Conservation Of Juwet ( Syzygium Cumini ) Plant Using In Vitro Culture Techniques

to determine the effect of the combination of 2,4-D and BAP growth regulators on the growth of and the effect of the combination and BAP on the growth of bud as a conservation effort. is experimental. Using a Completely Randomized Design (RAL) with combination 2,4-D (0; 0.5; 1; 1.5; 2; 2.5; 5) mg/L, and BAP (0; 0.25; 0.5; 0.75; 1) mg/L and combination NAA (0; 0.25; 0.5; 0.75; 1) mg/L and BAP (0; 0.5; 1; 1.5; 2) mg/L. Analysis by Two Way ANAVA test α = 5%. If there is a significant difference, the Duncan Multiple Range Test (DMRT) test with a significant level of 5%. Addition of 2.5 mg/L and 3 mg/L 2.4-D without BAP can induce intermediate callus, brownish yellow and there is a large cell nucleus in each cell. While the results of axillary bud growth is treatment in BAP 1 mg/L without NAA is the most effective interaction on the emergence of buds is 26.6 days after planting with the total of buds as much as 6.66, and the highest buds 5.37 cm and the highest total of leaves, namely 8.33 strands..


INTRODUCTION
Juwet is a local Indonesian plant, many people know it's a mystical plant because it grows wild and lush. However, juwet has the value of a local wisdom which can be utilized as one of them as traditional medicine, it can be seen in the Bune indigenous people (Katili, 2015).
Juwet plants can live in a high geographical range. Juwet is found in tropical and subtropical regions (Ayyanar & Subash-Babu, 2012). Juwet fruit (buni fruit, oval), young fruit is green, after cooking it is red purplish red in color and has a strong sour taste. Juwet seeds one in one fruit, oval, hard, white. Have taproots, sympodial branches and light brown stems (Dalimarta, 2003).
According to Mudiana (2007) states that the parts of bark, seeds, and leaves of Juwet plants are efficacious to reduce blood glucose levels in patients with type II diabetes mellitus. In addition, with the taste of sweet and sour fruit, it is cool, strong astringent, aromatic smelling efficacious to lubricate the organs of the lungs, stop coughing, laxative urine. Ayyanar & Subash-Babu, (2012) juwet contains antioxidants, one of which is anthocyanin and flavanioid.
So many benefit of juwet make this plant a high interest. But now in Indonesian the availability of juwets is low. because juwet plants belong to scarce plants, because of lack of attention and cultivation of the community (Dalimarta, 2003).
Cultivation through seeds will produce plants that have low viability compared to their parents. Whereas if using a graft or connecting technique requires seeds with large amounts and up to 90 days. Juwet seedlings can be planted during the rainy season. (Mudiana, 2007) is still little information about the growth of juwet. Not many juwet production centers or juwet cultivation areas. Knowledge of the growth and development of a type of plant is very necessary to know how to handle and maintain juwet species.
In vitro culture techniques one alternative that can solve this problem. In vitro culture is a technique to develop plant parts, both in the form of cells, tissues and organs in aseptic conditions in vitro. This technique is able to produce quality, uniform seedlings, identical to the parent and capable of multiplying plants in a relatively short time (Gunawan 1992).
Factors influence in the Vitro culture are media, growth regulating substances and explants used (Lestari, 2011). The growth regulator substances in In Vitro culture are auxin (2,4-D, NAA etc.) and cytokines (BAP, kinetin, IBA etc.).
One of the uses of In Vitro culture as plant propagation is embryogenic callus and multiplication of axillary buds (Admojo, 2014 andGunawan, 1992). Based on the existing problems, the research is carried out, it is hoped that it will increase juwet plants to be sustainable by utilizing biotechnology.
This study aims to determine the effect of the combination of 2,4-D and BAP to grow embryogenic callus and determine the effect of the combination of NAA and BAP on the growth of axillary buds. This is a form of plant conservation juwet (Syzygium cumini).

MATERIALS AND METHODS
This study included experiments and was conducted in March-August 2018 in the tissue culture laboratory in the Department of Biology, Maliki UIN Malang. Samples from plants that have been incubated at the Green House until new leaves appear. This research was started by taking explants, sterilizing the planets, planting, incubation, morphological and histological observations, then analyzing the data.
Sterilization for embryogenic callus formation, young leaves are taken and then washed, 5 minutes detergent, 30 minutes fungicide and 60 minutes of water flow. sterilization in laminar air flow using chlorok 30%, 20% and 10% each for 10 minutes and rinsed sterile water 3 times each for 5 minutes. then the leaves are cut 1x1 cm and soaked betadine for 1 minute. 1 bottle containing 3 explants, each treatment had 3 replications and incubated for 45 days.

Results
Result from this research would can see in Table 1.

DISCUSSION a. Embryogenic Callus Formation
Based on the ANAVA test that has an effect on embryogenic callus formation is the Callus emerge day. The concentration of 1 mg/L 2,4-D + 1 mg/L BAP has the most significant value. In this combination the Callus emerge day on the 23 day after planting. Rosyida (2014) the right and balanced combination of auxin and cytokinin treatment will be produce optimal callus. Quality embryogenic callus is a callus that has a crumb texture. However, in the observation results there were no crumby callus. At concentrations of 2.5 mg/L 2,4-D + 0 mg/L BAP and 3 mg/L 2,4-D + 0 mg/L BAP have intermediate textures, while at the other concentrations they are compact. So that intermediate textured callus is considered the best texture in embryogenic callus formation and is supported by histological observations. The best color observation results on embryogenic callus were at a concentration of 2.5 mg/L 2,4-D + 0 mg/L BAP and 3 mg/L 2,4-D + 0 mg/L BAP. At a concentration of 2.5 mg/L 2,4-D + 0 mg/L BAP has a yellowish-white color, at a concentration of 3 mg/L 2,4-D + 0 mg/L BAP has a brownish yellow color. According to Mahadi (2016) Characteristics of embryogenic callus are yellowish white. While according to Al-Gendi (2013) the characteristics of embryogenic callus are brownish yellow. The color of embryogenic callus produced by each different plant explant will have a different color. This is because the pigmentation, light intensity and plants are different.
The histology callus at a concentration of 2.5 mg/L 2,4-D + 0 mg/L BAP and 3 mg/L 2,4-D + 0 mg/L BAP showed embryogenic callus. At this concentration there is a large cell nucleus in each cell, there is a clear vacuole but its size is smaller than the cell nucleus and there is a  Gunawan (1987) added that there were solid cytoplasm, small vacuoles and starch grains.

b. induction Axillary Buds
Effect, so that all variables are followed by a 5% DMRT test. The concentration of 1 mg/L is the most effect concentration in all fariables. On the buds emerge day with an average of 26 HST, the number of axillary buds with an average value of 6.06, the height of axillary buds with an average value of 5.35 cm and the number of leaves in axillary buds with an average value of 7,33 barley.
The concentrations of 0.5 mg/L NAA + 0 mg/L BAP were the most significantly different concentrations of buds emerging with an average value of 24.66 days after planting. At a concentration of 0 mg/L NAA + 1 mg/L BAP gave the most effective results for the number of axillary buds with an average value of 6.66. At concentrations of 0.5 mg/L NAA + 1 mg/L BAP the most effective results for axillary buds height with an average value of 6.16 cm. And in the treatment of 0 mg/L NAA + 1 mg/L BAP gave the most effective results for the number of leaves in axillary buds with an average value of 8.33 strands.
Based on the picture (Figure 2) above, it can be seen that the most results are shown in the concentration treatment of NAA 0 mg/L + BAP 1 mg/L. The results of these statements, it is suspected that the high percentage of buds formation without addition of NAA growth regulator can grow, this is possible because physiologically the content of endogenous NAA from juvenile stem explants is sufficient for buds formation. So that the NAA treatment in low concentrations of explants was able to induce buds. Zulkarnain (2009) more cytokines than auxin will form buds, whereas if auxin is more than cytokinin, roots will form.

CONCLUSION
The addition of 2.5 mg/L and 3 mg/L 2,4-D without BAP can induce quality embryogenic callus with yellowish white and brownish yellow, intermediate texture, and is a large nucleus cell in each cell. The addition of BAP 1 mg/L without NAA was most effective against the buds emerge day which was 26.66 days after planting, number of buds 6.66, highest buds of 5.337 and the highest total of leaves was 8.33 strands.