Brassinolide (BR) is an important plant hormone that regulates the growth and development of plants and the formation of yield. The yield and quality of latex from Hevea brasiliensis are regulated by phytohormones. The understanding of gene network regulation mechanism of latex formation in rubber trees is still very limited. In this research, the rubber tree variety CATAS73397 was selected to analyze the relationship between BR, water deficit resistance, and latex yield. The results showed that BR improves the vitality of rubber trees under water deficit by increasing the rate of photosynthesis, reducing the seepage of osmotic regulatory substances, increasing the synthesis of energy substances, and improving the antioxidant system. Furthermore, BR increased the yield and quality of latex by reducing the plugging index and elevating the lutoid bursting index without decreasing mercaptan, sucrose, and inorganic phosphorus. This was confirmed by an increased expression of genes related to latex flow. RNA-seq analysis further indicated that DEG encoded proteins were enriched in the MAPK signaling pathway, plant hormone signal transduction and sucrose metabolism. Phytohormone content displayed significant differences, in that trans-Zeatin, ethylene, salicylic acid, kinetin, and cytokinin were induced by BR, whereas auxin, abscisic acid, and gibberellin were not. In summary, the current research lays a foundation for comprehending the molecular mechanism of latex formation in rubber trees and explores the potential candidate genes involved in natural rubber biosynthesis to provide useful information for further research in relevant areas.

The plant MYB transcription factor family featured as highly conserved DNA-binding domains consisting of 1 to 4 imperfect repeats (R). Increasing evidence indicates that MYB genes participates in growth, differentiation, metabolism, and biotic and abiotic stress responses. However, the functions of MYB genes in the rubber tree remain to be deeply elucidated, especially R2R3-MYB gene family. In this study, molecular biology, bioinformatics, and qRT-PCR were used to identify and analyze HbR2R3-MYB gene family members in the rubber tree. A total of 132 members of the R2R3-MYB gene family were identified in the rubber tree based on genome-wide level. Most of the HbR2R3-MYBs were mapped to 17 rubber tree chromosomes except four genes. A phylogenetic analysis divided all the HbR2R3-MYBs into 20 subgroups with Arabidopsis thaliana. MEME analysis showed that the protein of HbR2R3-MYBs was characterized by 9 conserved motifs. Twenty-six representative R2R3 HbMYBs from different subgroups were selected for expression profiles analysis and the results revealed that the HbR2R3-MYBs members showed various expression patterns in different tissues, powdery mildew-infected and ethylene treatment, implying the diversity of their functions in rubber trees. These results provide fundamental knowledge for further studying the response of the HbR2R3-MYB family to stress and regulation latex flow in rubber tree.

Brassinosteroids (BRs) are important for plant growth and development, with BRI1 and BAK1 kinases playing an important role in BR signal transduction. Latex from rubber trees is crucial for industry, medicine and defense use. Therefore, it is beneficial to characterize and analyze HbBRI1 and HbBAK1 genes to improve the quality of the resources obtained from Hevea brasiliensis (rubber tree). Based on bioinformatics predictions and rubber tree database, five HbBRI1s with four HbBAK1s were identified and named HbBRI1~HbBRL3 and HbBAK1a~HbBAK1d, respectively, which were clustered in two groups. HbBRI1 genes, except for HbBRL3, exclusively contain introns, which is convenient for responding to external factors, whereas HbBAK1b/c/d contain 10 introns and 11 exons, and HbBAK1a contains eight introns. Multiple sequence analysis showed that HbBRI1s include typical domains of the BRI1 kinase, indicating that HbBRI1s belong to BRI1. HbBAK1s that possess LRR and STK_BAK1_like domains illustrate that HbBAK1s belong to the BAK1 kinase. BRI1 and BAK1 play an important role in regulating plant hormone signal transduction. Analysis of the cis-element of all HbBRI1 and HbBAK1 genes identified hormone response, light regulation and abiotic stress elements in the promoters of HbBRI1s and HbBAK1s. The results of tissue expression patterns indicate that HbBRL1/2/3/4 and HbBAK1a/b/c are highly expressed in the flower, especially HbBRL2-1. The expression of HbBRL3 is extremely high in the stem, and the expression of HbBAK1d is extremely high in the root. Expression profiles with different hormones show that HbBRI1 and HbBAK1 genes are extremely induced by different hormone stimulates. These results provide theoretical foundations for further research on the functions of BR receptors, especially in response to hormone signals in the rubber tree.

Mildew resistance locus O (MLO) is a plant-specific gene family that plays an important role in the growth and development of plants and their interactions with the environment. However, the available information on this gene family in pear is limited. Here, 24 PbrMLO genes were identified and divided into five subfamilies (I, II, III, IV and V). Whole-genome duplication (WGD) and dispersed duplication contributed to the expansion of the PbrMLO family. In addition, gene expression analysis revealed that PbrMLO genes were distributed in various pear tissues, suggesting their diverse functions. We selected PbrMLO23 for further functional analysis. Expression profile analysis by qRT-PCR showed that PbrMLO23 was highly expressed in pollen. Subcellular localization analysis showed that PbrMLO23 was located on the plasma membrane. When the expression level of PbrMLO23 was knocked down by using antisense oligonucleotides, pollen tube lengths increased, indicating that PbrMLO23 plays a functional role in inhibiting pollen tube growth. In summary, these results provide evolutionary insight into PbrMLO and its functional characteristics and lay a foundation for further analysis of the functions of PbrMLO members in pear.

The yield and quality of rubber tree latex are affected by environmental stresses and plant hormone stimulation. Heat shock protein 90 (HSP90) is widely involved in various developmental processes and stress responses in plants, especially in drought stress. In this study, we cloned the HbHSP90.3 gene and characterized its expression pattern in different tissues and mechanical wounding treatments of the rubber tree and found that it is highly expressed in latex and responds to mechanical wounding treatment. To reveal the roles of plant hormones and HSP90.3 protein in the drought resistance process of rubber trees. Treatment with the specific HSP90 protein inhibitor geldanamycin (GDA) and yeast expression experiments demonstrated that HbHSP90.3 has a relieving effect on water deficit in rubber trees. The expression pattern showed that the HbHSP90.3 gene was closely related to hormone signaling, especially for Indole acid (IAA) and Zeatin (ZT) induction under different plant hormone treatments. Protein interaction analysis showed that HbHSP90.3 interacted with the suppressor of the G2 allele of skp1 (HbSGT1b). Taken together, HbHSP90.3 interacts with HbSGT1b in the nucleus and plays a key role in water deficit.

Natural rubber is synthesized in the laticifers of Hevea brasiliensis, but the transcriptional networks regulating latex-associated processes remain poorly understood. In particular, whether WRKY transcription factors participate in brassinosteroid (BR)-responsive regulation remains unclear. We identified the HbWRKY family using the haplotype A telomere-to-telomere reference genome of Hevea brasiliensis and integrated phylogeny, gene structure, duplication type, synteny, Ka/Ks, promoter cis-elements, tissue expression, and BR-treated transcriptomic data. The 109 HbWRKY proteins comprised 21 Group I, 72 Group II, and 14 Group III members, together with one atypical protein and one Group I-like protein. Whole-genome or segmental duplication accounted for 102 genes (93.6%), and Ka/Ks was below 1 for all 111 duplicated HbWRKY pairs, consistent with purifying selection. G-box elements occurred in 84 promoters (77.1%). Eighty-nine genes were expressed in bark and 37 in latex, with latex showing the highest value for nine genes. Seven HbWRKY genes were differentially expressed after BR treatment, and all were downregulated. HbWRKY7 (HbAchr10M0003572) and HbWRKY28 (HbAchr15M0012484) were prioritized because both combined differential expression following BR treatment, their highest recorded expression in bark among the seven tissues, and predicted promoter G-boxes. This updated family resource narrows the candidates for testing a BR-WRKY regulatory module in rubber tree.