Pertumbuhan Kultur Tunas Dahlia sp. Pada Media MS dengan Pengurangan Kadar Gula dan Tutup Tabung Berventilasi

 Dahlia sp. merupakan tanaman hias yang menghasilkan umbi mengandung inulin yang bermanfaat sebagai antioksidan. Kultur jaringan tanaman diaplikasikan untuk menghasilkan bibit dahlia yang seragam, dalam waktu relatif singkat, dan tidak dipengaruhi musim. Modifikasi media dan lingkungan in vitro sering dilakukan untuk meningkatkan pertumbuhan dan ketegaran. Penelitian ini bertujuan untuk mengetahui pengaruh pengurangan konsentrasi gula dan ventilasi pada tutup tabung kultur terhadap pertumbuhan tunas Dahlia sp. secara in vitro. Percobaan menggunakan rancangan acak lengkap faktorial dengan faktor yang diuji yakni gula dengan konsentrasi 10, 20 dan 30 g/l yang dikombinasikan dengan ventilasi tutup tabung berupa filter berjumlah 0, 1, 2 dan 4. Percobaan mempunyai 12 ulangan. Peubah yang diamati adalah tinggi tanaman, jumlah daun, jumlah tunas samping serta jumlah akar, diamati 2 kali seminggu dari umur 0 – 4 minggu. Pengukuran diameter batang, bobot basah dan bobot kering dilakukan pada umur 4 minggu. Hasil penelitian menunjukkan bahwa konsentrasi gula berpengaruh terhadap jumlah tunas samping sedangkan jumlah ventilasi filter berpengaruh terhadap tinggi tanaman dan jumlah daun. Tidak terdapat interaksi antara 2 faktor yang diujikan. Pada umur 4 minggu tinggi tanaman, jumlah daun, jumlah tunas samping serta jumlah akar yang tinggi terdapat pada perlakuan tanpa filter yang dikombinasikan dengan 10 dan 20 g/l gula, sedangkan terendah terdapat pada perlakuan ventilasi 4 filter yang dikombinasikan dengan 30 g/l gula. Pada perlakuan ventilasi 2 filter yang dikombinasikan dengan 20 g/l gula menghasilkan diameter batang tertinggi, berbeda nyata dengan perlakuan lainnya, pada perlakuan ini juga menghasilkan bobot basah dan bobot kering yang lebih tinggi.

Rudiyanto, R., Wulandari, D. R., & Ermayanti, T. M. (2018). Pertumbuhan kultur tunas Dahlia sp. pada media MS dengan pengurangan kadar gula dan tutup tabung berventilasi. Prosiding SEMNASTAN, 184-195.

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Somatic Embryo Germination of Jatropha curcas L in Presence of Sucrose and Poly Ethylene Glycol (PEG)

Jatropha curcas L. is a potential source of a non-edible biofuel. Conventional propagation of J. curcas technique has some limitations. Somatic embryo can produce a large number of embryos and obtain a large number of plants all year round. Treatment of sucrose in combination with polyethylene glycol (PEG) was proven to enhance germination of somatic embryos in many plant species. The aim of the study was to investigate the effect of sucrose in combination of PEG on somatic embryo germination in J. curcas. Globular somatic embryos at 0.025-0.030 g fresh weight having 0.4-0.5 cm in diameter were grown on MS medium solidified with 3 g/l of Gelzan supplemented with sucrose at 20, 30, 40, and 50 g/l in combination with PEG at 0, 2.5, 5, 10, and 15%. Results showed that the best medium for germination of J. curcas somatic embryo cultures was MS medium supplemented with 20 and 30 g/l of sucrose in combination with 5% of PEG. The numbers of germinated embryos per clump had significant enhancement on those medium compared with the control (PEG free treatment) (2.65 to 5.65) and (2.55 to 5.50). In addition, those treatments resulted in the highest percentage of clumps forming germinated embryos (100%), with an average of normal germinated embryos at 94.163 and 96.065%. The addition of 40 and 50 g/l of sucrose in combination with 15% of PEG caused all embryos to fail at germinating.

Rudiyanto, R., Efendi, D., & Ermayanti, T. M. (2014, June). Somatic embryo germination of Jatropha curcas L in presence of sucrose and poly ethylene glycol (PEG). In Annales Bogorienses (Vol. 18, No. 1, pp. 35-43).

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Media Dasar dalam Kultur Jaringan Tanaman

 

Secara kasat mata, orang awam melihat media dalam kultur jaringan hanya berupa agar yang dipadatkan di dalam botol. Namun agar tersebut tentu bukan sembarang agar, media tanam dalam kultur jariangan tersebut selain berisi agar juga berisi nutrisi yang diperlukan oleh tanaman agar dapat tumbuh dan berkembang dengan baik. Media dasar yang digunakan tersebut sebagai sumber nutrisi bagi sel atau jaringan tanaman yang akan dikulturkan secara in vitro (suci hama/pathogen). Media ini berisi nutrisi (hara makro dan mikro), sumber karbon (sukrosa), vitamin, bahan organik dan zat pengatur tumbuh yang dibutuhkan untuk pertumbuhan dan perkembangan sel atau jaringan tanaman.

Detection of Somaclonal Variation in Jatropha curcas Linn. Plantlets Regenerated from Somatic Embryo using ISSR Markers

Beberapa komponen penting dari media dasar kultur jaringan meliputi:

1.      Unsur Hara Makro: Nitrogen, fosfor, kalium, kalsium, magnesium, dan sulfur (makroelemen)

2.      Unsur Hara Mikro: Besi, mangan, boron, tembaga, seng, dan molibdenum (mikroelemen). Komposisi hara makro dan mikro ini berbeda-beda antara jenis media dasar yang digunakan

3.      Karbohidrat: Biasanya ditambahkan sukrosa (gula) sebagai sumber energi bagi sel tanaman. Dalam kultur jaringan tanaman konsentrasi gula yang digunakan umumnya sebanyak 30 g/L media.

4.      Vitamin: Beberapa vitamin penting bagi tanaman seperti tiamin (vitamin B1), piridoksin (vitamin B6), dan asam nikotinat untuk mendukung metabolisme sel.

5.      Zat Pengatur Tumbuh: Zat pengatur tumbuh yang sering digunakan antara lain auksin dan sitokinin untuk menginduksi pertumbuhan dan diferensiasi sel atau jaringan.

6.      Pemadat: Yakni menggunakan agar untuk membuat media menjadi padat sehingga eksplan dapat ditancapkan di atas permukaan media.

7.      pH Media: pH media biasanya diatur antara 5.6 hingga 5.8 agar ketersediaan hara dalam media dapat diserap oleh tanaman.

Teknik Dasar Kultur Jaringan Tanaman, Pengertian, Keunggulan dan Aplikasinya

Jenis media dasar yang biasanya digunakan dalam kultur jaringan antara lain media MS: Murashige & Skoog (1962); WPM: McCown & Lloyd (1981); DKW: Driver & Kuniyuki (1984); NN: Nitsch & Nitsch (1969); B5: Gamborg et al. (1968) dsb.

Sumber: Rudiyanto et al., 2021

Pengaruh Modifikasi KH2PO4, NH4NO3 dan Sukrosa terhadap Pertumbuhan Tunas serta Pembentukan Umbi Mikro Taka (Tacca leontopetaloides) secara In vitro

Catatan: Setiap jenis tanaman memerlukan optimasi media dasar agar dapat tumbuh dan berkembang dengan optimal

 

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Comparison of the Reduction Effect of Sucrose and Table Sugar Concentration on Growth Characteristics of Red Ginger (Zingiber officinale Rocs.) Cultured in Liquid Medium

The aim of this research was to compare the reduction effect of sucrose or table sugar concentration on growth  characteristics of red ginger cultured in MS liquid medium. Shoots of red ginger was cultured on MS liquid medium without addition of plant growth regulators, supplemented with 5, 10, and 20 g/l of sucrose or table sugar for 8 weeks. Resulted plantlets were acclimatized in a greenhouse to investigate their growth and survival rate. Numbers of stomata, chlorophyll concentration as well as cross section of leaves from plantlets grown in vitro were compared to those of transplants grown in the greenhouse. The results showed that the use of table sugar at concentration of 20 g/l gave the best growth of red ginger. Meanwhile, the reduction of table sugar from 20 to 10 g/l reduced growth and survival rate of in vitro shoots as well as that of transplants in the greenhouse. Only few shoots formed roots when they were grown on the medium containing 5 g/l of table sugar, and transplants failed to grow in the greenhouse. It found that the chlorophyll content of in vitro plantlets was lower than those of transplants grown in the glasshouse. However, the number of stomata of the in vitro plantlets was higher than that of transplants grown in the glasshouse. There was no anatomical abnormalities found on the cross section of leaves between in vitro plantlets and transplants grown in the greenhouse. The replacement of sucrose with table sugar may reduce the production cost of plantlets. 

Hapsari, B. W., Ermayanti, T. M., Rantau, D. E., & Rudiyanto, R. (2011, June). Comparison of the Reduction Effect of Sucrose and Table Sugar Concentration on Growth Characteristics of Red Ginger (Zingiber officinale Rocs.) Cultured in Liquid Medium. In Annales Bogorienses (Vol. 15, No. 1, pp. 15-20).

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The effect of sucrose concentrations and different types of tube cap on in vitro growth of Dahlia (Dahlia sp.) using vermiculite as substrate

Dahlia sp. is an ornamental plant belongs to Compositae. Plantlets produced through conventional in vitro culture have low growth ability during acclimatization. The use of ventilation on in vitro culture is expected to affect the condition of the plantlet micro environment. Reducing sucrose concentration will improve the physiological functions of the plant. This study aimed to investigate the effect of reduction of sucrose concentrations and the use of various culture tube caps on the growth of shoots cultures of Dahlia sp. The experiments used was factorial completely randomized design with two factors ie. concentrations of sucrose (10, 20, and 30 g/l) in combination with four different type of tube caps (Al foil, transparent plastic, filter 1 cm and 2 cm). The experiments used vermiculite as substrate and MS medium with 9 replicates. Parameters observed were plant height, number of leaves, number of nodes, and number of roots which observed every week up to 8 weeks of culture. At 8 weeks culture, fresh weight, and dry weight were also recorded. The results showed that reducing sucrose concentration in combination with tube cap significantly affected on the growth of shoot height, number of leaves, nodes and roots as well as length of root. The highest Dahlia shoot was found in the media containing 10 g/l sucrose in tubes with plastic cap and in 20 g/l sucrose with 2 cm filter cap. The highest number of leaves and nodes was found in media containing 30g/l of sucrose using Al foil and 1 cm filter caps, both were significantly different with others. The highest fresh weight was found in MS medium containing 30 g/l of sucrose with 1 cm filter caps. Highest number of roots were found in the 10 g/l of sucrose using 1cm filter cap and in 30 g/l sucrose with a plastic cap.

Rudiyanto, R., Hapsari, B. W., Rantau, D. E., & Ermayanti, T. M. (2022). The effect of sucrose concentrations and different types of tube cap on in vitro growth of Dahlia (Dahlia sp.) using vermiculite as substrate. Biogenesis: Jurnal Ilmiah Biologi, 10(2), 215-224.

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Pengaruh Modifikasi KH2PO4, NH4NO3 dan Sukrosa terhadap Pertumbuhan Tunas serta Pembentukan Umbi Mikro Taka (Tacca leontopetaloides) secara In vitro

Polynesian arrowroot (Tacca leontopetaloides (L.) Kuntze), which is one of the bulbous herbaceous plants, have high nutritional value. Modification of macro nutrients by reducing nitrogen content and increasing phosphorus on the medium gave affects on shoot growth and initiated micro tuber formation on in vitro cultures. The aim of this research was to determine the effect of modified macro nutrients in combination with the increase in sucrose concentrations on shoot growth and micro tuber formation of T. leontopetaloides. The experimental design was factorial completely randomized design.The factors tested were modifications of MS macro nutrients that were. M1 (170 mg/l KH2PO4 and 1650 mg/l NH4NO3; normal, control treatment); M2 (340 mg/l KH2PO4 and 825 mg/l NH4NO3); and M3 (680 mg/l KH2PO4 and 412.5 mg/l NH4NO3 in combination with 30 (S1) (control treatment), 40 (S2), 50 (S3) and 60 g/l of sucrose (S4). The variables tested were shoot height, number of leaves, number of roots and number of micro tuber which were observed weekly at 0-8 weeks after culturing. The results showed that the modification of macro nutrient in combination with sucrose concentration had significant effect on shoot height, number of leaves and number of roots but not significant on the number of tubers. The highest shoots were found in M1S3 treatment, the highest number of leaves was in M1S1 and M1S3 treatment and the highest number of roots was in M1S4 treatment. The number of tubers not significantly different between the treatments tested.

Rudiyanto, R., Hapsari, B. W., & Ermayanti, T. M. (2018). Pengaruh Modifikasi KH2PO4, NH4NO3 dan Sukrosa terhadap Pertumbuhan Tunas serta Pembentukan Umbi Mikro Taka (Tacca leontopetaloides) secara In vitro. JURNAL BIOLOGI INDONESIA, 14(1).

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