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 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 Establishment of Tissue Culture Regeneration System for Medicago ruthenica L. cv. ‘Zhilixing’

2021 ◽  
Author(s):  
Bo Xu ◽  
Rina Wu ◽  
Cuiping Gao ◽  
Fengling Shi
Keyword(s):  
Tissue Culture ◽  
Stress Resistance ◽  
Callus Induction ◽  
Combination Method ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Medicago Ruthenica ◽  
Rooting Medium ◽  
Drought And Salt Tolerance

Background: Medicago ruthenica L. ‘Zhilixing’ is a new variety with superior forage and seed yield compared to the wild type. The cold, drought and salt tolerance of Zhlixing are better than those of alfalfa, suggesting that this variety can serve as a high-quality genetic resource for improving the stress resistance of alfalfa. However, because of the lack of tissue culture regeneration system, it is difficult to perform genetic transformation studies on stress resistance genes. This study aimed to establish an efficient tissue culture regeneration system for Zhilixing variety. Methods: Three types of explants were selected and tested on four types of basal media supplemented with different combinations of auxin and cytokinin for callus induction and differentiation, based on orthogonal tests to select the combinations of auxin and cytokinin suitable for callus induction and differentiation. Two-factor combination method was used to formulate a suitable rooting medium. Result: The hypocotyledonary axis was found to be an excellent explant for callus induction on MS medium. The optimum callus induction medium contained thidiazuron (TDZ, 0.5 mg/L), 2,4-dichlorophenoxyacetic acid (2,4-D, 1.0 mg/L) and naphthaleneacetic acid (NAA, 0.5 mg/L) where the callus induction rate was 93.33%. The differentiation medium was supplemented with TDZ (0.75 mg/L), 2,4-D (0.25 mg/L) and 6-benzyladenine (6-BA, 1.5 mg/L) where the differentiation rate was 63.33%. Thidiazuron played the key role in both processes of callus induction and differentiation. Half-strength MS containing 0.1 mg/L of NAA was the most efficient rooting medium.

scholarly journals Response to Callus Induction and Regeneration of Newly Released BRRI Rice Varieties

2020 ◽  
Vol 23 (2) ◽  
pp. 17-25
Author(s):  
SD Joya ◽  
S Sultana ◽  
J Ferdous ◽  
MA Qayum ◽  
ME Hoque
Keyword(s):  
Callus Induction ◽  
Casein Hydrolysate ◽  
The Other ◽  
Induction Medium ◽  
Regeneration Ability ◽  
Regeneration System ◽  
Rice Varieties ◽  
Green Plants ◽  
Induction Frequency ◽  
The Media

A study was carried out for developing an efficient callus induction and regeneration system for three newly developed BRRI varieties namely BRRI dhan86, BRRI dhan87 and BRRI dhan89. Dehusked seeds were plated onto MS and N6 media with two hormone combinations for callus induction. Calli obtained from each callus induction medium were transferred to four different regeneration media. Callus induction frequency and regeneration ability were significantly influenced by rice varieties, and interactions of variety and media. Among the media compositions, the highest callus (59.44%) were obtained from C1 (MS+2mg/l 2,4-D) followed by C2 ( MS+2 mg/l 2,4-D+0.5 mg/l kinetin) , C3 ( N6+2 mg/l 2,4-D) and C4 (N6+2 mg/l 2,4-D+0.5 mg/l kinetin) medium. The highest regeneration (45.74%) was obtained from R2 (MS+4 mg/ml BAP+1.2 mg/ml kinetin+0.5 mg/ml NAA), followed by R3 (1 mg/ml BAP+1 mg/ml Kinetin+1 mg/ml NAA), R4 (2 mg/ml kinetin+1 mg/ml NAA+300 mg casein hydrolysate) and R1 (2 mg/ml BAP+1 mg/ml kinetin+1 mg/ml NAA). BRRI dhan86 showed the highest regeneration ability (53.06%) than the other two varieties. It is observed that all varieties performed better in C1 medium for callus induction and R2 medium for regeneration. This study also revealed that BRRI dhan86 was more responsive to callus induction and regeneration of green plants than the other two varieties. Bangladesh Rice j. 2019, 23(2): 17-25

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.

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