A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis

Wang, Wei; Vignani, Rita; Scali, Monica; Cresti, Mauro

doi:10.1002/elps.200500722

Cited by 493 publications

(409 citation statements)

References 11 publications

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“…Samples from different thalli were pooled to a total amount of 2 g. Liquid nitrogen was added to the pooled thalli, which were subsequently ground to a fine powder using mortar and pestle. Proteins were extracted as described by Wang et al (2006).…”

Section: Sample Preparation For Protein Extractionmentioning

confidence: 99%

Exploring functional contexts of symbiotic sustain within lichen-associated bacteria by comparative omics

et al. 2014

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Symbioses represent a frequent and successful lifestyle on earth and lichens are one of their classic examples. Recently, bacterial communities were identified as stable, specific and structurally integrated partners of the lichen symbiosis, but their role has remained largely elusive in comparison to the well-known functions of the fungal and algal partners. We have explored the metabolic potentials of the microbiome using the lung lichen Lobaria pulmonaria as the model. Metagenomic and proteomic data were comparatively assessed and visualized by Voronoi treemaps. The study was complemented with molecular, microscopic and physiological assays. We have found that more than 800 bacterial species have the ability to contribute multiple aspects to the symbiotic system, including essential functions such as (i) nutrient supply, especially nitrogen, phosphorous and sulfur, (ii) resistance against biotic stress factors (that is, pathogen defense), (iii) resistance against abiotic factors, (iv) support of photosynthesis by provision of vitamin B 12 , (v) fungal and algal growth support by provision of hormones, (vi) detoxification of metabolites, and (vii) degradation of older parts of the lichen thallus. Our findings showed the potential of lichenassociated bacteria to interact with the fungal as well as algal partner to support health, growth and fitness of their hosts. We developed a model of the symbiosis depicting the functional multi-player network of the participants, and argue that the strategy of functional diversification in lichens supports the longevity and persistence of lichens under extreme and changing ecological conditions.

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Section: Sample Preparation For Protein Extractionmentioning

confidence: 99%

Exploring functional contexts of symbiotic sustain within lichen-associated bacteria by comparative omics

et al. 2014

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“…Protein extraction using the hybrid technique, as carried out by Wang et al (2006) was performed along with several modifications using trichloroacetic acid (TCA), acetone and phenol extractions. Briefly, 100 mg of the freezedried samples were mixed with 1 mL 10% TCA: acetone (1:1) (v/v) and rotated at 16,000 rpm for 3 min at 4ºC.…”

Section: Total Protein Extraction and Determinationmentioning

confidence: 99%

Proteomic Analysis of Stored Core Oil Palm Trunk (COPT) Sap Identifying Proteins Related to Stress, Disease Resistance and Differential Gene/Protein Expression

Mostapha

Jahar

Azizan

et al. 2018

JSM

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“…Total protein was extracted according to the method described by Wang et al [22]. In brief, about 200 mg leaf tissue was ground to fine powder in liquid nitrogen and immediately suspended in 800 µl of ice cold 10% trichloroacetic acid (TCA) in acetone with 1% 2-mercaptoethanol (2-ME).…”

Section: Protein Extraction From B Monnieri Leavesmentioning

confidence: 99%

A Proteomic Approach to Evaluate the Effects of Endogenous Expression of Cryptogein Gene in Crypt-Transgenic Plants of Bacopa monnieri

Paul

Nandagopal

Agarwala

et al. 2017

JABB

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Cryptogein, a 10 kDa sterol scavenging protein secreted by the oomycete Phytophthora cryptogea [1], belongs to 'elicitin' class of elicitors [2]. Cryptogein has been used to study the underlying molecular mechanisms of disease resistance in tobacco. Application of purified cryptogein protein, even in nanomolar quantity causes hypersensitive response (HR)-like necrosis in tobacco plants [3] and also induces systemic acquired resistance (SAR) to P. parasitica var nicotianae [4].There are several reports on transformation with cryptogein (crypt) gene aimed to achieve the resistance against plant pathogens. Over expression of β-cryptogein induces HR in tobacco and triggers resistance against P. parasitica var nicotianae [5,6] and other pathogens. Effective increase in biomass and/ or secondary metabolites accumulation in crypt-transgenic cultures in a number of plant species have also been observed [7][8][9], where the crypt gene was either under the constitutive promoter [7] or under the inducible promoter [8]. AbstractCryptogein, a proteinaceous elicitor has been reported to affect growth and secondary metabolism in several plant species. We established two transgenic plant lines of Bacopa monnieri L., an indigenous medicinal species, following Agrobacterium tumefaciens mediated transformation. The transgenic plant lines obtained following transformation with strains LBA4404-nptII-crypt (plant line BmAt-ncrypt) and LBA4404-nptII (plant line BmAt-n), were maintained on selection media i.e. MS medium containing 100 mg/l Kanamycin. To evaluate the effects of endogenous expression of cryptogein gene in crypt-transgenic plants of B. monnieri, in the present study, a proteomic approach was employed. Using 2D-PAGE followed by MALDI TOF MS/MS, we found 54 spots with reproducible differences in abundance (≥ +2 or ≤ -0.5-fold) between the plant lines. Bioinformatics analyses helped their identification, functional annotation and categorization.Significantly upregulated gene expression (p≤ 0.5) of most of the key enzymes of triterpenoid saponin biosynthetic pathway (mevalonate kinase, mvk; mevalonate diphosphate decarboxylase, mdc; squalene synthase, sqs; β-amyrin synthase, bas and UDP-glycosyl transferase 2, ugt 2) in crypt-transgenic plant lines (BmAt-ncrypt) of B. Monnieri was observed as compared to non-transformed (Bm-NT) and empty vector control (BmAt-n) plant lines. Cryptogein-induced ROS production, a typical stress response of plant cells towards cryptogein, was significantly increased (p≤ 0.5) in specific activities and higher abundance of several anti-oxidant enzymes, such as SOD (3.0-5.0-fold), APX (2.3-3.0-fold) and CAT (2.6-fold) in BmAt-ncrypt. The upregulation of CaM (1.6-fold) and calcium-transporting ATPase-2 (2.0-fold) in BmAt-ncrypt indicate possible calcium-dependant cryptogein elicitation. A 3.0-fold increase in EIN3 levels with down-regulation of F-box protein, its regulator also indicate induction of JA-ET signalling pathway in crypt-transgenic plants. This study not only demonstrates involvement of ...

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A universal and rapid protocol for protein extraction from recalcitrant plant tissues for proteomic analysis

Cited by 493 publications

References 11 publications

Exploring functional contexts of symbiotic sustain within lichen-associated bacteria by comparative omics

Exploring functional contexts of symbiotic sustain within lichen-associated bacteria by comparative omics

Proteomic Analysis of Stored Core Oil Palm Trunk (COPT) Sap Identifying Proteins Related to Stress, Disease Resistance and Differential Gene/Protein Expression

A Proteomic Approach to Evaluate the Effects of Endogenous Expression of Cryptogein Gene in Crypt-Transgenic Plants of Bacopa monnieri

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