Induction and Proliferation of Moringa oleifera Somatic Embryo Callus using Solid Liquid and Temporary Immersion System

Moringa oleifera Lam., a promising functional food, can be propagated rapidly through somatic embryogenesis technique. This method has advantages, such as rapid seed generation, absence of a dormant phase, and bipolar cotyledons. The induction of M. oleifera callus involves plant growth regulators, the type of explants, and the media used. In this study, we investigated the effects of 0.0, 0.5, 1.0 and 2.0 mg/L of BAP and picloram on inducing embryonic callus in M. oleifera plants. Additionally, we examined the impact of amino acids such as proline, glutamine, and cysteine on the proliferation stage of the embryonic callus. The results showed that 1 mg/L BAP and picloram significantly enhanced callus induction, as indicated by increased callus diameter, weight, and volume. The growth of M. oleifera callus was also influenced by the concentration of amino acids. The optimal proliferation callus was achieved using 200 mg/L of proline in Driver and Kuniyuki Walnut (DKW) medium with the Temporary Immersion System (TIS). Furthermore, embryogenic callus was generated using 200 mg/L proline, 50 mg/L glutamine, and 50 and 100 mg/L cysteine. The addition of 200 mg/L proline to DKW yielded the best results in TIS.

Rudiyanto, Purwito A, Efendi D, Martin AF. 2024. Induction and proliferation of Moringa oleifera somatic embryo callus using solid liquid and temporary immersion system. IJAB 32(3):294-300.

 

Link DOI: https://doi.org/10.17957/IJAB/15.2204

 

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Pertumbuhan Dan Morfologi Kultur Tunas Sempur (Dillenia philippinensis Rolfe) Pada Media MS-BAP-NAA

Propagation of sempur (Dillenia philippinensis Rolfe) using conventional vegetative methods is considered ineffective due to slow process and low germination level. Propagation by tissue culture is considered more effective because it does not depend on the season and requires less plant material. It can benefit sempur conservation since it is categorized as threatened with extinction in 2020 on the IUCN red list. This study aimed to evaluate the effect of BAP and NAA on the growth of sempur’s shoots and to observe the morphology of shoot culture. The media used as a control was MS without growth regulators. The treatment medium was MS with the addition of BAP and NAA. Shoots were used as explants. Shoot growth and plantlet morphology were observed eight weeks after planting. The results indicated that the combination of 1 mg/l BAP and 0.5 mg/l NAA resulted in the highest total number of leaves, nodes and adventitious shoots. The combination of BAP and NAA, each 1 mg/l, resulted in the highest fresh weight and callus formation. The combination of 1 and 2 mg/l BAP and 0.5-1 mg/l NAA could not form roots. In comparison, the combination of BAP and NAA in culture media could change the shape and size of the leaves. The survival rate of growth of plantlets derived from MS medium was 50% at 22 weeks after acclimatization.

Rantau, D. E., Wulandari, D. R., Ermayanti, T. M., Rudiyanto, R., Hapsari, B. W., Wulansari, A. & Firdaus, H. L. Pertumbuhan Dan Morfologi Kultur Tunas Sempur (Dillenia philippinensis Rolfe) Pada Media MS-BAP-NAA. Jurnal Penelitian Hutan Tanaman, 18 (1):65-78. DOI: https://doi.org/10.20886/jpht.2021.18.1.65-78

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

Physic nut (Jatropha curcas Linn.) has the potential as a source of sustainable biofuels. Somatic embryo proliferation of J. curcas may cause somaclonal variations. This research aimed to investigate somaclonal variations of J. curcas somatic embryo derived-plantlet using ISSR markers. Somatic embryos of J. curcas at the globular phase were cultured on liquid MS medium supplemented with 0, 0.5, 1.0, 1.5, and 2.0 mg L-1 of 2,4-D. Parameter observed were embryos weight, embryos volume, colour, and size of embryos. After proliferation, the embryos were cultured on a germination medium until the cotyledonary phase. The plantlet’s leaves were used for DNA samples for ISSR analysis. The results showed that proliferation of J. curcas somatic embryos was optimal at MS medium supplemented with 1 mg L-1 2,4-D and produced the highest weight and volume of embryos. The furthest genetic distance occurred between the control and J. curcas plantlet which was regenerated from MS + 1 mg L-1 2,4-D which had 0.60 of similarity coefficient.

Rudiyanto, Efendi, D., Al-Hafiizh, E., & Ermayanti, T. M. (2021). DETEKSI VARIASI SOMAKLONAL PLANLET Jatropha curcas Linn. HASIL REGENERASI EMBRIO SOMATIK DENGAN MARKA MOLEKULAR ISSR. Jurnal Bioteknologi & Biosains Indonesia (JBBI), 8(1), 14-24.

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Growth and Proline Accumulation in Response to Osmotic Stress Induced by Polyethylene Glycol Treatment in Tacca leontopetaloides Cultured In Vitro

 

Tacca leontopetaloides known as Polynesian arrowroot is a wild tuberous perennial herb. In addition, T. leontopetaloides is a valuable plant not only as staple food but also as a medicinal herb. Nonetheless, there are very few literatures and research on the potential of this plant. In this study, the effect of polyethylene glycol (PEG) as stress osmoticum in T. leontopetaloides cultured in vitro on growth and proline accumulation was investigated. The shoot culture of T. leontopetaloides was subjected to six levels of PEG concentrations (2.5; 5; 7.5; 10; 12.5 and 15%). Growth parameter was evaluated by observing shoot height, number of shoots, number of leaves and number of roots for 6 weeks of culture. After 6 weeks of culture, fresh weight and proline content were determined. The results showed that number of shoots and number of leaves increased after 2.5% PEG treatment while number of roots increased after 5% PEG treatment. Conversely, higher concentrations of PEG inhibited growth. In line with this, culture fresh weight decreased along with the increase of PEG concentrations. To summarize, the growth of culture decreased along with the increase of PEG concentrations. In contrast, proline concentration increased along with the increase of PEG concentrations.

Martin, A. F., Hapsari, B. W., Rudiyanto, R., & Ermayanti, T. M. (2018). Growth and proline accumulation in response to osmotic stress induced by polyethylene glycol treatment in Tacca leontopetaloides cultured in vitro.

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