Genetic Diversity for Seed Mineral Composition in the Wild Legume Teramnus Labialis
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Plant Foods Hum Nutr (2008) 63:105–109 DOI 10.1007/s11130-008-0078-8 ORIGINAL PAPER Genetic Diversity for Seed Mineral Composition in the Wild Legume Teramnus labialis Michael A. Grusak Published online: 19 June 2008 # Springer Science + Business Media, LLC 2008 Abstract Teramnus labialis (L.) Spreng. is a wild, tropical protein, energy (in the form of carbohydrates or lipids), legume whose seeds are collected and used as a food source several vitamins, and many human essential minerals. by tribal populations. In order to assess the potential of this Various legume species have been harnessed for large-scale legume to provide dietary minerals for humans, fourteen cultivation in diverse climactic zones; however, several diverse accessions were grown under controlled, nutrient- wild legume species are also collected and consumed on a replete conditions and seeds were harvested for mineral smaller scale by rural or tribal populations [3–6]. The analysis. The germplasm originated from Indonesia, Africa, potential conversion of these wild legumes into cultivated the Caribbean, and South America. Seed concentrations of crops has received considerable attention in recent years, phosphorus (P), potassium (K), sodium (Na), iron (Fe), especially as efforts have expanded to increase the mineral copper (Cu), manganese (Mn), and zinc (Zn) were found to concentration of seed foods for humans [7]. Populations fall within the range of published values for several that are at-risk for micronutrient mineral deficiencies (e.g., cultivated grain legumes, while calcium (Ca) and magne- Fe, Zn, I) or macronutrient mineral deficiencies (e.g., Ca) sium (Mg) were higher in T. labialis seeds. Mineral could be helped with nutrient-dense, biofortified legume concentrations across the diverse accessions showed ranges and cereal grains [8]. Recent studies have assessed the of 1.3- to 2.3-fold for the macronutrient minerals (Ca, Mg, genetic diversity of seed mineral concentrations in several P, K) and 1.8- to 15.9-fold for the micronutrient minerals cultivated crops, in order to use selected lines for breeding (Fe, Cu, Mn, Zn, and Na). The existing genetic diversity in or investigative research purposes [9]. Germplasm of wild, this wild legume, especially for the essential minerals Ca edible species should also be studied [10], thereby and Mg, could be exploited to develop T. labialis as a new providing opportunities for developing these legumes either cultivated legume for tropical regions of the world. as new food crops themselves, or as biological models for the discovery of genes relevant to seed mineral accretion. Keywords Calcium . Germplasm . Magnesium . One such wild legume that has received only limited Seed minerals . Wild legume attention as a grain crop is Teramnus labialis (L.) Spreng. This legume grows in tropical areas of Asia, Africa, and the Americas [11], and has recently been reported to be Introduction consumed as a seed food by tribal sects in South India [12]. As with several other legumes, T. labialis is used as a Legume seeds are consumed worldwide as an important forage crop in certain parts of the world [13, 14]; thus, its source of dietary nutrients for humans [1, 2]. They provide current utilization by rural populations as forage makes it a good candidate for expansion into a cultivated, human food M. A. Grusak (*) source. Interestingly, the limited nutritional data on T. USDA/ARS Children’s Nutrition Research Center, labialis suggests that it is a very good source of essential Department of Pediatrics, Baylor College of Medicine, minerals, including Ca, Mg, and K. In particular, the 1100 Bates Street, Houston, TX 77030, USA reported seed concentration for Mg (812 mg/100 g dry e-mail: [email protected] weight) [12] is one of the highest seed Mg values we have 106 Plant Foods Hum Nutr (2008) 63:105–109 found in the literature, and is much higher than that of solution to each pot, three times a day. Sufficient solution existing, cultivated legume (or cereal) crops [15]. was delivered to ensure soil saturation; excess solution was In order to gain a better understanding of the seed allowed to drain from the pots. The nutrient solution mineral composition of this wild legume, fourteen diverse contained the following concentrations of mineral salts: T. labialis germplasm accessions were grown for seed 1.0 mM KNO3, 0.4 mM Ca(NO3)2, 0.1 mM MgSO4, harvest and analysis. Plants were maintained under con- 0.15 mM KH2PO4,25μM CaCl2,25µMH3BO3,2µm trolled environmental conditions, with continuous mineral MnSO4, 2 µM ZnSO4, 0.5 µM CuSO4, 0.5 µM H2MoO4, fertilization used to prevent mineral inadequacies. The data 0.1 µM NiSO4, and 1 µM Fe(Ш)-N, N′-ethylenebis[2-(2- from the diverse accessions are presented and compared hydroxyphenyl)-glycine] (Sprint 138; Becker-Underwood, with seed values for various cultivated and wild legumes. Inc., Ames, Iowa, USA). The potential for developing T. labialis into a cultivated The environmental conditions within the greenhouse legume is also discussed. were a temperature regime of 22±3 °C day and 20±3 °C night, with relative humidity ranging from 45% to 65% throughout the day/night cycle. Sunlight was supplemented Materials and Methods with metal halide lamps set to a 15-h day, 9-h night photoperiod (lights on at 700 h), and a minimum intensity Plant Material and Growth Conditions (i.e., lamps only) of photosynthetically active radiation of 200 μmol photons m−2 s−1 at the top of the plants. Fourteen accessions of T. labialis (L.) Spreng. (Fabaceae) were used in this study (Table 1). These were obtained from Seed Harvest and Tissue Analysis the USDA germplasm collection (USDA/ARS Plant Ge- netic Resources Conservation Unit, Griffin, Georgia, USA; Plants were grown for up to 15 months, which allowed all accessions labeled with PI numbers) or from the perennial accessions time to reach reproductive age and to enable Glycine germplasm collection (University of Illinois, adequate pod production for seed collection. Seeds were Champaign-Urbana, Illinois, USA) (both the CIAT identi- harvested as pods matured and were combined from all six fiers and the Champaign-Urbana [CU] identifiers are plants per accession. Seeds were removed from pods, provided). extraneous material was removed, and seeds were dried Plants were grown in 5-l black plastic pots filled with a for a minimum of 3 days in a 60 °C, forced-air drying oven. 2:1 (v/v) mixture of synthetic soil (Metro-Mix 360; Scotts- Average seed weights were calculated by averaging the Sierra Horticultural Products Co., Marysville, Ohio, USA) weight of three sets of 100 random seeds for each and vermiculite (Strong-Lite Medium Vermiculite, Sun Gro accession. Horticulture Co, Seneca, Illinois, USA). Plants of each For seed mineral analysis, a minimum of 3 g of dried accession were grown in two pots, with plants thinned to seeds for each accession was homogenized. Three aliquots three seedlings per pot, 4 d after emergence. Pots were (0.25 g each) of each accession were digested at 125 °C randomly assigned positions in a greenhouse. An automat- (3 h) and 200 °C (digestate taken to dryness) using trace- ed drip irrigation system was used to deliver nutrient metal grade hydrogen peroxide and nitric acid. Digestates Table 1 Source country and mean seed weights of Accession identifier Country of origin Mean seed weight (mg/100 seeds±standard error) Teramnus labialis germplasm used in this study CIAT 926, CU 409 Antigua and Barbuda 650±1 CIAT 4986, CU 411 Indonesia 615±2 CIAT 7442, CU 412 Cuba 745±5 CIAT 20072, CU 416 Colombia 267±0 PI 200233 Kenya 668±7 PI 277511 Virgin Islands 758±4 PI 365055 South Africa 841±5 PI 365056 South Africa 824±5 PI 365057 South Africa 796±2 PI 406170 Kenya 698±3 PI 490301 Kenya 763±4 PI 517204 Ethiopia 1,250±4 PI 538317 Virgin Islands 706±0 PI 538318 Virgin Islands 953±7 Plant Foods Hum Nutr (2008) 63:105–109 107 were resuspended in 2% ultra-pure nitric acid and analyzed from 1.74 to 11.99 mg/100 g dry weight; Zn varied 1.9- for Ca, Mg, K, P, Fe, Zn, Mn, Cu, and Na concentrations. fold, ranging from 4.48 to 8.53 mg/100 g dry weight; and Elemental analysis was performed using inductively cou- Na varied 6.1-fold, ranging from 0.48 to 2.95 mg/100 g dry pled plasma–optical emission spectrometry (CIROS ICP weight. Model FCE12; Spectro, Kleve, Germany). Certified rice flour standards (SRM 1568A; National Institute of Stand- ards and Technology, Gaithersburg, Maryland, USA) were Discussion digested and analyzed along with samples to verify the reliability of the procedures and analytical measurements. Although grown in a greenhouse with somewhat atypical conditions (i.e., non-tropical temperatures or humidity), all fourteen T. labialis accessions grew well and produced Results many pods in this study. Seed weights (Table 1) were also indicative of this species, suggesting that the plants were Fourteen T. labialis accessions obtained for this study healthy and producing normal seeds. Mineral nutrients were represented germplasm collected from Asia, Africa, the provided to the plants on a daily basis with the intent of Caribbean, and South America (Table 1). Inspection of leaf ensuring complete, non-limiting nutrient conditions. This characters, growth habit, pod phenotype (data not shown), was done to assess the full genetic potential of each accession and seed weight (Table 1) suggested that all the material with respect to the transport of minerals to developing seeds. was T. labialis germplasm, as described by Verdcourt [11]. All plants appeared healthy throughout the study with no Pods of all plants exhibited the elongated hook that is evidence of leaf chlorosis or necrotic lesions.