Location: Tropical Crops and Germplasm Research
Project Number: 6090-21000-051-03-T
Project Type: Trust Fund Cooperative Agreement
Start Date: Oct 1, 2013
End Date: Mar 30, 2015
To screen a large number of temperate and tropical bamboo germplasm accessions from multiple sites to quantify total protein, nutrient, and amino acid composition. In addition, a number miscellaneous underutilized crop species will be included in the exploratory evaluations on protein, nutrient and amino acid compositions.
Tropical and temperate bamboo genetic resources maintained at the USDA-ARS, Tropical Agriculture Research Station (TARS) in Mayaguez, Puerto Rico will be sampled. Additional genetic resources will be collected and evaluated from Govardhan Gardens, a privately owned tropical bamboo collection with collaborative ties to the TARS site. Leaf samples at similar physiological stages will be collected systematically from each accession in both locations. A representative number of other underutilized plants and plant products reported to be high in protein content will also be evaluated. Approximately 250 samples will be dried and ground prior to processing. Two procedures will be explored for the evaluation of crude protein. A modification of the micro-Kjeldahl method will be initially tested for its effectiveness in protein extraction and quantification. A P-PER Plant Protein Extraction Reagent in combination with a bis-cinchioninic acid (BCA) Protein Assay Kit from Thermo Fisher Scientific following manufacturer’s methodology will also be tested. A dry ashing method will be implemented for the determination of sample nutrient composition using a 7300 DV inductively coupled plasma (ICP) - spectrophotometer. Elemental analysis will include potassium, sodium, calcium, phosphorous magnesium, zinc, iron, manganese, copper, and molybdenum. Amino acid analysis will be carried out in three steps: crude protein purification, hydrolysis, and amino acid composition detection and quantification using high-performance liquid chromatography (HPLC). Acid hydrolysis will be used as the method for hydrolyzing protein peptide bonds and peptides for analysis.