Rubber tree (Hevea brasiliensis) is reproduced by bud grafting for commercial planting, but significant intraclonal variations exist in bud-grafted clones. DNA methylation changes related to grafting may be partly responsible for intraclonal variations. In the current study, whole-genome DNA methylation profiles of grafted rubber tree plants (GPs) and their donor plants (DPs) were evaluated by whole-genome bisulfite sequencing. Data showed that DNA methylation was downregulated and DNA methylations in CG, CHG, and CHH sequences were reprogrammed in GPs, suggesting that grafting induced the reprogramming of DNA methylation. A total of 5,939 differentially methylated genes (DMGs) were identified by comparing fractional methylation levels between GPs and DPs. Transcriptional analysis revealed that there were 9,798 differentially expressed genes (DEGs) in the DP and GP comparison. A total of 1,698 overlapping genes between DEGs and DMGs were identified. These overlapping genes were markedly enriched in the metabolic pathway and biosynthesis of secondary metabolites by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Global DNA methylation and transcriptional analyses revealed that reprogramming of DNA methylation is correlated with gene expression in grafted rubber trees. The study provides a whole-genome methylome of rubber trees and an insight into the molecular mechanisms underlying the intraclonal variations existing in the commercial planting of grafted rubber trees.

Natural rubber (NR) is utilized in more than 40,000 products, and the demand for NR is projected to reach $68.5 billion by 2026. The primary commercial source of NR is the latex of Hevea brasiliensis. NR is produced by the sequential cis-condensation of isopentenyl diphosphate (IPP) through a complex known as the rubber transferase (RTase) complex. This complex is associated with rubber particles, specialized organelles for NR synthesis. Despite numerous attempts to isolate, characterize, and study the RTase complex, definitive results have not yet been achieved. This review proposes an innovative approach to overcome this longstanding challenge. The suggested method involves isolating the RTase complex without using detergents, instead utilizing the native membrane lipids, referred to as \"natural nanodiscs\", and subsequently reconstituting the complex on liposomes. Additionally, we recommend the adaptation of large nanodiscs for the incorporation and reconstitution of the RTase complex, whether it is in vitro transcribed or present within the natural nanodiscs. These techniques show promise as a viable solution to the current obstacles. Based on our experimental experience and insights from published literature, we believe these refined methodologies can significantly enhance our understanding of the RTase complex and its role in in vitro NR synthesis.

Natural rubber (NR) latex derived from Hevea brasiliensis is a complex colloid comprising mainly rubber hydrocarbons (latex particles) and a multitude of minor non-rubber constituents such as non-rubber particles, proteins, lipids, carbohydrates, and soluble organic and inorganic substances. NR latex is susceptible to enzymatic attack after it leaves the trees. It is usually preserved with ammonia and, to a lesser extent, with other preservatives to enhance its colloidal stability during storage. Despite numerous studies in the literature on the influence of rubber proteins on NR latex stability, issues regarding the effect of protein hydrolysis in the presence of ammonia on latex stability during storage are still far from resolved. The present work aims to elucidate the interplay between protein hydrolysis and ammoniation in NR latex stability. Both high- and low-ammonia (with a secondary preservative) NR latexes were used to monitor the changes in their protein compositions during storage. High-ammonia (FNR-A) latex preserved with 0.6% (v/v) ammonia, a low 0.1% ammonia/TMTD/ZnO (FNR-TZ) latex, and a deproteinized NR (PDNR) latex were labeled with fluorescence agents and observed using confocal laser scanning microscopy to determine their protein composition. Protein hydrolysis was confirmed via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results revealed that protein hydrolysis increased with the storage duration. The change in protein composition accompanying hydrolysis also allows the spatial distribution of allergenic proteins to be estimated in the latex. Concurrently, the latex stability increased with the storage duration, as measured by the latex\'s mechanical stability time (MST) and the zeta potential of the latex particles. As monitored by AFM, the surface roughness of the NR latex film increased markedly during extended storage compared with that of the DPNR latex, which remained smooth. These results underscore the pivotal role of ammonia in bolstering NR latex stability brought on by protein hydrolysis, which greatly impacts latex film\'s formation behavior. NR latex stability underpins the quality of latex-dipped goods during manufacturing, particularly those for medical gloves.

Tapping panel dryness (TPD) results in a severe reduction in latex yield in Hevea brasiliensis. However, the molecular regulatory mechanisms of TPD occurrence are still largely unclear. In this study, whole-transcriptome sequencing was carried out on latex from TPD and healthy trees. In total, 7078 long noncoding RNAs (lncRNAs), 3077 circular RNAs (circRNAs), 4956 miRNAs, and 25041 mRNAs were identified in latex, among which 435 lncRNAs, 68 circRNAs, 320 miRNAs, and 1574 mRNAs were differentially expressed in the latex of TPD trees. GO and KEGG analyses indicated that plant hormone signal transduction, MAPK signaling pathway, and ubiquitin-mediated proteolysis were the key pathways associated with TPD onset. Phytohormone profiling revealed significant changes in the contents of 28 hormonal compounds, among which ACC, ABA, IAA, GA, and JA contents were increased, while SA content was reduced in TPD latex, suggesting that hormone homeostasis is disrupted in TPD trees. Furthermore, we constructed a TPD-related competitive endogenous RNA (ceRNA) regulatory network of lncRNA/circRNA-miRNA-mRNA with 561 edges and 434 nodes (188 lncRNAs, 5 circRNAs, 191 miRNAs, and 50 mRNAs) and identified two hub lncRNAs (MSTRG.11908.1 and MSTRG.8791.1) and four hub miRNAs (hbr-miR156, miR156-x, miRf10477-y, and novel-m0452-3p). Notably, the lncRNA-miR156/157-SPL module containing three hubs probably plays a crucial role in TPD onset. The expression of network hubs and the lncRNA-miR156/157-SPL module were further validated by qRT-PCR. Our results reveal the TPD-associated ceRNA regulatory network of lncRNA/circRNA-miRNA-mRNA in latex and lay a foundation for further investigation of molecular regulatory mechanisms for TPD onset in H. brasiliensis.

Cold stress poses significant limitations on the growth, latex yield, and ecological distribution of rubber trees (Hevea brasiliensis). The GSK3-like kinase plays a significant role in helping plants adapt to different biotic and abiotic stresses. However, the functions of GSK3-like kinase BR-INSENSITIVE 2 (BIN2) in Hevea brasiliensis remain elusive. Here, we identified HbBIN2s of Hevea brasiliensis and deciphered their roles in cold stress resistance. The transcript levels of HbBIN2s are upregulated by cold stress. In addition, HbBIN2s are present in both the nucleus and cytoplasm and have the ability to interact with the INDUCER OF CBF EXPRESSION1(HbICE1) transcription factor, a central component in cold signaling. HbBIN2 overexpression in Arabidopsis displays decreased tolerance to chilling stress with a lower survival rate and proline content but a higher level of electrolyte leakage (EL) and malondialdehyde (MDA) than wild type under cold stress. Meanwhile, HbBIN2 transgenic Arabidopsis treated with cold stress exhibits a significant increase in the accumulation of reactive oxygen species (ROS) and a decrease in the activity of antioxidant enzymes. Further investigation reveals that HbBIN2 inhibits the transcriptional activity of HbICE1, thereby attenuating the expression of C-REPEAT BINDING FACTOR (HbCBF1). Consistent with this, overexpression of HbBIN2 represses the expression of CBF pathway cold-regulated genes under cold stress. In conclusion, our findings indicate that HbBIN2 functions as a suppressor of cold stress resistance by modulating HbICE1 transcriptional activity and ROS homeostasis.

UNASSIGNED: La planta Hevea brasiliensis se utiliza ampliamente en la industria como fuente de extracción de caucho, un elemento empleado en diversas áreas comerciales y médicas. Los estudios inmunológicos de esta especie indican que es una fuente alergénica importante, que puede provocar sensibilización y alergia. Se han identificado diferentes componentes alergénicos de esta planta, con diversas propiedades inmunitarias y bioquímicas, y estudiado más de diez tipos diferentes de alérgenos, cada uno con distinta capacidad de inducir síntomas alérgicos. En esta revisión informamos los avances actuales en el estudio de Hevea brasiliensis.
UNASSIGNED: Hevea brasiliensis, a plant species used extensively for rubber extraction, is a common allergenic source that can cause sensitization and allergic reactions. Recent immunological studies have characterized various allergenic components of Hevea brasiliensis that possess diverse immune and biochemical properties. Over ten types of allergens have been identified, each with varying capacities to induce allergic symptoms. This review presents the current advances in the study of this allergenic source.

The cultivation of Hevea brasiliensis, the primary commercial source of natural rubber, is strongly impacted by South American leaf blight (SALB) disease, caused by the fungus Pseudocercospora ulei. Various management strategies have been implemented, including the selection of resistant genotypes and the identification of escape zones. This study evaluated the growth, early yield, and resistance to SALB of nine Colombian elite genotypes from the ECC-100 series and IAN 873 clone (control) in a large-scale clone trial in an area with low SALB pressure in the Colombian Amazon during 2017-2020. Favorable early performance was evident, although there was a significant increase in the severity and sporulation of P. ulei over time, especially in the ECC 35, ECC 60, and IAN 873 genotypes. However, these scores indicate low susceptibility. Genotypes with higher resistance to SALB demonstrated greater growth and early yield compared to more highly susceptible genotypes. The ECC 64, ECC 73, ECC 90, ECC 25, and ECC 29 genotypes were more desirable in low SALB pressure zones due to their higher resistance and early performance. It is important to highlight that this research contributes to the selection of new SALB-resistant Colombian genotypes of H. brasiliensis. However, it is also necessary to evaluate the productivity of these selections in the mature stage and long-term resistance to SALB before recommending and promoting their commercial adoption in the Colombian Amazon.

Tapping panel dryness (TPD) has become the mostimportant limiting factor for increasing natural rubber yield, whereas illuminating the molecular mechanisms underlying TPD is the prerequisite for solving the problem of TPD. However, molecular mechanisms underlying TPD are largely unknown. In this study, healthy and different stages of TPD-affected rubber trees were utilized to analyze TPD for the first time. We found that the changing tendencies of key latex physiological parameters were closely related to TPD occurrence and development. To reveal the molecular mechanisms underlying TPD, we sequenced and compared bark transcriptomes among healthy rubber tree, and TPD-affected ones at initial and advanced stages. In total, 8607 genes were identified as TPD-related genes in contrast to healthy rubber tree. According to gene expression profiles, the five samples were divided into three groups including healthy rubber tree, and TPD-affected rubber tree in the initial and advanced stages, which was consistent with the stages of TPD occurrence and development. Interestingly, only asmall proportionof the TPD-related genes were constantly down- or up-regulated with TPD occurrence and development. The TPD-related genes in KEGG pathways significantly enriched were closely associated with protein metabolism, cell division and differentiation, PCD, stress responses, terpene biosynthesis, and various metabolism processes. Moreover, overexpression of HbAPX2 identified as a TPD-related gene enhanced oxidative stress tolerance in S. cerevisiae. The typical symptoms of TPD, partial or complete dry zone (no latex flow) on tapping panel, might attribute to lower IPP available for rubber biosynthesis, and downregulation of the genes in post-IPP steps of rubber biosynthesis and the genes involved in latex flow. Our results not only provide new insights into molecular mechanisms underlying TPD occurrence and development but also contribute to developing effective measures to control TPD in rubber trees.

BACKGROUND: Aedes albopictus and Aedes aegypti are known for their potential as vectors of dengue (DENV) and chikungunya (CHIKV) viruses. However, entomological surveys are mostly carried out during epidemics. In Gabon where outbreaks of both viruses have occurred, there is no vector control program targeting these arboviruses. Therefore, we assessed the presence of Aedes species along a rural-urban gradient in Lambaréné (Gabon) and its surroundings and determined ecological factors associated to their presence.
METHODS: An entomological survey was conducted in Lambaréné and its surrounding rural areas. Mosquitoes were collected with aspirators around human dwellings, and ecological and environmental data were collected from each study area. Morphological identification keys were used to identify Aedes species. RNA was extracted from pools of female mosquitoes and amplified by RT-qPCR to detect the presence of DENV and CHIKV.
RESULTS: Overall, the most common vector collected was Aedes albopictus (97%, 4236/4367 specimens), followed by Aedes aegypti (3%, 131/4367). Albopictus vectors was more abundant in the rural area (Wilcoxon signed-rank test, Z = 627, P = 0.043) than in the urban area. In the urban area, a higher number of mosquitoes (45%) were recorded in the economic zone (zone 3) than in the historical and administrative zones (zone 1 and 2). In the rural area, the proportions of species numbers were significantly higher along the south rural transect (92%) compared to the north rural transect (Wilcoxon signed-rank test, Z = 43, P ˂ 0.016). We also noted a high abundance of vectors in environments characterized by monocultures of Hevea brasiliensis (Hevea) and Manihot esculenta (cassava) (Kruskal-Wallis H-test, H = 25.7, df = 2, P < 0.001). Finally, no mosquito pools were positive for either DENV or CHIKV.
CONCLUSIONS: Aedes albopictus was the dominant vector across the study sites due to its high invasiveness capacity. This presence re-affirms the potential for local transmission of both DENV and CHIKV, as indicated previously by serological surveys conducted in our study area, even though no transmission was detected during the current study. These findings underscore the need for regular arbovirus surveillance in the study region, with the aim of supporting vector control efforts in the event of outbreaks.

Unreduced megagametophytes via second-division restitution were confirmed through heterozygosity analysis, and four candidate physical centromeres of rubber were located for the first time. The evaluation of maternal heterozygosity restitution (MHR) is vital in identifying the mechanism of 2n gametogenesis and assessing the utilization value of 2n gametes. In this study, three full-sib triploid populations were employed to evaluate the MHR of 2n female gametes of rubber tree clone GT1 and to confirm their genetic derivation. The 2n female gametes of GT1 were derived from second-division restitution (SDR) and transmitted more than half of the parental heterozygosity. In addition, low recombination frequency markers were developed, and four candidate physical centromeres of rubber tree were located for the first time. The confirmation that 2n female gametes of rubber tree clone GT1 are derived from SDR provides insights into the molecular mechanisms of 2n gametogenesis. In addition, the identified centromere location will aid in the development of centromeric markers for the rapid identification of the 2n gametogenesis mechanism.