The vital roles of R2R3-MYB transcription factors (TFs) in regulating stress response and phytohormone signaling have been thoroughly studied in numerous plant species, but the functions of these TFs in rubber tree are poorly understood. Rubber tree is the most important source of natural rubber but often suffers from various abiotic and biotic stresses that cause severe yield losses each year. In this study, we reported a novel MYB44 gene in rubber tree (named HbMYB44) and revealed its biological function. HbMYB44 was highly similar to AtMYB44 and clustered into subgroup 22. Transient expression indicated that HbMYB44 is a nuclear localized protein and displays transactivation activity at the C-terminus. HbMYB44 was ubiquitously expressed in rubber tree, and its expression was strongly induced by multiple phytohormones, drought stress, wounding, and H₂O₂ treatments. Furthermore, overexpression of HbMYB44 in Arabidopsis (OE) demonstrated that OE plants significantly enhanced stress tolerance, i.e., salt stress, osmotic stress, and drought stress. Additionally, HbMYB44 promoted recovery from root growth inhibition of OE plants caused by exogenous phytohormones (including abscisic acid, methyl jasmonic acid, gibberellic acid 3, and salicylic acid), but the opposite effect was present in response to ethephon. Interestingly, HbMYB44 increased the expression of its homologous genes and interacting protein-encoding genes in OE plants. Overall, HbMYB44 plays versatile functions in modulating multiple phytohormone signaling pathways and stress tolerance.

Powdery mildew (Oidium heveae Steinm.) is one of the most important leaf diseases in rubber tree (Hevea brasiliensis). However, physiological and molecular processes associated with the Hevea-powdery mildew interaction are still unclear. In this study, effects of powdery mildew infection on the mitochondrial and chloroplast functions in rubber tree were comprehensively investigated. Powdery mildew damaged the structure and function of mitochondria prior to chloroplasts, causing inner and outer membranes disruption. The intact rate of mitochondria membrane was reduced from 70% in control leaves to 23.1% in the leaves at 5 days after inoculation (dai). Significant decreases in the activities of cytochrome c oxidase, NADH oxidation and malate dehydrogenase (MDH) were observed in the powdery mildew-infected leaves. Tricarboxylic acid cycle (TCA) and electron transfer capacity were seriously impaired after powdery mildew invasion. Chlorophyll (Chl) contents, maximal photochemical efficiency (Fv/Fm), actual photochemical efficiency of photosystem II (ΦPSII), and electron transport rate (ETR) were dramatically decreased in the infected leaves from 10 dai. Our results provided new insights into understanding the mechanism of Hevea-powdery mildew interaction in rubber tree.

Light is one of most important factors to plants because it is necessary for photosynthesis. In this study, physiological and gene expression analyses under different light intensities were performed in the seedlings of rubber tree (Hevea brasiliensis) clone GT1. When light intensity increased from 20 to 1000 µmol m⁻² s⁻¹, there was no effect on the maximal quantum yield of photosystem II (PSII) photochemistry (Fv/Fm), indicating that high light intensity did not damage the structure and function of PSII reaction center. However, the effective photochemical quantum yield of PSII (Y(II)), photochemical quenching coefficient (qP), electron transfer rate (ETR), and coefficient of photochemical fluorescence quenching assuming interconnected PSII antennae (qL) were increased significantly as the light intensity increased, reached a maximum at 200 µmol m⁻² s⁻¹, but decreased from 400 µmol m⁻² s⁻¹. These results suggested that the PSII photochemistry showed an optimum performance at 200 µmol m⁻² s⁻¹ light intensity. The chlorophyll content was increased along with the increase of light intensity when it was no more than 400 µmol m⁻² s⁻¹. Since increasing light intensity caused significant increase in H₂O₂ content and decreases in the per unit activity of antioxidant enzymes SOD and POD, but the malondialdehyde (MDA) content was preserved at a low level even under high light intensity of 1000 µmol m⁻² s⁻¹, suggesting that high light irradiation did not induce membrane lipid peroxidation in rubber tree. Moreover, expressions of antioxidant-related genes were significantly up-regulated with the increase of light intensity. They reached the maximum expression at 400 µmol m⁻² s⁻¹, but decreased at 1000 µmol m⁻² s⁻¹. In conclusion, rubber tree could endure strong light irradiation via a specific mechanism. Adaptation to high light intensity is a complex process by regulating antioxidant enzymes activities, chloroplast formation, and related genes expressions in rubber tree.

The rubber tree (Hevea brasiliensis) is a widely cultivated crop in tropical acidic soil that is tolerant to high concentration of aluminum and the aluminum-activated malate transporter (ALMT) plays an important role in plant aluminum detoxification. However, the effects of ALMT on rubber tree aluminum tolerance, growth performance, and latex production are unclear. In this study, 17 HbALMT genes were identified from the genome of rubber trees. The physiological and biochemical characteristics, phylogenetic relationships, gene structures, conserved motifs, cis-elements of promoter, and expression patterns of the identified HbALMT genes were studied. Phylogenetic relationships indicated that these genes were divided into four clusters and genes in the same cluster have similar gene structures and conserved motifs. The promoters of HbALMT genes contain many cis-elements associated with biotic stress and abiotic stress. Quantitative real-time PCR analysis revealed HbALMTs showed various expression patterns in different tissues, indicating the functional diversity of HbALMT genes in different tissues of rubber trees. Transcriptome analysis and qRT-PCR assay showed that most of the HbALMT genes responded to aluminum stress, and among the 17 HbALMTs, HbALMT1, HbALMT2, HbALMT13, and HbALMT15 displayed higher expression levels in roots after two or five days of Al treatments, indicating their potential involvement in aluminum detoxification. Taken together, this study laid a foundation for further understanding the molecular evolution of the ALMT genes and their involvement in rubber tree aluminum adaption.

Mildew resistance locus O (Mlo) gene was first found in barley as a powdery mildew susceptibility gene, and recessive mlo alleles confer durable resistance to barley powdery mildew. To identify candidate Mlo susceptibility genes in rubber tree, HbMlo12 was cloned from rubber tree clone CATAS7-33-97, which is susceptible to powdery mildew. Protein architecture analysis showed that HbMlo12 was a typical Mlo protein with seven transmembrane domains. Protein blast search in the Arabidopsis thaliana proteome database showed that HbMlo12 shared the highest similarity with AtMlo12, with 63% sequence identity. Furthermore, HbMlo12 together with the dicot powdery mildew susceptible Mlo proteins (including AtMlo2, AtMlo6, AtMlo12, tomato SlMlo1, pepper CaMlo2, pea PsMlo1, etc.) were grouped into clade V. Subcellular localization analysis in tobacco epidermal cells revealed that HbMlo12 was localized to the endoplasmic reticulum membrane. HbMlo12 was preferentially expressed in the flower and leaf of rubber tree. Moreover, its expression was significantly upregulated in response to powdery mildew inoculation. Application of exogenous ethephon caused a distinct increase in HbMlo12 expression. Additionally, HbMlo12 transcript was quickly induced by spraying salicylic acid and gibberellic acid and reached the maximum at 0.5 h after treatments. By contrast, HbMlo12 expression was downregulated by methyl jasmonate, abscisic acid, and drought stress treatments. There was no significant change in HbMlo12 expression after indole-3-acetic acid, H₂O₂, and wounding stimuli. Taken together, these results suggested that HbMlo12 might be a candidate Mlo gene conferring susceptibility to powdery mildew in rubber tree. The results of this study are vital in understanding Mlo gene evolution and developing new rubber tree varieties with powdery mildew resistance using reverse genetics.