scholarly journals An Effective Procedure for In Vitro Culture of Eleusine coracana (L.) and Its Application

ISRN Botany ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Alla I. Yemets ◽  
Galina Ya. Bayer ◽  
Yaroslav B. Blume

Efficient protocols for callus production, plantlet regeneration, protoplast isolation, and micronucleation of finger millet (Eleusine coracana (L.) Gaertn.) were developed. White nodulated calli were formed on medium with N6 macrosalts, MS microsalts, 2.4-dichlorophenoxyacetic acid (2 mg L−1), kinetin (0.4 mg L−1), 1-naphthalene acetic acid (2 mg L−1), and certain additives. It was found that appropriate supplementation leads to formation of numerous shoots. Healthy rooted plantlets formed on hormone-free media. Although different tested additives had no significant effect on percentage of callus formation, it affected callus quality that further dictated plant-forming capacities. Seedlings were better source tissues for protoplasts isolation compared to callus cultures. About protoplasts were isolated from one gram of seedling coleoptyles. Microcolonies were visible after 20–25 days' incubation on KM8p medium supplemented with glutamine (100 mg L−1) and proline (500 mg L−1). Here we also present a procedure of an efficient induction of micronuclei after chlorpropham (10 μM) and cytochalasin-B (20 μM) seedlings treatment with subsequent microprotoplasts isolation. This technique is discussed for the transfer of alien chromosomes and genes from finger millet by microprotoplast-mediated chromosome transfer.

2018 ◽  
Vol 22 ◽  
pp. 274-281
Author(s):  
N. B. Kravets ◽  
N. V. Tulaidan ◽  
M. Z. Mosula ◽  
N. M. Drobyk

Aim. The aim of the research was to choose the conditions for microclonal propagation and obtain callus cultures from Carlina аcaulis L., Carlina cirsioides Klok and Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl plants in vitro. Methods. For microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia we used rosettes of 2–3-month specimens and planted them on semi-solid Murashige and Skoog (MS) medium with decreased macro- and microsalts concentrations (MS/2) supplemented with kinetin (Кin) (from 1–3 mg/l) and 0.1 mg/l of 1-naphthaleneacetic acid (NAA). For induction of callus formation, we used root, stem explants from С. acaulis, C. cirsioides and C. onopordіfolia, and planted them on nutrient media MS, MS/2, and Gamborg and Eveleigh (В5) supplemented with different concentrations of cytokinins – 6-benzylaminopurine (BAP) or Кin and auxins – 2.4-dichlorophenoxyacetic acid (2.4-D) or NAA and indole-3-acetic acid (IAA). Results. MS/2 medium supplemented with growth regulators of NAA and Кin were the most efficient to provide the formation of microclones. For C. сirsioides plants, this indicator was 6.6–6.8 rosettes per graft after 6 months of cultivation and for С. acaulis and C. onopordіfolia – 4.2–5.0 and 4.8–5.2 respectively. To raise the percentage of rooting for microclones of Carlina species, it was expedient to steep them preliminarily in the solution of indole-3-butyric acid (IBA) with 1000 mg/l concentration for a minute. Optimal for obtaining callus tissue from Carlina plants was nutrient medium MS supplemented with 3 mg/l IAA, 0.5 mg/l NAA and 0.5 mg/l Kin and MS/2 with 0.1 mg/l BAP and 0.5 mg/l 2.4-D; under such conditions the percentage of callus induction exceeded 90 % for all types of explants. Conclusions. There were chosen the conditions for microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia and worked out the schemes for enrooting obtained microclones in vitro. Capable of growing rapidly callus cultures from root and stem explants of the investigated plant species were obtained. Keywords: Carlina аcaulis L., Carlina cirsioides  Klok, Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl, in vitro, microclonal propagation, callus induction.


2018 ◽  
Vol 1 (6) ◽  
pp. 106-116
Author(s):  
Huong Thanh Tran ◽  
Cuong Quoc Vo

In this paper, the roles of type and concentration of enzymes on protoplast isolation from in vitro leaves, multi-scalps (highly proliferating meristem culture), and young male flower of banana cv. cau man were studied. Respiration rate and content of plant hormones of these materials were analysed. Different techniques were used to culture these protoplasts. The development of protoplasts was observed under fluorescence microscope. The highest yield of protoplast (69.5 x 106 protoplasts / g fresh weight) was obtained from young male flowers after 16 hours treatment with 1.5 % cellulase, 0.25 % pectinase and 0.25 % hemicellulase. The combination of hanging drop cell technique (in 6 days), and carrot feeder layer cells in N6PKM medium supplemented with 0.2 mg/L 2,4- dichlorophenoxyacetic acid (2,4-D), 1 mg/L α- naphthalene acetic acid (NAA), and 0.5 mg/L zeatin are suitable for protoplast development. Protoplasts that were isolated from multi-scalps and young male flowers created the wall and divided when cultured by this method. The development of protoplasts from young male flower began with cell walls creation after 4 days, the cell division was after 6 days, and small colonies formation was after 28 days of culture. The differentiation and physiological activity of cells play an important role on quantity and quality of protoplasts, as on the well as protoplast development.


Biologia ◽  
2010 ◽  
Vol 65 (4) ◽  
Author(s):  
Sara Rostampour ◽  
Haleh Sohi ◽  
Ali Dehestani

AbstractPersian poppy (Papaver bracteatum Lindl.) is an important commercial source of medicinal opiates and related compounds. In this research, calli were induced from seeds, roots, cotyledons and hypocotyls of P. bracteatum at a high efficiency. The optimized callus induction media consisted of the Murashige and Skoog (MS) basic media supplemented with 1.0 mg/L 2, 4-dichlorophenoxyacetic acid (2,4-D), 0.1 mg/L kinetin and 15 mg/L ascorbic acid. The concentrations of 2,4-D and ascorbic acid were found critical to callus induction and proliferation. Subsequent subcultures resulted in excellent callus proliferation. Ascorbic acid at concentration 15 mg/L increased the callus proliferation significantly. Maximum callus growth was achieved when the explants were incubated at 25°C. MS salts at full strength were found inhibitory for callus induction, while ľ MS salts were found to favor callus induction. Shoot regeneration of calli in vitro was achieved on ľ MS medium containing 0.5 mg/L benzylamine purine and 1.0 mg/L naphthalene acetic acid. Analysis of alkaloid extracts from Persian poppy tissues by high-performance liquid chromatography showed that thebaine accumulated in the tissues of plants. The thebaine alkaloid profile of the Persian poppy is a well-defined model to evaluate the potential for metabolic engineering of thebaine production in P. bracteatum.


1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'


2015 ◽  
Vol 29 (1) ◽  
pp. 25-42
Author(s):  
Anna M. Domańska ◽  
Aldona Rennert

The clones of excised roots, leaves, petioles, cotylenods, hypocotyls and root calluses derived from the respective carrot fragments (cv. 'Perfekcja' commonly cultivated in Poland) were cultured <i>in vitro</i>. An influence of thiamine concentrations on the growth of root tissue was examined. Several various media were tested for callus cultures. Bee bread extract was also applied. The growth of isolated clones during early and later culture periods was compared.


1978 ◽  
Vol 56 (10) ◽  
pp. 1287-1290 ◽  
Author(s):  
Ming-Chin Liu ◽  
Wen-Huei Chen

Experiments have been performed to induce callus formation and organogenesis in anther culture of cassava (Manihot esculenta Crantz). Callusing was achieved on a modified Murashige and Skoog medium (MSB) supplemented with 4.44 μM 6-benzylaminopurine (BAP) and 4.52 μM 2,4-dichlorophenoxyacetic acid (2.4-D). No callus was formed from anthers pretreated at 4 °C for more than 48 h or on a medium containing 4g/ℓ activated charcoal. Callus on MSB with 4.44–8.88 μM BAP alone formed roots only. BAP (8.88 μM) in combination with α-naphthalene acetic acid (NAA) (10.74 μM) resulted in chlorophyll formation in callus. Abscisic acid (ABA) acted as an antagonist to NAA in reducing the frequency of callus greening when the latter was applied jointly with BAP. Chromosome counts of mitotic figures from callus cells ranged from 34 to 38 indicating that the calli were derived from the somatic tissues of the anthers.


2018 ◽  
Vol 47 (2) ◽  
pp. 538-543
Author(s):  
Rodrigo Kelson S. REZENDE ◽  
Ana Maria N. SCOTON ◽  
Maílson V. JESUS ◽  
Zeva V. PEREIRA ◽  
Fernanda PINTO

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.


2020 ◽  
Vol 8 (1) ◽  
pp. 1
Author(s):  
Sadia Afrin Jui ◽  
Md. Mijanur Rahman Rajib ◽  
M. Mofazzal Hossain ◽  
Sharmila Rani Mallik ◽  
Iffat Jahan Nur ◽  
...  

The experiment was designed to evaluate the effect of growth regulators on leaf explant of Gerbera for callus induction. Various kinds of plant growth regulators such as 6-Benzylaminopurine (BAP), α-Naphthalene acetic acid (NAA), 2, 4-Dichlorophenoxyacetic acid (2, 4-D), Indole-3-acetic acid (IAA) were used to initiate cultures. These were added to Murashige and Skoog medium in different combinations and concentrations. Leaf explants cultured on MS medium supplemented with BAP+ 2, 4-D+ IAA in T4 treatment & BAP+ 2,4-D in T5 treatment showed the best results for callus induction. On the other hand callus was induced early in the combination of BA+ 2,4-D + IAA hormone in T5, T9 & T8 treatment respectively. The rate of callus induction was very low in BA + NAA combinations but it was much earlier.   


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