Modifikasi sistem kultur in vitro untuk meningkatkan vigor planlet stevia (Stevia rebaudiana Bert.)

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]

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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

E-Journal Menara Perkebunan ◽

10.22302/ppbbi.jur.mp.v79i2.59 ◽

2016 ◽

Vol 79 (2) ◽

Author(s):

. SUMARYONO ◽

Masna Maya SINTA

Keyword(s):

Light Intensity ◽

Plant Growth ◽

Stevia Rebaudiana ◽

Shoot Multiplication ◽

Culture Technique ◽

Multiplication Rate ◽

Stevia Rebaudiana Bertoni ◽

Plant Grown ◽

Planting Materials

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.

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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

E-Journal Menara Perkebunan ◽

10.22302/iribb.jur.mp.v79i2.59 ◽

2016 ◽

Vol 79 (2) ◽

Author(s):

. SUMARYONO ◽

Masna Maya SINTA

Keyword(s):

Light Intensity ◽

Plant Growth ◽

Stevia Rebaudiana ◽

Shoot Multiplication ◽

Culture Technique ◽

Multiplication Rate ◽

Stevia Rebaudiana Bertoni ◽

Plant Grown ◽

Planting Materials

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.

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Pengaruh jenis penutup botol kultur terhadap pertumbuhan planlet kelapa sawit (Elaeis guineensis Jacq.) Effect of different culture vessel closures on the growth of oil palm (Elaeis guineensis Jacq.) plantlets

E-Journal Menara Perkebunan ◽

10.22302/ppbbi.jur.mp.v79i1.68 ◽

2016 ◽

Vol 79 (1) ◽

Author(s):

Masna Maya SINTA ◽

Imron RIYADI ◽

. UMARYONO

Keyword(s):

Light Intensity ◽

Oil Palm ◽

Room Temperature ◽

Elaeis Guineensis ◽

Aluminum Foil ◽

Culture Vessel ◽

Shoot Height ◽

Plantlet Growth ◽

Vessel Closure ◽

Elaeis Guineensis Jacq

AbstractMicroenvironment inside the culture vessel such astemperature, light intensity, relative humidity, and aerationaffect growth and development of plantlets. This experimentwas conducted to determine the effect of different culturevessel closures on microenvironmental conditions inside thevessel and on growth of plantlets of oil palm. Shoots of oilpalm derived from somatic embryos were cultured on DFmedium for eight weeks in transparent culture bottlescovered with five different vessel closures e.i. screw cap withplastic wrap, screw cap, plastic wrap, aluminum foil, andautoclavable plastic. The culture vessels were placed in theculture room with light intensity 20 µmol/m 2 /sec for 12 hoursphotoperiod, at room temperature 26°C. Parametersobserved on plantlet growth were shoot height, biomass freshweight, leaf number, and leaf color grade, while onmicroenvironment were temperature and light intensity. Atthe end of experiment, the volume and fresh weight of theremaining medium were measured to determine evaporationrate of each treatment. Results show that the use of differentculture vessel closures affected the microenvironment insidethe vessel, the volume of the remaining medium, and thegrowth of the plantlets. The closure increased thetemperature by 1.6 – 2.6°C and decreased the light intensityby 1.7 – 8.7 µmol/m 2 /sec inside the culture vessels dependson the culture vessel closures. Culture vessels with aluminumfoil closure had the lowest temperature (28.9°C) and thelowest light intensity (10.8 µmol/m 2 /sec) gave the best resultin the growth of the plantlets. Better plantlets growth wasalso observed in the culture vessel with autoclavable plasticclosure that less expensive, therefore it can be used as analternative vessel closure for the growth of oil palm plantlets.AbstrakLingkungan mikro di dalam botol kultur seperti suhu,intensitas cahaya, kelembaban nisbi dan aerasi mem-pengaruhi pertumbuhan dan perkembangan planlet.Penelitian ini dilakukan untuk mengetahui pengaruhpenggunaan penutup botol kultur yang berbeda terhadapkondisi lingkungan mikro di dalam botol kultur danpertumbuhan planlet kelapa sawit. Planlet kelapa sawit asalembrio somatik dikulturkan dalam botol kultur bening berisimedium DF selama delapan minggu dan ditutup mengguna-kan lima jenis penutup botol yang berbeda yaitu tutup ulirdengan plastik wrap, tutup ulir, plastik wrap, aluminium foildan plastik tahan diautoklaf. Kultur diletakkan dalam ruangkultur, di bawah lampu TL dengan intensitas cahaya20 µmol/m 2 /detik dan suhu ruang 26 o C. Parameterpertumbuhan planlet yang diamati adalah tinggi planlet,bobot basah, jumlah daun dan kelas warna daun, sedangkanlingkungan mikro adalah suhu dan intensitas cahaya. Padaakhir eksperimen, volume dan bobot basah medium yangtersisa diukur untuk mengetahui tingkat penguapan padasetiap perlakuan. Hasil penelitian menunjukkan bahwapenggunaan penutup botol yang berbeda berpengaruhterhadap lingkungan mikro, volume medium tersisa dalambotol kultur dan pertumbuhan planlet. Penutup botolmeningkatkan suhu 1,6 – 2,6 o C dan menurunkan intensitascahaya 1,7 – 8,7 µmol/m 2 /detik di dalam botol tergantungpada jenis penutup botol yang digunakan. Botol kulturdengan penutup berbahan aluminium foil mempunyaiintensitas cahaya terendah (10,8 µmol/m 2 /detik) dan suhuterendah (28,9 o C) memberikan hasil terbaik pada pembesaranplanlet kelapa sawit. Pertumbuhan planlet yang baik jugaterdapat pada botol kultur dengan penutup plastik tahandiautoklaf yang lebih murah, sehingga penutup ini dapatdigunakan sebagai pilihan untuk pembesaran planlet kelapasawit.

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Pengaruh jenis penutup botol kultur terhadap pertumbuhan planlet kelapa sawit (Elaeis guineensis Jacq.) Effect of different culture vessel closures on the growth of oil palm (Elaeis guineensis Jacq.) plantlets

E-Journal Menara Perkebunan ◽

10.22302/iribb.jur.mp.v79i1.68 ◽

2016 ◽

Vol 79 (1) ◽

Author(s):

Masna Maya SINTA ◽

Imron RIYADI ◽

. UMARYONO

Keyword(s):

Light Intensity ◽

Oil Palm ◽

Room Temperature ◽

Elaeis Guineensis ◽

Aluminum Foil ◽

Culture Vessel ◽

Shoot Height ◽

Plantlet Growth ◽

Vessel Closure ◽

Elaeis Guineensis Jacq

AbstractMicroenvironment inside the culture vessel such astemperature, light intensity, relative humidity, and aerationaffect growth and development of plantlets. This experimentwas conducted to determine the effect of different culturevessel closures on microenvironmental conditions inside thevessel and on growth of plantlets of oil palm. Shoots of oilpalm derived from somatic embryos were cultured on DFmedium for eight weeks in transparent culture bottlescovered with five different vessel closures e.i. screw cap withplastic wrap, screw cap, plastic wrap, aluminum foil, andautoclavable plastic. The culture vessels were placed in theculture room with light intensity 20 µmol/m 2 /sec for 12 hoursphotoperiod, at room temperature 26°C. Parametersobserved on plantlet growth were shoot height, biomass freshweight, leaf number, and leaf color grade, while onmicroenvironment were temperature and light intensity. Atthe end of experiment, the volume and fresh weight of theremaining medium were measured to determine evaporationrate of each treatment. Results show that the use of differentculture vessel closures affected the microenvironment insidethe vessel, the volume of the remaining medium, and thegrowth of the plantlets. The closure increased thetemperature by 1.6 – 2.6°C and decreased the light intensityby 1.7 – 8.7 µmol/m 2 /sec inside the culture vessels dependson the culture vessel closures. Culture vessels with aluminumfoil closure had the lowest temperature (28.9°C) and thelowest light intensity (10.8 µmol/m 2 /sec) gave the best resultin the growth of the plantlets. Better plantlets growth wasalso observed in the culture vessel with autoclavable plasticclosure that less expensive, therefore it can be used as analternative vessel closure for the growth of oil palm plantlets.AbstrakLingkungan mikro di dalam botol kultur seperti suhu,intensitas cahaya, kelembaban nisbi dan aerasi mem-pengaruhi pertumbuhan dan perkembangan planlet.Penelitian ini dilakukan untuk mengetahui pengaruhpenggunaan penutup botol kultur yang berbeda terhadapkondisi lingkungan mikro di dalam botol kultur danpertumbuhan planlet kelapa sawit. Planlet kelapa sawit asalembrio somatik dikulturkan dalam botol kultur bening berisimedium DF selama delapan minggu dan ditutup mengguna-kan lima jenis penutup botol yang berbeda yaitu tutup ulirdengan plastik wrap, tutup ulir, plastik wrap, aluminium foildan plastik tahan diautoklaf. Kultur diletakkan dalam ruangkultur, di bawah lampu TL dengan intensitas cahaya20 µmol/m 2 /detik dan suhu ruang 26 o C. Parameterpertumbuhan planlet yang diamati adalah tinggi planlet,bobot basah, jumlah daun dan kelas warna daun, sedangkanlingkungan mikro adalah suhu dan intensitas cahaya. Padaakhir eksperimen, volume dan bobot basah medium yangtersisa diukur untuk mengetahui tingkat penguapan padasetiap perlakuan. Hasil penelitian menunjukkan bahwapenggunaan penutup botol yang berbeda berpengaruhterhadap lingkungan mikro, volume medium tersisa dalambotol kultur dan pertumbuhan planlet. Penutup botolmeningkatkan suhu 1,6 – 2,6 o C dan menurunkan intensitascahaya 1,7 – 8,7 µmol/m 2 /detik di dalam botol tergantungpada jenis penutup botol yang digunakan. Botol kulturdengan penutup berbahan aluminium foil mempunyaiintensitas cahaya terendah (10,8 µmol/m 2 /detik) dan suhuterendah (28,9 o C) memberikan hasil terbaik pada pembesaranplanlet kelapa sawit. Pertumbuhan planlet yang baik jugaterdapat pada botol kultur dengan penutup plastik tahandiautoklaf yang lebih murah, sehingga penutup ini dapatdigunakan sebagai pilihan untuk pembesaran planlet kelapasawit.

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Plant Morphogenesis under Different Light Intensity

Defect and Diffusion Forum ◽

10.4028/www.scientific.net/ddf.386.201 ◽

2018 ◽

Vol 386 ◽

pp. 201-206 ◽

Cited By ~ 1

Author(s):

Yuriy Nikolaevich Kulchin ◽

Olga Valerievna Nakonechnaya ◽

Irina Victorovna Gafitskaya ◽

Olga Vadimovna Grishchenko ◽

Tatyana Yuryevna Epifanova ◽

...

Keyword(s):

Light Intensity ◽

Stevia Rebaudiana ◽

Illumination Intensity ◽

Soil Conditions ◽

Light Sources ◽

Cultivation Time ◽

Optimal Values ◽

The Cost ◽

Fluorescent Lighting

The innovative LED light source (Sun Box) with irradiation spectrum close to the sun spectrum in the wavelength range 440-660 nm was used in experiment for study the influence of light intensity (75, 135, 230 and 382 μmol/s*m2) on the growth and development of plants. Standard fluorescent lighting was used as a control. The experiments were carried out on plantlets ofStevia rebaudianaandSolanum tuberosum, cvs. Snegir, Rozhdestvenskiy and Kamchatskii)in vitro. The illumination intensity of 75 and 230 μmol/s*m2promoted development ofS. rebaudianaplantlets with optimal values of morphometric parameters and well developed roots, which is important for plantlet adaptation to soil conditions. ForS. tuberosumplantlets (Snegir and Rozhdestvenskiy cultivars), radiation intensity of 135 μmol/s*m2was optimal for micropropagation. The illumination intensity of 230 μmol/s*m2led to a formation of plantlets with the largest total fresh mass among experimental groups. Sun Box light with intensity of 75 μmol/s*m2could be applicated for micropropagation of these cultivars: plantlets were the highest with the largest internodes number. Thus, the plant response to different light intensity was species-spesific, and – in case of potato plantlets – cultivar-spesific. The use of artificial light sources with distinct PPFD level could be preferable forS. tuberosumandS. rebaudianaplantlet micropropagationin vitro, as it could shorten the cultivation time, accelerate cultivation time, and reduce the cost of electricity.

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