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AGRIVITA Journal of Agricultural Science 40 AGRIVITA Journal of Agricultural Science. 2019. 41(1): 40-47 AGRIVITA Journal of Agricultural Science www.agrivita.ub.ac.id Betacyanin and Growth of Beetroot (Beta vulgaris L.) in Response to Nitrogen Fertilization in a Tropical Condition S. M. Sitompul*) and Ajrina Puspita Zulfati Faculty of Agriculture, Universitas Brawijaya, Malang, East Java, Indonesia ARTICLE INFO ABSTRACT Keywords: A series of studies was carried out to explore the possibility of beetroot cul- Beetroots tivation at highlands in the tropics. This was initiated by a study on the effect Betacyanin of temperature and duration of beetroot storage on betacyanin content after Nitrogen determination of a wavelength that giving the maximum absorbance of beta- Wavelength cyanin extracts from beetroots. The second study was designed to exam- ine the response of betacyanin and plant growth to nitrogen (N) fertilization. Article History: A randomized block design was used to impose the treatment of N fertilizer Received: 24 October, 2018 (Urea) consisting of 0, 0.15, 0.30, 0.45, and 0.60 g N/plant with five repli- Accepted: 10 January, 2019 cates. The maximum absorbance of betacyanin extracts from fresh beet- roots was attained at the wavelength of 536 nm. Betacyanin content of beet- *) Corresponding author: roots increased from 229 ml/l to 639 ml/l during 7 days storage under room E-mail: [email protected] temperature (± 22 0C), and was declined after slight increases when stored at lower temperatures. Nitrogen fertilization with the dosage of 0.6 g N/plant reduced betacyanin content up to 25 % from 351.5 to 257.6 ml/l, while root yield and total dry weight (TDW) increased with the supply of N fertilizer. INTRODUCTION the beetroots. Beetroots contain compounds exceptionally rich in antioxidants (Clifford, Howatson, Beetroot (Beta vulgaris L.), also known West, & Stevenson, 2015) that capable of inhibiting as red beet, is a cool-weather crop that requires the oxidation of other substances such as blood temperatures between 15-20 oC for optimum vessel walls, protein molecules, carbohydrates, growth (ARC, 2013). Although leaves are edible DNA and lipids (Lobo, Patil, Phatak, & Chandra, and rich in nutrition, this vegetable crop is planted 2010; Yashin, Yashin, Wang, & Nemzer, 2013). for taproots (beetroots) particularly in large The antioxidants, inhibiting the process of oxidation scales as the major yield that are processed into through a set of different reactions, are different various products including pickles. In temperate from antiradicals that react directly with free radicals areas where this crop is widely cultivated, the so that molecules are protected from damage best planting seasons are during spring and (Lobo, Patil, Phatak, & Chandra, 2010; Tirzitis & autumn. The plant growth is highly affected by Bartosz, 2010). However, antioxidant or antiradical environmental conditions. A reduction in yield and activity is a general term used to measure the ability beetroot quality (flavor and texture) was observed to prevent the oxidation of molecules (Halliwell, to occur in addition to growth retardation especially Aeschbach, Löliger, & Aruoma, 1995; Khan, 2016). during prolong high temperature (ARC, 2013). The antioxidant capacity of beetroot juices, In recent years, the beetroots have attracted measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH•) increasing attention due to several reasons inter alia and ferric reducing antioxidant power (FRAP), a global trend in functional foods (Lau, Chan, Tan, was far above that of commonly vegetable juices & Kwek, 2013; Vicentini, Liberatore, & Mastrocola, such as tomato, carrot, and orange juices (Clifford, 2016) and medicinal herbs (Chandrika, Rao, Howatson, West, & Stevenson, 2015; Ryan & Sirisha, & Chandra, 2016) in addition to a better Prescott, 2010; Wootton-Beard, Moran, & Ryan, understanding of the health benefit of antioxidants, 2011). The dominant antioxidants in beetroots, and the presence of powerful antioxidants in ISSN: 0126-0537 Accredited First Grade by Ministry of Research, Technology and Higher Education of The Republic of Indonesia, Decree No: 30/E/KPT/2018 Cite this as: Sitompul, S. M., & Zulfati, A. P. (2019). Betacyanin and growth of beetroot (Beta vulgaris L.) in response to nitrogen fertilization in a tropical condition. AGRIVITA Journal of Agricultural Science, 41(1), 40-47 . https:// doi.org/10.17503/agrivita.v41i1.2050 41 S.M. Sitompul and Ajrina Puspita Zulfati: Betacyanin and Growth of Beetroot in Response to Nitrogen........................... based on analysis using an HPLC (high performance synthesized from amino acid tyrosine. Wang, liquid chromatography) and a spectrophotometer, Lopez-Nieves, Goldman, & Maeda (2017) carried were betalains that consist of 80 % of total pigments out comparative analyses of betalains and their in beetroots (Attia, Moussa, & Sheashea, 2013). precursor, tyrosine, in various beet genotypes to The value was equivalent to 3.80 mg/g fresh identify possible bottlenecks in betalain production. weight of betalains which is within the range of It was found that increased tyrosine levels were 2.50-8.50 mg/g fresh weight reported previously positively correlated with elevated betalain (Zakharova & Petrova, 1997). Betalains, found only accumulation among red but not yellow genotypes. in several families in the order of Caryophyllales, A hypothesis was then proposed that further are a large group of pigments classified as increase in tyrosine production will likely enhance yellow betaxanthins and red-violet betacyanin betacyanin accumulation in red beets (Wang, (Clifford, Howatson, West, & Stevenson, 2015). Lopez-Nieves, Goldman, & Maeda, 2017). In term Many studies have characterized betalains of the important role of protein Tyr phosphorylation as nitrogen-containing water-soluble compounds in many plant processes particularly developmental, derived from amino acid tyrosine and deposited in abiotic and biotic stress responses (Shankar, vacuoles, in term of quantity, antioxidant activity and Agrawal, Sharma, Pandey, & Pandey, 2015), it is other characteristics (Gandía-Herrero, Escribano, likely that an increase in nitrogen supply is followed & García-Carmona, 2016; Khan, 2016). In fact, by an increase in the level of tyrosine in plants. the antiradical activity of betalains characterized The present study was aimed at exploring for the first time in 1998 has led to a renewed a possibility of growing beetroot plants in the interest in beetroots and betalains (Khan, 2016). tropics on areas of high altitudes. The growth Studies conducted thereafter showed a correlation performance of plants, including yield (taproots), between radical−scavenging activity (RSA) and and betacyanin content were used as indicators. betalain content of crude plant extracts (Vulić et The study was designed especially to investigate al., 2014). Other study involving seven different the response of betacyanins and plant growth beetroot varieties cultivated in Upper Austria including yield to nitrogen fertilization. No studies, showed that betalain content, ranging between so far, have been conducted on the performance 0.8 and 1.3 g/l fresh juice, consisted 60 % of beetroots in the tropics particularly in relation betacyanins and 40 % betaxanthins (Wruss et al., to nitrogen fertilization. An initial study was 2015). Up to now, 75 betalains (32 betaxanthins, conducted on the spectrophotometric quantification and 43 betacyanins including betalamic acid of betacyanin involving the storage of beetroots. and betanidin) have been well-characterized MATERIALS AND METHODS (Khan, 2016). Betanin is the major component of betacyanin and may account for 75-95 % of the Study Sites red pigment (Chandran, Nisha, Singhal, & Pandit, A series of studies was carried out in 2016- 2014; Von Ei Be, Sy, Maing, & Gabelman, 1972). 2017 at the Laboratory of Plant Physiology, Faculty An effort to obtain fresh plant materials has of Agriculture, Universitas Brawijaya (UB) in Malang prompted an interest in the cultivation of beetroots (7057’ S, 1120 36’), and at the Agrotechnopark of in the tropics at highlands. These needs decent UB in Cangar (7°44’ S & 112°32’ E). Malang and study due to the lack of information on this crop in Cangar (about 50 km westward of Malang), is the respected region. An intriguing question to study located around 500 and 1700 m asl with daily is related to the response of betacyanin to nitrogen minimum-maximum temperature of about 16- fertilizer that may be mediated by the growth 29 0C and 10-26 0C respectively. During the day, response of plants bringing about a reduction in the air temperature at Cangar was estimated betacyanin content of beetroots. This is based on around 20 0C (Sitompul, Sitawati, & Sugito, 2013). the synthesis of secondary metabolites in general Studies that is suppressed in conditions favorable for plant The first study investigated the wavelength growth (Akula & Ravishankar, 2011). In turn, a giving the maximum absorbance which was positive relationship would be expected between followed by the examination of temperature and betacyanin content and the rate of nitrogen fertilizer duration of tuber storage in relation to betacyanin if the nitrogen supply influences directly betacyanins Copyright © 2019 Universitas Brawijaya 42 S.M. Sitompul and Ajrina Puspita Zulfati: Betacyanin and Growth of Beetroot in Response to Nitrogen........................... content in a factorial experiment. Fresh beetroots & Noranizan, 2011; Wong & Siow, 2015). The (tubers) after cleaning as necessary,
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