Snapshot of four mature quinoa (Chenopodium quinoa) seeds: a shotgun proteomics analysis with emphasis on seed maturation, reserves and early germination.

Chenopodium quinoa Willd. is a crop species domesticated over 5000 years ago. This species is highly diverse, with a geographical distribution that covers more than 5000 km from Colombia to Chile, going through a variety of edaphoclimatic conditions. Quinoa grains have great nutritional quality, raising interest at a worldwide level. In this work, by using shotgun proteomics and in silico analysis, we present an overview of mature quinoa seed proteins from a physiological context and considering the process of seed maturation and future seed germination. For this purpose, we selected grains from four contrasting quinoa cultivars (Amarilla de Maranganí, Chadmo, Sajama and Nariño) with different edaphoclimatic and geographical origins. The results give insight on the most important metabolic pathways for mature quinoa seeds including: starch synthesis, protein bodies and lipid bodies composition, reserves and their mobilization, redox homeostasis, and stress related proteins like heat-shock proteins (HSPs) and late embryogenesis abundant proteins (LEAs), as well as evidence for capped and uncapped mRNA translation. LEAs present in our analysis show a specific pattern of expression matching that of other species. Overall, this work presents a complete snapshot of quinoa seeds physiological context, providing a reference point for further studies. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01295-8.

Mitochondrial proteomics reveals new insights into embryogenic competence acquisition in Carica papaya L. callus.

Understanding the mechanisms that endow a somatic cell with the ability to differentiate into a somatic embryo, which could result in numerous biotechnological applications, is still a challenge. The objective of this work was to identify some of the molecular and physiological mechanisms responsible for the acquisition of embryogenic competence during somatic embryogenesis in Carica papaya L. We performed a broad characterization of embryogenic (EC) and nonembryogenic calli (NEC) of using global and mitochondrial proteomic approaches, histomorphology, histochemistry, respiratory activity, and endogenous hormonal and hydrogen peroxide (H2O2) contents. EC and NEC presented remarkable differences in anatomical and histochemical characteristics, with EC showing a higher reactivity for the presence of proteins and neutral polysaccharides. Our results demonstrate that mitochondrial metabolism affects the embryogenic competence of C. papaya callus. The EC presented higher participation of alternative oxidase (AOX) enzymes, higher total cell respiration and presented a stronger accumulation of mitochondrial stress response proteins. Differential accumulation of auxin-responsive Gretchen Hagen 3 (GH3) family proteins in EC was related to a decrease in the content of free 2,4-dichlorophenoxyacetic acid (2,4-D). EC also showed higher endogenous H2O2 contents. H2O2 is a promising molecule for further investigation in differentiation protocols for C. papaya somatic embryos. SIGNIFICANCE: To further advance the understanding of somatic embryogenesis, we performed a broad characterization of embryogenic and nonembryogenic callus, through global and mitochondrial proteomic approaches, histomorphology, histochemistry, respiratory activity, and endogenous hormonal and hydrogen peroxide contents. Based on these results, we propose a working model for the competence of papaya callus. This model suggests that GH3 proteins play an important role in the regulation of auxins. In addition, embryogenic callus showed a greater abundance of stress response proteins and folding proteins. Embryogenic callus respiration occurs predominantly via AOX, and the inhibition of its activity is capable of inhibiting callus differentiation. Although the embryogenic callus presented greater total respiration and a greater abundance of oxidative phosphorylation proteins, they had less COX participation and less coupling efficiency, indicating less ATP production.

Deciphering the major metabolic pathways associated with aluminum tolerance in popcorn roots using label-free quantitative proteomics.

MAIN CONCLUSION: Al responsive proteins are associated with starch, sucrose, and other carbohydrate metabolic pathways. Sucrose synthase is a candidate to Al tolerance. Al responses are regulated at transcriptional and post-transcriptional levels. Aluminum toxicity is one of the important abiotic stresses that affects worldwide crop production. The soluble form of aluminum (Al3+) inhibits root growth by altering water and nutrient uptake, a process that also reduces plant growth and development. Under long-term Al3+ exposure, plants can activate several tolerance mechanisms. To date, no reports of large-scale proteomic data concerning maize responses to this ion have been published. To investigate the post-transcriptional regulation in response to Al toxicity, we performed label-free quantitative proteomics for comparative analysis of two Al-contrasting popcorn inbred lines and an Al-tolerant commercial hybrid during 72 h under Al-stress conditions. A total of 489 differentially accumulated proteins (DAPs) were identified in the Al-sensitive inbred line, 491 in the Al-tolerant inbred line, and 277 in the commercial hybrid. Among them, 120 DAPs were co-expressed in both Al tolerant genotypes. Bioinformatics analysis indicated that starch, sucrose, and other components of carbohydrate metabolism and glycolysis/gluconeogenesis are the biochemical processes regulated in response to Al toxicity. Sucrose synthase accumulation and an increase in sucrose content and starch degradation suggest that these components may enhance popcorn tolerance to Al stress. The accumulation of citrate synthase suggests a key role for this enzyme in the detoxification process in the Al-tolerant inbred line. The integration of transcriptomic and proteomic data indicates that the Al tolerance response presents a complex regulatory network into the transcription and translation dynamics of popcorn root development.

Stage-specific protein regulation during somatic embryo development of Carica papaya L. 'Golden'.

Somatic embryogenesis is an important biotechnological technique for large-scale propagation of elite genotypes. Identifying stage-specific compounds associated with somatic embryo development can help elucidate the ontogenesis of Carica papaya L. somatic embryos and improve tissue culture protocols. To identify the stage-specific proteins that are present during the differentiation of C. papaya somatic embryos, proteomic analyses of embryos at the globular, heart, torpedo and cotyledonary developmental stages were performed. Mass spectrometry data have been deposited in the ProteomeXchange with the dataset identifier PXD021107. Comparative proteomic analyses revealed a total of 801 proteins, with 392 classified as differentially accumulated proteins in at least one of the developmental stages. The globular-staged presented a higher number of unique proteins (16), and 7 were isoforms of 60S ribosomal proteins, suggesting high translational activity at the beginning of somatic embryogenesis. Proteins related to mitochondrial metabolism accumulated to a high degree at the early developmental stages and then decreased with increasing development, and they contributed to cell homeostasis in early somatic embryos. A progressive increase in the accumulation of vicilin, late embryogenesis abundant proteins and chloroplastic proteins that lead to somatic embryo maturation was also observed. The differential accumulation of acetylornithine deacetylase and S-adenosylmethionine synthase 2 proteins was correlated with increases in putrescine and spermidine contents, which suggests that both polyamines should be tested to determine whether they increase the conversion rates of globular- to cotyledonary-staged somatic embryos. Taken together, the results showed that somatic embryo development in C. papaya is regulated by the differential accumulation of proteins, with ribosomal and mitochondrial proteins more abundant during the early somatic embryo stages and seed maturation proteins more abundant during the late stages.

LED lamps enhance somatic embryo maturation in association with the differential accumulation of proteins in the Carica papaya L. 'Golden' embryogenic callus.

The use of light-emitting diode (LED) lamps has been shown to be a promising approach for improving somatic embryo maturation during somatic embryogenesis. The aim of this work was to study the influence of the light source on somatic embryo differentiation and its relationship with the differential abundance of proteins in the Carica papaya L. 'Golden' embryogenic callus at 14 days of maturation. The white plus medium-blue (WmB) LED and fluorescent lamp treatments produced an average of 82.4 and 47.6 cotyledonary somatic embryos per callus, respectively. A shotgun proteomics analysis revealed 28 upaccumulated and 7 downaccumulated proteins. The proteins upaccumulated in the embryogenic callus matured under the WmB LED lamp compared with that matured under the fluorescent lamp included indole-3-acetic acid-amido synthetase (GH3) and actin-depolymerizing factor 2 (ADF2), which are involved in the regulation of auxin levels by auxin conjugation and transport. Additionally, proteins related to energy production (aconitate, ADH1, GAPCp, PKp and TPI), cell wall remodeling (PG and GLPs), and intracellular trafficking (NUP50A, IST1, small GTPases and H+-PPase) showed significantly higher abundance in the embryogenic callus incubated under the WmB LED lamp than in that incubated under the fluorescent lamp. The results showed that the WmB LED lamp improved somatic embryo maturation in association with the differential accumulation of proteins in the C. papaya 'Golden' embryogenic callus.

Morphological analyses and variation in carbohydrate content during the maturation of somatic embryos of Carica papaya.

Efficient protocols for somatic embryogenesis of papaya (Carica papaya L.) have great potential for selecting elite hybrid genotypes. Addition of polyethylene glycol (PEG), a nonplasmolyzing osmotic agent, to a maturation medium increases the production of somatic embryos in C. papaya. To study the effects of PEG on somatic embryogenesis of C. papaya, we analyzed somatic embryo development and carbohydrate profile changes during maturation treatments with PEG (6%) or without PEG (control). PEG treatment (6%) increased the number of normal mature somatic embryos followed by somatic plantlet production. In both control and PEG treatments, pro-embryogenic differentiation to the cotyledonary stage was observed and was significantly higher with PEG treatment. Histomorphological analysis of embryonic cultures with PEG revealed meristematic centers containing small isodiametric cells with dense cytoplasm and evident nuclei. Concomitant with the increase in the differentiation of somatic embryos in PEG cultures, there was an increase in the endogenous content of sucrose and starch, which appears to be related to a rising demand for energy, a key point in the conversion of C. papaya somatic embryos. The endogenous carbohydrate profile may be a valuable parameter for developing optimized protocols for the maturation of somatic embryos in papaya.