MADS-box protein PpDAM6 regulates chilling requirement-mediated dormancy and bud break in peach.

Bud dormancy is crucial for winter survival and is characterized by the inability of the bud meristem to respond to growth-promotive signals before the chilling requirement (CR) is met. However, our understanding of the genetic mechanism regulating CR and bud dormancy remains limited. This study identified PpDAM6 (DORMANCY-ASSOCIATED MADS-box) as a key gene for CR using a genome-wide association study analysis based on structural variations in 345 peach (Prunus persica (L.) Batsch) accessions. The function of PpDAM6 in CR regulation was demonstrated by transiently silencing the gene in peach buds and stably overexpressing the gene in transgenic apple (Malus × domestica) plants. The results showed an evolutionarily conserved function of PpDAM6 in regulating bud dormancy release, followed by vegetative growth and flowering, in peach and apple. The 30-bp deletion in the PpDAM6 promoter was substantially associated with reducing PpDAM6 expression in low-CR accessions. A PCR marker based on the 30-bp indel was developed to distinguish peach plants with non-low and low CR. Modification of the H3K27me3 marker at the PpDAM6 locus showed no apparent change across the dormancy process in low- and non-low- CR cultivars. Additionally, H3K27me3 modification occurred earlier in low-CR cultivars on a genome-wide scale. PpDAM6 could mediate cell-cell communication by inducing the expression of the downstream genes PpNCED1 (9-cis-epoxycarotenoid dioxygenase 1), encoding a key enzyme for ABA biosynthesis, and CALS (CALLOSE SYNTHASE), encoding callose synthase. We shed light on a gene regulatory network formed by PpDAM6-containing complexes that mediate CR underlying dormancy and bud break in peach. A better understanding of the genetic basis for natural variations of CR can help breeders develop cultivars with different CR for growing in different geographical regions.

The Impact of Urbanization on Urban Heat Island: Predictive Approach Using Google Earth Engine and CA-Markov Modelling (2005-2050) of Tianjin City, China.

Urbanization has adverse environmental effects, such as rising surface temperatures. This study analyzes the relationship between the urban heat island (UHI) intensity and Tianjin city's land cover characteristics. The land use cover change (LUCC) effects on the green areas and the land surface temperature (LST) were also studied. The land cover characteristics were divided into five categories: a built-up area, an agricultural area, a bare area, a forest, and water. The LST was calculated using the thermal bands of spatial images taken from 2005 to 2020. The increase in the built-up area was mainly caused by the agricultural area decreasing by 11.90%. The average land surface temperature of the study area increased from 23.50 to 36.51 °C, and the region moved to a high temperature that the built-up area's temperature increased by 1.5%. Still, the increase in vegetation cover was negative. From 2020 to 2050, the land surface temperature is expected to increase by 9.5 °C. The high-temperature areas moved into an aerial distribution, and the direction of urbanization determined their path. Urban heat island mitigation is best achieved through forests and water, and managers of urban areas should avoid developing bare land since they may suffer from degradation. The increase in the land surface temperature caused by the land cover change proves that the site is becoming more urbanized. The findings of this study provide valuable information on the various aspects of urbanization in Tianjin and other regions. In addition, future research should look into the public health issues associated with rapid urbanization.

Temperature-responsive module of OfAP1 and OfLFY regulates floral transition and floral organ identity in Osmanthus fragrans.

The MADS-box transcription factor APETELA1 (AP1) is crucially important for reproductive developmental processes. The function of AP1 and the classic LFY-AP1 interaction in woody plants are not widely known. Here, the OfAP1-a gene from the continuously flowering plant Osmanthus fragrans 'Sijigui' was characterized, and its roles in regulating flowering time, petal number robustness and floral organ identity were determined using overexpression in Arabidopsis thaliana and Nicotiana tabacum. The expression of OfAP1-a was significantly induced by low ambient temperature and was upregulated with the floral transition process. Ectopic expression OfAP1-a revealed its classic function in flowering and flower ABC models. The expression of OfAP1-a is inhibited by LEAFY (OfLFY) through direct promoter binding, as confirmed by yeast one-hybrid and dual luciferase assays. Arabidopsis plants overexpressing OfAP1-a exhibited accelerated flowering and altered floral organ identities. Moreover, OfAP1-a-overexpressing plants displayed variable petal numbers. Likewise, the overexpression of OfLFY in Arabidopsis and Nicotiana altered petal number robustness and inflorescence architecture, partially by regulating native AP1 in transformed plants. Furthermore, we performed RNA-seq analysis of transgenic Nicotiana plants. DEGs were identified by transcriptome analysis, and we found that the expression of several floral homeotic genes was altered in both OfAP1-a and OfLFY-overexpressing transgenic lines. Our results suggest that OfAP1-a may play important roles during floral transition and development in response to ambient temperature. OfAP1-a functions as a petal number modulator and may directly activate a subset of flowers to regulate floral organ formation. OfAP1-a and OfLFY mutually regulate the expression of each other and coregulate genes that might be involved in these phenotypes related to flowering. The results provide valuable data for understanding the function of the LFY-AP1 module in the reproductive process and shaping floral structures in woody plants.

Discovery of a key gene associated with fruit maturity date and analysis of its regulatory pathway in peach.

Fruit maturity is an important agronomic trait of fruit crops. Although in previous studies, several molecular markers are developed for the trait, the knowledge about its candidate genes is particularly limited. In this study, a total of 357 peach accessions were re-sequenced to obtain 949,638 SNPs. Combing with 3-year fruit maturity dates, a genome-wide association analysis was performed, and 5, 8, and 9 association loci were identified. To screen the candidate genes for those year-stable loci on chromosomes 4 and 5, two maturity date mutants were used for transcriptome sequencing. Gene expression analysis indicated that Prupe.4G186800 and Prupe.4G187100 on chromosome 4 were essential to fruit ripening in peaches. However, the expression analysis of different tissues showed that the first gene has no tissue-specific character, but transgenic studies showed that the latter is more likely to be a key candidate gene than the first for the maturity date in peach. The yeast two-hybrid assay showed that the proteins encoded by the two genes interacted and then regulated fruit ripening. Moreover, the previously identified 9 bp insertion in Prupe.4G186800 may affect their interaction ability. This research is of great significance for understanding the molecular mechanism of peach fruit ripening and developing practical molecular markers in a breeding program.

The Promoter Analysis of VvPR1 Gene: A Candidate Gene Identified through Transcriptional Profiling of Methyl Jasmonate Treated Grapevine (Vitis vinifera L.).

Methyl jasmonate (MeJA) plays a vital role in plant disease resistance and also induces the expression of disease resistance genes in plants. In this study, a transcriptome analysis was performed on grapevine leaves after 12, 24 and 48 h of MeJA-100 µM treatment. A total of 1242 differentially expressed genes (DEGs) were identified from the transcriptome data, and the analysis of the DEGs showed that genes related to phytohormone signal transduction, jasmonic acid-mediated defense, Mitogen-activated protein kinase (MAPK), and flavonoid biosynthetic pathways were upregulated. As Pathogenesis-related gene 1 (PR1) is an important marker gene in plant defense also upregulated by MeJA treatment in RNA-seq data, the VvPR1 gene was selected for a promoter analysis with ß-glucuronidase (GUS) through transient expression in tobacco leaves against abiotic stress. The results showed that the region from -1837 bp to -558 bp of the VvPR1 promoter is the key region in response to hormone and wound stress. In this study, we extended the available knowledge about induced defense by MeJA in a grapevine species that is susceptible to different diseases and identified the molecular mechanisms by which this defense might be mediated.

Identification of key gene networks controlling anthocyanin biosynthesis in peach flower.

Flavonoids, particularly anthocyanin is the main pigment that determined the red color of peach flowers, and help the plant to attract pollinators, protect the reproductive organs of flower from photo-oxidative effects of light and various non-communicable diseases. Through weightage gene coexpression network analysis (WGCNA) we identified a network of 15 hub genes that co-expressed throughout peach flower development including 5 genes coded for the key enzymes (CHI, F3'H, DFR, LAR and UFGT) of flavonoid biosynthetic pathway and 1 gene Prupe.1G111700 identified as R2R3 family transcription factor MYB108. Over expression of PpMYB108 significantly increased anthocyanin biosynthesis in Tobacco flowers. Moreover, the expression correlation between PpMYB108 and PpDFR, suggests that PpMYB108 play the role of transcriptional activator for PpDFR. This was further supported by a 6 bp insertion of MYB biding site in the core promoter region of PpDFR in red flower. The positive interaction of PpMYB108 with PpDFR promoter from red flower was confirmed in yeast one hybrid assay. These findings may be helpful in peach breeding programs as well as in identifying anthocyanin related genes in other species.

Multi-omics approaches identify a key gene, PpTST1, for organic acid accumulation in peach.

Organic acid content in fruit is an important determinant of peach organoleptic quality, which undergoes considerable variations during development and maturation. However, its molecular mechanism remains largely unclear. In this study, an integrative approach of genome-wide association studies and comparative transcriptome analysis were applied to identify candidate genes involved in organic acid accumulation in peach. A key gene PpTST1, encoding tonoplast sugar transporter, was identified and the genotype of PpTST1 with a single-base transversion (G1584T) in the third exon which leads to a single amino acid substitution (Q528H) was associated with low level of organic acid content in peach. Overexpression of PpTST1His resulted in reduced organic acid content along with increased sugar content both in peach and tomato fruits, suggesting its dual function in sugar accumulation and organic acid content reduction. Two V-type proton ATPases interact with PpTST1 in yeast two-hybridization assay. In addition, the G1584T transversion appeared and gradually accumulated during domestication and improvement, which indicated that PpTST1 was under selection. The identification and characterization of PpTST1 would facilitate the improvement of peach fruit quality.

Transcriptome analysis reveals pathogenesis-related gene 1 pathway against salicylic acid treatment in grapevine (Vitis vinifera L).

Salicylic acid (SA) is a well-studied phenolic plant hormone that plays an important role in plant defense against the hemi-biothrophic and biothrophic pathogens and depends on the living cells of host for the successful infection. In this study, a pathogenesis test was performed between Vitis davidii and V. vinifera cultivars against grape white rot disease (Coniella diplodiella). V. davidii was found to be resistant against this disease. SA contents were found to be higher in the resistant grape cultivar after different time points. RNA-seq analysis was conducted on susceptible grapevine cultivars after 12, 24, and 48 h of SA application with the hypothesis that SA may induce defense genes in susceptible cultivars. A total of 511 differentially expressed genes (DEGs) were identified from the RNA-seq data, including some important genes, VvWRKY1/2, VvNPR1, VvTGA2, and VvPR1, for the SA defense pathway. DEGs related to phytohormone signal transduction and flavonoid biosynthetic pathways were also upregulated. The quantitative real-time PCR (qRT-PCR) results of the significantly expressed transcripts were found to be consistent with the transcriptome data, with a high correlation between the two analyses. The pathogenesis-related gene 1 (VvPR1), which is an important marker gene for plant defense, was selected for further promoter analysis. The promoter sequence showed that it contains some important cis-elements (W-box, LS7, as-1, and TCA-element) to recruit the transcription factors VvWRKY, VvNPR1, and VvTGA2 to express the VvPR1 gene in response to SA treatment. Furthermore, the VvPR1 promoter was serially deleted into different fragments (-1,837, -1,443, -1,119, -864, -558, -436, and -192 ) bp and constructed vectors with the GUS reporter gene. Deletion analysis revealed that the VvPR1 promoter between -1837 bp to -558 bp induced significant GUS expression with respect to the control. On the basis of these results, the -558 bp region was assumed to be an important part of the VvPR1 promoter, and this region contained the important cis-elements related to SA, such as TCA-element (-1,472 bp), LS7 (-1,428 bp), and as-1 (-520 bp), that recruit the TFs and induce the expression of the VvPR1 gene. This study expanded the available information regarding SA-induced defense in susceptible grapes and recognized the molecular mechanisms through which this defense might be mediated.

Effect of Salinity Stress on Germination, Seedling Growth, Mineral Uptake and Chlorophyll Contents of Three Cucurbitaceae Species

Abstract This study was performed to screen out the various species of 'Cucurbitaceae' family, musk melon (Kalash and Durga), bottle gourd (Crystal Long and Nuefield) and squash (Green Round, and Squash Malika) against the salt stress. All genotypes were treated with five different levels of NaCl (T0 = control, T1 = 1.5 dS m-1, T2 = 3.0 dS m-1, T3 = 4.5 dS m-1 and T4 = 6.0 dS m-1) and half strength of Hoagland's nutrients solution as the base nutrient solution. Results showed that the bottle gourd varieties "Nuefield" and "Crystal Long" performed best by maintaining the highest germination (93.2% and 85.6%), number of leaves per plant (4.5 and 5.7), shoot length (16.84 cm and 16.14 cm), root length (13.48 cm and 13.00 cm), plant fresh weight (942.2 g and 918.6 g), plant dry weight (118.4 g and 107.5 g), leaf area (171.2 cm2 and 169.1 cm2), chlorophyll content (3.5 μg/cm-2 and 3.4 μg/cm-2) with low chloride (1.57 ppm and 1.59 ppm) and sodium content (0.47 ppm and 0.51 ppm) under salt stress followed by varieties of Squash (Green Round, and Squash Malika) and musk melon (Kalash and Durga). It was also found that a higher level of salinity (4.5 dS m-1 and 6.0 dS m-1) has more adverse effects on the performance of all selected genotypes. Conclusively, it can be recommended that as compared to all tested species, bottle gourd varieties "Nuefield" and "Crystal Long" have the ability to withstand against salinity stress and should be planted under salt stress conditions.