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Effect of the use of Potassium Fertilizer on the Resistance and Growth of Tomato to Bacterial Wilt caused by Ralstonia solanacearum

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The research aims to study the effect of sources and doses of potassium fertilizer on the resistance and growth of tomato to bacterial wilt caused by Ralstoniasolanacearum. We conduct experiment in a screen house in Faculty of Agriculture, Islamic University Malang. The research is conducted experimentally using completely randomized block design (RAK) arranged in factorial with three repetition. There are six combinations of treatment. Factor I: source of potassium fertilizer, consists of two levels: KCl and K2SO4. Factor II:dose of K2O, consists of three levels: 50 kg ha-1, 100 kg ha-1 and 200 kg ha-1. The inoculation of Ralstonia solanacearum is conducted a week after transplanting. There is significant effect on the use of different sources and doses of potassium fertilizer. As whole, the use of potassium fertilizer originated from K2SO4 is better than that of KCl and the magnitude of the increase depends on dose applied. The best result indicates by treatment of the use of K2SO4 with dose of 200 kg ha-1 K2O that able to extend the incubation period of 6,27 days, decrease the attack level of 73,15%, increase the uptake of potassium and leaf chlorophyll of 4,58% and 7,17%, respectively,and increase root lignin of 3%, whereas total phenol is decreased of 27,27% compare to the use of KCl in the same dose.
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    International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017 http://dx.doi.org/10.22161/ijeab/2.4.89 ISSN: 2456-1878 www.ijeab.com Page  | 2265   Effect of the use of Potassium Fertilizer on the Resistance and Growth of Tomato to Bacterial Wilt caused by Ralstonia solanacearum   Anis Rosyidah 1  , Indiyah Murwani 2 , Bambang Siswadi 3 1,2  Agrotechnology Department of Faculty Agriculture - Islamic University of Malang 65144 East Java, Indonesia 3 Agribusiness Department of Faculty Agriculture - Islamic University of Malang 65144 East Java, Indonesia  Abstract   —  The research aims to study the effect of  sources and doses of potassium fertilizer on the resistance and growth of tomato to bacterial wilt caused by  Ralstoniasolanacearum. We conduct experiment in a  screen house in Faculty of Agriculture, Islamic University  Malang. The research is conducted experimentally using completely randomized block design (RAK) arranged in  factorial with three repetition. There are six combinations of treatment. Factor I: source of potassium fertilizer, consists of two levels: KCl and K  2 SO 4 . Factor II:dose of  K  2 O, consists of three levels: 50 kg ha -1  , 100 kg ha -1 and 200 kg ha -1 . The inoculation of Ralstonia solanacearum is conducted a week after transplanting. There is significant effect on the use of different sources and doses of  potassium fertilizer. As whole, the use of potassium  fertilizer srcinated from K  2 SO 4  is better than that of KCl and the magnitude of the increase depends on dose applied. The best result indicates by treatment of the use of K  2 SO 4 with dose of 200 kg ha -1  K  2 O that able to extend the incubation period of 6,27 days, decrease the attack level of 73,15%, increase the uptake of potassium and leaf chlorophyll of 4,58% and 7,17%, respectively,and increase root lignin of 3%, whereas total phenol is decreased of 27,27% compare to the use of KCl in the  same dose. Keywords  —   Potassium fertilizer, Source, Dose, Plant resistance, Ralstonia solanacearum. I.   INTRODUCTION Tomato is one of superior horticultural commodities in Indonesia and has a promising economic prospect. Therefore, it needs serious handling, especially, in terms of increasing its yield and fruit quality. The projection of national tomato demand for 2014-2019 is around 970.499  –   1,107,168 ton, whereas tomato product by 2013 was only 922,780 ton with average productivity of 16.61 t.ha -1  (Bureau of Statistics and Directorate General of Horticulture, 2014). One of obstacles in the low production of tomato is the occurrence of wilt disease caused by  Ralstonia  solanacearum  bacteria, which is a threat for hot climate areas or areas with warm rainy season. Result of field observation shows that the disease has caused the loss of fresh fruit in approximately 7.1  –   63.7% (Rosyidah et al.,  2014). It is unfavorable for farmers since the investment for production cost is high. Various efforts of controlling the disease have been conducted, such as the use of organic material from chicken manure (Rosyidah, A., 2012), the use of cabbage family as bio-fumigant, and the use of resistance variety (Rosyidah et al  ., 2014). Another effort is the use of  potassium fertilizer with appropriate source and dose. This effort is another alternative to increase plant resistance against environmentally friendly disease. Tomato plant absorbs large amount of potassium element in approximately 1-5 % of plant’s dry weight (Chen and Gabelman, 2000). Potassium plays important role in plant metabolism (Farhad et al  ., 2010), helps in the formation of protein, carbohydrate, enzyme activity, regulation of osmotic, water use efficiency, translocation of  photosynthate (McKenzie, 2001), stimulate the development of root and increase the size of fruit (Marsono and Sigit, 2001), and increase the transportation of sugar and acid to storage organ (Bernardi et al.,  2013). The application of potassium could increase the formation of thick lignin compound; therefore, wall cell will be stronger and able to protect plant from pathogen interference (Fageria et al  ., 2009). Potassium fertilizer mostly used in Indonesia is muriate of  potash (KCl) containing about 60% of K  2 O. Recently, however, there is a development in the use of potassium sulphate (K  2 SO 4 ). Some research found that the incident of disease is higher when potassium fertilizer used is srcinated from KCl, whereas potassium sulphate is  proven to improve some characteristics of quality of various vegetable products (Gunadi, 2009). At present, the role of potassium in increasing plant resistance, especially in tomato, has not been studied. The aim of the research is to study the effect of combination of source and dose of potassium fertilizer on    International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017 http://dx.doi.org/10.22161/ijeab/2.4.89 ISSN: 2456-1878 www.ijeab.com Page  | 2266   the resistance and growth of tomato toward bacteria wilt caused by  Ralstonia solanacearum . II.   MATERIALS AND METHODS The research was conducted experimentally in a screen house in Faculty of Agriculture, Islamic University Malang in April  –   July, 2016. The altitude of the location was 460 above sea level. Type of soil is loam. Air temperature is around 22.5 0 C  –   25.5 0 C with air humidity of 80% - 86%. The research was conducted experimentally using completely randomized block design (RAK) arranged in factorial with three repetition. There were six combinations of treatment. Factor I: source of potassium fertilizer, consisted of two levels: KCl and K  2 SO 4 . Factor II was dose of K  2 O, consisted of three levels: 50 kg ha -1 , 100 kg ha -1 and 200 kg ha -1 . Each treatment consisted of ten sample plants. Tomato seeds from Lentana variety were planted in seeding basin with media of soil + sand + compost that  previously sterilized with hot steam for 3 hours with ratio of 1:1:1. At the age of 10 days, plant’s seedlings were transferred to seedling glass, one seedling per glass. Growing media used weresoil:sand:organic material of chicken manure (C/N = 12) (ratio of 2:1:1) that  previously sterilized with hot steam for 3 hours. Growing media of 8 kg was put into a polybag. The transplanting of tomato seedling was conducted when the seedling has height of 10 cm and 4 leaves. Inorganic fertilizers applied were SP-36 and it was applied on 3 days after transplanting with dose of 150 kg/Ha and Urea on 7 days after transplanting with dose of 150 kg Ha -1 . The application of KCl and K  2 SO 4  fertilizers was conducted at the age of 7 days after transplanting with dose in accordance with the treatment. The isolate of  R. solanacearum  used was the result of isolation of tomato plant attacked by  R. solanacearum in Donowarih Village, Karangploso, Malang. The  purification isolation and propagation of  R. solanacearum  was conducted using media of TZC (2,3,5-triphenyl tetrazolium chloride). Population density for inoculation was 2.78x10 8  cfu.mL -1 measured with spectrophotometer in OD 600. Plants at the age of 1 week after transplanting were inoculated with  R. solanacearum  with concentration of 10 8 cfu/ml of 20 ml by wounding the plant roots using scalpel. Observation was conducted on: incubation period of the disease, level of disease attack (Sinaga, 2003), potassium uptake in leaves (through extraction using NH 4 Oac), root lignin (Acid detergent fiber method), total phenol (Folin-Denis method), level of leaf chlorophyll (SPAD) and  plant height. In order to see the influence of treatments on observation conducted, data of observation result was statistically analyzed based on analysis of variance (ANOVA) and followed by Least Significance Difference test in confidence level of 95% to see their significances. III.   RESULT ANDDISCUSSION The status of soil fertility used in the experiment is  presented in Table 1. The content of carbon (C), nitrogen (N), and C/N ratio is classified as low. Phosphor (P) and  potassium (K) are classified as medium and low, respectively, whereas, cation exchange capacity (KTK) is classified as medium.  Incubation period and level of attack of R. solanacearum disease Based on research result (Table 2), it can be seen that there was a significant interaction (p < 0.05) in the treatment of the use of potassium fertilizer sources and doses on incubation period of the disease and the attack level of bacterial wilt disease. Based on Table 2, it is known that the use of different  potassium fertilizer sources and doses resulted in different disease incubation period and level of attack. The incubation period of the disease was ranged from 16.21 to 25.74 days after inoculation. Level of attack of the disease was ranged from 6.22% to 12.21% at the age of 35 days after transplanting. The application of potassium fertilizer of K  2 SO 4  is better than those of KCl. It is estimated that it is due to the content of SO 4  in potassium sulphate fertilizer since one of the functions of sulfur (S) is to reduce the attack of the disease (Tisdale et al. , 1990). The higher the dose of potassium fertilizer (200 kg ha -1 K  2 O) applied, the longer the incubation period and the lower the attack level caused by  R. solanacearum  pathogen. It happens because the initial content of K element in soil used for the experiment was low. With the addition of  potassium of 200 kg ha -1  K  2 O in the treatment gives sufficient nutrient and good plant resistance. One of the function of K element is to improve plant resistance by strengthening plant tissues and thickening epidermic wall.  Nurhayati (2008) stated that potassium in plant plays role in the formation of protein and carbohydrate as well as in the increasing of resistance against pathogen.  Potassium uptake, root lignin and total phenol Research result shows that there was significant interaction (p < 0.05) in the treatment of the use of  potassium fertilizer sources and doses on potassium uptake, root lignin and total phenol (Tabel 3). Based on Table 3, it is known that the use of different  potassium fertilizer sources and doses resulted in different  potassium uptake, root lignin level and total phenol level of plant. The application of potassium fertilizer K  2 SO 4  was better than that of KCl. In addition, the higher the dose of potassium fertilizer (200 kg ha -1  K  2 O) applied,    International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017 http://dx.doi.org/10.22161/ijeab/2.4.89 ISSN: 2456-1878 www.ijeab.com Page  | 2267   the higher the leaf potassium intake and root lignin level and the lower the level of total phenol. Result of observation on the level of potassium uptake in leaves shows that bigger potassium uptake in leaves will increase availability status of potassium in plant organs. The sufficiency of potassium has function in increasing the status of plant defense to improve damage caused by  pathogen since plant is able to increase the strength of its cell wall. Hardter, R (2003) and Pervez, H et al  ., (2007) add that the sufficient level of potassium in plant could increa se the strength of paddy’s stem and stalk due to the increase of its resistance. It is also explained that plant stomata and lenticel work well if sufficient potassium is exist. When pathogen invaded the plant, stomata and lenticel have the ability to close quickly. Potassium is also able to improve the work of enzyme for plant metabolism. The sufficiency of potassium in plant will increase the synthesis of molecular compounds with high molecular weight (protein, starch, cellulose) thus decreasing the synthesis of molecular compounds with low molecular weight, such as: organic acid, amino acid, and amide in  plant tissues. It is the decrease of the synthesis of compounds with low molecular weight that able to increase plant resistance against pathogen infection (Marschner, P., 2012; Mengel, K., 2001). Potassium element also plays role in lignification of sclerenchyma tissue (Fageria et al  ., 2009). Therefore, the sufficiency of  potassium could increase the formation of thicker lignin compound; thus, cell wall is stronger and able to protect  plant from external disturbance. Observation on total phenol shows that the increasing of  potassium dose applied will decrease the level of total  phenol. In other words, the lower the doses of potassium fertilizer applied, the bigger the level of attack; therefore, a tendency of the increase in phenol compound level. The increase in phenol compound is the reaction of plant toward infection of  R. Solanacearum  pathogen and root wounding before pathogen inoculation. Pieterse et al  ., (2009) stated that the increase of plant resistance through SAR (Systemic Acquired Resistance) occurs after local  pathogen infection in plant; the infected plant, then, activates genes that play role in the resistance to produce chemical compounds for plant resistance, such as salicylate. When the plant has the resistance, it will be able to protect itself if another pathogen exists; thus,  pathogen infection will not be developed. According to Goodman et al.,  (1986), plant tissue infected by pathogen indicates a change in metabolic pattern, including, activating peroxide and other phenoloxidase enzymes. It is in line with Matern et al  . (1995) and De Ascensao et al.  (2003) stated that great phenol synthesis will occur if  plant is attacked by pathogen. Agrios (2005) stated that  pathogen microorganism causing mechanical and chemical damages will stimulate plant to produce toxin compound against pathogen (phytoalexin). Plants need  peroxide enzyme to produce resistance compounds, such as lignin, chitin, and various compounds that build cell wall (Hallman, 2001). Further, Bruce et al. (1989) stated that peroxide is another component in the initial response of plant to pathogen attack and plays key role in the  biosynthesis of lignin that limit the area of pathogen distribution.  Leaf chlorophyll and plant height Research result shows that there was significant interaction (p<0.05) on the treatment of the use of  potassium fertilizer sources and doses on the content of leaf chlorophyll. Regarding observation on final plant height, it shows no interaction between potassium fertilizer sources and doses tested (Figure 1A and 1B). Treatment of the application of K  2 SO 4 shows more chlorophyll content than that of KCl. It is likely due to the content of sulfur in K  2 SO 4  fertilizer. Sulfur is the main element in the formation of leaf chlorophyll that closely related to photosynthesis process and takes part in various metabolism reactions, such as carbohydrate, fat and  protein (Tisdale et al.,  1990). The increase in potassium fertilizer dose applied will increase chlorophyll content. It is due to the sufficiency of potassium in the plant that will increase the work of enzymes thus increasing the activation of plastid in leaf, synthesis of protein,  photosynthesis and stomata movement. It results in the increase in the production of leaf chlorophyll. The optimum availability of potassium in leaf will make the leaf to be more efficient in the photosynthesis and plant will be more resistance and tolerance. IV.   CONCLUSIONS In the research, we report the effect of the use of  potassium fertilizer on the resistance and growth of tomato to bacterial wilt caused by  R. solanacearum . The result can be summarized as follow: the effect of the interaction is significant in the component of resistance  but not in the observation on plant height. Treatment of K  2 SO 4  and doses of potassium fertilizer (200 kg ha -1 K  2 O) is the best as indicated by longer incubation period of 6.27 days, decrease in the level of attack of 73.15%, increase in potassium uptake and chlorophyll in leaf of 4.58% and 7.17%, respectively and the increase in root lignin of 3% and decrease in phenol of 27.27% compare to the use of KCl in the same dose. ACKNOWLEDGEMENTS The authors would like to thank the Directorate General of Ministry of Research, technology and higher education for the support for Competitive Research Grant, Agrotechnology Department of Faculty Agriculture -    International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017 http://dx.doi.org/10.22161/ijeab/2.4.89 ISSN: 2456-1878 www.ijeab.com Page  | 2268   Islamic University of Malang, Laboratory of Biology - Faculty of Science Brawijaya University and all parties who help the research. REFERENCES [1] Agrios, G.N. 2005. Pengantar Ilmu Penyakit Tumbuhan. Gajahmada University Press. Yogyakarta. 713 p. [2] Bruce, RJ., West CA. 1989. Elicitation of Lignin Biosynthesis and Isoperoxidase Activity by Pectic Fragments in Suspension Cultures of Cluster Bean. Plant Physiol. 91: 889-897 [3] BPS dan Dirjen Hortikultura, 2014. Produksi Tomat. www.bps.go.id. Diakses tanggal 25 Oktober 2015. [4] Chen, J., W.H. Gabelman. 2000. Morphological and Physiological Characteristics of Tomato Roots Assosiated with Potassium-acquisition Efficiency. Scientia Horticulturae 83:213-255. [5] De Ascensao, A.F.R.D.C. and Dubrey, I.A. 2003. Soluble and Wall-Bound Phenolic Polymers in  Musa acuminata Roots Exposed to Elicitors From  Fusarium oxysporum f.sp. cubens . Phytochemistry 63, 679-686. [6] Fageria, NK, M.P.B. Filho, and J.H.C. Da Costa. 2009. Potassium in The Use of Nutrients in Crops Plant. CRC Press Taylor & Francis Group, Boca Raton. London. New York. Pp.131-163 [7] Farhad, I.S.M., M.N. Islam, S. Hoque, and M.S.I. Bhuiyan. 2010. Role of Potassium and Sulphur on The Growth, Yield, and Oil Content of Soybean ( Glycine max  L.) Ac. J. Plant Sci. 3(2):99-103 [8] Gunadi, N. 2009. Kalium Sulfat dan Kalium Klorida Sebagai Sumber Pupuk Kalium pada Bawang Merah. J. Hortikultura. 19(2):174-185 [9] Hardter, R. Potassium and Biotic Stress of Plants. 2003. In Feed the Soil to Feed the People: The Role of Potash in Sustainable Agriculture . Johnston, A.E., Ed.. International Potash Institute: Basel, Switzerland. pp. 345  –  362. [10] McKenzie, R. 2001. Potassium Fertilizer Application in Crop Production. http://www.agric.gov.ab.ca/universalpages/ includes/docheader.map. [14 Maret 2015]. [11] Marschner. 2012. Mineral Nutrition of Higher Plants , 3rd ed.; Academic Press: London. pp. 178  –  189. [12] Matern, U., Grimmig, B., and Kneusel, R.E. 1995. Plant Cell Wall Reinforcement in the Disease-Resistance Response: Molecular Composition and Regulation. Canandian Journal of Botany 73, p. 511-S517. [13 Mengel, K. 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Pemanfaatan Bahan Organik dan Trichoderma harzianum  dalam Bentuk Tepung untuk Mengendalikan Layu Bakteri Ralstonia solanacearum pada Kentang ( Solanum tuberosum  L.). J.Primordia. 8(2):144-153 [19] Tisdale, S. L, Nelson W. L.,and Beaton J. D. 1990. Soil fertility and fertilizers 4 th Ed. Maxwell Mc milan Publishing. Singapura. 52  –   92 Table.1: Status of soil fertility used as growing media  pHC NC/NPKKTK    (H2O)....%....(Bray)me/100gme/100g5,40,980,138417,10,2922,38  Source: Laboratory of Soil, Faculty of Agriculture, University of Brawijaya, 2016
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