RESUMEN
Pholidota chinensis Lindl. is an epiphytic or lithophytic perennial herb of Orchidaceae family used as a garden flower or medicinal plant to treat high blood pressure, dizziness and headache in traditional Chinese medicine. Gastrodin (GAS) is considered as a main bioactive ingredient of this herb but the biosynthetic pathway remains unclear in P. chinensis. To elucidate the GAS biosynthesis and identify the related genes in P. chinensis, a comprehensive analysis of transcriptome and metabolome of roots, rhizomes, pseudobulbs and leaves were performed by using PacBio SMART, Illumina Hiseq and Ultra Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS). A total of 1,156 metabolites were identified by UPLC-MS/MS, of which 345 differential metabolites were mainly enriched in phenylpropanoid/phenylalanine, flavone and flavonol biosynthesis. The pseudobulbs make up nearly half of the fresh weight of the whole plant, and the GAS content in the pseudobulbs was also the highest in four tissues. Up to 23,105 Unigenes were obtained and 22,029 transcripts were annotated in the transcriptome analysis. Compared to roots, 7,787, 8,376 and 9,146 differentially expressed genes (DEGs) were identified in rhizomes, pseudobulbs and leaves, respectively. And in total, 80 Unigenes encoding eight key enzymes for GAS biosynthesis, were identified. Particularly, glycosyltransferase, the key enzyme of the last step in the GAS biosynthetic pathway had 39 Unigenes candidates, of which, transcript28360/f2p0/1592, was putatively identified as the most likely candidate based on analysis of co-expression, phylogenetic analysis, and homologous searching. The metabolomics and transcriptomics of pseudobulbs versus roots showed that 8,376 DEGs and 345 DEMs had a substantial association based on the Pearson's correlation. This study notably enriched the metabolomic and transcriptomic data of P. chinensis, and it provides valuable information for GAS biosynthesis in the plant.
RESUMEN
Pinellia ternata is a perennial traditional Chinese medicinal plant that undergoes different phenological patterns of dormancy depending on where it is growing. Plants grown in central and southern China typically display two growth cycles every year before and after hot summer days, exhibiting a summer dormancy. However, germplasms from these areas do not go into a dormancy phase in northern China where the summer monthly average temperatures range from 29-31°C. The northern China herbal growers prefer plant stocks from central China due to their longer growing quality and better tuber harvests. Here, we introduced a heat responsive receptor-like kinase ERECTA (ER) gene into P. ternata to explore changes in the growth cycle which were aimed at disrupting the summer dormancy. The 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene was also co-transformed with ER to improve the commercial trait. For the thermo-tolerance evaluation, all plants were treated with high temperatures (35°C/40°C) in a growth chamber or grown in natural field temperature in an isolated field before measurement of different agricultural, biochemical and physiological traits. The transgenics showed significantly (P < 0.05) higher heat tolerance, maintaining healthy vegetative growth unlike the empty vector (EV) harboring controls that became chlorotic and necrotic. Better performance in some of the monitored physiological traits was evident for overexpression lines exposed to the heat stress. In open isolated field trials, the transgenic genotypes did not show a summer dormancy but had a survival rate of 84-95%. The tuber biomasses were also significantly (P < 0.05) higher for the transgenic lines as compared to the EV controls, except for line ER118. Metabolites analysis indicated that the HMGR overexpressing lines (HMGR orHMGR + ER) exhibited significantly higher amounts of bioactive compounds including aromadendrene-4, 10-diol and 4, 8, 13-cyclotetradecatriene-1, 3-diol, 1, 5, 9-trimethyl-12-(1-methylethyl). Our findings show that the summer dormancy of P. ternata which is a naturally evolved trait, can be removed by a single heat responsive gene. The study contributes to generating heat tolerant new Pinellia varieties with enhanced commercially valuable chemicals.
