| Nusantara Biosci | vol. 10 | no. 3 | pp. 137-202 | August 2018| | ISSN 2087-3948 | E-ISSN 2087-3956|

ISEA Journal of Bio l o g i c a l S c i e n c e s

The growth performances and the gut health parameters of Sentul chickens supplemented 137-141 with various dosage of turmeric powder INDRAWATI Y. ASMARA, TUTI WIDJASTUTI, IWAN SETIAWAN, ABUN, RUHYAT PARTASASMITA Short Communication: Tetrazolium test for evaluating viability of Capsicum annum seeds 142-145 ADITYA KUSUMAWARDANA, BAMBANG PUJIASMANTO, PARDONO The effect of combination of indigenous phosphate solubilizing bacteria of , on 146-150 the available phosphorus and phosphorus uptake of soybean LUFITANUR ALFIAH, DELITA ZUL, NELVIA NELVIA Male gametophyte development steps in Pistacia vera L. 151-158 ELAHE SADEGHIRAD, AHMAD MAJD, ALIREZA IRANBAKHSH, AMANOLLAH JAVANSHAH Antibacterial potency of simple fractions of ethyl acetate extract of Begonia baliensis 159-163 HARTUTININGSIH-M. SIREGAR, R.S. PURWANTORO, PRAPTIWI, A. AGUSTA Total phenolic content and antioxidant activity of ginger extract and SNEDDS with eel fish 164-169 bone oil (Anguilla spp.) IMAS IFRIAN WIJAYANTI, AGUNG BUDIHARJO, ARTINI PANGASTUTI, FEA PRIHAPSARA, ANIF NUR ARTANTI The potential of agroforestry as an adaptation strategy to mitigate the impacts of climate 170-177 change: A case study of Kiine Community, Kenya MUNENE ANNE NYARUAI, JOHN K. MUSINGI, BONIFACE N. WAMBUA Influence of NAA and coconut water with variation of soaking duration on the growth of 178-182 yellow bamboo branch cutting TIA SETIAWATI, AGINTA PUTRI REHULINA KELIAT, RULY BUDIONO, RUHYAT PARTASASMITA, JOHAN ISKANDAR Effect of sucrose and plant growth regulators on callogenesis and preliminary secondary 183-192 metabolic of different explant Myrmecodia tuberosa YANTI PUSPITA SARI, EKO KUSUMAWATI, CHAIRUL SALEH, WAWAN KUSTIAWAN, SUKARTINGSIH Incorporation of dietary palm date pits in all-male nile tilapia (Oreochromis niloticus ) diets 193-202 NEDAL MOHAMMED SIDDIG SWAR, ASAAD HASSAN WIDAA MOHAMED

Society for Indonesian Biodiversity

Sebelas Maret University Surakarta

ISSN 2087-3948 E-ISSN 2087-3956 Published quarterly PRINTED IN INDONESIA

| Nusantara Biosci | vol. 10 | no. 3 | pp. 137-202 | August 2018 |

| ISSN 2087-3948 | E-ISSN 2087-3956|

Rizaldo Arbet Rizaldo

photoby

Sentulchicken of Balitnak

| Nusantara Biosci | vol. 10 | no. 1 | pp. 137-202 |August 2018 | |ISSN 2087-3948| E-ISSN 2087-3956| ISEA Journal of Bio l o g i c a l S c i e n c e s

EDITORIAL BOARD: Editor-in-Chief, Sugiyarto, Surakarta, Indonesia ([email protected])

Editorial Advisory Boards: Agricultural Sciences, Muhammad Sarjan, Mataram University, Mataram, Indonesia ([email protected]) Agricultural Sciences, Dragan Znidarcic, University of Ljubljana, Slovenia, EU ([email protected]) Animal Sciences, Freddy Pattiselanno, State University of , , Indonesia ([email protected]) Biochemistry and Pharmacology, Mahendra K. Rai, SGB Amravati University, Amravati, India ([email protected]) Biochemistry, Vinod K. Sangwan, Eternal University, Baru Sahib (Sirmour), India ([email protected]) Bioinformatics and Computational Biology, Guojun Li, University of Georgia, Athens, USA ([email protected]) Biomedical Sciences, Afiono AgungPrasetyo, Sebelas Maret University, Surakarta, Indonesia ([email protected]) Biomedical Sciences, Hui Yang, Guangzhou Medical University, Guangzhou, China ([email protected]) Bioremediation, Surajit Das, National Institute of Technology, Rourkela, India ([email protected]) Ecology and Environmental Science, Cecep Kusmana, Bogor Agricultural University, Bogor, Indonesia ([email protected]) Ethnobiology, Luchman Hakim, , Malang, Indonesia ([email protected]) Forestry, Rajesh Kumar, Rain Forest Research Institute, Assam, India ([email protected]) Genetics and Evolutionary Biology, Sutarno, Sebelas Maret University, Surakarta, Indonesia ([email protected]) Human Sciences, Yi Li, Texas A&M University-Kingsville, Kingsville, USA ([email protected]) Medicinal and Aromatic Plants, Khalid A.K. Ahmed, National Research Centre, Cairo, Egypt ([email protected]) Micology, Rajesh K. Gupta, Biologics Quality & Regulatory Consultants, LLC, North Potomac, USA ([email protected]) Molecular Biology, Darlina Md. Naim, Universiti Sains Malaysia, Minden, Malaysia ([email protected]) Microbiology, Kateryna Kon,Kharkiv National Medical University, Kharkiv, Ukraine ([email protected]) Microbiology, Román Ramírez, Universidad Pedagógica y Tecnológica de Colombia, Tunja, Colombia ([email protected]) Molecular Communication and Nanonetworks, Baris Atakan, Izmir Institute of Technology, İzmir, Türkiye ([email protected]) Parasitology (Immuno-parasitology), Hossein Nahrevanian, Pasteur Institute of Iran,Tehran, Iran ([email protected]) Plant Breeding and Biotechnology, Danial Kahrizi, Razi University, Kermanshah, Iran ([email protected]) Plant Physiology, Qingmei Guan, University of Maryland, College Park, Maryland, USA ([email protected]) Plant Physiology, Xiuyun Zhao, Huazhong Agricultural University, Wuhan, China ([email protected]) Plant Science, Muhammad M. Aslam, Kohat University of Science & Technology, Kohat, Pakistan ([email protected]) Plant Science, Pudji Widodo, General Soedirman University, Purwokerto, Indonesia ([email protected]) Toxicology, Shaukat Ali, University of Azad Jammu and Kashmir, Muzaffarabad, Pakistan ([email protected])

Management Boards: Managing Editor, Ahmad D. Setyawan, Sebelas Maret University, Surakarta, Indonesia ([email protected]) Associated Editor (English Editor), Muchdar Davis, NAIST Indonesia Office, Indonesia ([email protected]) Associated Editor (English Editor), Suranto, Sebelas Maret University, Surakarta, Indonesia ([email protected]) Technical Editor, Ari Pitoyo, Sebelas Maret University, Surakarta, Indonesia ([email protected]) Business Manager, A. Widiastuti, Development Agency for Seed Quality Testing of Food and Horticulture Crops, Depok, Indonesia ([email protected])

PUBLISHER: Society for Indonesian Biodiversity

CO-PUBLISHER: School of Graduates, Sebelas Maret University, Surakarta

FIRST PUBLISHED: 2009

ADDRESS: Bioscience Program, School of Graduates, Sebelas Maret University Jl. Ir. Sutami 36A Surakarta 57126, Indonesia. Tel. & Fax.: +62-271-663375, email: [email protected]

ONLINE: biosains.mipa.uns.ac.id/nusbioscience smujo.id/nb Society for Indonesia Sebelas Maret

Biodiversity University Surakarta

Guidance for Authors

Aims and Scope Nusantara Bioscience (Nusantara Biosci) encourages author(s); the corresponding authors detailed postage and e-mail submission of manuscripts dealing with all aspects of biological sciences addresses, and phone and fax numbers. that emphasize issues germane to biological and nature conservation, References Author-year citations are required. In the text give the authors including agriculture and forestry, animal science, biochemistry and name followed by the year of publication and arrange from oldest to pharmacology, biomedical science, ecology and environmental science, newest and from A to Z. In citing an article written by two authors, both ethnobiology, genetics and evolutionary biology, marine and freshwater of them should be mentioned, however, for three and more authors only biology, microbiology, molecular biology, physiology, and plant science. the first author is mentioned followed by et al., for example: Saharjo and Article types The journal seeks original full-length research papers, short Nurhayati (2006) or (Boonkerd 2003a, b, c; Sugiyarto 2004; El-Bana and research papers (short communication), reviews, monograph and letters to Nijs 2005; Balagadde et al. 2008; Webb et al. 2008). Extent citation as the editor about material previously published; especially for the research shown with word “cit” should be avoided. Reference to unpublished data conducted in the Islands of the Southeast Asian reign or Nusantara, but and personal communication should not appear in the list but should be also from around the world. cited in the text only (e.g., Rifai MA 2007, personal communication; Submission The journal only accepts online submission, through open Setyawan AD 2007, unpublished data). In the reference list, the references journal system (https://smujo.id/nb/about/submissions). The manuscript should be listed in an alphabetical order. Names of journals should be must be accompanied with a cover letter containing the article title, the abbreviated. Always use the standard abbreviation of a journal’s name first name and last name of all the authors, a paragraph describing the according to the ISSN List of Title Word Abbreviations (www.issn.org/2- claimed novelty of the findings versus current knowledge, and a list of 22661-LTWA-online.php). The following examples are for guidance. five suggested international reviewers (title, name, postage address, email Journal: address). Reviewers must not be subject to a conflict of interest involving Saharjo BH, Nurhayati AD. 2006. Domination and composition structure the author(s) or manuscript(s). The editor is not obligated to use any change at hemic peat natural regeneration following burning; a case reviewer suggested by the author(s). study in Pelalawan, Riau Province. Biodiversitas7: 154-158. Acceptance The acceptance of a paper implies that it has been reviewed The usage of “et al.” in long author lists will also be accepted: and recommended by at least two reviewers, one of whom is usually from Smith J, Jones M Jr, Houghton L et al. 1999. Future of health insurance. N the Editorial Advisory Board. Authors will generally be notified of Engl J Med 965: 325–329 acceptance, rejection, or need for revision within 1 to 2 months of receipt. Article by DOI: Manuscript is rejected, if the content is not in line with the journal scope, Slifka MK, Whitton JL. 2000. Clinical implications of dysregulated does not meet the ethical standards (i.e. false authorship, plagiarism, cytokine production. J Mol Med. Doi:10.1007/s001090000086 duplicate publication, fabrication of data and citation manipulation),does not meet the required quality, written in inappropriate format, has Book: incorrect grammar, or ignores correspondence in three months. The Rai MK, Carpinella C. 2006. Naturally occurring bioactive compounds. primary criteria for publication are scientific quality and biological or Elsevier, Amsterdam. natural conservation significance. The accepted papers will be published Book Chapter: in a chronological order. This journal is published in May and November. Webb CO, Cannon CH, Davies SJ. 2008. Ecological organization, However, online publication was done previously. biogeography, and the phylogenetic structure of rainforest tree Ethics Author(s) must obedient to the law and/or ethics in treating the communities. In: Carson W, Schnitzer S (eds).Tropical Forest object of research and pay attention to the legality of material sources and Community Ecology. Wiley-Blackwell, New York. intellectual property rights. Author(s) needs to adhere to the ethical Abstract: standards as determined by the Committee on Publication Ethics (COPE). Assaeed AM. 2007. Seed production and dispersal of Rhazya stricta. The th Copyright Submission of a manuscript implies that the submitted work 50 AnnualSymposium of the International Association for Vegetation has not been published before (except as part of a thesis or report, or Science, Swansea, UK, 23-27 July 2007. abstract); that it is not under consideration for publication elsewhere; that Proceeding: its publication has been approved by all co-authors. If and when the Alikodra HS. 2000. Biodiversity for development of local autonomous manuscript is accepted for publication, the author(s) agree to transfer government. In: Setyawan AD, Sutarno (eds).Toward Mount Lawu copyright of the accepted manuscript to Nusantara Bioscience. Authors National Park; Proceeding of National Seminary and Workshop on shall no longer be allowed to publish manuscript without permission. Biodiversity Conservation to Protect and Save Germplasm in Authors or others are allowed to multiply article as long as not for Island. Sebelas Maret University, Surakarta, 17-20 July 2000. commercial purposes. For the new invention, authors are suggested to [Indonesian] manage its patent before published. Thesis, Dissertation: Open access The journal is committed to free-open access that does not Sugiyarto. 2004. Soil Macro-invertebrates Diversity and Inter-cropping charge readers or their institutions for access. Users are entitled to read, Plants Productivity in Agroforestry System based on Sengon. download, copy, distribute, print, search, or link to the full texts of [Dissertation]. Brawijaya University, Malang. [Indonesian] articles, as long as not for commercial purposes. Online document: Preparing the Manuscript Balagadde FK, Song H, Ozaki J, Collins CH, Barnet M, Arnold FH, Quake SR, You L. 2008.A synthetic Escherichia coli Please make sure before submitting that: The manuscript is proofread predator-prey ecosystem. Mol Syst Biol 4 : several times by the author(s); and is criticized by some colleagues. The 187.www.molecularsystemsbiology.com language is revised by a professional science editor or a native English speaker. The structure of the manuscript follows the guidelines (sections, Tables should be numbered consecutively and accompanied by a title at references, quality of the figures, etc). Abstract provides a clear view of the top. Illustrations Do not use figures that duplicate matter in tables. the content of the paper and attracts potential citers. The number of cited Figures can be supplied in digital format, or photographs and drawings, references complies with the limits set by Nusantara Biosci (around 20 for which can be ready for reproduction. Label each figure with figure research papers). All manuscripts must be written in clear and number consecutively. grammatically correct English (U.S.). Uncorrected proofs will be sent to the corresponding author by e-mail as Microsoft Word files are required for all manuscripts. The manuscript .doc or .rtf files for checking and correcting of typographical errors. To should be as short as possible, and no longer than 8000 words (except for avoid delay in publication, proofs should be returned in 7 days. review), with the abstract 200 words. For research paper, the manuscript A charge There is free of charge for members of Society for Indonesian should be arranged in the following sections and appear in order: Title, Biodiversity (SIB) and/or non-Indonesian author(s). But, the cost of each Abstract, Key words (arranged from A to Z), Running title (heading), manuscript is USD 250 for Indonesian non-SIB members (valid for Introduction, Materials and Methods, Results and Discussion, Conclusion, manuscript submissions starting on January 1, 2016). Acknowledgements, and References. Scientific language, nomenclature Reprints The sample journal reprint is only available by special request. and standard international units should be used. The title page should Additional copies may be purchased when ordering by sending back the include: title of the article, full name(s), institution(s) and address(es) of uncorrected proofs by e-mail. NOTIFICATION: All communications are strongly recommended to be undertaken through email.

THIS PAGE INTENTIONALLY LEFT BLANK

NUSANTARA BIOSCIENCE ISSN: 2087-3948 Vol. 10, No. 3, pp. 146-150 E-ISSN: 2087-3956 August 2018 DOI: 10.13057/nusbiosci/n100303

The effect of combination of indigenous phosphate solubilizing bacteria of Riau, Indonesia on the available phosphorus and phosphorus uptake of soybean

LUFITANUR ALFIAH1,♥, DELITA ZUL2, NELVIA NELVIA3 1Agrotechnology Department, Faculty of Agriculture, Universitas Pasir Pengaraian, Pasir Pengaraian, Rokan Hulu, Riau, Indonesia, 28557 ♥email: [email protected], Tel.: +62- 762-7392272 2Biology Department, Faculty of Mathematics and Natural Sciences, Universitas Riau, , Indonesia, 28293 3Agrotechnology Department, Faculty of Agriculture, Universitas Riau, Pekanbaru, Riau, Indonesia, 28293

Manuscript received: 20 April 2016. Revision accepted: 1 August 2018.

Abstract. Alfiah L, Zul D, Nelvia N. 2018. The effect of combination of indigenous phosphate solubilizing bacteria of Riau, Indonesia on the available phosphorus and phosphorus uptake of soybean. Nusantara Bioscience 10: 146-150. Despite the abundant amount of phosphorus (P) in the soil, P uptake by the plants is very limited. In acidic soil, phosphorus (P) is bound to aluminum (Al) and iron (Fe), whereas in the alkaline soil, phosphorus (P) is bound to calcium (Ca). The improvement of efficiency and availability of P to plants can be made by utilizing a group of solubilizing phosphate microorganisms. Potency test to investigate the P solubility by Phosphate Solubilizing Bacteria (PSB) has been conducted by isolating the bacteria from peat soil at Biosphere Reserves of Giam Siak Kecil Bukit Batu, Riau, Indonesia. The semi-quantitative test revealed that the PSB were able to dissolve Ca3 (PO4)2, FePo4 and phosphate rock. However, the adaptation ability and potency of PSB from indigenous Riau peat soil inoculated into soybean (Glycine max L. Merr) plants in the mineral soil have not yet been investigated. The present study was carried out from March to June 2015 on the alluvial soil in Babussalam Village, Rambah Sub-district, Rokan Hulu District, Riau. The aim of this study was to determine the effect of PSB inoculation on bacterial population and phosphatase activity. The study also aimed to determine the available P and P uptake and their impact on soybean growth and production. The study employed a factorial experiment laid out in a completely randomized design (CRD) consisted of two factors, i.e., soil treatment and PSB. The first factor comprised of two levels, i.e., T0: non-sterilized soil, T1: sterilized soil. The second factor consisted of 4 levels, i.e., B0: without PSB inoculation, B1: inoculation using 2 isolates of PSB (BB_UB6 and BB_K9), B2: inoculation using 3 isolates of PSB (BB_UB6, BB_K9 and BB_K2), and B3: inoculation using 4 isolates of PSB (BB_UB6, BB_K9, BB_K2, and BB_HS13). The results showed that inoculation of starter 3 had the highest phosphatase activity rate of 12.43 μg p NP g-1 hour-1. The highest available P was produced by starter 2, while the P uptake on non-sterilized soil was higher than that on the sterilized soil at 2.63 mg plant-1. PSB inoculation and soil sterilization did not significantly affect the population of phosphate solubilizing bacteria.

Keywords: Available P, P uptake, phosphate solubilizing bacteria, soybean

INTRODUCTION substance such as IAA and gibberellins acid that can increase plant growth. Phosphate availability remains the main issue in the Potency test to examine the P solubility by PSB isolated cultivation of soybean (Glycine max L. Merr) plants. from peat soil at Biosphere Reserves of Giam Siak Kecil Available P in the soil is abundant, but the concentration of Bukit Batu, Riau, Indonesia has been conducted using a P that can be absorbed by plants is very low. The semi-quantitative approach resulted that the PSB were able concentration of available P in the soil is affected by the to dissolve Ca3 (PO4)2, FePO4 and phosphate rock. soil pH and soil type. In acidic soil, P is bound to However, a study focusing on the ability of adaptation and aluminum (Al) and iron (Fe), whereas in the alkaline soil, P the potency of PSB originated from indigenous Riau peat is bound to calcium (Ca) and magnesium (Mg). The soil and inoculated into soybean plants in the mineral soil presence of these bonds causes P to turn into chelate have not been investigated yet. Thus, this study aimed at compounds absorbed with soil colloids; therefore, P is not determining the effect of combined PSB isolates in available for plants. The provision of P fertilizer becomes improving available P and P uptake of soybean plants. less effective and efficient because a significant amount of P is bound through the absorption process. One possible effort to solve this problem is by using microorganisms MATERIALS AND METHODS from the phosphate solubilizing bacterial group (PSB) which can help release the P bond to make it available to This study was performed in two stages, namely the plants. The release of P bond is facilitated by the laboratory work and field work during March to June 2015. production of organic acids and phosphatase enzymes by Laboratory work was conducted in the Microbiology PSB. Additionally, PSB also produces growth promoting Laboratory of Biology Department, Faculty of Mathematics ALFIAH et al. – Effect of phosphate solubilizing bacteria on soybean 147 and Natural Sciences and Soil Laboratory of Faculty of polybag-1. After one week, the growing media was ready to Agriculture, , Pekanbaru, Indonesia. be used. Field research was carried out on the alluvial soil in Babussalam Village, Rambah Sub-district, Rokan Hulu, Sterilization of seed surface, pre-germination, and District, Riau Province, Indonesia. inoculation The study employed a factorial experiment laid out in a The seeds were surfaced sterilized following the completely randomized design (CRD) consisted of two method of Hallmann et al. (1997). The soybean seeds were factors, where the first factor, the soil treatment included washed, and then immersed in 70% alcohol for one minute. two levels, i.e., T0: non-sterilized soil, T1: sterilized soil, The seeds were then immersed in NaOCl 2.5% for eight while the second factor, PSB inoculation, consisted of 4 minutes and rinsed with sterile water for three times. The levels, i.e., B0: without PSB inoculation, B1: inoculation sterilized seeds were then pre-germinated by placing the using 2 isolates of PSB (BB_UB6 and BB_K9), B2: seeds on the wet cotton until the seeds ruptured and inoculation using 3 isolates of PSB (BB_UB6, BB_K9 and germinated. Germinated seeds were then soaked with PSB BB_K2), and B3: inoculation using 4 isolates of PSB starter according to treatment for 2 hours, after which the (BB_UB6, BB_K9, BB_K2, and BB_HS13). seeds were ready to be planted.

Preparation of PSB starter Planting and maintenance Starter 1 (a mixture of 2 PSB isolates): Starter 1 was The soybean seeds were sown by making planting holes made from two PSB isolates (BB_UB6 and BB_K9) by on the prepared media in a three cm depth. Each hole was inoculating one ose of each bacterial isolate into 10 ml of then filled with 2 seeds of soybean. After the seeds grow, liquid Pikovskaya medium, which then incubated at room thinning was done and 1 plant polybag-1 was retained up to temperature for 24 hours with 150 rpm agitation. After the harvest. Polybag arranged with a distance of 40 cm x 40 incubation period, 10 ml of each PSB inoculum was cm. Fertilizer was added at a dose of 0.125 g urea polybag-1 inoculated into an Erlenmeyer containing 40 ml of liquid and 0.5 g KCl polybag-1 was done at 20-30 days after planting. Pikovskaya. The culture was then incubated for 24 hours at room temperature with 150 rpm agitation. After the Data analysis incubation period, the two inoculum were mixed so as to Observations were made to determine the bacterial produce inoculum 1 in 100 ml volume consisting of a population, phosphatase activity rate, soil available P, and mixture of 2 isolates with a ratio of 1: 1. P uptake. Obta8ned data were subjected to Analysis of Starter 2 (a mixture of 3 PSB isolates): Starter 2 was Variance (ANOVA) using SAS 9.1 program. The treatment made of three PSB isolates (BB_UB6, BB_K9, and means were separated using a Duncan Multiple Range Test BB_K2) by inoculating one ose of each bacterial isolate at  5% significance level. into 10 ml of liquid Pikovskaya medium, then was incubated at room temperature for 24 hours with 150 rpm agitation. After the incubation period, 5 ml of each PSB inoculum RESULTS AND DISCUSSION was inoculated into Erlenmeyer containing 45 ml of liquid Pikovskaya medium. The culture was then incubated for 24 PSB population tended to increase by the inoculations hours at room temperature with 150 rpm agitation. After using starter 1, 2, and 3 as compared to no inoculation the incubation period, the three inoculum were mixed so as treatment both in sterilized and non-sterilized soils. PSB to produce Starter 2 in 120 ml volume consisting of a population in non-sterilized soil tended to be two times mixture of 3 PSB isolates with a ratio of 1: 1: 1. higher than that in the sterilized soil. PSB population in Starter 3 (a mixture of 4 isolate of PSB): Starter 3 was PSB inoculation using starter 1 tended to be higher, increased made following the same procedures of starter 1 and starter by nine times of the control treatment (Figure 1). 2, except that at the end of the inoculum preparation process, is the four PSB inoculum were mixed so as to produce Starter 3 in 100 ml volume consisting of 4 PSB isolates with a ratio of 1: 1: 1: 1.

Preparation of growing media Topsoil was hoed for approximately 30 cm depth and then collected. The composite soil sample was then mashed using a shovel and sieved. The non-sterilized soil was put into the polybags as much as 5 kg polybag-1. The sterilized soil was prepared by employing the Tyndallization method, 0 by gradually evaporating soil at 100 C for 1 hour and repeating the same process three times within 24 hours. Starter 1 = Mixture of 2 PSB isolates (BB_UB6 and BB_K9) The non-sterilized and sterilized soils were then let sit for Starter 2 = Mixture of 3 PSB isolates(BB_UB6, BB_K9 and BB_K2) Starter 3 = Mixture of 4 PSB isolates(BB_UB6, BB_K9, BB_K2 and BB_HS13) about one day and then added with phosphate rocks at a dose of 0.75 g polybag-1 and sterilized manure of 25 g Figure 1. The PSB population on non-sterilized and sterilized soil 30 days after inoculation

148 NUSANTARA BIOSCIENCE 10 (3): 146-150, August 2018

Sterilized and non-sterilized soil media influenced the the colony proved that the bacteria dissolved P through the variation of PSB population in each treatment. On non- production of phosphatase enzymes. The area of the clear sterilized soil, inoculated PSB synergized positively with zone qualitatively shows the level of bacteria's ability to indigenous soil bacteria; therefore, the population of PSB dissolve P from the insoluble phosphate (Rachmiati 1995). on this type of soil was more abundant than that on the The highest level of phosphatase activity in this sterilized soil. A study by Cahyani (2009) revealed that soil experiment was produced by inoculation treatment using sterilization significantly influenced the population of soil Starter 3 (12.43 μgpNP g-1 hour-1). The result of the present bacteria. Soil microbes affect one another resulting in very study is higher than that reported by Suliasih and Rahmat dynamic conditions in the rhizosphere (Lines-Kelly 2005). (2007), where the activity of inoculated PSB phosphatase Bacteria within a community convey their presence to each only ranged from 1.818 to 1.947 μg pNP g-1 hour-1. On the other by producing, detecting, and responding to small other hand, is the present results of phosphatase activity are signal molecules called autoinducers, and the process of lower than that reported by Tamad et al. (2013) at 30 intercommunication is called quorum sensing (Miller and μgpNP g-1 hour-1. Fitriatin et al. (2009) reported that the Bassler 2001). Quorum sensing enables the starter bacteria highest phosphatase activity was obtained in the and indigenous bacteria to interact positively, thereby inoculation of combined solubilizing phosphate bacteria causes the increase of bacterial activity. This study also with a phosphate solubilizing function of 201.50 μg pNP g- indicated that PSB originating from peat soil was able to 1 hour-1. adapt to the new environment on mineral soils. The tendency of PSB population increase may affect The PSB population found in this study is higher than the increase of phosphatase enzyme activity. The that obtained by Fitriatin et al. (2009), where the correlation between PSB population and phosphatase population was only 1.70 x 106. On the other hand, the enzyme activity is presented in Figure 2. present study results are lower than the results obtained by The presented scatter plot shows that there was a Lestari et al. (2011) which ranged at 41-72 x 108 at the end positive correlation between the two variables, which of the incubation period. Bacteria have the ability to grow means the higher population of bacteria; the higher is and adapt according to their growing conditions and can enzyme phosphatase activity. This condition proves that the utilize the nutrient sources contained in the substrate on the increase in PSB population will also increase phosphatase growth medium (Lestari et al. 2011). enzyme activity. This finding also indicates the occurrence of quorum sensing where the behavior of bacteria depends Phosphatase enzyme activity on the population. Bacteria, in high population, convey Soil sterilization has a significant effect on phosphatase their abilities, including phosphate solubilizing bacteria enzyme activity in soil (Table 1). Phosphatase enzyme (PSB) (DeAngelis 2006). This notion is also in line with activity on non-sterilized soil increased 1.5 times higher than that on the sterilized soil. These results indicate that Table 1. Phosphatase enzyme activity on non-sterilized and non-sterilized soil has higher phosphatase enzyme activity sterilized soils with PSB inoculation as a result of indigenous soil bacteria allowing the positive interaction among bacteria. Phosphatase enzyme activity (µg pNP g-1 hour-1) Mean phosphatase enzyme activity on sterilized soil Soil type Mean was found lower than on the non-sterilized soil. The soil Without Starter Starter Starter PSB 1 2 3 heating might have caused this during the sterilization a a a a a process which affected the phosphatase enzyme in the soil. Non-sterilized 8.72 13.60 9.87 16.43 12.15 Sterilized 5.80a 9.91a 8.32a 8.43a 8.11b Nelson and Cox (2000) suggested that during enzymatic c ab bc a Mean 7.26 11.75 9.09 12.43 reactions, the rise in temperature increases the kinetic Note: The numbers in the same rows and columns which is energy of reacting molecules; thereby it accelerates followed the same lowercase letter are not significantly different collisions among molecules. Collisions will facilitate the at α 5% DMRT. formation of substrate enzymes so that the product formed will be much more. At an optimum temperature, the collision between the enzyme and the substrate is very effective, so that the formation of the enzyme-substrate is easier and more products are formed. However, at a too high temperature, the collision will accelerate the damage to the formation of active enzyme groups (enzyme denaturation) in interacting with the substrate and the enzyme catalytic activity will decrease (Kilara and Harwalkar, 1996). Santosa (2009) also mentioned that soil sterilization has a significant effect on PSB activities. Phosphatase enzyme activity was found higher in PSB- inoculated growing media than that in non- inoculated growing media. This indicates that the PSB-inoculated growing media were capable of producing phosphatase Figure 2. The correlation between PSB population and enzymes. The formation of a clear zone (holozone) around phosphatase activity.

ALFIAH et al. – Effect of phosphate solubilizing bacteria on soybean 149

Table 2. Available P (ppm) and P uptake (mg.plant-1) on was the result of higher bacterial diversity. However, the sterilized and non-sterilized soils 30 days after inoculation role of indigenous microbes in providing soil P is insufficient to increase soil P uptake. The high available P Treatment found in this study was not in line with the high absorption Soil type Without Starter Starter Starter Mean of P by the soybean plants, which is indicated by the low PSB 1 2 3 mean P absorption by soybean plants due to sterilization -1 Available P (ppm) that ranged from 0.51 to 2.63 mg plant . Saidi (2002) Non-sterilized 20.23a 24.88a 36.94a 36.30a 29.59a stated that soil P is closely related to P uptake; thus the Sterilized 8.41a 26.34a 29.25a 22.49a 21.62a higher is the soil P, the higher is the P uptake in plants. Mean 14.32b 25.61ab 33.10a 29.40a Jones et al. (1991) suggested that P uptake is categorized as low in the range of 1.6 to 2.6 mg plant-1. P uptake (mg plant-1) The low absorption of P may be caused by the nutrient a a a a a Non-sterilized 1.90 1.52 0.69 6.42 2.63 limiting factor in the soil. The low content of sodium and Sterilized 0.51a 0.67a 0.51a 0.36a 0.51b a a a a magnesium in the soil before the study caused the Mean 1.21 1.10 0.60 3.39 inhibition of P absorption by plants. Besides, Mg is a Note: The numbers in the same rows and columns which is followed the same lowercase letter are not significantly different crucial nutrient that plays a role in the photosynthesis at α 5% DMR test. process. Syafii (2008) stated that Mg has a vital role in chlorophyll synthesis. The low content of Mg element may decrease the level of chlorophyll and, hence the photosynthesis rates; thus the photosynthesis products are the notion of Tamad et al. (2013) which stated that the PSB not optimally produced. The P absorption by soybean population was positively correlated with phosphatase plants in this study was lower than that reported by Lestari activity. Phosphatase enzyme activity in the soil is closely et al. (2011) (58.3 mg plant-1). We concluded from the related to microbial activity because microbial biomass is present study findings that PBS inoculation and soil the primary source of enzymes in the soil. Suliasih et al. sterilization can increase soil phosphatase activity and (2010) reported that phosphatase enzyme activity on available P. The results also indicate that the two variables tomato-growing soil increased in line with the increasing have no significant effect on bacterial population and P population of solubilizing phosphate bacteria. uptake of soybeans.

Available P PSB inoculation increased the available P of the soil as ACKNOWLEDGEMENTS much as twice higher than that without inoculation (Table 2). Starter 2 treatment produced the highest available P by The authors appreciate and thank Nia Sufika Indriani 33.10 ppm. However, the major effect of soil sterilization for the significant assistance provided to the author during and the interaction of the two treatments did not this research. significantly increase the soil available P. Non-sterilized soils tended to contain higher available P compared to sterilized soil. REFERENCES A significant increase of available P indicates that PSB from Bukit Batu peat soil are able to adapt well to mineral Cahyani VR. 2009. Influence of soil sterilization methods on nutrient soil in Rokan Hulu District. The increase in soil available P status, microbiota population, potential of mycorrhizal infection and in this study is much higher than the increase in available P plant growth. Soil Science 6 (1): 43-52 De Angelis KM. 2006. Microbial Community Ecology and Bacterial from the adding of PSB in Sei Garo (Lestari et al. 2011) at Quorum Sensing as Control Points in Rhizosphere Nitrogen Cycling. 16.2 ppm. Bacterial isolates have different adaptability to [Dissertation]. The University of California. Berkeley, CA. their newly grown environment. Widyati (2007) stated that Fitriatin BN, Joy B, Subroto T. 2009. The Influence of Organic the ability of bacteria in dissolving P depends on the Phosphorous Substrate on Phosphatase Activity of Soil Microbes, Proceeding International Seminar of Chemistry, Universitas process of their own isolate metabolism. PSB activity in Padjadjaran, Jatinangor, Indonesia, 30-31 Oktober 2008. dissolving P is highly dependent on soil temperature, soil Hallmann J, Quadt-Hallmann A, Mahaffee WF, Kloepper JW. 1997. moisture, pH, food supply and environmental conditions Bacterial endophytes in agricultural crops. J Microbiol 43(10): 895- during their growth (Widawati and Suliasih 2006). 914. Jones JB, Wolf B, Mills HA. 1991. Plant Analysis Handbook.Micro- Table 2 shows that the main effect of soil sterilization Macro Publishing, Inc. Georgia. USA. increased P uptake in soybean plants. It may be concluded Kilara A, Harwalkar VR. 1996. Denaturation. In Food Proteins: Properties that soybean plants were able to absorb P better in non- and Characterization. VCH Publishers. New York. sterilized soil than in sterilized soil. Available P in non- Lestari, Wahyu TM, Linda, Martina A. 2011. The ability of seigaro's phosphate solubilizing bacteria in provision of dissolved phosphate sterilized soil was five times higher than that in sterilized and its absorption on soybean plants. Biospecies 4 (2): 1-5. soil. However, inoculated PSB isolate mixture and Lines-Kelly R. 2005. Defend The Rhizosphere and Root Against treatment interaction did not significantly increase the P Pathogenic Microorganism. Department of Primary Industries, State uptake by soybean. of New South Wales, Sydney. Miller MB, Bassler BL. 2001. Quorum sensing in bacteria. Ann Rev This condition indicates the influence of indigenous soil Microbiol 55: 99-165. microbes in providing P nutrient to the non-sterilized soil Nelson DL, Cox MM. 2000. Lehninger Principles of Biochemistry. 3rd ed. Worth Publisher, New York.

150 NUSANTARA BIOSCIENCE 10 (3): 146-150, August 2018

Rachmiati Y. 1995. Phosphate solubilizing bacteria from plant rhizosphere Syafi S. 2008. Morphological and Physiological Response of Different and its ability in dissolving phosphate. Proceedings of the National Genotypes Seeds of Jatropha (Jatropha curcas L.) to Drought Stress. Congress VI HITI Volume 1. Land Use as a Spatial Planning Tool to [Thesis]. Bogor Agricultural University, Bogor. [Indonesia] Improve Human's Welfare, Soil Science Association of Indonesia. Tamad, Ma’as A, Radjagukguk B, Hanudin E, Widada J. 2013. Jakarta, 12-15 December 1995. Availability of phosphorus in andisol soil for maize (Zea mays L.) by Saidi D. 2002. Isolation and characterization of cellulolytic bacteria and phosphate solubilizing bacterial inoculum. J Agron. 41 (2): 112-117 phosphate solubilizer from andisol as biological fertilizer agent. Widawati S, Suliasih. 2006. Augmentation of potential Phosphate Habitat 8 (4): 201 - 211. Solubilizing Bacteria (PSB) for growth of spur caysim (Brasicaca Santosa E. 2009. The activity of Some Bacteria Solubilizing Phosphate ventis Oed.) in marginal land. Biodiversitas 7 (1): 10-14. Isolatesat Various Levels C-Organic in Ultisols Soil. Soil Research Widyati E. 2007. Utilization of sulfate-reducing bacteria for Institute. Bogor. bioremediation of coal mine former land. Biodiversitas 8 (4): 283- Suliasih R. 2007. Phosphatase activity and calcium phosphate dissolution 286. by some Phosphate Solubilizing Bacteria. Biodiversitas 8: 23-26.