In-depth assembly of organ and development dissected Picrorhiza kurroa proteome map using mass spectrometry

Abstract

Background Picrorhiza kurroa Royle ex Benth. being a rich source of phytochemicals, is a promising high altitude medicinal herb of Himalaya. The medicinal potential is attributed to picrosides i.e. iridoid glycosides, which synthesized in organ-specific manner through highly complex pathways. Here, we present a large-scale proteome reference map of P. kurroa, consisting of four morphologically differentiated organs and two developmental stages. Results We were able to identify 5186 protein accessions (FDR < 1%) providing a deep coverage of protein abundance array, spanning around six orders of magnitude. Most of the identified proteins are associated with metabolic processes, response to abiotic stimuli and cellular processes. Organ specific sub-proteomes highlights organ specialized functions that would offer insights to explore tissue profile for specific protein classes. With reference to P. kurroa development, vegetative phase is enriched with growth related processes, however generative phase harvests more energy in secondary metabolic pathways. Furthermore, stress-responsive proteins, RNA binding proteins (RBPs) and post-translational modifications (PTMs), particularly phosphorylation and ADP-ribosylation play an important role in P. kurroa adaptation to alpine environment. The proteins involved in the synthesis of secondary metabolites are well represented in P. kurroa proteome. The phytochemical analysis revealed that marker compounds were highly accumulated in rhizome and overall, during the late stage of development. Conclusions This report represents first extensive proteomic description of organ and developmental dissected P. kurroa, providing a platform for future studies related to stress tolerance and medical applications.

Functional trait correlation network and proteomic analysis reveal multifactorial adaptation mechanisms to a climatic gradient associated with high altitude in the Himalayan region

“…This study was conducted at Rohtang Pass (32°22′N, 77°15′E) of Indian western Himalaya in July 2017, 2019 and 2021 along an altitudinal gradient at 3400, 3800 and 4100 masl as described earlier (Kumari et al, 2020, 2021). These sites were varied on average air temperature (AT) (8–20°C), relative humidity (RH) (40%–56%), soil temperature (ST) (7.55–13.21°C), solar (570–649 W/m 2 ) and UV radiation (51.8–65.2 W/m 2 ) recorded at the time of sample collection (Table 1).…”

“…Western blot analysis was performed using pooled proteins (20 µg) with polyclonal antibodies raised against RA (Rubisco activase; Agrisera AS10 700), PsbA (D1 protein of PSII; Agrisera AS05 084), Lhca1 (PSI type I chlorophyll a/b‐binding protein; Agrisera AS01 005), Lhcb1 (LHCII type I chlorophyll a/b‐binding protein; Agrisera AS01 004), heat shock protein (HSP) 70 (Heat shock protein 70; Agrisera AS08 371), Tic40 (Inner envelope membrane translocon complex protein chloroplast; Agrisera AS10 709), SUS1 (Sucrose synthase 1; Agrisera AS15 2830), 14‐3‐3 (General regulatory element; Agrisera AS12 2119), Cat (Catalase; Agrisera AS09 501), AtpB (Beta subunit of ATP synthase; AS05 085) and ACT (Actin; Agrisera AS13 2640) as described previously (Kumari et al, 2021). Three independent western experiments were performed for each antibody.…”

“…This study was conducted at Rohtang Pass (32°22′N, 77°15′E) of Indian western Himalaya in July 2017, 2019 and 2021 along an altitudinal gradient at 3400, 3800 and 4100 masl as described earlier (Kumari et al, 2020(Kumari et al, , 2021. These sites were varied on average air 1).…”

“…For peptide identification, the raw data were analysed with Agrisera AS13 2640) as described previously (Kumari et al, 2021).…”

“…; a high‐altitude medicinal herb distributed 3000–5000 meters above sea level (masl) in the Himalayas. Owing to the unique composition of picrosides, P. kurroa is widely used in the treatment of liver disorders, fever, asthma and jaundice (Kumari et al, 2021). Our initial metabolomics study shed light on various biochemical processes and specialised metabolites associated with P. kurroa adaptation to an altitudinal gradient (Kumari et al, 2020).…”