Disease-resistant transgenic adzuki bean plants obtained through an efficient transformation system

2012 ◽  
Vol 63 (12) ◽  
pp. 1090 ◽  
Author(s):  
Huatao Chen ◽  
Xin Chen ◽  
Heping Gu ◽  
Xingxing Yuan ◽  
Hongmei Zhang ◽  
...  
Keyword(s):  
Soybean Mosaic Virus ◽  
Shoot Elongation ◽  
Zeatin Riboside ◽  
Medium Composition ◽  
Adventitious Bud ◽  
Adzuki Bean ◽  
Transformation System ◽  
Bean Plants ◽  
Bud Induction

An efficient regeneration and transformation system was established and optimised for adzuki bean (Vigna angularis (Willd.) Ohwi & Ohashi). 6-Benzylaminopurine at 5 mg L–1 was used to increase adventitious bud induction frequency. The highest frequency of shoot elongation was 92.8% when using a medium composition of MS salts combined with 0.1 mg L–1 of IAA, 0.5 mg L–1 of GA3, 1.0 mg L–1 of zeatin-riboside, 50 mg L–1 of aspartic acid, and 50 mg L–1 of glutamic acid. In vitro rooting was 100% when shoots were cultured on the solid MS medium supplemented with 1.0 mg L–1 of NAA. Reproducible transformation of epicotyl explants was developed using the A. tumefaciens EHA105 strain. Using a concentration of 40 mg L–1 of acetosyringone, 20 mm MES, and 5 mg L–1 of 6-benzylaminopurine in the co-cultivation medium, a transformation efficiency of 12.6% was attained. Using this transformation protocol, we obtained transgenic adzuki bean plants resistant to soybean mosaic virus by introducing the V. angularis VaPR3 gene.

scholarly journals HISTOLOGICAL EXAMINATION OF IN VITRO ADVENTITIOUS BUD DEVELOPMENT ON HYPOCOTYLS OF FRASER FIR

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1102a-1102
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson
Keyword(s):  
Growth Regulators ◽  
Induction Medium ◽  
Adventitious Bud ◽  
Naphthaleneacetic Acid ◽  
Abies Fraseri ◽  
Fraser Fir ◽  
Cell Clusters ◽  
Cell Divisions ◽  
Bud Induction

Hypocotyls of Fraser fir (Abies fraseri (Pursh) Poir.) were excised from seeds germination 9 days and placed on bud induction medium containing 10 mg/liter benzyladenine (BA) and 0.01 mg/liter naphthaleneacetic acid (NAA) or medium without growth regulators. After 3 days on medium containing growth regulators, cell divisions were localized in epidermal and subepidermal layers of the hypocotyl while similar cell divisions were not observed in control-treated hypocotyls. Cell clusters consisting of two to five cells were present after 7 days in hypocotyls placed on bud induction medium. In control-treated hypocotyls, stomata continued to develop and cells within the cortex became vacuolated during the first 2 weeks in culture. All hypocotyls were transferred to secondary medium after 3 weeks. Cell clusters continued to enlarge into meristemoids in hypocotyls initially placed on bud induction medium. Gradually, meristemoids developed into buds and cataphylls were observed covering bud meristems.

scholarly journals Basal stem cluster bud induction and efficient regeneration for the Tibetan endemic medicinal plant Swertia conaensis

2021 ◽  
Vol 49 (2) ◽  
pp. 12152
Author(s):  
Yin-Kai XI ◽  
Heng-Yu HUANG
Keyword(s):  
Plant Regeneration ◽  
Medicinal Plant ◽  
Orthogonal Experiment ◽  
Occurrence Rate ◽  
Adventitious Bud ◽  
Ms Medium ◽  
Regeneration System ◽  
Bud Induction ◽  
Endemic Medicinal Plant

The artificial rapid propagation system for Swertia conaensis T. N. Ho et S. W. Liu was explored to screen the appropriate plant regeneration method and to provide an efficient propagation mode, useful for artificial breeding technology or for further research and development of the Tibetan endemic medicinal plant. In this study, the most suitable explant and hormone were chosen according to single factor test. Next, the effects of different hormone combinations on basal stem cluster bud induction, callus induction, adventitious bud occurrence and plant regeneration were investigated by using complete combination and orthogonal experiment. The obtained results showed that the explants suitable for in vitro of S. conaensis were stem tips with leaves, which were regenerated through the method of basal stem cluster bud occurrence in the MS medium with 2.0 mg∙L-1 6-BA, 0.5 mg∙L-1 NAA, but the proliferation coefficient was low, only 3.16 after 40 days of culture. Subsequently, the proliferation coefficient failed to improve, irrespective of change of the concentration ratio of 6-BA and NAA. Therefore, in the orthogonal experiment of adding ZT, the MS medium with 1.0 mg∙L-1 ZT, 0.5 mg∙L-1 NAA and 2.5 mg∙L-1 6-BA induced a large number of callus green and compact, with 86.30% callus occurrence rate. After 40 days of culture, the rate of adventitious bud occurrence was 96.55% and the proliferation coefficient was high (10.37). The rooting rate was 100% in the 1/2MS medium with 0.5 mg∙L-1 NAA. The survival rate of regenerated plants was more than 95%. Indirect organogenesis was more efficient than direct organogenesis in in vitro culture of S. conaensis. In this study, the efficient and stable regeneration system of S. conaensis was achieved through the method of explant to callus to adventitious buds, which provided an effective way to an endangered species.

Factors affecting adventitious bud induction in Pinus elliottii (Engelm.) embryos cultured in vitro

1987 ◽  
Vol 11 (1) ◽  
pp. 25-35 ◽  
Author(s):  
Pedro P�rez-Berm�dez ◽  
Harry E. Sommer
Keyword(s):  
Pinus Elliottii ◽  
Adventitious Bud ◽  
Factors Affecting ◽  
Bud Induction ◽  
Adventitious Bud Induction

scholarly journals In Vitro Adventitious Shoot Formation on Cotyledons of Pinus pinea

HortScience ◽  
1998 ◽  
Vol 33 (4) ◽  
pp. 749-750 ◽  
Author(s):  
María Victoria González ◽  
Manuel Rey ◽  
Raffaela Tavazza ◽  
Stefano La Malfa ◽  
Luigi Cuozzo ◽  
...  
Keyword(s):  
Adventitious Shoots ◽  
Shoot Multiplication ◽  
Adventitious Shoot ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Pinus Pinea ◽  
Induction Medium ◽  
Adventitious Shoot Formation ◽  
Bud Induction

Plant regeneration was obtained from adventitious buds induced in isolated cotyledons of Italian stone pine (Pinus pinea L.). The best results for bud induction were obtained by using half-strength LePoivre medium with 4.5 μM 6-benzyladenine for 30 days. Shoot elongation was achieved in the same medium without growth regulators but with the addition of 0.5% activated charcoal. The induction medium was the best also for shoot multiplication, but it was necessary to include subcultures on elongation medium. The slow elongation rate of adventitious shoots remains the greatest obstacle to multiplication. Root formation (15%) after 5 months was observed when shoots were cultured on elongation medium for long periods.

scholarly journals Histology of In Vitro Adventitious Bud Development on Cotyledons and Hypocotyls of Fraser Fir

1993 ◽  
Vol 118 (1) ◽  
pp. 163-167 ◽  
Author(s):  
Carole H. Saravitz ◽  
Frank A. Blazich ◽  
Henry V. Amerson
Keyword(s):  
Growth Regulators ◽  
Induction Medium ◽  
Adventitious Bud ◽  
Naphthaleneacetic Acid ◽  
Bud Development ◽  
Fraser Fir ◽  
Cell Clusters ◽  
Cell Divisions ◽  
Bud Induction

Cotyledons and hypocotyls of Fraser fir [Abies fraseri (Pursh) Poir.] were excised from seeds treated with H2 O2 for 9 days and placed on bud induction medium containing 10 mg BA/liter and 0.01 mg NAA/liter or medium without growth regulators. Although adventitious buds did not develop, cotyledons exposed to growth regulators responded differently than cotyledons placed on medium lacking growth regulators. Cotyledons and hypocotyls responded similarly to growth regulators during the initial phase in culture, but cell divisions ceased in cotyledons, thus preventing meristemoid and subsequent bud development. After 3 days on medium containing growth regulators cell divisions were localized in epidermal and subjacent layers of hypocotyls, whereas similar cell divisions were' not observed in hypocotyls placed on medium without growth regulators. Cell clusters consisting of two to five cells (promeristemoids) were present after 7 days on hypocotyls placed on bud induction medium. In hypocotyls placed on medium without growth regulators, stomata continued to develop and cells within the cortex became vacuolated during the first 2 weeks in culture. All explants were transferred to secondary medium after 3 weeks. Cell clusters continued to enlarge into meristemoids on hypocotyls initially placed on bud induction medium. Gradually, meristemoids developed into buds and cataphylls were observed covering bud meristems. Chemical names used: N -(phenylmethyl)-1 H -purine-6-amine (BA), 1-naphthaleneacetic acid (NAA).

scholarly journals Optimization of in vitro organogenesis in passion fruit (Passiflora edulis f. flavicarpa)

2005 ◽  
Vol 62 (4) ◽  
pp. 346-350 ◽  
Author(s):  
Flavio Trevisan ◽  
Beatriz Madalena Januzzi Mendes
Keyword(s):  
Culture Media ◽  
Shoot Elongation ◽  
Shoot Development ◽  
Passion Fruit ◽  
Coconut Water ◽  
In Vitro Organogenesis ◽  
Shoot Bud ◽  
Bud Induction ◽  
Water Media

In vitro organogenesis of passion fruit was studied by the induction of adventitious buds from leaf discs in culture media supplemented with benzyladenine (BAP) or thidiazuron (TDZ). To minimize adverse effects of ethylene accumulation on shoot development, silver nitrate (AgNO3) was added to the induction media. Both BAP (0; 2.2; 4.4; 6.6 µmol L-1) and TDZ (0; 1.1; 2.2; 3.4 µmol L-1) were effective in promoting shoot development. Although no significant differences were detected using AgNO3 (23.5 µmol L-1), buds grown in AgNO3-supplemented media were more vigorous. The number of explants with buds obtained using TDZ and AgNO3-supplemented media (5.6) were higher than those obtained using BAP and AgNO3 (3.0). MSM + giberrellic acid (GA3), MSM + coconut water, and ½ MSM culture media were tested for shoot bud elongation, incubated in flasks covered with either non-vented or vented lids. Best results were obtained by culturing buds in MSM + coconut water media in flasks covered with vented lids. Plantlets transferred to MSM + indol butyric acid (IBA) media rooted in a 30-day period. Passion fruit organogenesis was enhanced by using TDZ and AgNO3 for bud induction. Transferring the buds to MSM + coconut water media and incubating in flasks with vented lids favored shoot elongation and plantlet development.

scholarly journals In vitro organogenesis optimization and plantlet regeneration in Citrus sinensis and C. limonia

2002 ◽  
Vol 59 (1) ◽  
pp. 35-40 ◽  
Author(s):  
Weliton Antonio Bastos de Almeida ◽  
Francisco de Assis Alves Mourão Filho ◽  
Beatriz Madalena Januzzi Mendes ◽  
Adriana Pinheiro Martinelli Rodriguez
Keyword(s):  
Genetic Transformation ◽  
Citrus Sinensis ◽  
Sweet Orange ◽  
Adventitious Bud ◽  
Transformation Techniques ◽  
Rangpur Lime ◽  
Bud Induction ◽  
Bud Growth ◽  
Exogenous Genes

Exogenous genes can be introduced in plants by genetic transformation techniques. However, an efficient tissue culture system with high rates of plant recovery is necessary for gene introduction. This work aimed to define organogenesis and plant regeneration protocols for sweet orange varieties Natal, Valencia and Hamlin (Citrus sinensis L. Osbeck) and Rangpur lime (Citrus limonia L. Osbeck) which can be used in plant transformation experiments. Seeds of which teguments were removed, were germinated in vitro and maintained in the dark for three weeks, followed by one week at 16-h photoperiod (40 µmol m-2 s-1) and 27 ± 2°C. Organogenesis induction was done by introducing epicotyl segments in MT medium with 25 g L-1 sucrose and different BAP concentrations. After adventitious bud growth, the shoots were transferred to MT medium with either NAA or IBA (1 mg L-1), or absence of auxin, for rooting. The best results were obtained with 1 mg L-1 BAP for bud induction and 1 mg L-1 IBA for rooting for all three sweet orange cultivars. The use of 0.5-2.5 mg L-1 BAP, followed by 1 mg L-1 IBA were the best growth regulator combinations for bud induction and rooting, respectively, for 'Rangpur' lime. The protocols presented in this work are suitable for associations with genetic transformation experiments for these cultivars.

scholarly journals A Two-step Procedure for Adventitious Shoot Regeneration from in vitro-derived Lingonberry Leaves: Shoot Induction with TDZ and Shoot Elongation Using Zeatin

HortScience ◽  
2005 ◽  
Vol 40 (1) ◽  
pp. 189-192 ◽  
Author(s):  
Samir C. Debnath
Keyword(s):  
Survival Rate ◽  
Shoot Regeneration ◽  
Adventitious Shoot ◽  
Shoot Elongation ◽  
Adventitious Bud ◽  
Shoot Induction ◽  
Adventitious Shoot Regeneration ◽  
Explant Orientation ◽  
Step Procedure

The effects of TDZ (0, 0.1, 1, 5 and 10 μm) and explant orientation on adventitious shoot regeneration of `Erntedank' lingonberry were studied. Moderate concentration (1 to 5 μm) of TDZ supported bud and shoot regeneration, but strongly inhibited shoot elongation. TDZ initiated cultures were transferred to medium containing 1-2 μm zeatin and produced usable shoots after one additional subculture. Adventitious bud and shoot regeneration was greatly influenced by explant orientation. Elongated shoots were rooted on a 2 peat: 1 perlite (v/v) medium, and the plantlets were acclimatized and eventually established in the greenhouse with 80% to 90% survival rate.

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