scholarly journals PENGARUH JENIS EKSPLAN DAN KOMBINASI ZAT PENGATUR TUMBUH (ZPT) TERHADAP INDUKSI KALUS Begonia bimaensis Undaharta & Ardaka SECARA IN VITRO

2020 ◽  
Vol 23 (1) ◽  
pp. 82-90
Author(s):  
Ema Hendriyani ◽  
Tri Warseno ◽  
Ni Kadek Erosi Undaharta

Begonia bimaensis Undaharta & Ardaka is a potential ornamental plant, and currently known only from one population in Sumbawa. Propagation programs, both conventional and in vitro culture are necessary to ensure its conservation. The aim of this research is to observe the effects of explant types and plant growth regulator combination (2,4-D and kinetin) in inducing callus from B. bimaensis leaf in vitro. Callus induction was initiated from three parts of leaf explant, namely petiole, leaf base, and leaf lamina. The explants were planted on Murashige & Skoog (MS) medium with addition of 2,4-D and kinetin. Concentrations of 2,4-D were 0, 0.5, and 1 ppm, while kinetin concentrations were 0, 1, and 2 ppm. Each treatment was replicated 10 times. Results showed that leaf base was the best explant used for callus induction. Medium D1K2 (MS + 1 ppm kinetin) showed the fastest time for callus induction that was at 20 days after planting. The highest percentage of callus production (100%) was found on D1K3 (MS + 2ppm kinetin); D2K2 (MS + 0.5ppm 2,4-D + 1 ppm kinetin); D2K3 (MS + 0.5ppm 2,4-D + 2ppm kinetin) and D3K2 (MS + 1ppm 2,4-D + 1ppm kinetin).

2011 ◽  
Vol 63 (1) ◽  
pp. 209-215 ◽  
Author(s):  
S. Uranbey

A high frequency of bulblet regeneration was achieved for the endemic and endangered ornamental plant Muscari azureum using immature embryos. Immature embryos of M. azureum were cultured on a callus induction medium consisting of N6 mineral salts and vitamins, 400 gL-1 casein + 40 gL-1 sucrose + 2 mgL-1 L-proline, 2 mgL-1 2,4-D and 2 gL-1 Gelrite. Then the embryogenic callus clusters were transferred to a bulblet induction medium consisting of MS mineral salts and vitamins containing different concentrations and combinations of BAP, KIN, TDZ, Zeatin, IAA, NAA, 30 gL-1 sucrose and 7 gL-1 agar. Prolific bulblet multiplication (over 13 bulblets/embryo) was achieved from immature embryos after 5-6 months of culture initiation. Well-developed bulblets were excised and individually rooted on ? strength MS medium supplemented with 1 mgL-1 IBA, 0.5 gL-1activated charcoal, 20 gL-1sucrose and 6 gL-1agar and acclimatized. <br><br><font color="red"><b> This article has been retracted. Link to the retraction <u><a href="http://dx.doi.org/10.2298/ABS150608072E">10.2298/ABS150608072E</a><u></b></font>


2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  

Dalbergia sissoo Roxb. ex DC. (Sissoo) is a native forest tree species in Pakistan. Many ecological and economical uses are associated with this premier timber species, but dieback disease is of major concern. The objective of this study was to develop a protocol for in vitro regeneration of Sissoo that could serve as target material for genetic transformation, in order to improve this species. Callus formation and plantlet regeneration was achieved by culturing cotyledons, immature seeds, and mature embryos on a modified Murashige and Skoog (1962) (MS) medium supplemented with plant growth regulators. Callus induction medium containing 2.71 ?M 2, 4-dichlorophenoxyacetic acid (2,4-D) and 0.93 ?M kinetin produced better callus on all explants tested compared to other treatments, such as 8.88 ?M 6-benzylaminopurine (BA) and 2.69 ?M ?-naphthalene acetic acid (NAA), or 2.71 ?M 2, 4-D and 2.69 ?M NAA. Shoot regeneration was best on MS medium containing 1.4 ?M NAA and 8.88 ?M BA compared to other treatments, such as 1.4 ?M NAA and 9.9 ?M kinetin, or 2.86 ?M indole-3-acetic acid and 8.88 ?M BA. Murashige and Skoog medium containing 1.4 NAA ?M and 8.88 ?M BA was better in general for regeneration regardless of callus induction medium and the type of explant used. Rooting was best on half-strength MS medium with 7.35 ?M indole-3-butyric acid. Regenerated plantlets were acclimatized for plantation in the field. Preliminary genetic transformation potential of D. sissoo was evaluated by particle bombardment of callus explants with a pUbiGus vector. The bombarded tissue showed transient Gus activity 1week after bombardment. Transformation of this woody tree is possible provided excellent regeneration protocols. The best combination for regeneration explained in this study is one of such protocols.


2015 ◽  
Vol 804 ◽  
pp. 259-262
Author(s):  
Chonnikarn Khunchuay ◽  
Kanokporn Sompornpailin

The optimum ratios of auxin and cytokinin are necessary for callus induction and plant regeneration. This ratio is a key function involving in the promoting cell division and proliferation in tissue culture. The axillary buds of in vitro plantlets fromVetiveria nemoralisA. Camuscv. Roiet were used as explants for the callus induction experiment. These explants were cultured on Murashige & Skoog (MS) medium [1] supplemented with various combinations of auxins and cytokinins. Under this experimental study, the highest frequency of callus induction was found on MS medium supplemented with 2 mgL-1α-naphthalene acetic acid (NAA) and 1 mgL-12-furanylmethyl-1H-purine-6-amine (kinetin) (62.5%). On the other hand the combination of 2, 4-dichlorophenoxyacetic acid (2, 4-D) and 6-benzylaminopurine (BAP) was toxicity to this explants. All culturing explants were dead and no calli appearance. The calli derived from each medium were transferred into the same regeneration medium (MS with 1 mgL-1NAA and 2 mgL-1BAP). After culturing on regeneration medium, calli induced from the highest callus induction medium have shown high frequencies of regeneration and also shoot number per callus (58.33% and 7.1 shoots).


2015 ◽  
Vol 7 (1) ◽  
pp. 30-37 ◽  
Author(s):  
Sunil Kumar ◽  
Virendra Kumar ◽  
Manoj Kumar Sharma ◽  
Narendra Kumar ◽  
Anil Kumar ◽  
...  

Jatropha (Jatropha curcas L.) is an oil bearing crop growing in tropical and subtropical parts of the world. The present study was undertaken to investigate the effects of different plant growth regulators on in vitro callus induction in physic nut (J. curcus). In the present study, it was observed that all the explants viz., leaf lamina, petioles, nodal segments and cotyledonary nodes showed good callus induction responses on various culture media thus tried. Leaf lamina and petioles showed 100.0% callus induction responses on different MS media supplemented with auxins and cytokinins alone or in combinations whereas, nodal segments and cotyledonary nodes showed maximum 89.6% and 83.9% callus induction respectively. The presence of 2, 4-D in culture media with auxins or cytokinins was essential for good callus growth. Among different explants tried, leaf lamina was the best responding explants and MS-13 media supplemented with 5×10-6 M NAA and 10-5 M 2, 4-D is the best callusing and growth supporting medium. However, the regenerative competence of the callus tissues can differ depending on the type of explants used because certain types of plant tissues have more favorable regeneration responses than others. Callus induction rate from all explant types was highest than other reports. The results obtained in the present study would facilitate the high callus induction and regeneration responses in J. curcus for its improvement using biotechnological tools.


Author(s):  
Vi Thi Tuong Nguyen ◽  
Trinh Le Diem Ho ◽  
Kim Thi A Phan

Codonopsis javania (Blume) Hook.f. et Thomson a traditional medicine plant and now an endangered species in Vietnam is grown for roots. The research was carried out to establish the plant propagation for the purpose of concerving and exploting this endangered medicinal herbs. In vitro shoot tip explants (1 – 1.5 cm) were induced to form callus on MS medium containing NAA (0.5 – 2 mg /L) with TDZ 0.1 mg/L. After four weeks of culture, in the MS medium combine with NAA 1 mg/L and TDZ 0.1 mg/L the explant induced compact callus (green, solid) wsa achieved 85.33%. The callus induction to form shoots on medium MS containing BA (0.5 – 2.0 mg/L) with NAA 0.2 mg/L. After 4 weeks of culture, shoot formation was higher in the MS medium containing BA 1.0 mg /L and NAA 0.2 mg/L and achieved of 82.67 % with 9.92 shoots/explant. The best shoot proliferation (2 – 3 cm) was excised and transferred to a medium shoot multiplication with the same composition as the shoot induction medium in which NAA 0.2 mg/L was replaced by NAA 0.5 mg/L. When compared the shoot multiplication between the two mediums at the same BA concentration (2 mg/L), all shoots increased and reached 5.87 times after 60 days cultured. On rooting MS medium with IBA 1 mg/L, 88.67 % in vitro rooting was observed with the average root yield of 4.33 roots/shoot and the length of 8.27 cm. Root length and their yield quality were highly improved when using of coconut fiber (30 %) and earthworms compost (70 %) (v/v) in the transfer medium after acclimatisation stages.


2021 ◽  
Vol 20 (5) ◽  
pp. 119-126
Author(s):  
Burcu Tuncer

Rheum ribes L. is a perennial wild species. Young shoots and flower bunches are freshly consumed, and root and rhizomes are generally used for medicinal purposes. The aim of the present study was to improve the callus proliferation protocol for R. ribes L. under in vitro conditions. For callus induction, hypocotyl explants taken from 14-day old plantlets germinated in Murashige and Skoog (MS) media were cultured in MS media with 9 plant growth regulator (PGR) combinations containing 6-benzylaminopurine (BAP) (2, 3, and 4 mg/L) + naphthylacetic acid (NAA) (0.1, 0.5, and 1 mg/L). Then, for callus proliferation, 4 PGR combinations containing NAA (0.2 mg/L) + thidiazuron (TDZ) (0.5, 1, 2, and 3 mg) were used in the first set of experiments, and 36 PGR combinations containing BAP (1, 2, 3, and 4 mg/L) + indole-3-butyric acid (IBA) (0.2, 0.5, and 1 mg/L), BAP (1, 2, 3, and 4 mg/L) + NAA (0.2, 0.5, and 1 mg/L), and TDZ (1, 2, 3, and 4 mg/L) + NAA (0.2, 0.5, and 1 mg/L) were used in the second set of experiments. At the end of the second set of experiments, the greatest callus regeneration ratios were obtained due to the combinations including BAP and IBA as well as the low-dose TDZ- (especially 1 mg/L) and NAA- (0.2, 0.5, 1 mg/L) combinations. Regarding callus fresh weights, TDZ + NAA combinations were found to be more successful. The greatest callus fresh weight (12.7 ±0.4 g) was obtained from MS medium supplemented with 2 mg/L TDZ and 0.2 mg/L NAA.


2014 ◽  
Vol 26 (2) ◽  
pp. 155-162 ◽  
Author(s):  
Jaime A. Teixeira da Silva

ABSTRACT The mid- to long-term preservation of papaya (Carica papaya L.) would allow for the safeguarding of important germplasm. In this study, soft friable callus (SFC) and hard callus (HC) were induced from the first two true leaves of 10-day-old seedlings containing a midrib derived from the germinated seed of two cultivars (‘Rainbow’ and ‘Sunrise Solo’). Following germination on a Murashige and Skoog (MS) medium that contained 3% sucrose and was free of plant growth regulators (PGRs), sections of the first true leaves from 10-day-old seedlings were exposed to seven published callus or somatic embryogenesis protocols for zygotic embryos, leaves or hypocotyls. Optimal SFC and HC induction was carried out on a half-strength MS medium following the Fitch (1993) or the Ascêncio-Cabral et al. (2008) protocol, respectively. SFC formed shoots that could then convert to plants when transferred to a full-strength MS medium devoid of PGRs. Plantlets 10-cm tall were acclimatised in two steps: first by in vitro acclimatisation in aerated vessels, the Vitron, under CO2-enriched (3000 ppm CO2), then by the transfer of individually rooted plantlets in Rockwool® blocks to a substrate of soil: pine bark : perlite (1:1:1, v/v/v). SFC and HC were then encapsulated in alginate beads, which were exposed to low temperature storage (LTS) at 10°C and 15°C, and also cryopreserved for 30 days. All encapsulated alginate beads that contained SFC, HC or leaf tissue that had been stored under LTS or cryopreserved were able to regenerate callus when placed on an optimal callus induction medium. Plants derived from the control, LTS and cryopreservation protocols, either from SFC or HC, were successfully acclimatised.


2021 ◽  
Vol 5 (1) ◽  
pp. 61-67
Author(s):  
Sitti Inderiati ◽  
FNU Yanti ◽  
Eka Ria Mentari

In vitro propagation is a method to produce massive healthy new planting materials quickly. An experiment was carried out for morphogenic callus induction and regeneration of a domestic sugarcane variety. The Explants used was an inert folded leaf and incubated on modified MS medium augmented with 1 mg/l, 2.5 mg/l, and 5 mg/l of 2,4-D for callus induction. The leaf calluses were subcultured on MS medium enriched with different growth regulators for shoot initiation and multiplication. The highest percentage of callus formation was achieved in the medium containing 2.5 mg/l of 2,4-D, while the fastest callus initiation was noticed in MS medium supplemented with 5 mg/l 2,4-D, and maximum proliferation and the morphogenic response of callus were obtained in 3rd subculture. Two types of callus observed on the induction medium were dry nodular friable and smooth compact. This highly morphogenic callus was white to white creamy in color and easy to separate.   The highest shoot proliferation rate was found on the medium containing 2 mg/l Kinetin + 1 mg/l IAA and no growth was noticed on the medium containing Kinetin alone. Therefore, the study suggests that the growth hormone of cytokinin in combination with auxin is necessary for in vitro regeneration of sugarcane callus culture.


2009 ◽  
Vol 24 ◽  
pp. 82-88 ◽  
Author(s):  
Saraswoti Aryal ◽  
Sanu Devi Joshi

Rauvolfia serpentina (L.) ex. Kurz is an important medicinal plant. Callus induction and regeneration was studied from stem explant of in-vitro grown plant of Rauvolfia serpentina(L.) Benth. ex Kurz (Apocynaceae) on Murashige Skoog (1962) medium supplemented with 1mg/l 2,4-Dichlorophenocy acetic acid (2,4-D) and 1mg/l Kinetin (Kn). Vigorous growth of callus occurs after 4 weeks of culture. Callus was sub-cultured on Murashige and Skoog (MS) medium supplemented with different concentration of 2, 4-D (0.5-3.0 mg/l) and 10% coconut milk. Regeneration of plantlets occurred on MS medium containing 3 mg/1 of 2, 4-D and 10% coconut milk. These plantlets were rooted on MS medium supplemented with 1 mg/l IAA .The regenerated plantlets were able to grow on soil after short period ofacclimatization. Key words: Explant; In-vitro culture; MS medium;  2, 4 Dichlorophenoxy acetic acid; Kinetin; Callus; Tissue culture; Coconut milk. Journal of Natural History Museum Vol. 24, 2009 Page: 82-88


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