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Studies on micropropagation of gerbera (Gerbera jamesonii Bolus)
... Diện tích trồng lạc tăng nhanh trong gần 30 năm trở lại đây, đạt khoảng 246.000 ha (Thủy và ctv., 2013). Cây lạc quan trọng về mặt kinh tế trong nền nông nghiệp thế giới, sự chú trọng trong sản xuất đã chuyển từ đầu vào tối thiểu sang đầu vào cường độ cao, dẫn đến năng suất cao và chất lượng hạt giống được cải thiện (Bhavya et al., 2013). ...
Nghiên cứu này được thực hiện với nguồn vật liệu từ cuống lá, phiến lá, rễ mầm và trục thượng diệp của giống lạc L14. Kết quả nghiên cứu cho thấy công thức khử trùng hiệu quả đối với mẫu hạt lạc là khử trùng kép bằng dung dịch NaOCl 2,5% và 0,5ml Tween20 trong thời gian lần 1 là 5 phút, lần 2 là 15 phút, giữa 2 lần mẫu được rửa sạch bằng nước cất tiệt trùng ít nhất 3 lần. Sau 3 tuần nuôi cấy trên nền môi trường MS có bổ sung 10 mg/L 2,4-D, mẫu cấy từ phiến lá cảm ứng tạo mô sẹo cao nhất: 100%. Môi trường 2,4-D nồng độ 5 mg/L cho mô sẹo dạng chắc và xanh, mô sẹo cảm ứng từ phiến lá tạo chồi cao nhất trên môi trường bổ sung 2,4-D 1,25 g/L và BAP 1 mg/L. Tỉ lệ tạo rễ từ chồi cảm ứng từ mô sẹo phiến lá cao nhất trên môi trường bổ sung NAA 0,2 mg/L và với hàm lượng đa lượng MS 100%.
... A tissue culture method developed for the production of disease-free, high quality planting material and for the rapid production of many uniform plants is now possible to provide clean and uniform planting materials, in plantations like banana, pine, date, rubber tree, etc., field crops like sugarcane, pineapple, tomato, potato etc., root crops like cassava, yam, sweet potato etc. (Ogero et al. 2012 and agronomical crops like sugarcane , ornamental plants like orchids such as Coelogyne stricta (through pseudobulb segment culture) (Basker and Bai, 2006), Dendrobiwn crysanthum (Rao and Barman, 2014), anthuriums, gerbera (Gerbera jamesonii) (Bhargava et al. 2013), and Begonia rubrovenia var. meisneri which is a rare and endemic plant of Meghalaya (Mittal and Decker, 2013). ...
... ,Paduchuri et al. (2010),Son et al. (2011), Akter et al. (2012,Samanthi et al. (2013) andBhargava et al. (2013). ...
An experiment was undertaken to develop an improved in vitro regeneration protocol in gerbera. Murashige and Skoog (MS) medium was supplemented with various growth regulators at different concentrations for callus induction and organogenesis. Newly emerging leaves of Gerbera cv. Rosalin were used as explants. Experimental results showed that maximum rate (74.07%) of formation of callus with good growth was recorded on MS medium supplemented with 2.0mgL-1 2,4-D + BAP 0.5mgL-1. Best shoot regeneration (57.8 %) with maximum shoot number (12.0) was achieved on with BAP 2.0mgL-1 + NAA 0.5mgL-1 fortified MS medium. Maximum (66.7 %) and earliest (12.3 days) root formation in shoots was recorded on IBA 3.0mgL-1 is ½MS media. Survival rate of regenerated plantlets was maximum (73.33 %) in the potting mixture containing garden soil, sand and vermicompost (1:1:1).
... Almost similar results with 95% survivability and 114 survival plantlets were reported on G. jamesonii 'Black Jack' (Winarto and Yufdy, 2017). While other gerbera's studies, 50-60% survivability of plantlets were established in plastic pots (10 cm) containing a mixture of Farmyard manure (FYM): sand (1:1) (Kumar and Kanwar, 2006), 70-80% survivability was established in a mixture of sand and barnyard manure (1:1, v/v) (Kumar and Kanwar, 2007), 92% in autoclaved sand and perlite (1:1 ratio) (Shabanpour et al., 2012), 90% on cocopeat (Bhargava et al., 2013), 66.7-90.0% on a mixture of soil and peat (Shylaja et al., 2014), 84% survival rate in peat + perlite mixture (2:1) (Gök et al., 2016). ...
A new route of in vitro mass propagation protocol of Gerbera jamesonii (H. Bolus ex Bolus f.) derived from application of
thin cell layers (TCL) and adenine sulphate (AS) was successfully developed and established. Shoot tip explants and halfstrength
MS medium containing 0.25 mg/l N6-benzylaminopurine (BAP), 20 g/l sucrose and 7 g/l Swallow agar were used as
explant source and basic medium. Different TCL of transversal TCL (tTCL) and longitudinal TCL (lTCL) in four slicing
positions of 1, 2, 3 and 4; varieties and clones i.e. G. jamesonii ‘Black Jack’, ‘Carambole’, ‘Nuance’, ‘Violente’, 01.098 and 11.46
clone; AS concentrations viz. 0, 20, 40, 60, 80 and 100 mg/l were tested in the study. Each step of in vitro culture established
had unique and specific results. In the initiation stage, first slicing position of ‘Black Jack’ shoot tip tTCL was the most optimal
combination treatment to produce 7.0 shoots per explant with 13.5 leaves. The first slicing position on shoot tip explants of
01.098 clone tTCL and 20 mg/l AS in half-strength MS medium containing 0.25 mg/l BAP were the most optimal
combination treatment in obtaining the highest number of shoots produced per shoot up to 9.4 shoots per shoot with 34.1
leaves and 2.37 cm length of leaves in the proliferation stage, however the treatment did not give significant effect compared to
control. Under periodical subcultures on the basic medium, number of shoots and leaves increased gradually from the initial
culture with 3-6 shoots per shoot and 9.4-11.6 leaves till the fourth subculture with 6-11 shoots per shoot and 16.7-28.8 leaves
and declined thereafter. Subculturing of shoots in accordance to produce qualified shoots for planting materials could be
carried out till sixth to seventh subculture. The highest shoot multiplication rate (SMR) was established on 01.098 clone with
as high as 7.3. The well shoots were easily rooted on half-strength MS medium supplemented with 0.1 mg/l BAP, 0.05 mg/l
NAA and 1.5 g/l AC. Plantlets were then transferred to ex vitro condition for acclimatization on a mixture of burned-rice husk
and organic manure (1:1, v/v) with 85-100% survivability. The ‘Black Jack’ and 11.46 clone were the best genotypes on the
acclimatization stage with 100% survivability of plantlets. Results of the study have implication that first slicing position of
shoot tip tTCL can be applied in establishing of in vitro propagation protocol for other gerberas.
... Hasil penelitian Artha (2014) menunjukkan bahwa terdapat kecenderungan peningkatan pertumbuhan pucuk bibit melinjo dan meranti dengan penambahan 10-30% cocopeat pada media tanam. Lebih lanjut penelitian Bhargava et al. (2013) Berdasarkan Tabel 4 diketahui bahwa pengaruh interaksi antara media tanam dan varietas pada tinggi kanopi menunjukkan bahwa varietas Black Jack merupakan varietas terbaik pada media cocopeat dengan tinggi kanopi tanaman10.10 cm, akan tetapi tidak berpengaruh nyata jika dibandingkan dengan varietas lain pada taraf 5%. ...
... Almost similar results with 95% survivability and 114 survival plantlets were reported on G. jamesonii 'Black Jack' (Winarto and Yufdy, 2017). While other gerbera's studies, 50-60% survivability of plantlets were established in plastic pots (10 cm) containing a mixture of Farmyard manure (FYM): sand (1:1) (Kumar and Kanwar, 2006), 70-80% survivability was established in a mixture of sand and barnyard manure (1:1, v/v) (Kumar and Kanwar, 2007), 92% in autoclaved sand and perlite (1:1 ratio) (Shabanpour et al., 2012), 90% on cocopeat (Bhargava et al., 2013), 66.7-90.0% on a mixture of soil and peat (Shylaja et al., 2014), 84% survival rate in peat + perlite mixture (2:1) (Gök et al., 2016). ...
A new route of in vitro mass propagation protocol of Gerbera jamesonii (H. Bolus ex Bolus f.) derived from application of thin cell layers (TCL) and adenine sulphate (AS) was successfully developed and established. Shoot tip explants and half-strength MS medium containing 0.25 mg/l N6-benzylaminopurine (BAP), 20 g/l sucrose and 7 g/l Swallow agar were used as explant source and basic medium. Different TCL of transversal TCL (tTCL) and longitudinal TCL (lTCL) in four slicing positions of 1, 2, 3 and 4; varieties and clones i.e. G. jamesonii ‘Black Jack’, ‘Carambole’, ‘Nuance’, ‘Violente’, 01.098 and 11.46 clone; AS concentrations viz. 0, 20, 40, 60, 80 and 100 mg/l were tested in the study. Each step of in vitro culture established had unique and specific results. In the initiation stage, first slicing position of ‘Black Jack’ shoot tip tTCL was the most optimal combination treatment to produce 7.0 shoots per explant with 13.5 leaves. The first slicing position on shoot tip explants of 01.098 clone tTCL and 20 mg/l AS in half-strength MS medium containing 0.25 mg/l BAP were the most optimal combination treatment in obtaining the highest number of shoots produced per shoot up to 9.4 shoots per shoot with 34.1 leaves and 2.37 cm length of leaves in the proliferation stage, however the treatment did not give significant effect compared to control. Under periodical subcultures on the basic medium, number of shoots and leaves increased gradually from the initial culture with 3-6 shoots per shoot and 9.4-11.6 leaves till the fourth subculture with 6-11 shoots per shoot and 16.7-28.8 leaves and declined thereafter. Subculturing of shoots in accordance to produce qualified shoots for planting materials could be carried out till sixth to seventh subculture. The highest shoot multiplication rate (SMR) was established on 01.098 clone with as high as 7.3. The well shoots were easily rooted on half-strength MS medium supplemented with 0.1 mg/l BAP, 0.05 mg/l NAA and 1.5 g/l AC. Plantlets were then transferred to ex vitro condition for acclimatization on a mixture of burned-rice husk and organic manure (1:1, v/v) with 85-100% survivability. The ‘Black Jack’ and 11.46 clone were the best genotypes on the acclimatization stage with 100% survivability of plantlets. Results of the study have implication that first slicing position of shoot tip tTCL can be applied in establishing of in vitro propagation protocol for other gerberas.
... This problem was also reported by Mishra and Datta (2001). Bhargava et al. (2013) also reported significant improvement in shoot proliferation with the use of growth hormones and linear increase in number of shoots with increased concentration of cytokinins and their combinations. They also reported that MS medium supplemented with 4mgl -1 BAP and 2mgl -1 kinetin was found optimum for proliferation of quality shoots in gerbera. ...
... Many researchers have reported successful shoot regeneration from capitulum explant (Hussein et al. 2008;Bhatia et al. 2011). Similar to the 123 results obtained in this study, Warar et al. (2008) and Bhargava et al. (2013) have also reported efficiency of BA and IAA in in vitro shoot induction from capitulum explant. The dose of radiation at which 50% survival or mortality is obtained is called lethal dose LD 50 . ...
In this study, the effect of gamma irradiation in inducing resistance/tolerance towards powdery mildew disease was investigated in Gerbera jamesonii cv. ‘Harley’. In vitro shoot cultures were established through capitulum explants on Murashige and Skoog medium supplemented with 22.2 µM 6-benzyladenine (BA) and 2.53 µM indole acetic acid (IAA), followed by gamma irradiation of regenerated shoots (3–5 cm). Activity of four antioxidant enzymes i.e. superoxide dismutase, ascorbate peroxidase, catalase and glutathione reductase increased significantly as compared to the control and reached to highest level at the most stringent doses of mutagen. Ninety randomly selected irradiated plants (6 months old) and 100 control plants were inoculated with fungal conidial suspension, to screen for tolerance/resistance against powdery mildew. The severity of the disease was recorded on 0–4 scale with ‘0’ indicating highly resistant; ‘1’ indicating resistant; ‘2’ indicating medium resistance; ‘3’ indicating susceptible and ‘4’ indicating highly susceptible. Three plants (3.33%) irradiated with 5 Gy were found to be tolerant to powdery mildew as these plants showed slight and delayed development of fungal colonies on the leaves. The random amplified polymorphic DNA characterization showed that the irradiated plants had DNA patterns that were different from the control and mother plants.
... In the commercial production of gerberas, micropropagation enables the large-scale production of plants in a limited space, as well as the obtaining of disease-and pest-free uniform plants (Bhargava et al., 2013). This technique also ensures precision in production schedules and product quality, regarding plant homogeneity and vigor. ...
Micropropagation requires controlling contamination that might compromise the success of the process. Thermal sterilization is traditionally used; however, costs deriving from equipment acquisition and maintenance render this technique costly. With the purpose of finding an alternative to thermal sterilization, this research aimed at assessing the efficiency and ideal concentration of sodium hypochlorite for sterilization of culture media and glassware used during rooting of micropropagated Gerbera hybrida cv. Essandre. Two experiments were carried out. In the first one, treatments consisted of control I (no sterilization), control II (thermal sterilization), and total active chlorine concentrations of 0.0005, 0.001, 0.002 and 0.003%. In the second experiment, based on the results observed in the first experiment, treatments consisted of control I (thermal sterilization) and II (chemical sterilization), and total active chlorine concentrations of 0.002, 0.0025 and 0.003%. Plant behavior was assessed based on the length of aerial part and roots, number of roots, and dry biomass of plants. Results showed that the addition of an active chlorine concentration of 0.003% to culture media provided total control of contaminants, and there were no significant differences regarding the variables analyzed between plants obtained with thermal sterilization and with sodium hypochlorite sterilization. Thus, chemical sterilization can be used as a replacement for thermal sterilization of nutrition media for rooting of gerbera in vitro. Key words : Sodium hypochlorite (NaOCl), tissue culture, contamination, chemical sterilization, autoclaving.
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