Translate Bery.docx

The Effect of IBA (Indole Butyric Acid) on The WPM Media (Wood Plant Media) to Induction of Sandalwood’s (Santalum album L.) Shoots of Planlet Root by In Vitro by : BERI ADIMAS ARYANTO GINTNG 12620020 INTRODUCTION Background The plant is one of the organisms that played an important role in human life. The role of the plant including food and clothing, as the Board. Allah said in surat al-Syu'ara (26): 7;

. ٍ‫ن ُك ٍِل َز ْوجٍ ك َِريم‬ ٍْ ‫ض َك ٍْم أ َ ْنبَتْنَا فِي َها ِم‬ ٍ ِ ‫أ َ َولَ ٍْم يَ َر ْوا ِإلَى ْاْل َ ْر‬ Meaning: do they not look at the Earth, how much is the number we grow on earth that a variety of herbs that are good? (QS ash-Syuara/26:7) Based on the word of Allah SWT in the Quran surat as-Syu'ara (26): 7, said Karim (ٍ‫)ك َِريم‬ means either/noble. As for the word al-karam in Arabic is al-fadl (virtue). These words refer to the word Anbatsna (‫ )أ َ ْن َبتْنَا‬which means to grow crops. God Almighty has a good crop of menumbuhan i.e. plants that grow in the fertile and beneficial (Katsir, 2004; al-Qurthubi, 2008). One of the plants is a good crop of sandalwood. Sandalwood (Santalum album L.) is the plants that grow naturally in Indonesia particularly in the region of East Nusa Tenggara (NTT). Sandalwood’s plant has a variety of benefits, namely wood and oil is widely used for ceremonies and funerals. While the oil is widely used in the world of pharmaceuticals, cosmetics and utilized as a perfume (Herawan, 2012). Sandalwood’s oil content can reach 10% of the weight of the root and 1.5%-2.10% of its weight or average ranged from 4.5%-6.25% (Rahayu, et al., 2002). Countries that export oil and sandalwood are the European countries, the USA, China, Hongkong, Korea, Taiwan, and Japan (Herawan dkk, 2015). For the needs of the world's sandalwood oil is estimated at around 200 tonnes each year. The majority of needs are supplied from india around 100 tonnes, while Indonesia, Australian, New Caledonia and Fuji around 20 tons so a shortage of about 80 tons each year (Masyhud, 2009). According to Surata, and Idris (2001) the success of the Sandalwood plant assessed low is less than 50%. The death of sandalwood’s seeds quite high with success rate grows between 20-40% when grown using techniques of planting directly ditanah, (Surata, 2006). These conditions encourage to make a breakthrough providing Sandalwood plant seed massively. The provision of the Sandalwood plant seed massively can be done in a vegetative or generative. In generative can be done through seed, but with the scarcity of plants can be difficult in getting seed with sufficient quantity and quality (Herawan, 2012). To resolve the issue, then the vegetative breeding techniques with tissue culture is expected to fulfill the provision of seedlings massively in short time.

Redundancy of plant breeding techniques with sandalwood in vegetative tissue culture can be done with induce rooting of sandalwood. One of the factors that play a role in inducing roots in plant reproduction in vitro is a medium (Yusnita, 2003). The media is a determine factor in the success of activities in tissue culture. In addition to the use of the media, then planting media combinations with the growing regulatory substances (ZPT) was also very influential in stimulating of the roots.

One of the growing regulatory substances play a role in stimulating of the roots is Auxin (Hendaryono and Wijayani, 1994). The working mechanism of Auxin in root formation begins when the Auxin slow down the activation of the compound that affects the formation of calcium pektat, causing the cell wall becomes more elastic (Hastuti et al, 2002). Cell wall elastic causes ions such as H +, K +, Ca + can enter into cells. As a result, the cytoplasm is easier to push outwards cell walls and cell volume expand. In addition, Auxin led to exchanges between H + ions with the K + ions. The K + ions will enter into the cytoplasm and spur the absorption of water into the cytoplasm to maintain turgor pressure within the cell. After experiencing swelling of the cell wall then it will be a hard back due to metabolic activities occur in the form of the absorption of Ca + ions out of the cell, which perfected the composition of calcium pektat in cell wall (Hasanah and Nintya, 2007). Water and ions into the cytoplasm activates the enzyme growth, so that the cells terinisiasi by Auxin hormones going through enlargement. Enlargement of cells that occur on the trunk of sandalwood which was given hormone Auxin will grow into roots that serves to absorb the nutrient substances. On the basis of the types of Auxin, Auxin is IBA is more stable and low toxicity compared to NAA and IAA (George at al., 2007). According to Wattimena (1992), IBA is a type of Auxin in the most frequently used type of Auxin induces root than others, because it has a high ability in controlling the initiation of root (Wattimena, 1992). Janarthanam et al., (2011) in his research states that the use of MS media with a combination of ZPT 0.5 mg/L IBA and NAA 0.25 mg/L-1 manufactured 4.2 ± 0.10 cm root with high average 4.8 ± 0.2 cm after dikultur for six weeks at the plant Santalum album. For the success of the acclimatization up to 70%. Based on the above description, then the necessary research on the influence of addition of IBA (Indole Butyric Acid) on media WPM (Wood Plant media) against the induction of root buds planlet sandalwood (Santalum album l.) by in vitro. In this study, the use of astringent balance grow IBA created a diverse concentration of aims to get the concentration of the substance in the appropriate growing balance induces root buds planlet sandalwood by in vitro.

Formulation of the problem in this research is... The purpose in this research are: 1. To know the influence of addition of IBA (Indole Butyric Acid) on media WPM (Wood Plant Media) against the induction of root buds planlet sandalwood (Santalum album l.) by in vitro.

2. To know the concentration of the addition of IBA (Indole Butyric Acid) on media WPM (Wood Plant Media) are most effective in inducing roots planlet shoots sandalwood (Santalum album l.) by in vitro. The limitations of problem in this research mentioned above as follows: 1. using planlet from the results of culture of stem shoots sandalwood (Santalum album L.) by Imaniah Bazlina (2016). Age planlet ± 1 month and have to germinate with the length of 3-4 cm of shoots and leaves ± 5 leaves. 2. The medium used is the media WPM (Wood Plant Media). 3. is used ZPT IBA (Indole Butryric Acid) with a concentration of 0 mg/l; 1.5 mg/l; 1.75 mg/l and 2 mg/l. 4. The observations made during the 56 days after planting (HST) 5. The parameters observed were: (a) the speed of the rise of the root (b) average number of the root (c) average length of root and (d) moisture content root includes the weight of the wet and dry weight of roots. For observations of contamination is not loaded in the observation data (just do observations only)

BAB II Sandalwood in the classification not much cause dissent among the botanist. The number of species in Indonesian only one, that of Santalum album L. Classification of sandalwood according to holmes (1983) is as follows: 2.1.2.1 Climate This type of precipitation areas grew at an average of 625-1625 mm/year, The average temperature ranges between 10-35oC in the daytime

To produce good growth this type need fertile soil, nests, good drainage, soil reaction of alkalis and thin soil within the solum. This tree species grow in p. Timor with height 0-1200m from the surface of the sea. a. Stem Sandalwood tree has a distinctive architecture, monopodial stem: arthotropis (points up), continuous growth b. Leaf Sandalwood is a single leaf leaves that grow on twigs, face special, green leaves are shiny and hairless sandalwood elip, form the edge of the flat, tapered tip but sometimes there are blunt or rounded (Sindhu, 2010). c. Flowers and Seeds Inflorescence is terminal or Sandalwood eksiler, recimus, the flower pedicel articulatus 3-5 cm, bald, tube-shaped perigonium campanulatus, length of 3 mm and a diameter of approximately 2 mm, has 4 lobes perigonium, triangular shape, blunt at the tip and second surface hairless (Sindhu, 2010). d. Fruit Sandalwood has a stone fruit and round, time to cook the meat of the fruit skin is black, has a layer of eksocarp, mesocarp fleshy, hard endocarp with a line from the tip to the base of the e. Root Sandalwood is the root rooting system good clear roots with many strong branches.

BAB III 3.1 The design of Research This research was performed using Random Design Complete the single factor that is administering the treatment type ZPT IBA media WPM. As for the granting of the IBA ZPT treatment i.e. W0:0 mg/l as control, W1:1.5 mg/l, W2:1.75 mg/l and W3:2 mg/l. Research consists of 4 treatments and each treatment was repeated as many as 8 repeats and repeats each contain 1 planlet. 3.2 Research Variables The variables used in this study are as follows: 1. free: substance concentration Variable regulator grow IBA i.e. 0 mg/l; 1.5 mg/l; 1.75 mg/l; 2 mg/l. 2. Variable: speed the emergence of the roots, the average number of average length, root root, root water content include the weight of the wet and dry weight of roots. 3. The variables controlled include: room temperature, pH of media, media, media creation, time planlet, planting planlet and length of observation. 3.3 time and place This research was conducted in the laboratory of Plant tissue culture Department of Biology Faculty of science and technology in the Islamic State University of Malang Maulana Malik Ibrahim on April until June 2017. 3.4 The tools and materials 3.4.1 Tools The tools used in this research is measuring cup, erlenmeyer flask, petri dish, stir bar, bottle kultur, dissection tools (scalpel, tweezers, scissors), LAF (Laminar Air Flow), analytic scales, oven, autoclave, bunsen, sprayers alcohol (sprayer), pH meter (pH indicator), refrigerator, shelf kultur, AC (Air Conditioner), lights, hot plate and magnetic stirrer, tissue culture, rack, aluminum foil, plastic wrap, paper labels, rubber , plastic pot, stove and heaters. 3.4.2 Materials The main material used is the result of culture induction Sandalwood’s shoots (Santalum album L.) were used as eksplan which will be planted in the media. The material used is the media WPM (Wood Plant Media) with product code instant Cat # 30630054-1, gelatin swallow white, sugar sand. Growing regulatory substances (ZPT) used are IBA. 1. Sterilization of Room Planting 2. Sterilisation of tools 3. The manufacture of Hormones Stock 4.Manufacture of Basic Media Work steps in the creation of as many as 600 WPM media 1 ml consist of 4 and 8 treatment of Deuteronomy, each treatment is divided into 125 ml) are as follows: 1. measure the media with Weighted as follows: Woody Plant medium (WPM) 1.3812 grams, sugars 18 g, so 1 gram/125 ml media 2. Homogenized media WPM and sugar with stirer above hot plate. 3. media dihomogenkan Divided into 4 parts. Each divided as much as 125 ml consist of media W0, W1, W2, W3. 4. Added ZPT IBA W1:1.5 mg/l, W2:1.75 mg/l and W3:2.0 mg/l then the respective media homogenized with the above stirrer hot plate. 5. Each measured pH media W0, W1, W2 and W3 amounted to 5.8 with indicators of pH. If the pH is less then 5.8 added 0.1 N NaOH and if more 5.8 then added HCl 0.1 N. 6. Added so that 1 gram in each respective media.

7. the media is heated and stirred up to the boil. 8. Put each media (W0, W1, W2 and W3) into small size bottle of culture as much as 12.5 ml media that has been cooked. 9. Closed bottle containing culture media with plastic and bound with rubber. 10. media culture that has been Sterilized by diautoklaf at a temperature of 1210C with pressure 17.5 psi for 15 minutes. 11. Saved the media that have been sterilized dirak media in sterile room for one week prior to the initiation of the plant. 3.5.2 Phase Implementation 1. Planting the shoots Induction of sandalwood (Initiation) Planting (initiation) planlet in the LAF (Laminar Air Flow). As for the working steps in the implementation of planting (initiation) is as follows: 1. hand-Sprayed with alcohol 70% outside the Laminar Air Flow (LAF), further cleaned with Kleenex and alcohol 70%. 2. Marinate tools like tweezers, scalpel, scissors are needed in kultur in 96% alcohol and burned with the fire of bunsen. 3. Put on the top of the lid stainless steel box (put into sterile aquades) and allowed to cool. 4. the members of the incoming body Sprayed within the LAF with alcohol 70%. 5. the induction results Taken planlet sandalwood shoots (± 1 month old and have to germinate with the length of 3-4 cm of shoots and leaves ± 5 leaves). 6. Put planlet in petri dish. 7. Cut ± 0.3 cm bottom planlet sandalwood. 8. Planted of planlet (1 bottle contains 1 planlet) into a wide range of media concentration on ZPT WPM using tweezers. 9. Closed bottle culture who have initiated planlet with plastic wrap, heat-resistant plastic and bound with rubber. 10. Incubated bottles that have been planted on culture in space planlet temperature 21-270C. 11. the Observed once every 2 weeks after HST (days after planting) the early emergence of the root, the number of roots and root length (use a ruler) as well as other things such as crop contamination, contamination of the media and the changes that occur at the plant. After 8 weeks, root water content calculated that includes wet and dry weight of roots. 3.6 Observations 3.6.1 Observations the speed the emergence of the roots Observations about the speed the emergence of the roots are observed every day until the end of the observation in the day-to-56 after planting (HST) are marked by protuberances on the ends of the rod. 3.6.2 Observation speed of emergence of the roots, the average number of roots, root length and average water content of roots (BB and BK). 1. Final Observations made at day 56 observations after planting. The observation parameters include the following: (a) the speed of the rise of the observed root every day until the end of the observation; (b) the average number of roots; (c) the average length of root and (d) moisture content root include wet and dry weight of roots. As for the formula used in these observations is as follows:

3.5 techniques of Data analysis The data obtained are then tested its homogeneity and normality using the software SPSS 16.0. After that, to know their effect was then tested using the Anava (Variant Analysis) one line. If there is a real difference, then calculated first coefficients of diversity. As a result, if the data have a great diversity of coefficient of at least 10% in a homogeneous condition can be tested using Duncan, if their coefficients are between 5-10% in a homogeneous condition can be tested further using BNT (the smallest Real Difference) and in a diversity of small coefficient up to 5% in a homogeneous condition can be tested further using BNJ (Unlike a real honest) (Hanafiah, 2014).