Abstract
Pigeonpea is an important legume crop with high protein content. However, it is often subjected to various abiotic and biotic stresses. Proteomics is a state-of-the-art technique used to analyze the protein profiling of a tissue for deciphering the molecular entities that could be manipulated for developing crops resistant to these stresses. In this context, developing a comprehensive proteome profile from different vegetative and reproductive tissues has become mandatory. Although several protein extraction protocols from different tissues of diverse plant species have been reported, there is no report for pigeonpea. Here, we report tissue-specific protein extraction protocols representing vegetative (young leaves), and reproductive (flowers and seeds) organs and their subsequent analysis on 2-dimensional gel electrophoresis. The study explicitly demonstrated that the efficacy of a particular protein extraction protocol is dependent on the different tissues, such as leaves, flowers and seeds that differ in their structure and metabolic constituents. For instance, phenol-based protocol showed an efficacy toward higher protein yield, better spot resolution and a minimal streaking on 2-DE gel for both leaves and flowers. Protein extraction from seeds was best achieved by employing phosphate-TCA-acetone protocol.
Highlights
India is the largest consumer and producer of Pigeonpea (Cajanus cajan), a tropical legume, for its dietary properties
The flow diagram (Figure 1) presented here showed different steps involved for proteomics comprising protein precipitation, quantification and iso-electric focusing, 2-D gel electrophoresis, gel staining and scanning and image analysis
Other than commonly used trichloro acetic acid (TCA)/acetone precipitation method (Cho et al, 2006), phenol extraction followed by methanol/ammonium acetate precipitation has been shown to efficient (Hurkman and Tanaka, 1986)
Summary
India is the largest consumer and producer of Pigeonpea (Cajanus cajan), a tropical legume, for its dietary properties. Due to high protein content (18–30%) and its ease of digestibility (68%), it is a major source of proteins for a large section of Indian population that strictly depends on vegetarian meals (Reddy et al, 1979; Chitra et al, 1996; Sharma et al, 2011) In this context pigeonpea has become a priority crop in India with an effort toward developing high yielding varieties by conventional breeding approach and/or biotechnological interventions. Pigeonpea is the first seed legume to Comparative method for protein extraction have its complete genome sequenced through global research consortium (Singh et al, 2012; Varshney et al, 2012) This has provided further fillip toward developing this legume that would be tolerant to various biotic and/or abiotic stresses and exhibiting high yield potential under adverse agro-climatic conditions
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