Search Results (32)

Pinus massoniana Lamb. is an economically important conifer native to China. However, it is highly susceptible to the pine wood nematode (Bursaphelenchus xylophilus, PWN), the causal agent of pine wilt disease (PWD), resulting in substantial ecological and economic losses. To elucidate potential molecular defense mechanisms, 50 NAC (NAM, ATAF1/2, and CUC2) transcription factors (PmNACs) were identified in the P. massoniana genome. Phylogenetic analysis divided these PmNACs into seven subfamilies, and motif analysis identified ten conserved motifs associated with stress responses. Twenty-three genes were selected for expression analysis in various tissues and under exogenous salicylic acid (SA), methyl jasmonate (MeJA), and PWN infection. Six genes (PmNAC1, PmNAC8, PmNAC9, PmNAC17, PmNAC18, and PmNAC20) were significantly up-regulated by both hormonal treatment and PWN infection, implying their involvement in JA/SA-mediated immune pathways. Functional characterization showed PmNAC8 is a nuclear-localized transcription factor with autoactivation activity. Furthermore, transient overexpression of PmNAC8 in Nicotiana benthamiana induced reactive oxygen species (ROS) accumulation and necrotic lesions. Collectively, these results elucidate NAC-mediated defense responses to PWN infection in P. massoniana and identify candidate genes for developing PWD-resistant pine varieties. Full article

The endemic Patagonian conifer, Austrocedrus chilensis, is threatened by the pathogen Phytophthora austrocedri. This study presents the first histological and histochemical analysis of A. chilensis affected by this pathogen. We examined the stem tissues of naturally infected adult trees (over 30 years old) and artificially inoculated saplings (8–12 years old) to identify the pathogen’s colonization strategies and the tree’s histological responses. Using light and scanning electronic microscopy along with several histochemical techniques (Lugol, toluidine blue, vanillin-HCl, Phloroglucinol, Calcofluor white, and aniline blue), we found that P. austrocedri can grow in all active tissues, leading to cambium and parenchyma necrosis. The pathogen spreads through sieve cells and tracheids, moving to the adjacent cells via sieve plates and bordered pits and colonizing nearby parenchyma cells. We observed loss of starch in necrotic tissues. In contrast, starch accumulation and an increase in the number of polyphenolic cells occur in the healthy areas adjacent to the margins of the lesion, indicating a tree’s induced defense mechanisms. The tree’s responses include cambium reprogramming, which leads to the formation of traumatic resin ducts, alterations in cell shape and size, and the deposition of phenolic compounds. We analyze the tree responses and discuss their potential relationship with a methyl jasmonate-induced defense and a hypersensitive-like response. Full article

Most fir species in the Mediterranean have small to medium-sized distributions, are often endemic and endangered, and are mainly found in relict areas, except for Abies alba. The IUCN Red List of Threatened Species identified Abies nebrodensis as the rarest conifer in the world, with only 30 adult trees remaining. Additionally, Abies pinsapo is threatened and limited to five fragmented locations in Spain and Morocco. This study aimed to characterize the seed terpene profiles of Mediterranean Abies species, such as A. nebrodensis, A. pinsapo, and A. alba, since morphological results showed minimal variation among the Abies populations examined. Terpenes were extracted using n-heptane and then analyzed by GC-MS. The chemical composition revealed the dominance of limonene and α-pinene as the main monoterpenes in all the species, while A. nebrodensis reported the considerable presence of germacrene D-4-ol and selina-6-en-4-ol as sesquiterpenes. The relative contents of most of the terpenes were significantly different among the species, and subsequent statistical multivariate analysis showed clear discrimination among three distinct groups. These results confirmed the suitability of the terpene profile as a potential tool to study chemotaxonomic differences between species from the same family. Moreover, the compounds identified can be interesting for further studies on plant defense against biotic stress to reduce the risk of species extinction caused by pests and diseases. Full article

Terpenoids, abundant and structurally diverse secondary metabolites in plants, especially in conifer species, play crucial roles in the plant defense mechanism and plant growth and development. In Pinus massoniana, terpenoids’ biosynthesis relies on both the mevalonate (MVA) pathway and the 2-methyl-D-erythritol-4-phosphate (MEP) pathway, with 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase (HDS) catalyzing the sixth step of the MEP pathway. In this study, we cloned and conducted bioinformatics analysis of the PmHDS gene from P. massoniana. The results showed that PmHDS shares homology with HDS proteins from other species. Analysis of tissue expression patterns indicated that PmHDS exhibits the highest expression level in xylem tissue, followed by stems, with significantly lowest expression in the apical meristem. Treatment with NaCl, abscisic acid (ABA), ethylene (ETH), methyl jasmonate (MeJA), and salicylic acid (SA) upregulated the expression of PmHDS. Furthermore, we successfully cloned the PmHDS promoter (about 2220 bp) and integrated it into a GUS reporter vector, which resulted in GUS activity being observed in various tissues of Arabidopsis thaliana. Overexpression of the PmHDS gene in A. thaliana significantly increased the content of carotenoids, chlorophylls a and b, and related enzyme activities, as well as the levels of terpenoid derivatives such as cytokinin (CTK), gibberellic acid (GA), and ABA, thereby enhancing the resistance to those abiotic stresses. These findings suggest that PmHDS plays an important role in the terpenoid synthesis pathway. This study provides a theoretical basis for understanding the biosynthesis of terpenoids and lays a foundation for future research on the regulation of terpene synthesis and resistance in molecular breeding. Full article

Background/objectives: Methyl jasmonate is a plant signaling molecule involved in a wide range of functions, including stress responses. This study investigates the relative differential expression of microRNAs and their target genes in response to methyl jasmonate treatment of Scots pine needles. Methods: A combined strategy of high-throughput sequencing and in silico prediction of potential target genes was implemented. Results: a total of 58 differentially expressed (DE) microRNAs (miRNAs) (43 up-regulated and 15 down-regulated), belonging to 29 miRNA families, were identified. The 41 DE miRNAs from 17 families were conifer-specific miRNA families—miR946, miR947, miR950, miR1312, miR1313, miR1314, miR3693, miR3107, miR11452, miR11466, miR11487, miR11490, miR11504, miR11511, miR11532, miR11544, and miR11551. The other DE miRNAs (miR159, miR164, miR169, miR396, miR397, miR398, miR408, miR535) were conserved miRNAs, which are also found in angiosperm species. Transcriptome analysis identified 389 gene transcripts with 562 miRNA-target sites targeted by 57 of the 58 DE miRNAs. Of these, 250 target genes with 138 different GO annotations were found for the 41 DE conifer-specific conserved miRNAs. Conclusions: The 26 DE miRNAs from 14 DE miRNA families, of which almost all (12 families, 24 miRNAs) are conifer specific, and were associated with 68 disease resistance and TMV resistance proteins, TIR-NBS-LRR, LRR receptor-like serine/threonine-protein kinase, putative CC-NBS-LRR protein, and putative NBS-LRR protein target transcripts with 29 target gene GO term descriptions. Some of the genes targeted by conifer-specific miRNAs have been previously reported to be targeted by other miRNAs in angiosperms, indicating that the miRNA-target gene regulation system can vary between species. Full article

The study was carried out to assess the effect of the larch wooly adelgid Adelges laricis Vallot (Hemiptera: Adelgidae) infestation on its secondary host, the European larch Larix decidua Mill. Morphology and anatomy of adelgid-infested needles, and content of defense phenolic compounds including individual flavonoids isorhamnetin, kaempferol, quercetin, rutin, catechin, epicatechin, apigenin, ampelopsin and taxifolin, were analyzed. The amount of total phenols in needles from adelgid-infested twigs of L. decidua increased following the development of the A. laricis population, from the end of April until the end of June. The most abundant among flavonoids were the flavanols, mainly catechin and epicatechin, which predominated in the larch needles during the whole period of adelgid infestation. The content of catechin and epicatechin increased following the increase in the adelgid population number. An increase in content occurred also in flavanonols ampelopsin and taxifolin, while the content of the flavonol kaempferol decreased as the population number of A. laricis increased. The analysis of the anatomical structure of needles showed changes in the shape of the needles, the presence of a thicker layer of epicuticular waxes, and a higher number of mesophyll layers as a result of adelgid feeding. Full article

In the context of climate change, methods to improve the resistance of coniferous trees to biotic and abiotic stress are in great demand. The common plant response to exposure to vastly different stressors is the generation of reactive oxygen species (ROS) followed by activation of the defensive antioxidant system. We aimed to evaluate whether seed treatment with physical stressors can activate the activity of antioxidant enzymes and radical scavenging activity in young Picea abies (L.) H. Karst seedlings. For this, we applied seed treatment with cold plasma (CP) and electromagnetic field (EMF) and compared the response in ten different half-sib families of Norway spruce. The impact of the treatments with CP (1 min—CP1; 2 min—CP2) and EMF (2 min) on one-year-old and two-year-old P. abies seedlings was determined by the emergence rate, parameters of growth, and spectrophotometric assessment of antioxidant capacity (enzyme activity; DPPH and ABTS scavenging) in needles. The results indicated that the impact of seed treatment is strongly dependent on the genetic family. In the 577 half-sib family, the activity of antioxidant enzymes catalase (CAT), ascorbate peroxidase (APX), peroxidase (POX), and glutathione reductase (GR) increased after EMF-treatment in one-year-old seedlings, while similar effects in 477 half-sib family were induced by CP2 treatment. In two-year-old seedlings, CP1-treatment increased CAT, APX, POX, GR, SOD, DPPH, and ABTS activity in the 457 half-sib family. However, no significant impact of the treatment with CP1 was determined in one-year-old seedlings in this family. The application of novel technologies and the consideration of the combinatory impact of genetic and physical factors could have the potential to improve the accumulation of compounds that play an essential role in the defense mechanisms of P. abies. Nevertheless, for different resistance and responses to stressors of plants, their genetic properties play an essential role. A comprehensive analysis of interactions among the stress factors (CP and EMF), genetic properties, and changes induced in the antioxidant system can be of importance both for the practical application of seed treatment in forestry and for understanding fundamental adaptation mechanisms in conifers. Full article

Climate change-induced hazards, such as drought, threaten forest resilience, particularly in vulnerable regions such as the Mediterranean Basin. Maritime pine (Pinus pinaster Aiton), a model species in Western Europe, plays a crucial role in the Mediterranean forest due to its genetic diversity and ecological plasticity. This study characterizes transcriptional profiles of scion and rootstock stems of four P. pinaster graft combinations grown under well-watered conditions. Our grafting scheme combined drought-sensitive and drought-tolerant genotypes for scions (GAL1056: drought-sensitive scion; and Oria6: drought-tolerant scion) and rootstocks (R1S: drought-sensitive rootstock; and R18T: drought-tolerant rootstock). Transcriptomic analysis revealed expression patterns shaped by genotype provenance and graft combination. The accumulation of differentially expressed genes (DEGs) encoding proteins, involved in defense mechanisms and pathogen recognition, was higher in drought-sensitive scion stems and also increased when grafted onto drought-sensitive rootstocks. DEGs involved in drought tolerance mechanisms were identified in drought-tolerant genotypes as well as in drought-sensitive scions grafted onto drought-tolerant rootstocks, suggesting their establishment prior to drought. These mechanisms were associated with ABA metabolism and signaling. They were also involved in the activation of the ROS-scavenging pathways, which included the regulation of flavonoid and terpenoid metabolisms. Our results reveal DEGs potentially associated with the conifer response to drought and point out differences in drought tolerance strategies. These findings suggest genetic trade-offs between pine growth and defense, which could be relevant in selecting more drought-tolerant Pinus pinaster trees. Full article

As one of the largest and most diverse classes of specialized metabolites in plants, terpenoids (oprenoid compounds, a type of bio-based material) are widely used in the fields of medicine and light chemical products. They are the most important secondary metabolites in coniferous species and play an important role in the defense system of conifers. Terpene synthesis can be promoted by regulating the expressions of terpene synthase genes, and the terpene biosynthesis pathway has basically been clarified in Pinus massoniana, in which there are multiple rate-limiting enzymes and the rate-limiting steps are difficult to determine, so the terpene synthase gene regulation mechanism has become a hot spot in research. Herein, we amplified a PmDXR gene (GenBank accession no. MK969119.1) of the MEP pathway (methyl-erythritol 4-phosphate) from Pinus massoniana. The DXR enzyme activity and chlorophyll a, chlorophyll b and carotenoid contents of overexpressed Arabidopsis showed positive regulation. The PmDXR gene promoter was a tissue-specific promoter and can respond to ABA, MeJA and GA stresses to drive the expression of the GUS reporter gene in N. benthamiana. The DXR enzyme was identified as a key rate-limiting enzyme in the MEP pathway and an effective target for terpene synthesis regulation in coniferous species, which can further lay the theoretical foundation for the molecularly assisted selection of high-yielding lipid germplasm of P. massoniana, as well as provide help in the pathogenesis of pine wood nematode disease. Full article

The 19 species of Dendroctonus bark beetles native to North America include several of the most significant biotic mortality agents of conifers in this region, and they have impacted millions of hectares of forest in recent decades. Semiochemical attractants play several vital roles in the biology of these insects including host finding, host assessment, and mate finding. They are key to the virulence of these insects, since they mediate the mass attacks that enable killing and reproduction within healthy trees. These attractants consist of combinations of thirteen identified pheromone components in three primary chemical classes and at least ten host-associated compounds dominated by monoterpene hydrocarbons in host defensive resin. Due to their potential for use in pest management technologies, semiochemical attractants for Dendroctonus have received extensive research into their chemical compositions and effects on insect behavior. This paper is intended as a synthesis of this research over the past 60 years as well as a critical examination of approaches to investigation of this topic and interpretation of experimental results. Its purpose is to assist practitioners in formulating suitable attractive lures for use in applications and identify gaps in knowledge of the semiochemistry of Dendroctonus that should be addressed if the practical potential of attractants is to be fully realized. Full article

Conifers have evolved sophisticated terpenoid defenses for protection against herbivores and pathogens. Pinus massoniana Lamb. is the most widely distributed pioneer afforestation and resin tree species in China, but is seriously harmed by pine wilt disease. Monochamus alternatus is the main vector of pine wilt disease in China. Monoterpenes, sesquiterpenes and diterpenes, the main secondary defensive compounds of P. massoniana, are catalyzed by different terpene synthases (TPSs), which participate in the important defense pathways against external biotic and abiotic stresses. Here, we aimed to identify the terpene synthases (TPSs) in P. massoniana, responding to the feeding of M. alternatus, and to characterize the functions and products of the mono-TPSs. We identified six differentially expressed TPS genes in the P. massoniana fed upon by M. alternatus, including four mono-TPS and two sesqui-TPS genes. The functions of the four mono-TPSs were verified by analysis of the main product and by-products of these mono-TPSs. (+)-α-Pinene, (−)-α-pinene, and limonene were the major products of TPS (+)-α-pinene, TPS (−)-α-pinene, and TPS limonene, respectively, but TPS (−)-β-pinene only catalyzed a trace amount of (−)-β-pinene in the products. Our findings shed light on the potential relationships between the structure of terpene synthases and their corresponding products. Full article

Due to climate warming, the occurrence of Lymantria monacha outbreaks is predicted to become more frequent, causing repeated and severe damage to conifer trees. Currently, the most effective way to control the outbreaks is aerial spraying with the bioinsecticide Foray 76B. The present study aimed to determine the impact of both: (i) L. monacha outbreaks and (ii) treatment with Foray 76B on tree resistance through the synthesis of polyphenols (TPC), flavonoids (TFC), photosynthetic pigments (chlorophyll a and b, carotenoids), lipid peroxidation (MDA), and soluble sugars (TSS) in Pinus sylvestris needles. Samples were collected from visually healthy (control), damaged/untreated, and damaged/Foray 76B-treated plots in 2020 and 2021 (following year after the outbreaks). The results revealed that L. monacha outbreaks contributed to the increase in TPC by 34.1% in 2020 and 26.7% in 2021. TFC negatively correlated with TPC, resulting in 17.6% and 11.1% lower concentrations in L. monacha-damaged plots in 2020 and 2021, respectively. A decrease in MDA was found in the damaged plots in both 2020 and 2021 (10.2% and 23.3%, respectively), which was associated with the increased synthesis of photosynthetic pigments in 2021. The research results also showed that in the following year after the outbreaks, the increase in the synthesis of photosynthetic pigments was also affected by the treatment with Foray 76B. Moreover, the increase in the synthesis of TPC and photosynthetic pigments in the damaged plots in 2021 illustrates the ability of pines to keep an activated defense system to fight biotic stress. Meanwhile, a higher synthesis of photosynthetic pigments in Foray 76B-treated plots indicates a possible effect of the treatment on faster tree growth and forest recovery after L. monacha outbreaks. Full article

Alpha-pinene is one of the main defensive components in conifers. Monochamus alternatus (Coleoptera: Cerambycidae), a wood borer feeding on Pinaceae plants, relies on its detoxifying enzymes to resist the defensive terpenoids. Here, we assayed the peroxide level and GST activity of M. alternatus larvae treated with different concentrations of α-pinene. Meanwhile, a gst gene (MaGSTe3) was isolated and analyzed. We determined its expression level and verified its function. The results showed that α-pinene treatment led to membrane lipid peroxidation and thus increased the GST activity. Expression of MaGSTe3 was significantly upregulated in guts following exposure to α-pinene, which has a similar pattern with the malonaldehyde level. In vitro expression and disk diffusion assay showed that the MaGSTe3 protein had high antioxidant capacity. However, RNAi treatment of MaGSTe3 did not reduce the hydrogen peroxide and malonaldehyde levels, while GST activity was significantly reduced. These results suggested MaGSTe3 takes part in α-pinene adaptation, but it does not play a great role in the resistance of M. alternatus larvae to α-pinene. Full article

Pine wilt disease (PWD) is a devastating disease that threatens pine forests worldwide, and breeding resistant pines is an important management strategy used to reduce its impact. A batch of resistant seeds of P. thunbergii was introduced from Japan. Based on the resistant materials, we obtained somatic plants through somatic embryogenesis. In this study, we performed transcriptome analysis to further understand the defense response of resistant somatic plants of P. thunbergii to PWD. The results showed that, after pine wood nematode (PWN) infection, resistant P. thunbergii stimulated more differential expression genes (DEGs) and involved more regulatory pathways than did susceptible P. thunbergii. For the first time, the alpha-linolenic acid metabolism and linoleic acid metabolism were intensively observed in pines resisting PWN infection. The related genes disease resistance protein RPS2 (SUMM2) and pathogenesis-related genes (PR1), as well as reactive oxygen species (ROS)-related genes were significantly up-expressed in order to contribute to protection against PWN inoculation in P. thunbergii. In addition, the diterpenoid biosynthesis pathway was significantly enriched only in resistant P. thunbergii. These findings provided valuable genetic information for future breeding of resistant conifers, and could contribute to the development of new diagnostic tools for early screening of resistant pine seedlings based on specific PWN-tolerance-related markers. Full article

Trees are unique in terms of development, sustainability and longevity. Some species have a record lifespan in the living world, reaching several millennia. The aim of this review is to summarize the available data on the genetic and epigenetic mechanisms of longevity in forest trees. In this review, we have focused on the genetic aspects of longevity of a few well-studied forest tree species, such as Quercus robur, Ginkgo biloba, Ficus benghalensis and F. religiosa, Populus, Welwitschia and Dracaena, as well as on interspecific genetic traits associated with plant longevity. A key trait associated with plant longevity is the enhanced immune defense, with the increase in gene families such as RLK, RLP and NLR in Quercus robur, the expansion of the CC-NBS-LRR disease resistance families in Ficus species and the steady expression of R-genes in Ginkgo biloba. A high copy number ratio of the PARP1 family genes involved in DNA repair and defense response was found in Pseudotsuga menziesii, Pinus sylvestris and Malus domestica. An increase in the number of copies of the epigenetic regulators BRU1/TSK/MGO3 (maintenance of meristems and genome integrity) and SDE3 (antiviral protection) was also found in long-lived trees. CHG methylation gradually declines in the DAL 1 gene in Pinus tabuliformis, a conservative age biomarker in conifers, as the age increases. It was shown in Larix kaempferi that grafting, cutting and pruning change the expression of age-related genes and rejuvenate plants. Thus, the main genetic and epigenetic mechanisms of longevity in forest trees were considered, among which there are both general and individual processes. Full article