View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Directory of Open Access Journals Diversity of cacao fat in germplasm from Rondônia, Brazil Crop Breeding and Applied Biotechnology 8: 141-148, 2008 Brazilian Society of Plant Breeding. Printed in Brazil Diversity of cacao fat in germplasm from Rondônia, Brazil Caio Márcio Vasconcellos Cordeiro de Almeida1, Valdir Alves Facundo2, Henrique Otávio Braga Brazil2, Luiz Antônio dos Santos Dias3, José Raymundo Pires Medeiros4, and Jorge Luiz Andrade Costa3 Received 15 November 2007 Accepted 05 February 2008 ABSTRACT - Chocolate is food of high nutritional value made from the fat extracted from cacao beans. Fat content and melting point are features of utmost importance for the chocolate, pharmaceutical and cosmetics industries. This paper quantified the cacao germplasm diversity from Rondônia for these two attributes. Expressive diversity was detected among the 102 evaluated clones in April/May 2003 for fat content and melting point. The mean fat content was 55.53% (51.04 to 58.19%) and the mean fat melting point 29.55 ºC (25.60 to 32.53 ºC). The accessions from wild populations in Rondônia were outstanding for both attributes. The two periods (September/October 2001 and April/May 2003) affected the evaluated clones differently. The plot of principal components analysis showed expressive variability and was, to a certain extension, able to separate wild from domesticated accessions. Key words: cacao germplasm, diversity, fat content and melting point, genetic breeding. INTRODUCTION Cacao fat, extracted from the beans, is used predominantly in chocolate production. It is the most Cacao (Theobroma cacao L.) for chocolate valuable product of the fruit of the cacao. Fat of cocoa fabrication has been cultivated in the Brazilian Amazon is the most relevant feature for food, pharmaceutical region since 1670 (Alden 1974). It is highly nutritive and cosmetics industries. It is therefore of economic food owing to its high sugar, fat and protein contents. interest to deepen the understanding of the chemical Today’s cacao agribusiness in the Amazon is composition and physical characteristics of cacao fat, represented by around 106 thousand hectares of cacao particularly in the case of chocolate bars. When made stands, of which 75% are concentrated in Pará and of fat with a low melting point the bars soften up on Rondônia according to Mendes (1998). It was first warm days. Different genotypes and seasonal climatic planted in 1971, in the state of Rondônia, which is third variations can additionally affect the fat composition, in dry bean yield (17.8 thousand t year-1). Around 615 whose content in the beans is fairly variable. Pires et al. accessions, mostly wild populations from the state itself, (2003), evaluated 490 cacao accessions and observed are maintained in the genebank of the Experimental great variability in fat content (45.4 to 60.3%) and station of Ouro Preto. Their qualitative characteristics possibilities of progress with selection. In the south of still need to be evaluated. Bahia, higher temperatures during fruit development 1 CEPLAC/SUPOC, Avenida Governador Jorge Teixeira, 86, Bairro Nova Porto Velho, 78.906-100, Porto Velho, RO, Brasil 2 Departamento de Química, Universidade Federal de Rondônia (UFR), BR 364, Km 9,5, 78.900-500, Porto Velho, RO, Brasil 3 Departamento de Fitotecnia, Universidade Federal de Viçosa (UFV), 36.570-000, Viçosa, MG, Brasil. *E-mail: [email protected] 4 Estação Experimental Ouro Preto - ESTEX-OP/CEPLAC, BR 364, Km 325, 78.950-000, Ouro Preto do Oeste, RO, Brasil Crop Breeding and Applied Biotechnology 8: 141-148, 2008 141 CMVC Almeida et al. contributed to a higher saturation and, consequently, developed without water stress and; September/October hardness of the fat. Fruits that developed in the colder – when fruits developed under occurrence of water months had a lower fat melting point than fruits grown stress Almeida et. al (2005). Samples of September/ under higher temperatures. Powell (1984) observed that October 2001 from 36 accessions and samples of April/ the climatic conditions can be determinant for the May 2003 from 102 accessions were evaluated for fat chemical and physical properties of cacao fat. Indeed, content (%) and fat melting point (ºC), resulting in 29 the fat content is strongly affected by environmental common clones in the two periods. influence, presenting low narrow-sense heritability Laboratory analyses (12.5%), as estimated by Pardo and Enriquez (1988). In the Brazilian Amazon region, studies showed For each sample, 50 normal unfermented seeds the effect of the genotype on fat content and melting were taken randomly from the fruits. The pulp was point. Lambert et al. (1999a), analyzing cacao Amazon extracted in a blender and dried to constant weight at genotypes, evidenced a great variability in fat content 105 ºC. The beans were then peeled and their cotyledons (49.1 to 59.4%). The studies further showed that sent to the Chemistry Department of the Federal commercial cacao from the Amazon region meets the University of Rondônia - UNIR, in Porto Velho, for chocolate industry’s standards (Ribeiro 1988, Ávila and laboratory analyses. Each sample was ground to Dias 1993). The present study aimed at a quantification homogenous cacao powder and thereafter dried at 105 of the diversity in germplasm from Rondônia for cacao ºC for 20 hours. A sub-sample of 5 g was digested in fat content and melting point. This information is crucial HCL 4 N solution, filtered and washed with warm water. for the generation of superior clonal varieties and as a The digested material was placed in a Soxhlet flask and guideline in genetic improvement of the species. subjected to extraction with petroleum ether under reflux for 5 hours in a Soxhlet extractor in three replications per sample. The results were expressed in percentage MATERIALS AND METHODS of fat of dry matter. The melting point was determined Clone Accessions by the open capillary method (BSI 1958), with three replications per sample. The evaluated clonal accessions were from the cacao genebank Experimental Station ESTEX (lat 10º 44’ S, long Statistical analyses 62º 13’ W, and alt 280 m asl), in Ouro Preto do Oeste, The two evaluated characteristics were analyzed Rondonia. According to the Köppen classification, for normal distribution. The existing slight deviations Rondonia has a rainy tropical climate of the Aw type–with from normality would not justify the use of mean temperatures above 18 ºC in the coldest month and a transformation functions. Consequently, the mean well-defined dry period. The region of Ouro Preto do Oeste original data were subjected to analyses of variance in presents mean annual precipitation values of 1940 mm, a a factorial scheme, considering the interaction clones x mean temperature of 24.6 ºC and relative air humidity of periods as error. The accession means were compared 81%. The soil is predominantly podzolic mesotrophic and by the Scott-Knott test (Scott-Knott1974). This kind of of intermediate natural fertility. mean test allows to analyse a great number of treatment Of the 615 accessions established, 207 were and clustered them in groups of means similar selected for fruit and seed characterization Almeida et. statistically. Analyses of principal components were also al (2005). One hundred and two of these accessions performed with the mean original data. A plot was were evaluated in the present study, of which 2 were constructed with the two components for the multivariate accessions of series BE, 81 of series CAB, 14 of series analysis of diversity Dias (2006). All analyses were run on EEOP, 2 of series MA, 2 of series POUND and IMC 67 softwares SAS (1989) and Statistica (1997). (Table 1). The criterion used for the selection of these accessions was the presence of ripe and healthy fruits, that is yellow and free-diseased synthom fruits, for the RESULTS AND DISCUSSION characterization of defined periods regarding the water Highly significant (P<0.01) differences were availability: April/May – period in which fruits detected among the 102 evaluated clones for fat content 142 Crop Breeding and Applied Biotechnology 8: 141-148, 2008 Diversity of cacao fat in germplasm from Rondônia, Brazil Table 1. Code and origin of the accessions Code Origin BE Belém, district. Accessions collected in 1965 in the surroundings of Belém, PA CAB Cacao from the Brazilian Amazon: i) CAB 8 to 13 - accessions collected in 1976 in the Ji-Paraná river basin amongst wild populations on the Muqui farm, Presidente Médici, RO; ii) CAB 35 to 42 - accessions collected in the region of Santarém, PA; iii) CAB 218 to 411 - accessions collected in 1981 and 82 in the Jamari river basin, amongst wild populations in the district of Ariquemes, RO; iv) CAB 417 to 473, accessions collected in 1983 in the Ji-Paraná river basin, amongst wild populations in the districts of Jaru, Ouro Preto do Oeste, Ji-Paraná, and Mirante da Serra, RO EEOP Experimental station in Ouro Preto. Accessions selected in 1977, on a plantation of hybrid varieties in José Wensing’ farm, Ouro Preto do Oeste, RO, considering the criterion witches’ broom tolerance IMC Iquitos Mixed Calabacillo. Accession selected in 1938 by JF Pound, in Iquitos, Peru MA Manaus, district. Accessions collected in 1965 on the Careiro island, Careiro, AM POUND Family name of JF Pound. Accessions selected in 1943 by JF Pound in Iquitos, Peru and melting point collected in the period April/May 2003, Table 2. Summary of the analyses of variance for fat content and melting point in 102 cacao accessions, evaluated in April/May indicating a significant diversity in the accessions (Table 2003 2) The coefficients of variation (CV) presented very low Mean Squares values (0.21 and 0.22% for fat content and melting point, Sources of df Fat Melting respectively), as expected in a laboratory study under variation content (%) point (oC) uniform experimental conditions.