Identification and analysis of UGT genes associated with triterpenoid saponin in soapberry (Sapindus mukorossi Gaertn.).

BACKGROUND: Soapberry (Sapindus mukorossi) is an economically important multifunctional tree species. Triterpenoid saponins have many functions in soapberry. However, the types of uridine diphosphate (UDP) glucosyltransferases (UGTs) involved in the synthesis of triterpenoid saponins in soapberry have not been clarified. RESULTS: In this study, 42 SmUGTs were identified in soapberry, which were unevenly distributed on 12 chromosomes and had sequence lengths of 450 bp to 1638 bp, with an average of 1388 bp. The number of amino acids in SmUGTs was 149 to 545, with an average of 462. Most SmUGTs were acidic and hydrophilic unstable proteins, and their secondary structures were mainly α-helices and random coils. All had conserved UDPGT and PSPG-box domains. Phylogenetic analysis divided them into four subclasses, which glycosylated different carbon atoms. Prediction of cis-acting elements suggested roles of SmUGTs in plant development and responses to environmental stresses. The expression patterns of SmUGTs differed according to the developmental stage of fruits, as determined by transcriptomics and RT-qPCR. Co-expression network analysis of SmUGTs and related genes/transcription factors in the triterpenoid saponin synthesis pathway was also performed. The results indicated potential roles for many transcription factors, such as SmERFs, SmGATAs and SmMYBs. A correlation analysis showed that 42 SmUGTs were crucial in saponin synthesis in soapberry. CONCLUSIONS: Our findings suggest optimal targets for manipulating glycosylation in soapberry triterpenoid saponin biosynthesis; they also provide a theoretical foundation for further evaluation of the functions of SmUGTs and analyses of their biosynthetic mechanisms.

Physiology and transcriptome of Sapindus mukorossi seeds at different germination stages.

Sapindus mukorossi has a wide distribution range, high application value, and broad developmental potential. Previous studies have mostly focused on the medicinal and economic value of soapberry; however, few studies have been conducted on its seed germination. This study measured the physiological indicators and hormone content of soapberry seeds at different germination stages and preliminarily determined that abscisic acid (ABA) and indole-3-acetic acid (IAA) are the key hormones that affect the germination of soapberry seeds. Both Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG enrichment) analyses detected hormone transduction pathways, further confirming the key role of plant hormones in the germination process of soapberry seeds. Through transcriptome analysis, we speculated that CYP707A and IPA are key genes in the ABA and IAA synthesis pathways, respectively. This study revealed the close relationship between plant hormones and soapberry seed germination and provided new ideas for further exploration of the germination mechanism of soapberry seeds.

Metabolome and Transcriptome Analysis Reveals the Transcriptional Regulatory Mechanism of Triterpenoid Saponin Biosynthesis in Soapberry (Sapindus mukorossi Gaertn.).

Soapberry (Sapindus mukorossi Gaertn.) pericarps are rich in valuable bioactive triterpenoid saponins. However, the saponin content dynamics and the molecular regulatory network of saponin biosynthesis in soapberry pericarps remain largely unclear. Here, we performed combined metabolite profiling and transcriptome analysis to identify saponin accumulation kinetic patterns, investigate gene networks, and characterize key candidate genes and transcription factors (TFs) involved in saponin biosynthesis in soapberry pericarps. A total of 54 saponins were tentatively identified, including 25 that were differentially accumulated. Furthermore, 49 genes putatively involved in sapogenin backbone biosynthesis and some candidate genes assumed to be responsible for the backbone modification, including 41 cytochrome P450s and 45 glycosyltransferases, were identified. Saponin-specific clusters/modules were identified by Mfuzz clustering and weighted gene coexpression network analysis, and one TF-gene regulatory network underlying saponin biosynthesis was proposed. The results of yeast one-hybrid assay and electrophoretic mobility shift assay suggested that SmbHLH2, SmTCP4, and SmWRKY27 may play important roles in the triterpenoid saponin biosynthesis by directly regulating the transcription of SmCYP71D-3 in the soapberry pericarp. Overall, these findings provide valuable information for understanding the molecular regulatory mechanism of saponin biosynthesis, enriching the gene resources, and guiding further research on triterpenoid saponin accumulation in soapberry pericarps.

Rhizosphere bacteriome of the medicinal plant Sapindus saponaria L. revealed by pyrosequencing.

Sapindus saponaria L. of Sapindaceae family is popularly known as soldier soap and is found in Central and South America. A study of such medicinal plants might reveal a more complex diversity of microorganisms as compared to non-medicinal plants, considering their metabolic potential and the chemical communication between their natural microbiota. Rhizosphere is a highly diverse microbial habitat with respect to both the diversity of species and the size of the community. Rhizosphere bacteriome associated with medicinal plant S. saponaria is still poorly known. The objective of this study was to assess the rhizosphere microbiome of the medicinal plant S. saponaria using pyrosequencing, a culture-independent approach that is increasingly being used to estimate the number of bacterial species present in different environments. In their rhizosphere microbiome, 26 phyla were identified from 5089 sequences of 16S rRNA gene, with a predominance of Actinobacteria (33.54%), Acidobacteria (22.62%), and Proteobacteria (24.72%). The rarefaction curve showed a linear increase, with 2660 operational taxonomic units at 3% distance sequence dissimilarity, indicating that the rhizosphere microbiome associated with S. saponaria was highly diverse with groups of bacteria important for soil management, which could be further exploited for agricultural and biotechnological purposes.

The complete chloroplast genome sequence of Sapindus mukorossi.

The complete chloroplast genome sequence of Sapindus mukorossi, a critical Chinese medicine, was reported here. The total length of the chloroplast genome is 160,481 bp long with 37.7% overall GC content. A pair of IRs (inverted repeats) of 27,979 bp were separated by SSC (18,873 bp) and LSC (85,650 bp). It contains 78 protein-coding genes, 30 tRNA genes and four rRNA genes. Sixteen genes contain one or two introns.

Genetic variability and population structure in Sapindus emarginatus Vahl from India.

Sapindus emarginatus is an economically important tropical tree species sparsely distributed in different geographical provinces like Gangetic Plains, Western Ghats, and Deccan Plateau in India. In the present paper estimation of genetic variability within and among 41 accessions representing five populations was carried out using 3 single primer amplification reaction (SPAR) methods viz. RAPD, DAMD and ISSR. The cumulative data analysis was carried out for all three SPAR methods, and showed 82.32% polymorphism across all the accessions of S. emarginatus. Jaccard's similarity values among 41 accessions ranged from 0.15 to 0.49 with an average value of 0.37. The intra-population genetic diversity revealed highest values of Nei's genetic diversity (0.19,) Shannon information index (0.29) and polymorphic loci (55.18%), among the accessions of Gujarat (GJ) population, while the corresponding lowest values were (0.10), (0.15) and (26.40%) respectively among the accessions of Rajasthan (RJ) population. The maximum inter-population average genetic distance (0.20) was between Karnataka (KA) and RJ, while the corresponding least genetic distance (0.06) was between Allahabad (AL) and Varanasi (VS) populations. The analysis of molecular variance (AMOVA) revealed maximum percentage of variation among individuals of populations (72%) followed by 16% among regions and 12% among populations. Principal coordinate analysis (PCA) of cumulative data also supported the clustering pattern in the UPGMA dendrogram. These results suggest that genetic diversity is corroborating with the geographical diversity. Mantel's test was performed which revealed a highly significant correlation between cumulative vs RAPD, and showed the maximum (0.93) correlation coefficient, followed by cumulative vs ISSR (0.78) and cumulative vs DAMD (0.91) respectively, and this clearly indicates that the SPAR methods (RAPD, DAMD and ISSR) are sufficiently informative and are suitable to analyze the genetic variability within and among the populations of S. emarginatus.

Molecular characterization of high plant species using PCR with primers designed from consensus branch point signal sequences.

A novel method is introduced for producing molecular markers in plants using single 15- to 18-mer PCR primers designed from the short conserved consensus branch point signal sequences and standard agarose gel electrophoresis. This method was tested on cultivated peanut and verified to give good fingerprinting results in other plant species (mango, banana, and longan). These single primers, designed from relatively conserved branch point signal sequences within gene introns, should be universal across other plant species. The method is rapid, simple, and efficient, and it requires no sequence information of the plant genome of interest. It could be used in conjunction with, or as a substitute for, conventional RAPD or ISSR techniques for applications including genetic diversity analysis, phylogenetic tree construction, and quantitative trait locus mapping. This technique provides a new way to develop molecular markers for assessing genetic diversity of germplasm in diverse species based on conserved branch point signal sequences.