scholarly journals Transformation of callus sugarcane (Saccharum officinarum L.) using Agrobacterium tumefaciens – Mediated method and regeneration of transgenic shoot

2013 ◽  
Vol 16 (1) ◽  
pp. 69-76
Author(s):  
Ha Van Tran ◽  
Phuong Hoang Phi Cung ◽  
Le Van Bui

Sugarcane (Saccharum officinarum L.) is a major industrial crop in the world. The creation of transgenic sugacane varieties contributed to overcome the disadvantages of traditional farming methods. Immature cutting cane stems after 10 days of putting on MS medium (Murashige and Skoog) supplemented 2,4-dichlorophenoxyacetic acid (2,4-D) 3 mg/L triggered the highest rate of explants inducing callus (91.11%). Calli were cultured on MS medium supplemented 6- Benzyladenin (BA) with concentrations of 0, 0.5, 1.0, 1.5, 2.0, 2.5 mg/L for inducing shoot. All six concentrations of BA could induce shoot, but the concentration of BA 1 mg/L was the most suitable. The 4-to-5-week old calli were introduced desired gene by using Agrobacterium tumefaciens, which was affected by acetosyringone concentration, inoculation and co-culture time. Consequently, transformation with acetosyringone 100 μM or 150 μM could be successful and regenerate transgenic shoots with assumed rate 25%.

2018 ◽  
Vol 54 (No. 1) ◽  
pp. 9-16 ◽  
Author(s):  
E.S. Kurtar ◽  
A. Balkaya ◽  
M. Ozbakir Ozer

Although haploids were successfully produced via irradiated pollen technique and anther culture in Cucurbita maxima and Cucurbita moschata, the haploidization efficiency is still low due to genotype dependence. Thus, as an alternative technique, the efficacy of the ovule culture was investigated. Ovules were extracted at different flowering time and then cultured on a solid MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D), benzylaminopurine (BAP), thidiazuron (TDZ), and naphthaleneacetic acid (NAA) to induce callogenesis and plant regeneration. The gynogenic response was influenced by the combination of plant growth regulators, genotype and culture time. The medium containing of 4.0 mg/l BAP + 0.05 mg/l NAA + 0.1 mg/l TDZ provided the highest response at anthesis time. Plantlets were rooted and elongated on a solid MS medium supplemented with 0.01 mg/l indole-3-acetic acid (IAA) + 1.0 mg/l BAP. The ploidy observations of 122 plants revealed that 70 plants were haploid, 46 plants were diploid and the others were mixoploid.  


2003 ◽  
Vol 55 (3-4) ◽  
pp. 77-80 ◽  
Author(s):  
Aneta Bijelovic ◽  
Marko Sabovljevic

Callus induction of moss species Aloina aloides (Schultz) Kindb. was obtained on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) or with 1.0 mg/L 2,4-D and 1.0 mg/L kinetin (KIN) or with 0.2 mg/L indole-3-butyric acid (IBA) and 2.0 mg/L 6-benzylaminopurine (BAP) or with 7.5 g/L of sucrose or with 15 g/L of sucrose or hormone - free and sugar free MS basal medium. The callus can be maintained for a long period of time without bud formation subcultured on the above media, at 16 h day/8 h night, 25 ? 2?C, 60-70% air humidity and irradiance of 50 ?mol m-2s-1. To obtain plant regeneration pieces, calli were transferred onto MS media supplemented with different concentrations of auxins and cytokinins (1.0 mg/L 2,4-D and 2 mg/L KIN; 0.2 mg/L IBA and 2 mg/L KIN; or 0.2 mg/L IAA and 2 mg/L BAP). In these media after subculturing, callus enlarges and turns to gametophytes with buds. Except for a smaller size, the plants obtained on the callus did not differ morphoanatomically from the shoots in the nature.


2020 ◽  
Vol 12 (3) ◽  
pp. 673-682
Author(s):  
Yasmina BENABDESSLEM ◽  
Kadda HACHEM ◽  
Samia GHOMARI

The chickpea (Cicer arietinum) is one of the leguminous species most appreciated by consumers in the Mediterranean basin, while being an important source of protein. Nevertheless, its crop yields are greatly limited by several biotic and abiotic stresses, the main one being Ascochyta rabiei, the causal agent of anthracnose. As traditional breeding methods have proved to be ineffective in controlling this pathogen, resorting to biotechnological methods is necessary. Therefore, in this study, the callogenic capacity of stem and leaflet explants from three genotypes of chickpea, namely ‘FLIP 84-92 C’, ‘ILC 32-97’, and ‘ILC 263’, cultured on Murashige and Skoog (MS) medium with different hormonal balances of auxins (indole-3-acetic acid [IAA] and 2,4-dichlorophenoxyacetic acid [2,4-D]) and cytokinin (kinetin), was determined. For all the genotypes, high percentages of callogenesis were recorded in the different explants grown on an MS medium with 2 mg of both IAA and kinetin. Then, a patho-system of Cicer arietinum calluses with Ascochyta rabiei was investigated, followed by a histological assessment of this interaction. The presence of the fruiting bodies of the pathogen was revealed in the calluses of the ‘ILC 32-97’ and ‘ILC 263’ genotypes. Notably, the latter showed a high sensitivity to the pathogen, as indicated by an abundance of pycnidia in its tissues. As for the ‘FLIP 84-92 C’ genotype, the histological sections showed a total absence of inter- and intracellular fruiting bodies of the pathogen in the callus tissues. Therefore, this genotype was considered as resistant to Ascochyta rabiei.


2014 ◽  
Vol 34 (6) ◽  
pp. 523-528 ◽  
Author(s):  
Ana F. Vigário ◽  
Simone M.T. Sabóia-Morais

The 2,4-dichlorophenoxyacetic acid, usually named 2,4-D is one of the most widely used herbicides in the world. Acute toxicity of 2,4-D herbicide was investigated through its effects on guppies (Poecilia vivipara Bloch et Schneider 1801). Fish were exposed to the herbicide at concentrations of 10, 20 and 40µl per liter of water for 24 hours to determine its effects on gills and liver epithelia. The estimated LC50 was 34.64µl of 2,4-D per liter of water. Histochemical analyses and Feulgen's reaction were conducted to detect glycoconjugates and DNA, respectively, in gills and liver epithelia. Histochemistry revealed qualitative variations of glycoconjugates present on mucous cells and granules. The four types of mucous cells contained neutral granules, acids, or both. Increasing amounts of syalomucins were observed from the control group to the group exposed to the highest concentration of 2,4-D, suggesting increased mucous viscosity and the formation of plaques that could inhibit gas exchange and osmoregulation. Lamellar fusion observed in the group exposed to 40µl of 2,4-D suggests a defense mechanism. Hepatocytes showed vacuolization in the 10 and 20µl/L groups. The 40 µl/L group showed normal hepatocytes as well as changed ones, many Ito cells, micronuclei, and nuclear swelling. These effects may be associated with toxicity or adaptative processes to cellular stress. The data from this study indicates the importance of assessing similar risks to aquatic species and suggests that Poecilia vivipara is an adequate biological model for analysis of environmental contamination.


1996 ◽  
Vol 44 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Perumal Venkatachalam ◽  
Narayanasamypillai Jayabalan

High yields of protoplasts were obtained from immature leaves of aseptically grown plants of Arachis hypogaea using an enzyme solution containing cellulase 2.0% (w/v) and Macerozyme 1.0% (w/v) in 0.6 M mannitol. Isolated protoplasts were cultured in Kao's medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and mini calli in 4 weeks. After 4 weeks, protoplast colonies were transferred to the Murashige and Skoog (MS) medium supplemented with a-naphthalene acetic acid (NAA) and BAP. Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones differentiated roots on the same medium. Alternative methods for plant regeneration from protoplast derived callus cultures were tried through somatic embryogenesis. Protoplast-derived calli treated with 2,4-D and BAP formed somatic embryos. Somatic embryogenesis began in the proembryo stage and proceeded from globular to dicotyledonary stage. Embryos were then transferred onto hormone-free MS medium for germination. Five to ten percent of these embryoids germinated and grew to plantlets. Regenerated plants were transferred to plastic cups and grown to maturity.


2019 ◽  
Vol 7 (4.14) ◽  
pp. 107
Author(s):  
D S M Soder ◽  
D N A A Khalid ◽  
A Saleh ◽  
F Pardi ◽  
N J Sidik

Barringtonia racemosa is mangroves type of plant which had been extensively utilized in conventional practices for relieving ailments of pain and inflammation. Many studies have been done on ethnobotanical profiles, pharmacological activities and chemical compounds in Barringtonia racemosa. However, there is a limited study on callogenesis of this plant particularly from different maturity stage of fruits. The present study is to identify the callogenesis of Barringtonia racemosa from endosperm explants of immature and mature fruits in MS medium supplemented with different concentrations of hormones 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 0.5, 1.0, 1.5 and 2.0 mg/L) and Kinetin (KIN) (0, 0.5, 1.0, 1.5 and 2.0 mg/L). The optimum hormone combination was found in callus grown on endosperm of immature fruits in MS medium supplemented with 1.5 mg/L 2,4-D and 1.0 mg/L KIN. It was also found that the callus in this treatment grew profusely with highest fresh weight (0.513 ± 0.022 g), 100% callus induction and friable callus texture. The callus fresh weight on endosperm explants was higher in immature fruits compared to mature fruits for all the hormone combinations. Therefore, callogenesis were found more efficient from endosperm explant of immature fruits in Barringtonia racemosa species.   


2017 ◽  
Vol 1 (2) ◽  
Author(s):  
Nining Intan Toharah ◽  
Dwi Soelistya Dyah Jekti ◽  
Lalu Zulkifli

This study aims to determine the concentration of growth regulators BAP and   2,4-D which have the highest effect in stimulating the formation of callus melon plants (Cucumis melo) Mai 119 variety. Completely randomized design (CRD) was used in this research. Media used on callus induction was MS medium with addition of several concentration of BAP  (0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L) and 2,4-D (0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L) either alone or in a combination of both. Parameters measured were the time appearing of callus, callus diameter, callus texture, and callus color. Anova followed by Tukey's test was used to the analyse of time appearing of callus. Data of callus diameter was analyzed using Kruskal Wallis test followed by Mann-Whitney test. In the analysis of parameter related to the callus texture and callus color, descriptive test were used. The results showed that there were differences in the effect of growth regulators on the callus formation. The fastest callus induction and the largest diameter of callus were obtained on media with concentration of 2 mg/L BAP and 3 mg/L BAP.Keywords: BAP (benzyl amino purine), 2,4-D (2,4-dichlorophenoxyacetic acid), callus induction, melon (Cucumis melo) varieties Mai 119


2011 ◽  
Vol 3 (1) ◽  
pp. 79-86 ◽  
Author(s):  
Abd El-Moneim Mohamed Radwan AFIFY ◽  
Hossam Saad EL-BELTAGI ◽  
Anwer Abd El-Aziz HAMMAMA ◽  
Mahassen Mohamed SIDKY ◽  
Omneya Farouk Ahmed MOSTAFA

In the present study, seeds from local cultivar of fennel were germinated on Murashige and Skoog medium (MS) without plant growth regulators. Different types of explants from the growing seedling such as cotyledonal leaves, hypocotyls, epicotyls and roots were cultured on MS medium, contained different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) either alone or with kinetin. Differential responses in the essential oil constituents were observed in the induction and development of callus. The major components of essential oils includes estragole, trans-anethole, limonene and fenchone were studied under different conditions to find out the best methods which could be used to reduce the amount of estragole (not favorite for human consumption) and increase the amount of trans-anethole.


1995 ◽  
Vol 43 (4) ◽  
pp. 385-390 ◽  
Author(s):  
S. Kulothungan ◽  
A. Ganapathi ◽  
A. Shajahan ◽  
K. Kathiravan

Embryogenic callus was induced from seedling leaf explants of cowpea (Vigna unguiculata (L.) Walp. cv. C152 on Murashige and Skoog (MS) medium containing 2.0 mg 1−1 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of somatic embryogenesis was noticed when this callus was transferred to MS liquid medium supplemented with 2 mg 1−1 2,4-D. Further studies on ontogeny of somatic embryos showed that the cells destined to become somatic embryos divided into spherical or filamentous proembryos. Subsequent divisions in the proembryo led to globular, heart, torpedo-shaped, and cotyledonary-stage somatic embryos. Tiny plantlets were obtained by transferring the cotyledonary-stage somatic embryos to MS liquid medium containing 0.5 mg 1−1 2,4-D.


2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.


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