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 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.

Plant tissue cultures from four Tuscan globe artichoke cultivars

2012 ◽  
Vol 7 (4) ◽  
pp. 680-689 ◽  
Author(s):  
Laura Bedini ◽  
Mariella Lucchesini ◽  
Francesco Bertozzi ◽  
Alberto Graifenberg
Keyword(s):  
Central Italy ◽  
Basal Medium ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Globe Artichoke ◽  
Ms Medium ◽  
Indole Butyric Acid ◽  
Double Phase ◽  
Rooting Medium ◽  
Nitrate Concentrations

AbstractThe aim of the study was to examine the possibility of propagating in vitro four of the most common cultivars in Tuscany (central Italy): Terom, Violetto di Toscana, Chiusure and Empolese. The first three belong to the “Violetti” group, while cv Empolese belongs to the “Romaneschi” group. Explants were cultured on an induction medium (IM), which is a modified MS medium consisting of nitrate concentrations reduced by one quarter, 0.8 mg L−1 6-benzylaminopurine (BA) and 0.2 mg L−1 3-indole butyric acid (IBA). Explants were then transferred to a proliferation medium (PM) consisting of the same basal medium together with 0.03 mg L−1 BA and 0.05 mg L−1 gibberellic acid (GA3). A rooting double-phase was then established. The pre-rooting medium (PRM), consisting of a basal MS medium with half strength nitrate concentrations, 0.5 mg L−1 indole-3-acetic acid (IAA) and 1 mg L−1 paclobutrazol (PBZ) was used for two weeks. Over the next four weeks, a rooting medium (MR) was used, consisting of a basal MS medium with 2 mg L−1 β-cyclodextrin and 2 mg L−1 α-naphthaleneacetic acid sodium salt (NAA). The cv Empolese provided the highest number of proliferated explants and rooted plantlets using the method described.

scholarly journals Establishment of a Rapid and Efficient Micropropagation System for Succulent Plant Haworthia turgida Haw.

HortScience ◽  
2017 ◽  
Vol 52 (9) ◽  
pp. 1278-1282 ◽  
Author(s):  
Boling Liu ◽  
Hongzhou Fang ◽  
Chaorong Meng ◽  
Ming Chen ◽  
Qingdong Chai ◽  
...  
Keyword(s):  
Callus Induction ◽  
Adventitious Shoots ◽  
Germplasm Conservation ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Succulent Plant

In the present study, the effect of plant growth regulators (PGRs) on callus regeneration, adventitious shoot differentiation, and root formation of Haworthia turgida Haw. was investigated. The greatest callus induction percentage (95.6%) was achieved with leaf explants inoculated on Murashige and Skoog (MS) medium with 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 1-naphthaleneacetic acid (NAA), and this callus induction medium supplemented with 2.5 mg·L−1 thidiazuron (TDZ) was optimal for callus proliferation. The maximum number of shoots (25.7) was obtained when the callus was cultured on MS medium supplemented with 1.0 mg·L−1 BA and 0.2 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The highest number of roots per shoot (6.2) and highest rooting frequency (82.0%) were obtained when adventitious shoots were inoculated on MS medium with 0.05 mg·L−1 NAA. Regenerated plantlets were transferred to a mixture of vermiculite and soil and acclimated in a greenhouse. The survival rate of the transplanted plantlets was about 91.6%. The rate of ex vitro rooting was 83.3%, indicating that this technique is effective for root induction in H. turgida. This study has established a rapid and efficient micropropagation system that can be beneficial for commercial cultivation and germplasm conservation of H. turgida.

scholarly journals Development of an Efficient Plant Regeneration System for the Selenium-hyperaccumulator Astragalus racemosus and the Nonaccumulator Astragalus canadensis

HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2138-2142 ◽  
Author(s):  
Chiu-Yueh Hung ◽  
Jiahua Xie
Keyword(s):  
Plant Regeneration ◽  
Basal Medium ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Naphthaleneacetic Acid ◽  
Shoot Induction ◽  
Regeneration System ◽  
Rooting Medium ◽  
Significant Difference

A method of in vitro plant regeneration for both the selenium-hyperaccumulator Astragalus racemosus ‘Cream Milkvetch’ and the nonaccumulator Astragalus canadensis ‘Canadian Milkvetch’ was developed with two induction media, M1 and M2. The M1 and M2 contain Murashige and Skoog basal medium plus vitamins, 8.07 μm N-(2-chloro-4-pyridyl)-N′-phenylurea, 2.5% (w·v−1) sucrose, 0.7% (w·v−1) agar (pH 5.7), and 0.89 μm or 3.12 μm a-naphthaleneacetic acid, respectively. In vitro cultures were initiated on these two types of media with three types of explants: cotyledons, hypocotyls, and roots. More than 93% of cultured explants from both species could form calli or calli with shoots. With regard to shoot formation, A. canadensis could produce multiple shoots from all types of explants more efficiently than A. racemosus. The highest shoot induction was approximately three shoots per explant in A. racemosus, whereas A. canadensis could reach ≈10 shoots per explant. M1 could induce more shoots than M2 no matter what type of explant was used, but the overall induction rates were no significant difference. Among the three types of explants used, the cotyledons were the best explants for shoot induction in A. canadensis, whereas hypocotyls were the best in A. racemosus. In A. racemosus, shoots could also be obtained from calli on the rooting medium containing Murashige and Skoog basal plus vitamins, 2.84 μm indole-3 acetic acid, 2.5% (w·v−1) sucrose, and 0.7% (w·v−1) agar (pH 5.7). Approximately 43% of A. canadensis shoots and 19% of A. racemosus shoots could be rooted on the rooting medium.

scholarly journals Optimization of in vitro regeneration protocol for a popular Indica rice (Oryza sativa L. cv Swarna)

2016 ◽  
Vol 5 (08) ◽  
pp. 1395 ◽  
Author(s):  
Vijaya Naresh Juturu ◽  
Gopala Krishna Mekala ◽  
Mallikarjuna Garladinne ◽  
Puli Chandra Obul Reddy ◽  
Akila Chandra Sekhar*
Keyword(s):  
Callus Induction ◽  
Oryza Sativa L ◽  
Indica Rice ◽  
In Vitro Regeneration ◽  
Regeneration System ◽  
Embryogenic Calli ◽  
Rice Varieties ◽  
Regenerated Plants ◽  
Rooting Medium

Though regeneration system in rice has been very well established compare to other crop plants, the fact remains that, most of the indica rice varieties are still recalcitrant for regeneration and genetic transformation. Therefore, refinement of tissue culture protocol for generation of embryogenic calli and regeneration of the fertile plants from a single cell should be a pre requisite event for development of transgenic plants. Here, in this study we reported high frequency robust regeneration protocols for a popular Indica cultivar Swarna.Mature seeds were used as initial material as explants. Highest callus induction % was observed in MSCIMP medium containing 2.0 mg-1 2,4, D + 0.5 mg-1 Kn as phytohormonal combinations. In addition, maximum regeneration was observed in 2.0 mg-l KN + 0.5 mg-l NAA. Regenerated plants were shifted to rooting medium followed by polyhouse for hardening. The callus induction and regeneration reported in this study were well suited for transformation agronomical important genes or functional genomics studies.

Establishment of a highly efficient regeneration system for in vitro culture of young wheat spikes

2006 ◽  
Vol 3 (2) ◽  
pp. 147-154
Author(s):  
Zhao Lin-Shu ◽  
Liu Lu-Xiang ◽  
Wang Jing ◽  
Zheng Qi-Cheng ◽  
Guo Hui-Jun ◽  
...  
Keyword(s):  
Callus Induction ◽  
Shoot Formation ◽  
Immature Embryos ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration System ◽  
Differentiation Medium ◽  
Efficient Regeneration ◽  
Young Spike

AbstractThis study used three winter wheat (Triticum aestivum L.) genotypes (H6756, H311 and SP8581) to compare the effects of sampling time, callus induction media, differentiation media and rooting media on in vitro culture of young spikes in wheat. In all these three genotypes, the frequencies of green plantlet differentiation were high when their young spikes were cultured between the stages of protective glume primordium formation and pistil and stamen primordium formation, but low at other stages. The optimum medium for callus induction was Murashige and Skoog (MS) medium+2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). The optimum green plantlet differentiation medium was MS medium. Some abnormal plantlets regenerated from calli. When these plantlets were transferred to another differentiation medium [MS+1.0 mg/l 1-naphthaleneacetic acid (NAA)+0.2 mg/l 6-benzylaminopurine (6-BA)], shoot formation and elongation were induced. This allowed 90.91% of them to develop into normal green plantlets. The optimum rooting medium was 1/2MS+0.2 mg/l 3-Indolylacetonitrile (IAA)+80 g/l sucrose. An efficient regeneration system for young spike culture of wheat was set up based on such methods. Using this wheat-regeneration system, young spikes and immature embryos of 17 genotypes of wheat were in vitro cultured to study and compare the callus induction frequencies and green plantlet differentiation frequencies. The results of two successive years showed that in 15 out of the 17 genotypes (88.24%) the green plantlet differentiation frequencies were higher than those of immature embryos by 6.2–65.1%. These results showed that the regeneration system established in this trial for young spike culture of wheat was effective.

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 Callus induction and betacyanin quantification by HPLC/MS-MS in Alternanthera brasiliana (L.) Kuntze

Hoehnea ◽  
2017 ◽  
Vol 44 (1) ◽  
pp. 90-95 ◽  
Author(s):  
Andressa Reis ◽  
Alítcia Moraes Kleinowski ◽  
Fátima Rosane Schuquel Klein ◽  
Renata Trevizan Telles de Souza ◽  
Luciano do Amarante ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Callus Induction ◽  
High Performance ◽  
Large Scale ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Scale Production ◽  
Future Studies ◽  
Large Scale Production ◽  
Equilibrium Concentrations

ABSTRACT The objective of this study was to establish a protocol for callus induction and betacyanin production in plants of Alternanthera brasiliana. Explants of A. brasiliana and five combinations of cytokinin and auxin were used for callus induction. Calli were transferred to a Betacyanin Induction Medium (MIB), composed of MS, with 0.5 mg L-1 of thidiazuron (TDZ) and 1 mg L-1 of naphthaleneacetic acid (NAA), and kept in the light for 45 days. The aspect and intensity of pigments were assessed and total betacyanins were quantified in high performance liquid chromatography (HPLC). The combination of internodal segments and a medium containing equilibrium concentrations of auxins and cytokinins was the most efficient metod to induce calli and increased production of betacyanins. The presence of amaranthine in calli of A. brasiliana justifies its medical use and the consequent need for future studies for the large-scale production of this molecule.

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 Tissue Culture Propagation of Yam in Puerto Rico

1969 ◽  
Vol 67 (4) ◽  
pp. 419-428
Author(s):  
Amelia Cortés-Monllor ◽  
Lii-Jang Liu
Keyword(s):  
Tissue Culture ◽  
Puerto Rico ◽  
Indoleacetic Acid ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Dioscorea Rotundata ◽  
Skoog Medium ◽  
Rooting Medium ◽  
Murashige And Skoog Medium ◽  
Tissue Culture Propagation

Rapid propagation of yam (Dioscorea rotundata cv. Habanero) was obtained from nodal segments cultured in modified Murashige and Skoog medium containing indoleacetic acid (IAA) and kinetin as establishment medium, and naphthaleneacetic acid (NAA) as rooting medium. The proliferation cycle, which takes approximately 2-3 months, increased four times the production of yam plantlets. These plantlets were successfully transferred to potting mixture and soil. This procedure is extremely useful for regenerating virusfree plantlets suitable for producing healthy tubers for planting.

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