Long-Term Subculture Affects Rooting Competence via Changes in the Hormones and Protein Profiles in Cedrela Fissilis Vell. (Meliaceae) Shoots

Abstract

Long-term subculture plays an essential role in the large-scale multiplication and production of somatic plantlets. We investigated the effects of long-term subculture on in vitro shoot development and ex vitro rooting associated with changes in the hormones and protein profiles in C. fissilis. The number of subcultures of shoots induced a decrease in the ex vitro rooting response. The reduction in adventitious root (AR) formation was associated with decreases in the contents of indole-3-acetic acid (IAA), abscisic acid (ABA), 12-oxo phytodienoic acid (OPDA), putrescine (Put), and spermine and increases in jasmonic acid (JA), jasmonoyl-isoleucine, trans-cinnamic acid, and salicylic acid contents in shoots at the fourth subculture compared to the first. The ornithine decarboxylase enzyme preferentially functions in the Put biosynthesis pathway and was related to the highest AR formation in shoots at the first subculture. Down-accumulation of the auxin-binding protein ABP19a in shoots from the fourth subculture compared to the first subculture was related to a decrease in both IAA contents and AR formation. In addition, down-accumulation of glucose-6-phosphate isomerase, glutamine synthetase leaf isozyme chloroplastic, 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase, L-ascorbate peroxidase, cytosolic, monodehydroascorbate reductase, and 2-Cys peroxiredoxin BAS1-like, chloroplastic and up-accumulation of caffeoyl-CoA O-methyltransferase 1 and isoforms of peroxidase 4 proteins in shoots from the fourth relative to the first subculture were associated with a reduction in AR formation. These results showed that the understanding of hormonal and molecular mechanisms related to the potential of AR formation in shoots under successive subcultures is relevant to improving large-scale plantlet production in C. fissilis.

Reduction of heat stress pressure and activation of photosystem II repairing system are crucial for citrus tolerance to multiple abiotic stress combination

“…Comparative proteomic analyses were performed with three biological replicate leaf samples (300 mg of fresh weight per sample) of Carrizo citrange and Cleopatra mandarin plants subjected to individual stresses (WS, HL or HS) and to their factorial combinations (HL + HS, WS + HL, WS + HS and WS + HL + HS). Total protein extraction was performed according to Oliveira et al ( 2022 ).…”

“…Spectral processing and database searching were performed using the ProteinLynx Global Server (PLGS; version 3.0.2; Waters) as described in Oliveira et al ( 2022 ). For protein identifications, the sweet orange ( C. sinensis ) protein sequence database from National Center for Biotechnology Information Search database was used (NCBI; https://www.ncbi.nlm.nih.gov ).…”

Germinación de semillas, micropropagación y conservación de germoplasma in vitro de Cedrela odorata (Meliaceae) en el norte del Perú

“…Asimismo, un avance importante fue la micropropagación de C. odorata a partir de brotes juveniles en medio suplementado con BAP 2 mg L -1 y ANA 3 mg L -1 utilizando nudos y también se estimuló el enraizamiento en el mismo medio de micropropagación (García-Gonzáles et al 2011). Por otro lado, en brotes de Cedrela fissilis Vell., especie estrechamente relacionada con C. odorata, los explantes (ápices caulinares y segmentos nodales cotiledonales), obtenidos de plántulas de 60 días, enraizaron en medio de cultivo MS suplementado con BA 0,6 mg L -1 , aunque el subcultivo a largo plazo afectó la competencia de enraizamiento (Oliveira et al 2021). En el presente estudio, el medio de cultivo utilizado fue MS suplementado con AIA 0,02 mg L -1 y AG 3 0,02 mg L -1 , denominado "medio de cultivo de Piper" puesto que es ampliamente utilizado en la micropropagación de numerosas especies del género Piper (familia Piperaceae), particularmente leñosas En los primeros intentos de conservar el germoplasma in vitro de C. odorata se utilizó la criopreservación en nitrógeno líquido (Engelmann 2011).…”