Optimization of callus induction medium for Hymenocallis littoralis (Melong kecil) using root and bulb explants

2012 ◽  
Vol 6 (12) ◽  
Author(s):  
Rosli Noormi
Keyword(s):  
Callus Induction ◽  
Induction Medium ◽  
Callus Induction Medium

scholarly journals In vitro Regeneration of Dalbergia sissoo Roxb. and the Potential for Genetic Transformation

2012 ◽  
Vol 40 (2) ◽  
pp. 140 ◽  
Author(s):  
Hafiz Mamoon REHMAN ◽  
Iqrar Ahmad RANA ◽  
Siddra IJAZ ◽  
Ghulam MUSTAFA ◽  
Faiz Ahmad JOYIA ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Genetic Transformation ◽  
Callus Induction ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Native Forest ◽  
Ms Medium ◽  
Dalbergia Sissoo ◽  
Callus Induction Medium

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.

scholarly journals The Effect of Daminozide, Dark/Light Schedule and Copper Sulphate in Tissue Culture of Triticum timopheevii

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2620
Author(s):  
Dmitry Miroshnichenko ◽  
Anna Klementyeva ◽  
Sergey Dolgov
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Copper Ions ◽  
Induction Medium ◽  
Tissue Cultures ◽  
Triticum Timopheevii ◽  
Dark Light ◽  
Green Plants ◽  
Callus Induction Medium

Triticum timopheevii Zhuk. is a tetraploid wheat that is utilized worldwide as a valuable breeding source for wheat improvement. Gene-based biotechnologies can contribute to this field; however, T. timopheevii exhibits recalcitrance and albinism in tissue cultures, making this species of little use for manipulation through genetic engineering and genome editing. This study tested various approaches to increasing in vitro somatic embryogenesis and plant regeneration, while reducing the portion of albinos in cultures derived from immature embryos (IEs) of T. timopheevii. They included (i) adjusting the balance between 2,4-D and daminozide in callus induction medium; (ii) cultivation using various darkness/illumination schedules; and (iii) inclusion of additional concentrations of copper ions in the tissue culture medium. We achieved a 2.5-fold increase in somatic embryogenesis (up to 80%) when 50 mg L−1 daminozide was included in the callus induction medium together with 3 mg L−1 2,4-D. It was found that the dark cultivation for 20–30 days was superior in terms of achieving maximum culture efficiency; moreover, switching to light in under 2 weeks from culture initiation significantly increased the number of albino plants, suppressed somatic embryogenesis, and decreased the regeneration of green plants. Media containing higher levels of copper ions did not have a positive effect on the regeneration of green plants; contrarily, the elevated concentrations caused albinism in plantlets. The results and relevant conclusions of the present study might be valuable for establishing an improved protocol for the regeneration of green plants in tissue cultures of T. timopheevii.

scholarly journals Plant Regeneration from Protocorm-derived Callus of Five Paphiopedilum Hybrids

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1101D-1101
Author(s):  
Michael Compton
Keyword(s):  
Plant Regeneration ◽  
Growth Regulator ◽  
Callus Induction ◽  
Callus Formation ◽  
Induction Medium ◽  
Regeneration Medium ◽  
Petri Dishes ◽  
Callus Induction Medium

Callus was induced from protocorms of five Paphiopedilum hybrids (Paph. 03-1, Paph. 03-4, Paph. 03-5, Paph. 03-6, and Paph. 03-7) on callus induction medium [MS inorganics (412.5 mg NH4NO3 instead of 1650 mg and 475 mg KNO3 instead of 1900 mg) and vitamins plus (per liter) 0.1 g myo-inositol, 30 g sucrose, and 2.5 g Gelrite; pH 5.5] containing various concentrations and combinations of thidiazuron (TDZ; 4.5 and 45 μm) and 2,4-D (4.5 and 45 μm). Callus formation was greatest for protocorms of Paph. 03-1, Paph. 03-4, Paph. 03-6, and Paph. 03-7. Among the most competent hybrids, callus formation was greatest among protocorms induced in medium containing 4.5 μm 2,4-D and 4.5 to 45 μm TDZ. Induced calli were transferred to 100 × 15 mm petri dishes containing 25 mL of PLB and plant regeneration medium (similar to callus induction medium) containing various concentrations of either benzyladenine (BA; 0.5, 5, or 10 μm), TDZ (0.25, 2.5, or 5 μm) or no growth regulator (control). PLB and plant formation was greatest on medium containing BA.

scholarly journals Development of a protocol for the micropropagation of mature Eucalyptus grandis clones through somatic embryogenesis

2001 ◽  
Author(s):  
◽  
Andiswa Tsewana
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Callus Induction ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Induction Medium ◽  
Mature Trees ◽  
The Media ◽  
Callus Induction Medium

Biotechnology techniques such as micropropagation VIa somatic embryogenesis offer potential significant advances in the improvement of forest species, which could sustain forest production in South Africa, as well as globally, without increased use of land. In order to apply such techniques to commercial breeding and clonal programmes of E. grandis species, it is necessary to develop reliable and efficient protocols applicable to explants of proven superior genotypes. Most of the research on E. grandis somatic embryogenesis has used the genetically variable embryos or seedlings as explant sources, which results in the propagation of material of unproven genetic value. In order to exploit somatic embryogenesis maximally for cloning of superior trees, somatic embryos have to be induced from highly selected and, hence, mature trees. The aim of this investigation was to develop such a protocol for E. grandis and to test its applicability to various E. grandis hybrids. Somatic embryos were induced from buds, stems, leaves and petioles, with petioles and buds giving the best results. Thus, these were selected for further studies which involved testing the effect of medium composition on embryogenic callus induction. Media used for this purpose contained MS or B5 nutrients, 1 mg.l' 2,4-D, 0.5 g.r! glutamine, 0.5 g.r! casein hydrolysate, 4 g.r! Gelrite and 30 or 50 g.rl sucrose. All the media tested were able to support induction of embryogenic callus, although the number of explants producing embryogenic calli was affected significantly by the media composition (10-91 %). Callus induction media with B5 nutrients seemed to have a significant effect onn the developmental stage of embryos in the callus induction medium. Presence of 50 g.r! sucrose in the callus induction medium reduced the embryo yield, but the progress of embryo development was enhanced. The callus induction medium containing B5, 1 mg.l' 2,4-D, 0.5 g.rl glutamine, 0.5 g.r! casein hydrolysate, 4 g.r! Gelrite and 30 g.l' sucrose was chosen for subsequent studies. Of all the media tested for embryo development, the medium with B5, 2.5 mg.l' 2iP, 0.5 g.r! glutamine, 0.5 g.r! casein hydrolysate, 4 g.r! Gelrite and 50 g.r! sucrose was found to be the most suitable for embryo development to the cotyledonary stage. Experiments involving incorporation of both ABA and 2iP aiming at maturation of E. grandis somatic embryos led to an increase in size of the cotyledonary embryos formed but not to germination.

Construction of Potato DM1-3-516-R44 (DM) Transgenic System Based on Agrobacterium Transformation

2020 ◽  
Author(s):  
Chao Zhang ◽  
Dongdong Wang ◽  
Yongzhi Yang ◽  
Qin Chen
Keyword(s):  
Gene Function ◽  
Callus Induction ◽  
System Optimization ◽  
Induction Medium ◽  
Transformation System ◽  
Regeneration System ◽  
Shoot Differentiation ◽  
Pcr Technology ◽  
Callus Induction Medium ◽  
Transformation Condition

Abstract Background: The sequencing potato DM1-3-516-R44 played an irreplaceable role in the study of gene function. So far, no one research the transformation system about DM. Therefore, our experiment was studied from three aspects: plant regeneration system, optimization of agrobacterium infection conditions and the effect of hygromycin on DM. Results: A relatively suitable method for genetic transformation of DM was obtained: 1) The stem callus induction medium was MS + IAA 0.5 mg/L + 6-BA 2.0 mg/L, the leaf callus induction medium was MS + NAA 1.0mg/L + 6-BA 0.5mg/L and the shoot differentiation medium was MS + 6-BA 3.0 mg/L + ZT 0.5 mg/L. 2) The specific transformation condition was the agrobacterium concentration kept the OD600 = 0.3, and co-culture time consisted 3 days in the dark. The hygromycin concentration chose 8 mg/L to screen the transgenic plants. 3) Using hygromycin to screen about 100 transgenic shoots, 75 shoots were obtained and 53 strains were identified had target stripe by PCR technology. Conclusion: The efficiency of the transformation system we created was over 50%. It provided a good basis for the study of potato gene function.

scholarly journals Adaptation of in vitro regeneration protocol for Brazilian wheat genotypes

2018 ◽  
Vol 48 (11) ◽  
Author(s):  
Eduardo André Roesler ◽  
Ernandes Manfroi ◽  
Andréa Morás ◽  
Dielli Aparecida Didoné ◽  
Magali Ferrari Grando ◽  
...  
Keyword(s):  
Callus Induction ◽  
Somatic Embryos ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Regeneration Capacity ◽  
Target Region ◽  
Wheat Genotypes ◽  
Regeneration Efficiency ◽  
Callus Induction Medium

ABSTRACT: The availability of an efficient protocol for in vitro regeneration is imperative for genetic transformation. Using genotypes adapted to the target region as a transgenic platform accelerates the development of cultivars. Therefore, this study aimed to adapt an in vitro regeneration protocol for Brazilian wheat genotypes. For this purpose, the in vitro regeneration capacity of immature embryos from six Brazilian wheat genotypes using two protocols of regeneration of somatic embryos was analysed. Furthermore, combinations of 2,4-D and picloram in the callus induction medium were tested in order to improve regeneration efficiency. Genotypes with higher regeneration efficiency were BR18-Terena and PF020037, yielding 0.42 and 1.13 shoots per explant using the Hu and the Wu protocol, respectively. Adding 1mgL-1 2,4-D in the callus induction medium was the most favourable, producing 3.73 and 3.07 shoots per explant for PF020037 and BR18-Terena, respectively. In conclusion, a protocol for regeneration for two Brazilian wheat genotypes recommended and developed to be cultivated at the Cerrado region has been adapted.

scholarly journals Pengaruh Komposisi Media Dasar dan Jenis Eksplan terhadap Pembentukan Embrio Somatik Kakao

2016 ◽  
Vol 3 (3) ◽  
pp. 127
Author(s):  
Nur Ajijah
Keyword(s):  
Somatic Embryo ◽  
Callus Induction ◽  
Callus Formation ◽  
Basal Medium ◽  
Induction Medium ◽  
Explant Type ◽  
Salt Medium ◽  
Basal Salt Medium ◽  
Primary Callus ◽  
Callus Induction Medium

<em>The composition of basal medium determines the regeneration success of in vitro culture. The study aimed to evaluate the effect of basal medium in the primary callus induction medium and explant type on the formation of cacao somatic embryo. The research was conducted at the Tissue Culture Laboratory of IAARD, Bogor, from June 2014 to December 2015. Primary callus induction derived from staminoid and petal explants of ICCRI 4 clone used two types of basal medium, i.e. DKW+ 9 µM 2.4-D + 1.16 µM kinetin or WPM + 9 µM 2.4-D + 1.16 µM kinetin.</em> <em>After 14 days, callus was subcultured onto secondary callus induction medium (WPM + 2.4-D 9 </em><em>μM + kinetin 0.58 μM), and then onto DKW medium without growth regulators to induce the formation of somatic embryo. The research was designed in two-factor factorial design with five replications. The first factor was the type of basal medium on the primary callus induction medium (DKW and WPM) and the second factor was the type of explants (petal and staminoid). The results showed significant interaction effect between basal medium type and explant type on the formation of callus and somatic embryo of cacao. The highest percentage of callus formation was derived from staminoid explants on the DKW basal salt medium (92.5%). However, the highest percentage of somatic embryo formation and the number of somatic embryo per explant were obtained from DKW basal salt medium with petal explants (36.5% and 2.3).</em> <em>Therefore, t</em><em>he use of DKW basal salt medium and petal explant were recommended for the induction of somatic embryo of the ICCRI 4 clone.</em>

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