A haplotype-resolved chromosome-scale genome for Quercus rubra L. provides insights into the genetics of adaptive traits for red oak species.

Northern red oak (Quercus rubra L.) is an ecologically and economically important forest tree native to North America. We present a chromosome-scale genome of Q. rubra generated by the combination of PacBio sequences and chromatin conformation capture (Hi-C) scaffolding. This is the first reference genome from the red oak clade (section Lobatae). The Q. rubra assembly spans 739 Mb with 95.27% of the genome in 12 chromosomes and 33,333 protein-coding genes. Comparisons to the genomes of Quercus lobata and Quercus mongolica revealed high collinearity, with intrachromosomal structural variants present. Orthologous gene family analysis with other tree species revealed that gene families associated with defense response were expanding and contracting simultaneously across the Q. rubra genome. Quercus rubra had the most CC-NBS-LRR and TIR-NBS-LRR resistance genes out of the 9 species analyzed. Terpene synthase gene family comparisons further reveal tandem gene duplications in TPS-b subfamily, similar to Quercus robur. Phylogenetic analysis also identified 4 subfamilies of the IGT/LAZY gene family in Q. rubra important for plant structure. Single major QTL regions were identified for vegetative bud break and marcescence, which contain candidate genes for further research, including a putative ortholog of the circadian clock constituent cryptochrome (CRY2) and 8 tandemly duplicated genes for serine protease inhibitors, respectively. Genome-environment associations across natural populations identified candidate abiotic stress tolerance genes and predicted performance in a common garden. This high-quality red oak genome represents an essential resource to the oak genomic community, which will expedite comparative genomics and biological studies in Quercus species.

Changes in Metabolite Profiling and Expression Levels of Key Genes Involved in the Terpenoid Biosynthesis Pathway in Garden Sage (Salvia officinalis) under the Effect of Hydrazine Hydrate.

Mutagenesis is a highly efficient tool for establishing genetic variation and is widely used for genetic enhancement in various plants. The key benefit of mutation breeding is the prospect of enhancing one or several characteristics of a variety without altering the genetic background. In this study, we exposed the seeds of Salvia officinalis to four concentrations of hydrazine hydrate (HZ), i.e., (0%, 0.1%, 0.2%, and 0.3%) for 6 h. The contents of terpenoid compounds in the S. officinalis plantlets driven from the HZ-treated seeds were determined by GC-MS, which resulted in the identification of a total of 340 phytochemical compounds; 163 (87.48%), 145 (84.49%), 65 (97.45%), and 62 (98.32%), from the four concentrations of HZ (0%, 0.1%, 0.2%, and 0.3%), respectively. Furthermore, we used the qRT-PCR system to disclose the "transcriptional control" for twelve TPS genes related to terpenoid and terpene biosynthesis, namely, SoGPS, SoMYRS, SoNEOD, SoCINS, SoSABS, SoLINS, SoFPPS, SoHUMS, SoTPS6, SoSQUS, SoGGPS, and SoGA2. Altogether, results are likely to ensure some positive relationship between the concentrations of the chemical mutagen HZ used for treating the seeds, the type and amount of the produced terpenes, and the expression of their corresponding genes.

Integrated transcriptomic and metabolomic analysis reveals the changes in monoterpene compounds during the development of Muscat Hamburg (Vitis vinifera L.) grape berries.

Monoterpenes are important compounds that influence the aromas of grapes and wines. The molecular mechanisms underlying the changes in monoterpenes during the grape ripening period have not been thoroughly characterized. In this study, the free and bound monoterpene profiles in Muscat Hamburg grape berries at different phenological stages were investigated at the transcriptomic and metabolomic levels. Principal component analyses indicated that the free and bound monoterpene profiles were affected by the developmental stages. Most monoterpenes were produced slowly before veraison, but they accumulated rapidly during the veraison period, after which their contents decreased slightly in mature berries. The transcriptomic analysis revealed 35 differentially expressed genes involved in the monoterpene synthesis pathway. The VIT_04s0008g04970, VIT_03s0063g02030 and VIT_15s0024g00850 expression levels were consistent with the changes in the accumulation of monoterpene compounds. The quantitative real-time polymerase chain reaction analysis of eight key differentially expressed genes in the monoterpenoid pathway confirmed the RNA-seq data were reliable. Our findings provide new insights into Muscat Hamburg grape aroma development. Further research on the period with the highest aroma potential may lead to enhanced grape berry aroma qualities.

Genome-wide analysis of terpene synthase gene family to explore candidate genes related to disease resistance in Prunus persica.

In plants, a family of terpene synthases (TPSs) is responsible for the biosynthesis of terpenes and contributes to species-specific diversity of volatile organic compounds, which play essential roles in fitness of plants. However, little is known about the TPS gene family in peach and/or nectarine (Prunus persica L.). In this study, we identified 40 PpTPS genes in peach genome v2.0. Although these PpTPSs could be clustered into five classes, they distribute in several gene clusters of three chromosomes, share conserved exon-intron organizations, and code similar protein motifs. Thirty-five PpTPSs, especially PpTPS2, PpTPS23, PpTPS17, PpTPS18, and PpTPS19, altered their transcript levels after inoculation with Botryosphaeria dothidea, a cause of peach gummosis, compared to the mock treatments, which might further affect the contents of 133 terpenoids at 48 hours and/or 84 hours post inoculations in the current-year shoots of 'Huyou018', a highly susceptible nectarine cultivar. Moreover, about fifteen PpTPSs, such as PpTPS1, PpTPS2, PpTPS3, and PpTPS5, showed distinct expression patterns during fruit development and ripening in two peach cultivars, yellow-fleshed 'Jinchun' and white-fleshed 'Hikawa Hakuho'. Among them, the transcription level of chloroplast-localized PpTPS3 was obviously related to the content of linalool in fruit pulps. In addition, elevated concentrations (0.1 g/L to 1.0 g/L) of linalool showed antifungal activities in PDA medium. These results improve our understanding of peach PpTPS genes and their potential roles in defense responses against pathogens.

Liverwort oil bodies: diversity, biochemistry, and molecular cell biology of the earliest secretory structure of land plants.

Liverworts are known for their large chemical diversity. Much of this diversity is synthesized and enclosed within oil bodies (OBs), a synapomorphy of the lineage. OBs contain the enzymes to biosynthesize and store large quantities of sesquiterpenoids and other compounds while limiting their cytotoxicity. Recent important biochemical and molecular discoveries related to OB formation, diversity, and biochemistry allow comparison with other secretory structures of land plants from an evo-devo perspective. This review addresses and discusses the most recent advances in OB origin, development, and function towards understanding the importance of these organelles in liverwort physiology and adaptation to changing environments. Our mapping of OB types and chemical compounds to the current liverwort phylogeny suggests that OBs were present in the most recent common ancestor of liverworts, supporting that OBs evolved as the first secretory structures in land plants. Yet, we require better sampling to define the macroevolutionary pattern governing the ancestral type of OB. We conclude that current efforts to find molecular mechanisms responsible for the morphological and chemical diversity of secretory structures will help understand the evolution of each major group of land plants, and open new avenues in biochemical research on bioactive compounds in bryophytes and vascular plants.

The Draft Genome of a Flat Peach (Prunus persica L. cv. '124 Pan') Provides Insights into Its Good Fruit Flavor Traits.

The flat peach has become more and more popular worldwide for its fruit quality with relatively low acidity, high sugar content and rich flavor. However, the draft genome assembly of flat peach is still unavailable and the genetic basis for its fruit flavor remains unclear. In this study, the draft genome of a flat peach cultivar '124 Pan' was assembled by using a hybrid assembly algorithm. The final assembly resulted in a total size of 206 Mb with a N50 of 26.3 Mb containing eight chromosomes and seven scaffolds. Genome annotation revealed that a total of 25,233 protein-coding genes were predicted with comparable gene abundance among the sequenced peach species. The phylogenetic tree and divergence times inferred from 572 single copy genes of 13 plant species confirmed that Prunus ferganensis was the ancestor of the domesticated peach. By comparing with the genomes of Prunus persica (Lovell) and Prunus ferganensis, the expansion of genes encoding enzymes involved in terpene biosynthesis was found, which might contribute to the good fruit flavor traits of '124 Pan'. The flat peach draft genome assembly obtained in this study will provide a valuable genomic resource for peach improvement and molecular breeding.

Early transcriptional response of terpenoid metabolism to Colletotrichum gloeosporioides in a resistant wild strawberry Fragaria nilgerrensis.

Modern strawberry production is often threatened by microbe pathogens. Anthracnose is among the most prominent fungal disease caused mainly by Colletotrichum gloeosporioides and leads to large-scale losses both in quality and yield. Little is known regarding the mechanisms underlying the genetics in the strawberry-C. gloeosporioides interaction. In the current research, a wild accession 'Fragaria nilgerrensis' is used as a resistant model to study the roles of terpenoid and terpene genes in leaf response to C. gloeosporioides. We found that several terpenoids and terpene genes were up-regulated at early time points after challenged with C. gloeosporioides. Among the metabolites detected, sesquiterpenes were the most significantly accumulated compounds, increasing up to ~12-fold at 18 h post infection (hpi), followed by monoterpenes which showed a slight increase upon infection. Consistently, the time-resolved transcriptome data revealed that genes pertaining to terpenoid metabolism were rapidly up-regulated and co-expressed with signaling pathway genes relevant to defense response. Notably, quantitative real-time PCR confirmed that the expression of five terpene synthase genes (TPS) were greatly enhanced, by a factor of one to three orders of magnitude at 3-6 hpi. Our results reveal a possible link between rapidly induced terpenoid metabolism and the autoimmunity underlying anthracnose resistance in a wild strawberry species.

Dynamic changes in monoterpene accumulation and biosynthesis during grape ripening in three Vitis vinifera L. cultivars.

Monoterpenes are important aroma components in grapes and wines. We analyzed the free and bound monoterpene profiles and the transcript levels of terpenoid biosynthesis genes in Vitis Vinifera cvs. Muscat Hamburg, Riesling, and Sauvignon Blanc grapes at five ripening stages. Principal component analyses revealed that the three cultivars had different free monoterpene profiles at harvest and the early stage of ripening. In all cultivars, the total bound monoterpene contents were higher than the free monoterpene contents during grape ripening. The changes in monoterpene profiles in different grape varieties were correlated with the transcript levels of some VviTPS and VviGT genes. In Riesling, the VviGT14 and VviUGT88A1L1 transcript levels were related to geraniol glucoside accumulation. In Muscat Hamburg, the VviPNLGl1, VviPNLGl2, and VviPNLGl4 transcript levels were related to linalool accumulation. Understanding the dynamic changes in monoterpene accumulation and biosynthesis will allow winemakers to devise strategies to improve grape and wine aromas.

Effect of cluster zone leaf removal on monoterpene profiles of Sauvignon Blanc grapes and wines.

Monoterpenes contribute to the varietal aromas of grapes and wines. We determined the effects of cluster zone leaf removal on the monoterpene profiles of Sauvignon Blanc grape berries and wines, and on the expression of key genes in the terpenoid pathway. Leaf removal at two intensities (half basic, 50%; full basic, 100%) was conducted at two weeks before veraison, veraison, and two weeks after veraison. Half basic leaf removal increased the pH and decreased the tartaric acid content in grapes and wines. The concentrations of most free- and bound-form monoterpenes in grapes were increased by early leaf removal. The total monoterpene contents were increased in wines in the defoliation treatments, but were significantly lower in wines from the full basic leaf removal treatments than in wines from the half basic leaf removal treatments. The defoliation treatments resulted in increased transcript levels of some key genes in terpene biosynthesis (VvPNLinNer1, VvPNLinNer2, VvPNLNGl1, VvPNLNGl2, and VvUGT88A1L1).

Effect of methyl jasmonate on the aroma of Sangiovese grapes and wines.

Methyl jasmonate (MeJA) was applied in a vineyard on leaves and grape clusters of cv Sangiovese to test its ability to stimulate the production of aromas and identify the main genes involved in the biosynthetic pathways switched on by the elicitor. MeJA application led to a delay in grape technological maturity and a significant increase in the concentration of several berry aroma classes (about twice the total aroma: from around 3 to 6µg/g of berry). Of these, monoterpenes showed the most significant increase. An analysis of the expression of terpenoid biosynthesis genes confirmed that the MeJA application activated the related biosynthetic pathway. The expression of all the TPS genes analyzedwas higher in samples treated with MeJA. Also the wines produced by microvinification of Sangiovese treated and untreated grapes showed a rise in the aroma concentration as in berries, with an important impact on longevity and sensorial characters of wines.

Expression of terpene synthase genes associated with the formation of volatiles in different organs of Vitis vinifera.

Plants produce a plethora of volatile organic compounds (VOCs) which are important in determining the quality and nutraceutical properties of horticultural food products, including the taste and aroma of wine. Given that some of the most prevalent grape aroma constituents are terpenoids, we investigated the possible variations in the relative expression of terpene synthase (TPS) genes that depend on the organ. We thus analysed mature leaves, young leaves, stems, young stems, roots, rachis, tendrils, peduncles, bud flowers, flowers and berries of cv Moscato bianco in terms of their VOC content and the expression of 23 TPS genes. In terms of the volatile characterization of the organs by SPME/GC-MS analysis, flower buds and open flowers appeared to be clearly distinct from all the other organs analysed in terms of their high VOC concentration. Qualitatively detected VOCs clearly separated all the vegetative organs from flowers and berries, then the roots and rachis from other vegetative organs and flowers from berries, which confirms the specialization in volatile production among different organs. Our real-time RT-PCR results revealed that the majority of TPS genes analysed exhibited detectable transcripts in all the organs investigated, while only some were found to be expressed specifically in one or just a few organs. In most cases, we found that the known products of the in vitro assay of VvTPS enzymes corresponded well to the terpenes found in the organs in which the encoding gene was expressed, as in the case of (E)-ß-caryophyllene synthases, α-terpineol synthase and α-farnesene synthase. In addition, we found groups of homologous TPS genes, such as (E)-ß-caryophyllene and ß-ocimene synthases, expressed distinctively in the various tissues. This thus confirmed the subfunctionalization events and a specialization on the basis of the organs in which they are mostly expressed.