scholarly journals Peningkatan laju multiplikasi tunas dan keragaan planlet Stevia rebaudiana pada kultur in vitro Increasing shoot multiplication rate and plantlet vigor of Stevia rebaudiana in vitro culture

2016 ◽  
Vol 79 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Masna Maya SINTA

AbstractStevia (Stevia rebaudiana Bertoni) is a natural zero-calorie sweetener plant grown in a high population density.Tissue culture technique is useful for rapid mass propagationof plants to provide superior planting materials. Experimentswere conducted to increase growth and multiplication ofshoots and vigor of plantlets of stevia. Explants used wereapical and axillary buds from plantlets grown on MS mediumwithout plant growth regulators. Combinations of BA andIAA at different concentrations were used for shoot growthand multiplication, whereas plant growth retardants(ancymidol and paclobutrazol) and light intensity were usedfor plantlet vigor. The results showed that stevia explantscultured on MS medium without plant growth regulatorsproduced the highest shoots (4.5 cm) with two shoots perexplant. The best multiplication rate of shoots were found onMS medium added with 1.13 mg/L BA combined with0.35 mg/L IAA which produced on average 4.5 shoots and11.9 nodes per initial explant. Ancymidol and paclobutrazolconcentrations affected significantly growth and vigor ofstevia plantlets. Increasing the concentration of ancymidoland paclobutrazol decreased plantlet height and biomassfresh weight, but increased stem diameter. Paclobutrazol at0.1 mg/L was the best treatment to increase the vigor ofstevia plantlets. Light intensity at 20 µmol/m 2 /s gave betterplantlet vigor than other light intensities. It can be concludedthat multiplication of stevia shoots should be grown on MSmedium supplemented with 1.13 mg/L BA + 0.35 mg/L IAAand the vigor of the shoots can be increased by culturing onMS medium containing 0.1 mg/L paclobutrazol underfluorescence lamps with 20 µmol/m 2 /s light intensity.AbstrakStevia (Stevia rebaudiana Bertoni) adalah tanamanpemanis alami nir-kalori yang ditanam dengan kerapatanpopulasi yang sangat tinggi. Teknik kultur jaringan dapatdigunakan untuk perbanyakan tanaman secara massal dancepat untuk menyediakan bahan tanam unggul. Penelitiantelah dilakukan untuk meningkatkan pertumbuhan danmultiplikasi tunas dan keragaan planlet stevia. Eksplan yangdigunakan adalah tunas pucuk dan tunas samping dari planletyang ditumbuhkan pada medium MS tanpa zat pengaturtumbuh. Kombinasi BA dan IAA dengan konsentrasi yangberbeda digunakan untuk pertumbuhan dan multiplikasitunas, sedangkan zat penghambat tumbuh (ansimidol danpaklobutrazol) serta intensitas cahaya digunakan untukkeragaan planlet. Hasil penelitian menunjukkan bahwaeksplan stevia yang ditumbuhkan pada medium MS tanpa zatpengatur tumbuh menghasilkan tunas paling tinggi (4,5 cm)dengan dua tunas per eksplan. Multiplikasi tunas terbaikdiperoleh pada medium dengan BA 1,13 mg/L yangdikombinasikan dengan IAA 0,35 mg/L yang menghasilkan4,5 tunas dan 11,9 ruas per eksplan awal. Konsentrasiansimidol dan paklobutrazol berpengaruh nyata terhadappertumbuhan dan keragaan planlet stevia. Meningkatnyakonsentrasi ansimidol dan paklobutrazol menurunkan tinggiplanlet dan bobot basah biomassa, tetapi meningkatkandiameter batang. Paklobutrazol pada konsentrasi 0,1 mg/Lmerupakan perlakuan terbaik untuk meningkatkan keragaanplanlet stevia. Intensitas cahaya pada 20 µmol/m 2 /detikmemberikan keragaan planlet yang lebih baik dibandingkanintensitas cahaya yang lain. Dapat disimpulkan bahwamultiplikasi tunas stevia sebaiknya dilakukan pada mediumMS ditambah BA 1,13 mg/L + IAA 0,35 mg/L dan keragaanplanlet dapat ditingkatkan dengan menanam planlet padamedium MS ditambah paklobutrazol 0,1 mg/L di bawahlampu fluoresen dengan intensitas cahaya 20 µmol/m 2 /detik.

2016 ◽  
Vol 79 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Masna Maya SINTA

AbstractStevia (Stevia rebaudiana Bertoni) is a natural zero-calorie sweetener plant grown in a high population density.Tissue culture technique is useful for rapid mass propagationof plants to provide superior planting materials. Experimentswere conducted to increase growth and multiplication ofshoots and vigor of plantlets of stevia. Explants used wereapical and axillary buds from plantlets grown on MS mediumwithout plant growth regulators. Combinations of BA andIAA at different concentrations were used for shoot growthand multiplication, whereas plant growth retardants(ancymidol and paclobutrazol) and light intensity were usedfor plantlet vigor. The results showed that stevia explantscultured on MS medium without plant growth regulatorsproduced the highest shoots (4.5 cm) with two shoots perexplant. The best multiplication rate of shoots were found onMS medium added with 1.13 mg/L BA combined with0.35 mg/L IAA which produced on average 4.5 shoots and11.9 nodes per initial explant. Ancymidol and paclobutrazolconcentrations affected significantly growth and vigor ofstevia plantlets. Increasing the concentration of ancymidoland paclobutrazol decreased plantlet height and biomassfresh weight, but increased stem diameter. Paclobutrazol at0.1 mg/L was the best treatment to increase the vigor ofstevia plantlets. Light intensity at 20 µmol/m 2 /s gave betterplantlet vigor than other light intensities. It can be concludedthat multiplication of stevia shoots should be grown on MSmedium supplemented with 1.13 mg/L BA + 0.35 mg/L IAAand the vigor of the shoots can be increased by culturing onMS medium containing 0.1 mg/L paclobutrazol underfluorescence lamps with 20 µmol/m 2 /s light intensity.AbstrakStevia (Stevia rebaudiana Bertoni) adalah tanamanpemanis alami nir-kalori yang ditanam dengan kerapatanpopulasi yang sangat tinggi. Teknik kultur jaringan dapatdigunakan untuk perbanyakan tanaman secara massal dancepat untuk menyediakan bahan tanam unggul. Penelitiantelah dilakukan untuk meningkatkan pertumbuhan danmultiplikasi tunas dan keragaan planlet stevia. Eksplan yangdigunakan adalah tunas pucuk dan tunas samping dari planletyang ditumbuhkan pada medium MS tanpa zat pengaturtumbuh. Kombinasi BA dan IAA dengan konsentrasi yangberbeda digunakan untuk pertumbuhan dan multiplikasitunas, sedangkan zat penghambat tumbuh (ansimidol danpaklobutrazol) serta intensitas cahaya digunakan untukkeragaan planlet. Hasil penelitian menunjukkan bahwaeksplan stevia yang ditumbuhkan pada medium MS tanpa zatpengatur tumbuh menghasilkan tunas paling tinggi (4,5 cm)dengan dua tunas per eksplan. Multiplikasi tunas terbaikdiperoleh pada medium dengan BA 1,13 mg/L yangdikombinasikan dengan IAA 0,35 mg/L yang menghasilkan4,5 tunas dan 11,9 ruas per eksplan awal. Konsentrasiansimidol dan paklobutrazol berpengaruh nyata terhadappertumbuhan dan keragaan planlet stevia. Meningkatnyakonsentrasi ansimidol dan paklobutrazol menurunkan tinggiplanlet dan bobot basah biomassa, tetapi meningkatkandiameter batang. Paklobutrazol pada konsentrasi 0,1 mg/Lmerupakan perlakuan terbaik untuk meningkatkan keragaanplanlet stevia. Intensitas cahaya pada 20 µmol/m 2 /detikmemberikan keragaan planlet yang lebih baik dibandingkanintensitas cahaya yang lain. Dapat disimpulkan bahwamultiplikasi tunas stevia sebaiknya dilakukan pada mediumMS ditambah BA 1,13 mg/L + IAA 0,35 mg/L dan keragaanplanlet dapat ditingkatkan dengan menanam planlet padamedium MS ditambah paklobutrazol 0,1 mg/L di bawahlampu fluoresen dengan intensitas cahaya 20 µmol/m 2 /detik.


2017 ◽  
Vol 48 (5) ◽  
Author(s):  
Al-Obaidy & Khierallah

This research was conducted to study the effect of some plant growth regulators on in vitro shoots multiplication of stevia (Stevia rebaudiana Bertoni). The experiments included tests of various combinations of KIN with IBA or IAA in the shoot multiplication. Results indicated that KIN at 1.0 mg. L-1 plus 0.3 mg. L-1 of IBA produced the highest number of shoots (3.5 shoots) while KIN at 1.5 mg. L-1 plus IBA at 1.0 mg. L-1 produced the lowest shoot length (1.14 cm).  Hormone free medium produced the highest rate of the leaves number reached 28.56 leaves. KIN and IBA interaction increased fresh and dry weight significantly.   Treatment contained 2.0 mg -1 KIN plus 0.3 mg. L-1 IBA produced the highest fresh weight (1.739 g) while 0.5 mg. L-1 KIN and 0.3 mg. L-1 IBA produced the highest dry weight (0.822 g). As for the effect of interaction between the IAA and KIN it was significant in the number of shoots formed. Interaction between 1.0 mg. L-1 KIN with 0.1 mg. L-1IAA produced the highest number of shoots (3.8 shoots). Shoots length reached 8.10 cm in the media with 0.3 mg. L-1 IAA only. The highest fresh weight (1.267 g) was achieved with the interaction between 1.0 mg. L-1 KIN and 0.3 mg. L-1 IAA while 0.5 mg. L-1IAA without KIN produced the highest dry weight reached 0.138 g.  Shoots multiplication was improved by incorporation of the cytokinin TDZ in culture media. Shoots number, fresh and dry weights were increased significantly by adding 0.05 mg. L-1 of TDZ at present of 0.3 mg. L-1 of IBA giving 6.6 shoots, 0.974 g and 0.144 g respectively while shoots length decreased significantly as media without TDZ produced the highest shoots length reached 9.32 cm. The above results can adopt for the successful in vitro shoot multiplication of Stevia plants. 


2019 ◽  
Vol 1 (4) ◽  
pp. 261-269
Author(s):  
Thuy Linh Thi Nguyen ◽  
Ngoc Thi Pham ◽  
Thao Thi Ninh ◽  
Phuong Thao Thi Nguyen

This study was conducted to establish the procedure for in vitro propagation of Japanese cherry (Prunus sp.) to produce large quantity of plantlets and initial planting materials for climate adaptation research of this plant in Hanoi. Single nodal stems were used as the primary explants and initially produced shoots on MS medium supplemented with 1 mg L-1 BA. The highest shoot multiplication rate (9.57 times) was obtained on MS medium containing 1 mg L-1 BA and 0.25 mg L-1 a-NAA after 8 weeks of culture. 100% of the shoots produced roots with a mean of 10.10 roots per plant within 4 weeks on ½ MSM medium with 4 mg L-1 IBA. The survival rate of in vitro derived plantlets after a 6 to 7-week-period of rooting during acclimatization using a soil: coco peat: smoked rice husks (2:2:1, v/v/v) substrate was 100% and acclimatized plantlets showed good growth and development. This is the first report on a practical and efficient in vitro multiplication protocol for Japanese cherry in Vietnam, starting from shoot initiation to establishment of plants under greenhouse conditions.


2020 ◽  
Vol 50 (1) ◽  
Author(s):  
Marta Teresa Rokosa ◽  
Danuta Kulpa

ABSTRACT: The aim of the study was to develop optimum composition of plant growth regulators in media for the propagation and rooting of shoots of stevia (Stevia rebaudiana Bertoni) in in vitro cultures. Single-node shoot fragments obtained from plants propagated on MS medium were placed onto media supplemented with: BAP, 2iP and KIN at concentrations: 0.5, 1, 2 and 5 mg∙dm-3, whereas at the rooting stage with addition of: IAA, IBA and NAA at concentrations 1, 2, 4 and 8 mg∙dm-3. The highest number of shoots and leaves was reported for plants propagated on MS medium enriched with 0.5 mg∙dm-3 BAP. The greatest number of the longest roots was developed by stevia on the MS medium enriched with 1 mg∙dm-3 IAA.


2016 ◽  
Vol 72 (1) ◽  
Author(s):  
. JOKO-SANTOSO ◽  
Nurita TORUAN-MATHIUS ◽  
U SASTRAPRAWIRA ◽  
G SURYATMANA ◽  
D SAODAH

SummaryCinchona ledgeriana (Ledger) and C. succirubra (Succi) were industrial commodities which their barks of the trunk  contain alkaloid   used as raw materials in pharmaceutical, food, drug and beverages and chemical industries. The problem  faced in conventional plant propagation are. incompatibilities, high numbers of death caused by transportation, limited numbers and time consume in  plant materials  production. These problems may be  overcome by axillary buds multiplication.  The aim of the experiment were to find out propagation technology of Ledger and Succi by  tissue culture technique.  Experiments were conducted in three consecutive steps, viz the effect of (i) BAP on multiplication and growth of axillary’s bud of Ledger and Succi in vitro culture, (ii) IBA on root initiation and growth, (iii)  growth medium on the growth of plantlets in  acclimatization.The design of the experiments were Complete Randomized Design with 15 (i & ii) and four (iii) replications. The treatments were (i) 0,1,2,3,4, dan 5 mg/L BAP, (ii) 0.0; 0.5; 1.0; 1.5; 2.0; dan 2.5 mg/L IBA, and (iii) mixture of soil and rice husk charcoal (1:1), mixture of soil and compost (1:1),  mixture of soil, rice husk charcoal, and  compost  (1:1:1).  Parameters measured in the experiments were (i) the initiation of buds multiplication rate twice at axillary buds at subculture.  (ii) initiation  and  roots vigor. (iii) numbers of survived  plants and plants vigor. The explant source used derived from two-month old axillary buds cultured in Murashige and Skoog (MS) medium without growth regulator. Results of the experiment showed  that the best shoot multiplication of Ledger  and Succi  was obtained from the application of 3 mg/L BAP, with buds multipli-cation rate 7 buds/explant/month for Ledger, and 3-4 buds/explants/month for succi. The best root initation and root growth were found from the application of 2 mg/L IBA. The highest percentage of survived plantlets (100%) in acclimatization was obtained from mixture of soil and rice husk charcoal (1:1) medium.  Therefore it is  concluded that tissue culture technique could be used for planlet  mass propagation    of  elite C. Ledgeriana and C. Succirubra through axillary bud multiplication.Ringkasan Tanaman kina Cinchona ledgeriana (Ledger) dan C. succirubra (Succi)  merupakan tanaman industri yang mengandung alkaloid di dalam kulit batangnya dan berguna dalam bidang industri farmasi, makanan, minuman dan kimia. Kendala yang dihadapi dalam perbanyakan tanaman kina secara konvensional dengan sistem sambung  adalah inkompatibilitas,    kematian akibat pengangkutan cukup tinggi, jumlah bahan tanam yang diproduksi sangat terbatas dan waktu penyediaan yang cukup lama. Masalah tersebut dapat diatasi dengan menggunakan teknik kultur jaringan. Penelitian ini bertujuan untuk men-dapatkan teknologi perbanyakan tanaman kina Ledger dan Succi dengan teknik kultur jaringan. Penelitian terdiri atas (i) pengaruh BAP terhadap inisiasi dan penggandaan  tunas aksilar, (ii) pengaruh IBA terhadap inisiasi serta pertum-buhan akar planlet,   dan (iii) pengaruh beberapa medium terhadap pertumbuhan planlet dalam aklimatisasi. Percobaan menggunakan Rancangan Acak Lengkap, masing-masing diulang 15 (i & ii)  dan (iii) empat kali. Peubah yang diukur untuk percobaan (i) adalah waktu inisiasi tunas dan laju penggandaan tunas aksiler pada dua  kali  subkulur. (ii)  Waktu  inisiasi  dan vigor akar. (iii) Jumlah tanaman yang bertahan hidup setelah aklimatisasi, serta vigor tanaman. Sumber eksplan yang digunakan adalah tunas aksilar dari kecambah terpilih berumur dua bulan yang dikulturkan dalam medium Murashige dan Skoog tanpa zat pengatur tumbuh. Perlakuan untuk percobaan (i) adalah 0,0; 1,0; 2,0; 3,0; 4,0 dan  5,0 mg/L BAP, (ii) adalah 0,0; 0,5; 1,0; 1,5; 2,0; dan 2,5 mg/L IBA, sedang (iii) adalah medium tanam tanah, tanah : arang sekam (1:1),  tanah : kompos (1:1), tanah : arang sekam : kompos (1:1:1). Hasil yang diperoleh menunjukkan bahwa konsentrasi BAP terbaik untuk inisiasi dan penggandaan tunas tanaman kina Ledger dan Succi adalah 3 mg/L BAP, dengan laju penggandaan tujuh tunas/eksplan/bulan untuk Ledger dan 3-4 tunas/eksplan/bulan untuk Succi. Sedang untuk perakaran diperoleh dari medium MS dengan penambahan 2 mg/L IBA. Persentase tertinggi planlet (100%) yang mampu bertahan hidup pada aklimatisasi diperoleh dari medium campuran tanah : arang sekam (1:1). Berdasarkan hasil tersebut di atas dapat disimpulkan bahwa perbanyakan tanaman kina secara in vitro untuk menghasilkan bibit bermutu dapat dilakukan melalui teknik penggandaan tunas aksiler


2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Rizka Tamania SAPTARI ◽  
. SUMARYONO

 Stevia (Stevia rebaudiana Bert.), a sweetener plant, has been mass propagated by tissue culture technique. Optimal conditions to increase vigor of stevia plantlets are needed to support the sustainability of in vitro plantlet stocks and increase plantlet survival rate during acclimatization. The aim of this research was to investigate the effect of different media, culture vessel sizes, and vessel closure types on the vigor of stevia plantlets. The plant material was derived from apical shoot cuttings of sterile stevia plantlets grown on WP medium without growth regulator. Several treatments used in this study were solid or double layer media; short or tall culture vessel; and polypropile screw cap or plastic film closures.  Growth of plantlets was determined after 3 weeks of culture. Temperature and light intensity inside the vessels were also observed. The results showed that the best treatment to increase the vigor of stevia plantlets was a double-layer medium in a tall culture vessel (diameter 7 cm and height 11 cm) with either screw cap or plastic film. It was exhibited by significantly bigger stem diameter, more and bigger leaves, longer roots, and higher biomass fresh weight than those of other treatments. Higher temperature was observed on tall culture vessel, whereas all treatments did not significantly affect light intensity inside the vessels.[Keywords: stevia, plantlet vigor, double-layer medium, culture vessel size, vessel closure]AbstrakStevia (Stevia rebaudiana Bert.), tanaman pemanis, telah diperbanyak melalui teknologi kultur jaringan. Kondisi kultur optimal untuk meningkatkan vigor planlet stevia masih diperlukan untuk mendukung keberlanjutan tanaman stock in vitro dan untuk meningkatkan daya hidup planlet ketika diaklimatisasi. Penelitian yang dilakukan bertujuan untuk menentukan pengaruh penggunaan jenis media, ukuran botol kultur, dan jenis penutup botol yang berbeda terhadap vigor planlet stevia. Material tanaman yang digunakan didapat dari potongan tunas apikal plantlet stevia steril yang ditumbuhkan pada media WP tanpa zat pengatur tumbuh. Perlakuan jenis media terdiri atas media padat dan media dua-lapis (double-layer), ukuran botol pendek dan tinggi, serta jenis tutup ulir berbahan polipropilen dan lembaran plastik transparan. Pengamatan pertumbuhan planlet dilakukan setelah 3 minggu, juga dilakukan pengamatan terhadap suhu dan intensitas cahaya di dalam botol kultur. Hasil penelitian memperlihatkan bahwa perlakuan terbaik untuk meningkatkan vigor planlet stevia adalah dengan menggunakan media dua-lapis dalam botol kultur (diameter 7 cm, dan tinggi 11 cm), baik dengan tutup ulir maupun plastik. Hal ini ditunjukkan dari diameter batang lebih besar, daun lebih banyak dan besar, akar lebih panjang, serta bobot segar biomassa lebih tinggi dibandingkan dengan perlakuan lainnya. Suhu lebih tinggi terukur pada perlakuan botol tinggi, sedangkan semua perlakuan tidak mempengaruhi secara nyata intensitas cahaya di dalam botol kultur.[Kata kunci: stevia, vigor, media dua-lapis, ukuran botol kultur, tutup botol]


2019 ◽  
Vol 18 (6) ◽  
pp. 47-56
Author(s):  
Begum Kaplan ◽  
Selda Duraklioglu ◽  
Kenan Turgut

Stevia rebaudiana Bertoni is a perennial plant belonging to Asteraceae family and its leaves contain steviol glycosides (SGs) that are 150 to 300 times sweeter than sucrose. The sweeteners obtained from S. rebaudiana can be safely used by diabetics as insulin secretion is not required during digestion of this sweetener. As it has zero calories, it is also used in diet products. Adaptation studies for Stevia conducted in Antalya, Turkey have shown that the stevia plant could easily be cultivated as a perennial. However, the lack of a sustainable vegetative propagation method creates a significant problem for stevia production. In the generatively populations, homogeneity and therefore quality are decreased because of cross-pollination. Stevia, as a self-incompatible and cross-pollinated species, has been shown to have very high genetic diversity. Therefore, development of a sustainable in vitro propagation method to prevent genetic heterogeneity of selected varieties is crucial for stevia cultivation. The aim of this study was to evaluate 2 different gelling agents (plant agar and Gelrite) and 20 different growth regulators combinations. The results demonstrated an approximately 200-fold multiplication rate obtained within 13 weeks using MS medium supplemented with 0.5 mg·dm–3 BAP and 0.25 mg·dm–3 kinetin and solidified with Gelrite. Average stevioside and rebaudioside A contents in in vitro propagated plant samples were found to be 8.1% and 8.6%, respectively.


2016 ◽  
Vol 72 (1) ◽  
Author(s):  
. JOKO-SANTOSO ◽  
Nurita TORUAN-MATHIUS ◽  
U SASTRAPRAWIRA ◽  
G SURYATMANA ◽  
D SAODAH

SummaryCinchona ledgeriana (Ledger) and C. succirubra (Succi) were industrial commodities which their barks of the trunk  contain alkaloid   used as raw materials in pharmaceutical, food, drug and beverages and chemical industries. The problem  faced in conventional plant propagation are. incompatibilities, high numbers of death caused by transportation, limited numbers and time consume in  plant materials  production. These problems may be  overcome by axillary buds multiplication.  The aim of the experiment were to find out propagation technology of Ledger and Succi by  tissue culture technique.  Experiments were conducted in three consecutive steps, viz the effect of (i) BAP on multiplication and growth of axillary’s bud of Ledger and Succi in vitro culture, (ii) IBA on root initiation and growth, (iii)  growth medium on the growth of plantlets in  acclimatization.The design of the experiments were Complete Randomized Design with 15 (i & ii) and four (iii) replications. The treatments were (i) 0,1,2,3,4, dan 5 mg/L BAP, (ii) 0.0; 0.5; 1.0; 1.5; 2.0; dan 2.5 mg/L IBA, and (iii) mixture of soil and rice husk charcoal (1:1), mixture of soil and compost (1:1),  mixture of soil, rice husk charcoal, and  compost  (1:1:1).  Parameters measured in the experiments were (i) the initiation of buds multiplication rate twice at axillary buds at subculture.  (ii) initiation  and  roots vigor. (iii) numbers of survived  plants and plants vigor. The explant source used derived from two-month old axillary buds cultured in Murashige and Skoog (MS) medium without growth regulator. Results of the experiment showed  that the best shoot multiplication of Ledger  and Succi  was obtained from the application of 3 mg/L BAP, with buds multipli-cation rate 7 buds/explant/month for Ledger, and 3-4 buds/explants/month for succi. The best root initation and root growth were found from the application of 2 mg/L IBA. The highest percentage of survived plantlets (100%) in acclimatization was obtained from mixture of soil and rice husk charcoal (1:1) medium.  Therefore it is  concluded that tissue culture technique could be used for planlet  mass propagation    of  elite C. Ledgeriana and C. Succirubra through axillary bud multiplication.Ringkasan Tanaman kina Cinchona ledgeriana (Ledger) dan C. succirubra (Succi)  merupakan tanaman industri yang mengandung alkaloid di dalam kulit batangnya dan berguna dalam bidang industri farmasi, makanan, minuman dan kimia. Kendala yang dihadapi dalam perbanyakan tanaman kina secara konvensional dengan sistem sambung  adalah inkompatibilitas,    kematian akibat pengangkutan cukup tinggi, jumlah bahan tanam yang diproduksi sangat terbatas dan waktu penyediaan yang cukup lama. Masalah tersebut dapat diatasi dengan menggunakan teknik kultur jaringan. Penelitian ini bertujuan untuk men-dapatkan teknologi perbanyakan tanaman kina Ledger dan Succi dengan teknik kultur jaringan. Penelitian terdiri atas (i) pengaruh BAP terhadap inisiasi dan penggandaan  tunas aksilar, (ii) pengaruh IBA terhadap inisiasi serta pertum-buhan akar planlet,   dan (iii) pengaruh beberapa medium terhadap pertumbuhan planlet dalam aklimatisasi. Percobaan menggunakan Rancangan Acak Lengkap, masing-masing diulang 15 (i & ii)  dan (iii) empat kali. Peubah yang diukur untuk percobaan (i) adalah waktu inisiasi tunas dan laju penggandaan tunas aksiler pada dua  kali  subkulur. (ii)  Waktu  inisiasi  dan vigor akar. (iii) Jumlah tanaman yang bertahan hidup setelah aklimatisasi, serta vigor tanaman. Sumber eksplan yang digunakan adalah tunas aksilar dari kecambah terpilih berumur dua bulan yang dikulturkan dalam medium Murashige dan Skoog tanpa zat pengatur tumbuh. Perlakuan untuk percobaan (i) adalah 0,0; 1,0; 2,0; 3,0; 4,0 dan  5,0 mg/L BAP, (ii) adalah 0,0; 0,5; 1,0; 1,5; 2,0; dan 2,5 mg/L IBA, sedang (iii) adalah medium tanam tanah, tanah : arang sekam (1:1),  tanah : kompos (1:1), tanah : arang sekam : kompos (1:1:1). Hasil yang diperoleh menunjukkan bahwa konsentrasi BAP terbaik untuk inisiasi dan penggandaan tunas tanaman kina Ledger dan Succi adalah 3 mg/L BAP, dengan laju penggandaan tujuh tunas/eksplan/bulan untuk Ledger dan 3-4 tunas/eksplan/bulan untuk Succi. Sedang untuk perakaran diperoleh dari medium MS dengan penambahan 2 mg/L IBA. Persentase tertinggi planlet (100%) yang mampu bertahan hidup pada aklimatisasi diperoleh dari medium campuran tanah : arang sekam (1:1). Berdasarkan hasil tersebut di atas dapat disimpulkan bahwa perbanyakan tanaman kina secara in vitro untuk menghasilkan bibit bermutu dapat dilakukan melalui teknik penggandaan tunas aksiler


2017 ◽  
Vol 84 (2) ◽  
Author(s):  
Rizka Tamania SAPTARI ◽  
. SUMARYONO

AbstractStevia (Stevia rebaudiana Bert.), a sweetener plant, has been mass propagated by tissue culture technique. Optimal conditions to increase vigor of stevia plantlets are needed to support the sustainability of in vitro plantlet stocks and increase plantlet survival rate during acclimatization. The aim of this research was to investigate the effect of different media, culture vessel sizes, and vessel closure types on the vigor of stevia plantlets. The plant material was derived from apical shoot cuttings of sterile stevia plantlets grown on WP medium without growth regulator. Several treatments used in this study were solid or double layer media; short or tall culture vessel; and polypropile screw cap or plastic film closures.  Growth of plantlets was determined after 3 weeks of culture. Temperature and light intensity inside the vessels were also observed. The results showed that the best treatment to increase the vigor of stevia plantlets was a double-layer medium in a tall culture vessel (diameter 7 cm and height 11 cm) with either screw cap or plastic film. It was exhibited by significantly bigger stem diameter, more and bigger leaves, longer roots, and higher biomass fresh weight than those of other treatments. Higher temperature was observed on tall culture vessel, whereas all treatments did not significantly affect light intensity inside the vessels.[Keywords: stevia, plantlet vigor, double-layer medium, culture vessel size, vessel closure]AbstrakStevia (Stevia rebaudiana Bert.), tanaman pemanis, telah diperbanyak melalui teknologi kultur jaringan. Kondisi kultur optimal untuk meningkatkan vigor planlet stevia masih diperlukan untuk mendukung keberlanjutan tanaman stock in vitro dan untuk meningkatkan daya hidup planlet ketika diaklimatisasi. Penelitian yang dilakukan bertujuan untuk menentukan pengaruh penggunaan jenis media, ukuran botol kultur, dan jenis penutup botol yang berbeda terhadap vigor planlet stevia. Material tanaman yang digunakan didapat dari potongan tunas apikal plantlet stevia steril yang ditumbuhkan pada media WP tanpa zat pengatur tumbuh. Perlakuan jenis media terdiri atas media padat dan media dua-lapis (double-layer), ukuran botol pendek dan tinggi, serta jenis tutup ulir berbahan polipropilen dan lembaran plastik transparan. Pengamatan pertumbuhan planlet dilakukan setelah 3 minggu, juga dilakukan pengamatan terhadap suhu dan intensitas cahaya di dalam botol kultur. Hasil penelitian memperlihatkan bahwa perlakuan terbaik untuk meningkatkan vigor planlet stevia adalah dengan menggunakan media dua-lapis dalam botol kultur (diameter 7 cm, dan tinggi 11 cm), baik dengan tutup ulir maupun plastik. Hal ini ditunjukkan dari diameter batang lebih besar, daun lebih banyak dan besar, akar lebih panjang, serta bobot segar biomassa lebih tinggi dibandingkan perlakuan lainnya. Suhu lebih tinggi terukur pada perlakuan botol tinggi, sedangkan semua perlakuan tidak mempengaruhi secara nyata intensitas cahaya di dalam botol kultur.[Kata kunci: stevia, vigor, media dua-lapis, ukuran botol kultur, tutup botol] 


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