Effect of benzyl adenine and sucrose for in vitro microrhizome production in curcuma aromatica salisb

AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> <br /> <br /> <br /> EFFECT OF BENZYL ADENINE AND SUCROSE FOR IN VITRO MICRORHIZOME<br /> PRODUCTION IN CURCUMA AROMATICA SALISB<br /> <br /> Nguyen Thi Thuy Diem1<br /> 1<br /> An Giang University, VNU - HCM<br /> <br /> Information: ABSTRACT<br /> Received: 04/10/2018<br /> The aim of this research was carried out to rapid multiplication of Curcuma<br /> Accepted: 13/08/2019<br /> aromatica salisb with large quantities, uniform and good quality, stored and<br /> Published: 11/2019<br /> transported easily which are advantageous for in vitro multiplication. Experiment<br /> Keywords: comprised: the effect of tree concentration Benzyl adenin (3, 5, 7 mg/L) and six<br /> Sucrose, microrhizomes, in concentration sucrose (20, 40, 60, 80, 100, 120 mg/L) on the in vitro<br /> vitro, Curcuma aromatica, microrhizome production were investigated. The experiment was arranged in a<br /> Benzyl adenin. completely randomized form, two-factor, 18 treatments and 5 replications. The<br /> results indicated that shoots of Curcuma aromatica salisb cultured on MS medium<br /> supplemented with 80 g/L sucrose and BA 3 - 5 mg/L for to be the most suitable<br /> medium for in vitro microrhizomes induction of Curcuma aromatica with time<br /> sooner, the rate of in vitro tuber formation, the number of tubers, tuber weight and<br /> tuber diameter reached the highest after 16 weeks of culture.<br /> <br /> <br /> 1. INTRODUCTION types. On the other hand, white turmeric was<br /> White turmeric (Curcuma aromatic Salisb) is a demonstrated diffent from essenttial oil into white<br /> rare medicinal herb plant species, in Zinger family turmeric plant such as anti- inflammatory, anti<br /> – Zingiberaceae, this is projecting to develop platelet aggregation, cough suppression and<br /> herbs plant source in An Giang province from kidney protection (Sikha et al., 2015).<br /> now onwards to the year 2020. Studies show the In nature, white turmeric occurs from rhizome,<br /> biological active substances extracted from after about 210 day to establish the rhizome breed<br /> rhizome in white turmeric contain group acticve (Ravindran et al., 2007). The rhizome has a long<br /> antioxidant such as curcuminoid, flavonoid, domance and germinates the rainy season. Beside,<br /> terpenoid. In the adopted antioxidant mechanism, the work to maintain source species of white<br /> the manufatured product from white turmeric turmeric annually is both costly and effort. Often,<br /> have biological effects such as anti- inflammatory the disease on the plant such as rhizome is a<br /> stomach medication from wine (Jeon et al., 2010). negative ordor (by Pythium sp.) and spot leaf by<br /> The situation artery antherogenesis is reduced Taphrina species and Collectrichum are very<br /> (Lee et al., 2010), prevent to injury damaging to the preservation process breed in<br /> carcinogenesis on skin by ultraviolet ray (Panich field causing the lack of the breed resource in the<br /> et al., 2010). The power antioxidant mechanism, culture (Nayak & Naik, 2006). Beside, white<br /> white turmeric plant has potential to become turmeric is building plan for planting area in An<br /> effect medicine treatment assist for many cancer Giang province to make material area. This object<br /> <br /> <br /> <br /> <br /> 17<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> is new so the white turmeric breed source is seen disease medicine for 60 minutes in clean rice husk<br /> early. So, the cultural technology is set up to ash. After 15 – 20 days of incubation, the shoots<br /> produce breed and quality guaranteed to support of white turmeric began to appear.<br /> seedlings free from disease, uniform to request The shoots were divided from rhizome washed,<br /> breed produce. roots cut and leaves cleaned, 1/2 leaf cut and<br /> In making microrhizomes in vitro by method clean washed. Next, the shoots were soaked in<br /> micropropagation to processing and good light soap after immature buds were cleaned with<br /> achieved on object such as Solanum tuberosum L., running tap water. Explants were then immersed<br /> Dioscorea composita, Gladiolus spp., Zingiber in 70% ethanol for 1 min and the explants were<br /> officinale Rosc, Curcuma longa,…(Gopal et al., rinsed three times with distilled water. Next, the<br /> 1998; Alizade et al., 1998; Dương Tấn Nhựt et explants were soak with 20% Ca(OCl)2 for 15<br /> al., 2007; Sharma & Singh, 1995; Sunitibala et minutes, after the explants were rinsed three times<br /> al., 2001). Currently, the studies on microrhizome with distilled water. The explants were cut,<br /> in vitro have not been processed in Viet Nam divided to take the shoot top and culture in bottle<br /> through the seedling source for production to contain medium MS not supplemented plant<br /> support the farmer. growth regulator to create a material source to use<br /> 2. MATERIALS AND METHODS for experimentation in making microrhizome.<br /> 2.1 Plant Materials 2.3.2 Experimental arrangement<br /> The white turmeric rhizome (Curcuma aromatica) Excised buds of Curcuma aromatica were high<br /> to take on Tinh Bien Oriental medicine congress about 4-5 cm, initially cultured on MS basal<br /> in An Giang province and take it from the lab of medium. After 14 days of culture, the shoots<br /> Agriculture and Natural Resources, An Giang became root formations and were transfered to a<br /> University to nursery. culture medium MS supplement with BA and<br /> 2.2 Medium and culture conditions sucrose in different concentrations for<br /> microrhizome induction.<br /> The medium were used MS basal medium<br /> The experiment was arranged in Completely<br /> (Musrashige & Sokol, 1962) supplemented with<br /> Block Design (CBD), two factor, including 3<br /> agar (8 g/l), Myo - Inositol (0,1 g/l) and 1 mg/L<br /> concentrations of BA (3, 5 and 7 mg/L) and 6<br /> NAA. Depending on how the experiment<br /> concentrations of sucrose (20, 40, 60, 80, 100 and<br /> supplemented with benzyl adenin (BA) and<br /> 120 g/L). The experiment consists of 18<br /> sucrose in different concentrations. The pH of the<br /> treatments, each treatment was repeated 5 times,<br /> media were adjusted to 5.8 using 1M NaOH or<br /> each time was 5 the culture bottle (1 explant/ the<br /> 1M HCl. The culture bottles were steam sterilized<br /> culture bottle).<br /> in an autoclaved at 1210C, 1 atm for 30 min.<br /> 2.3.3 Observations recorded for experiments<br /> Cultural conditions: The explants were maintained<br /> in a growth room under white fluorescent light for The time microrhizome induction (weeks after<br /> a daily photoperiod of 16 hours, with temperature cultured - WAC), rate tuber formation (%), the<br /> 24 ± 2 0C. number average tuber/explants, the weight<br /> microrhizome (g), diameter microrhizome (cm).<br /> 2.3 Methods<br /> Observations were done every week starting from<br /> 2.3.1 Culture to make source sample starting the first week after culture initiation to 16 weeks<br /> Curcuma aromatica rhizomes were selected from after culture initiation.<br /> 7 to 8 months of age, uniform and not diseased. 2.3.4 Data analysis<br /> Samples were treating with Antracol 70WP<br /> <br /> 18<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> Data were analyzed using the Statistical SPSS The time set up microrhizome in vitro had interact<br /> software version 20.0 and analysis of variance between BA concentration and sucrose. The time<br /> (ANOVA) was used to investigate if there is any set up microrhizome induction in vitro earliest on<br /> significant difference. A mean separation test was two experiment to supplemented 3 mg/L BA<br /> conducted using Duncan multiple range. combine 80 mg/L sucrose and 5 mg/L BA<br /> 3. RESULT AND DISCUSSION combine 60 mg/L sucrose in 5 weeks after being<br /> culturation (Fig 1.), it was statistically different in<br /> 3.1 The time microrhizome induction in vitro of<br /> significance at 1% compared to the remaining<br /> white turmeric shoots<br /> treatments.<br /> <br /> Table 1. Effect of concentration of BA and sucrose on the time microrhizome white turmeric induction in vitro<br /> <br /> Sucrose Concentrations of BA (mg/L) Average<br /> concentrations (Sucrose)<br /> (g/L) 3 5 7<br /> 20 11.00 d 10.00 e 12.00 c 11.00 b<br /> 40 9.00 f 8.00 g 9.00 f 8.67 c<br /> 60 6.00 i 5.00 k 7.00 h 6.00 e<br /> 80 5.00 k 5.67 j 6.00 i 5.56 f<br /> 100 7.00 h 7.00 h 9.00 f 7.67 d<br /> 120 13.00 b 13.00 b 15.00 a 13.67 a<br /> Average (BA) 8.50 b 8.11 c 9.67 a<br /> F (Experiment) **<br /> F (BA) **<br /> F (Sucrose) **<br /> F (BA x Sucrose) **<br /> CV (%) 1.54<br /> Note: The following letters were the same, the difference was no significant statistically: ** = the<br /> difference was statistically significant at 1%<br /> <br /> <br /> <br /> <br /> Fig 1. In vitro microrhizomes induction of white turmeric after 5 weeks of incubation on culture medium.<br /> <br /> 19<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> The effect of BA concentration to time induction When the sucrose concentration were increase<br /> to microrhizome production in vitro. In from 20 – 80 g/L then the time induction<br /> concentrate 5 mg/L BA has time to show microrhizome production to shorten from 11<br /> microrhizome in earliest (8.11 weeks after weeks after cultured to decrease 5.56 weeks after<br /> cultured), the difference was statistically cultured. But, when the sucrose concentration<br /> significant at 1% compared with 3 mg/L BA and 7 were increased continuous up 100 – 120 g/L then<br /> mg/L BA concentration in a culture medium. It the time induction microrhizome production from<br /> suggest that the time induction microrhizome of 7.67 weeks after cultured to increase 13.67 weeks<br /> BA early or later then depended on BA after cultured show new microrhizome.<br /> concentration in supplemented in a culture In general, the time induction microrhizome<br /> medium. In the BA concentration suitable for production were MS medium supplement 1 mg/L<br /> induction microrhizome formation in vitro were 5 NAA combine 3 – 5 mg/L BA and 60 -80 g/L<br /> mg/L, the microrhizome were formation earlier, sucrose in earliest, the microrhizome was show 5<br /> average about 8.11 weeks after cultured, the – 6 weeks after culturation.<br /> shoots start set up microrhizome in 3 mg/ L BA<br /> concentration and 7 mg/L BA concentration after 3.2 The rate explant were established<br /> 8.50- 9.67 weeks after cultured the new microrhizome of shoot white turmeric<br /> microrhizome was establish. The results on table 2, the rate of explant to<br /> The sucrose concentration to effect come on the microrhizome established in 16 weeks after<br /> time induction microrhizome production in vitro. cultured to have interact between BA<br /> The sucrose on 80 g/L concentration were time concentration and sucrose. The explant culture<br /> induction production earliest was 5.56 weeks after rate were highest in experiment MS supplemented<br /> cultured, the difference was statistically 1 mg/L NAA with 3 mg/L BA and 80 g/L<br /> significant at 1% compared the sucrose sucroser to achieve 91.67%, the difference was<br /> concentration in the other in a culture medium. statistically significant at 1%.<br /> <br /> Table 2. Effect of BA concentration and sucrose on the explant rate microrhizome formation (%) in white turmeric<br /> in vitro in the 16 weeks after cultured<br /> <br /> Concentrations of BA (mg/L) Average<br /> Sucrose oncentration (Sucrose)<br /> (g/L) 3 5 7<br /> <br /> 20 25.00 d 25.00 d 25.00 d 25.00 c<br /> 40 25.00 d 25.00 d 41.67 d 30.56 c<br /> 60 41.67 d 66.67 bc 25.00 d 44.44 b<br /> 80 91.67 a 75.00 b 58.33 c 75.00 a<br /> 100 41.67 d 75.00 b 33.33 d 50.00 b<br /> 120 25.00 d 25.00 d 25.00 d 25.00 c<br /> Average (BA) 41.67 b 48.61 a 34.72 c<br /> F (Experiment) **<br /> F (BA) **<br /> F (Sucrose g) **<br /> F (BA x Sucrose) **<br /> CV (%) 21.60<br /> Note: The following letters were the same, the difference was not statistically significant: ** = the<br /> difference was statistically significant at 1%<br /> <br /> 20<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> The sucrose concentration to effect come the time BA concentration to effect to the number of<br /> induction microrhizome production in vitro. In the microrhizome. In 5 mg/L BA concentration for<br /> sucrose 80 g/L concentration for the explant rate the number of microrhizome to achieve highest<br /> microrhizome formation achieved highest 75%, was 1.94 microrhizome/explants, the difference<br /> the difference was statistically significant at 1% was statistically significant at 1% as compared to<br /> compared the sucrose concentration different in BA concentration 3 mg/L and 7 mg/L in the<br /> the culture medium. culture medium.<br /> The BA concentration to effect come the explants The sucrose concentration to effect come number<br /> rate microrhizome formation in vitro. On the 5 microrhizome in vitro. The sucrose concentration<br /> mg/L BA concentration for the explants rate was 80g/L achive then the number highest baby<br /> microrhizome formation were highest (48.61%), microrhizome was 3 microrhizome/explants, the<br /> the difference was statistically significant at 1% difference was statistically significant at 1% as<br /> compared with MS medium supplemented 3 mg/L compared the sucrose concentration other in the<br /> BA and 7 mg/L BA in the culture medium. culture medium.<br /> In the result to table 2, when BA concentration to The compared between experiment, the results on<br /> increase from 3 mg/L to 5 mg/L, the explants rate table 3, the number of microrhizome to highest<br /> microrhizome formation to increase from 41.67% increase in 3 mg/L BA concentration with 80 g/L<br /> to 48.61%. When BA concentration increase on 7 sucrose then achieved 3.67 microrhizome/<br /> mg/L, the explants rate microrhizome formation explants, the difference was statistically<br /> to go down 34.72%. In the sucrose concentration, significant at 1% as compared the other<br /> when the sucroser concentration to increase from experiment.<br /> 20 to 80 g/L then the explants rate microrhizome In general, the results in table 3, when increased<br /> formation to increase from 25% to 75%. But when BA concentration from 3 mg/L to 5 mg/ L, the<br /> the sucrose concentrate increased on 100 -120 g/L number of microrhizome to increase from 1.67<br /> then the explant rate microrhizome formation go microrhizome/ explants to 1.94 microrhizome/<br /> to down form 50% to 25%. It suggest that, the BA explants. When increase BA concentration up to 7<br /> concentration and high sucrose were inhibition to mg/L, the number of microrhizome in vitro<br /> microrhizome formation in vitro. become go to down only achieve 1.39<br /> Finally, white turmeric shoots to culture on the microrhizome/ explants. For sucrose, when the<br /> MS medium to have 1 mg/L NAA supplemented sucrose concentration to increase from 20 to 80 g/<br /> 3 mg/L – 5 mg/L BA combine with 80 g/L L then the number of microrhizome to increase<br /> sucrose for the explants rate microrhizome from 1 microrhizome/explants to 3 microrhizome/<br /> formation was highest to achieve from 75% to explants. But when the sucrose content up to 100 -<br /> 91.67%. 120 g/L then the number microrhizome formation<br /> 3.3 The number of shoot white turmeric in vitro go to down from 2 microrhizome/ explants to 1<br /> to increase microrhizome/explants. So, the white turmeric<br /> shoots to culture on MS medium contain 1 mg/L<br /> In the 16 weeks after cultured, the results on table NAA to supplemented 3 – 5 mg/L BA<br /> 3, the number of microrhizome to effected of BA concentration combine with 80 g/L sucrose are<br /> concentration and sucroser content. microrhizome to achieve highest (Fig.2).<br /> <br /> <br /> <br /> <br /> 21<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> <br /> <br /> <br /> Fig 2. The development of microrhizome in vitro on the white turmeric shoot on medium<br /> supplemented 3 -5 mg/ L BA and 80 g/ L sucrose at 16 weeks after cultured<br /> <br /> Table 3. Effect of BA concentration and sucrose on the number of to increase on microrhizome in<br /> vitro after 16 weeks after cultured<br /> <br /> Sucrose BA concentration (mg/L) Average<br /> concentration (g/L) (Sucrose)<br /> 3 5 7<br /> 20 1.00 d 1.00 d 1.00 d 1.00 c<br /> 40 1.00 d 1.00 d 1.67 d 1.22 c<br /> 60 1.67 d 2.67 bc 1.00 d 1.78 b<br /> 80 3.67 a 3.00 b 2.33 c 3.00 a<br /> 100 1.67 d 3.00 b 1.33 d 2.00 b<br /> 120 1.00 d 1.00 d 1.00 d 1.00 c<br /> Average (BA) 1.67 b 1.94 a 1.39 c<br /> F (Experiment) **<br /> F (BA) **<br /> F (Sucrose) **<br /> F (BA x Sucrose) **<br /> CV (%) 21.63<br /> <br /> Note: The following letters were the same, the difference was no significant statistically: ** = the difference was<br /> statistically significant at 1%<br /> <br /> <br /> <br /> 22<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> 3.4 The weight of microrhizome white turmeric statistically significant at 1% as compared the<br /> in vitro other experiment. Between on the experiment to<br /> The results on table 4 represent the 16 weeks contain 80g/ L sucrose combine with BA<br /> after cultured, the sucrose content to have effect concentration 3 and 5 mg/L then microrhizome in<br /> on weight increase of microrhizome in vitro. The vitro weight to achieve 0.93 gram/ microrhizome,<br /> sucrose content on 80 g/L to achieve highest the different was not statistically significant.<br /> weight is 0.98 gram/microrhizome, the difference The microrhizome in vitro weight on BA<br /> was statistically significant at 1% as compared the concentration (3, 5 and 7) the different was<br /> sucrose other in the cultural medium. The weight statistically significant, the microrhizome in vitro<br /> microrhizome in vitro on sucrose content to 40 average weight archive 0.55 gram/microrhizome<br /> mg/L and 100 mg/L not the different was after 16 weeks cultured. In general, the BA<br /> statistically significant. concentration not effect to the weight increase of<br /> The results were statistically significant about microrhizome in vitro.<br /> rhizome in vitro weight to increase all So, the experiment to supplemented 80 g/ L<br /> experiments. The rhizome in vitro weight to sucrose on MS medium with 1 mg/L NAA<br /> highest achieve on MS medium supplemented 3 combine BA concentration 3 -7 mg/ L for<br /> mg/L BA and 80 g/L sucrose (1,08 microrhizome in vitro weight was highest.<br /> gram/microrhizome), the different was<br /> <br /> Table 4. Effect of BA concentration and sucrose on up to weight (gram/microrhizome)<br /> white turmeric in vitro after 16 weeks cultured.<br /> <br /> Sucrose BA concentration (mg/L) Average<br /> concentration (g/L) (Sucrose)<br /> 3 5 7<br /> <br /> 20 0.50 fg 0.39 gh 0.29 hi 0.39 d<br /> <br /> 40 0.49 fg 0.64 e 0.59 ef 0.57 c<br /> <br /> 60 0.79 c 0.77 cd 0.59 ef 0.72 b<br /> <br /> 80 1.08 a 0.93 b 0.93 b 0.98 a<br /> <br /> 100 0.41 g 0.43 g 0.66 de 0.50 c<br /> <br /> 120 0.13 j 0.19 ij 0.08 j 0.13 e<br /> <br /> Average (BA) 0.57 0.56 0.52<br /> <br /> F (Experiment) **<br /> F (BA) ns<br /> F (Sucrose) **<br /> F (BA x sucrose) **<br /> CV (%) 12.88<br /> Note: The following letters were the same, the difference was no significant statistically: ** = the difference was<br /> statistically significant at 1%; ns= the difference was not statistically significant.<br /> <br /> <br /> <br /> 23<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> 3.5 The diameter to increase of white turmeric The sucrose content to effect on increase diameter<br /> microrhizome microrhizome in vitro. The sucrose content in<br /> In the results on table 5, the BA concentration and 80g/L to have diameter highest gain was 10.05<br /> sucrose content to interact on the increase of mm, the difference was statistically significant at<br /> diameter of white turmeric in vitro microrhizome. 1% as compared the sucrose content other in the<br /> culture medium.<br /> The BA concentration to effect on the diameter<br /> increase of rhizome in vitro. In the BA The result of table 5 suggest that the different was<br /> concentration 3 mg/L, the diameter of statistically significant about to increase diameter<br /> microrhizome in vitro achieve highest was 8.10 microrhizome in vitro all experiment. The<br /> mm, the difference was statistically significant on diameter of microrhizome in vitro highest gain on<br /> 1% as compared the BA concentration 7 mg/L, MS medium to supplement 3 mg/L BA and 80g/<br /> but the difference was not statistically significant L sucrose (13.7 mm), the difference was<br /> than 5 mg/ L BA to achieve 7.8 mm. statistically significant at 1% as compared the<br /> experiment content other in the culture medium.<br /> Table 5. The effect of BA concentration and sucrose diameter to increase of microrhizome (mm) on white turmeric<br /> in vitro after 16 weeks after cultured<br /> <br /> Sucrose BA concentration (mg/L) Average<br /> concentration (g/L) (Sucrose)<br /> 3 5 7<br /> <br /> 20 8.10 cde 8.00 de 5.60 gh 7.20 c<br /> 40 6.40 fg 8.30 bcd 8.40 bcd 7.70 c<br /> 60 9.30 b 8.60 bcd 6.90 f 8.20 b<br /> 80 13.70 a 9.10 bc 8.80 bcd 10.50 a<br /> 100 7.30 ef 8.50 bcd 6.50 fg 7.40 c<br /> 120 3.90 i 4.00 i 5.10 h 4.30 d<br /> <br /> Average (BA) 8.10 a 7.80 a 6.90 b<br /> <br /> F (Experiment) **<br /> F (BA) **<br /> F (Sucrose) **<br /> F (BA x sucrose) **<br /> CV (%) 7.23<br /> Note: The following letters were the same, the difference was no significant statistically: ** = the difference was<br /> statistically significant at 1%.<br /> In general, the experiments was supplemented 3 – microrhizome formation of white turmeric in in<br /> 5 mg/ L BA with 80g/L sucrose in MS medium vitro condition to effect of BA concentration and<br /> culture to contain 1 mg/L NAA for the diameter sucrose content interact. The white turmeric<br /> of microrhizome to biggest gain after 16 weeks shoots were culture on MS medium to<br /> after cultured (Fig 3.). supplemented 80g/L BA combine 3 – 5 mg/L BA<br /> In summary, the results of target about and 1 mg/L NAA then the time microrhizome<br /> microrhizome production in vitro suggest that the induction was earlier, at the same time the<br /> <br /> 24<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> microrhizome in vitro formation rate, the number microrhizome induction, the microrhizome in<br /> of microrhizome production and microrhizome vitro formation rate, the number of microrhizome<br /> diameter were highest. When the BA production and microrhizome diameter were<br /> concentration to increase 7 mg/L combine 100 decreased.<br /> mg/L and 120 mg/L sucrose then the time<br /> <br /> <br /> <br /> <br /> Fig 3. The microrhizome diameter of white turmeric shoot were growth on the medium supplemented 3-5 mg/L<br /> BA and 80 g/L sucrose in 16 weeks after 9 (cultured)<br /> The result of the experiment suggests that the BA sucrose from 6 – 8% (w/v) was the most<br /> concentration and sucrose content to effect important ingredient for formation in vitro tuber<br /> interact on microrhizome in vitro formation. On on Diascoria composita. It suggests that the study<br /> the MS medium contain 1 mg/L NAA to topic results concluded that, in the study to show<br /> supplemented 80 g/L sucrose with 3-5 mg/L BA the medium MS supplemented 80 g/L sucrose for<br /> for the time microrhizome induction, the the number of microrhizome in vitro was most.<br /> microrhizome formation rate, the number of The number of microrhizome in vitro was shortest<br /> microrhizome productions, the weight and on the MS medium contain 20 g/L and 120 g/L<br /> diameter of microrhizome were highest. It sucrose. The sucrose was role to promote<br /> suggests that this BA concentration to effect on formation organ store in plant. When the sucrose<br /> transport carbohydrate content in the white concentrate from 20 g/L to 80 g/L to show the<br /> turmeric shoots. The result that similar of Nanak microrhizome in vitro formation to rate, the<br /> (200), the number of microrhizome in vitro to number microrhizome, the weight microrhizome<br /> many formation on the medium supplemented and diameter microrhizome are increase. It<br /> 5mg/ L BA on C.aromatica Salisb. suggests that the show of carbon content in<br /> In Curcuma sp species, the shoots were rhizome sucrose and the white turmeric ingredients contain<br /> formation to view when the medium to supply carbohydrates and sucrose. The results of the<br /> sucrose concentration was 60 – 90 g/L (Nayak experiment showed the sucrose concentration in<br /> (2000); Sunitabala et al. (2001). In addition, the the medium to effect on the formation<br /> study about the formation zinger in vitro rhizome microrhizome in in vitro condition.<br /> of Sharma và Singh (1995), the sucrose In summary, the results showed the plant growth<br /> concentration was 75 g/L to effect in rhizome regulator was role on cell division, dimensions of<br /> formation. Sedigeh et al. (1998) to believe that the shoot germination and the effect on size of<br /> <br /> 25<br /> AGU International Journal of Sciences – 2019, Vol 7 (3), 17 – 27<br /> <br /> rhizome. The sucrose was effect in the growth moisture was provided by water spray twice a day<br /> process of microrhizome in vitro, the development (humid 70 – 80%). The temperature was 28 – 30<br /> of baby rhizome when the sucrose was highest, 0<br /> C in the nursery step. The results showed, after 2<br /> the sucrose supplemented nutrient to growth weeks plant on rice husk ash all the white<br /> rhizome and rhizome induction, the rhizome was turmeric rhizome in vitro show shoots, leaves and<br /> formation and development. roots ( Fig. 5A). After 30 days planted on pot to<br /> 3.6 The Acclimatization of white turmeric in show the white turmeric from rhizomes in vitro<br /> vitro the root system was developed and deep in the<br /> ground, the nursery plant was developed with 3 –<br /> The white turmeric rhizome in vitro were 4 leaves, the plant was moved out off the pot and<br /> harvested on the medium rhizome production, planted in black plastic (Fig. 5B).<br /> clean wash agar and nursery on rice husk ash. The<br /> <br /> <br /> <br /> <br /> A B<br /> Fig 5. The white turmeric rhizome in vitro to show shoot after 2 weeks cultured (A); the white turmeric from<br /> microrhizome in vitro (B) after 30 days planted into pot.<br /> 4. CONCLUSION Gopal, J., Minocha, J. L., & Dhaliwal, H. S.<br /> The medium microrhizome production in vitro (1998). Microtuberization in potato (Solanum<br /> was MS medium supplemented 3 mg/ L or 5 tuberosum L.). Plant Cell Rep, 17: 794-798.<br /> mg/L BA combine with 80 g/L sucrose. Jeon, W. Y., Lee, M. Y., Shin, I. S., Jin, S. E., &<br /> 5. RECOMMENDATION Ha, H. (2015). Curcuma aromatica Water<br /> Extract Attenuates Ethanol-Induced Gastritis<br /> Continue evaluation of the growth and<br /> via Enhancement of Antioxidant Status.<br /> development of rhizome in vitro on nursery stage<br /> Eviddence based Complementary and<br /> of the white turmeric.<br /> Alternative Medicine, 1.<br /> REFERENCE http://dx.doi.org/10.1155/2015/582496<br /> Alizadeh, S., Mantell, S. H., & Viana, A. M. Lee, H. S., Lee, M. J., Kim, H., & Choi, S. K.<br /> (1998). In vitro shoot culture and microtuber (2010). 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