This proved the feasibility and workability of this novel fertilization strategy. The nutritional requirements of grapevine during the flowering and berry developmental phases were summarized in terms of gene expression levels, in which grapevine needs more P at the flowering stage, more N at the first berry expansion stage, less nutrient at the seed stone hardening stage, and more P and K at the second berry expansion stage and the veraison stage. The present study is one of the novel and initial findings regarding the application of fertilizers in vineyards for better cultivation of grapevine during common cultural practices.