Improvement of the in vitro regeneration and Agrobacterium-mediated genetic transformation of Medicago sativa L.

Nofouzı, Farzad; Oğuz, Muhammet Çağrı; Khabbazi, Saber Delpasand; Ergül, Ali

doi:10.3906/tar-1804-52

Cited by 8 publications

(5 citation statements)

References 26 publications

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“…The results of the study emphasize that hydropriming positively facilitated in detaching zygotic embryos sandwiched between cotyledons. The results are in agreement with my previous observations (Nofouzi et al, 2019). All explants irrespective of the concentration of TDZ swelled before regeneration.…”

Section: Discussionsupporting

confidence: 93%

“…The results are not in agreement with findings from another group (Bahgat et al, 2009). Nofouzi et al (2019) Obtained the highest mean number of regenerated shoots for Alfalfa was achieved when explants were subjected to 0.55 and 0.15 mg/L TDZ, respectively. They regenerated somatic embryos on two Egyptian broad bean cultivars Giza 2 & Hyto using 10 mg l -1 BAP+0.5 mg l -1 NAA followed by transfer of the calli to 1/2 B5 medium.…”

Section: Discussioncontrasting

confidence: 86%

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Establishment of A Novel Plant Micropropagation System From Mature Hydroprimed Seeds Of Two Turkish Broad Bean Cultivars

Turan

Khawar

2022

Türk Tarım Ve Doğa Bilimleri Dergisi

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Leguminous Broad bean (Vicia faba L.) is an excellent source of minerals, vitamins and proteins that makes it highly valuable food for human and feed for animal consumption. The plant has an important characteristic to fix atmospheric Nitrogen and play an important role to establish a natural balance of N in the atmosphere. It is a highly self-pollinated plant and has problem of low variation with limited genetic pool. Moreover, the previous studies identify problem of recalcitrance in broad bean. Therefore, there is need to establish a repeatable micropropagation protocol that could ensure an increase in genetic variability to overwhelm problems in breeding. This system must also be able for efficient gene delivery and could be integrated with the conventional breeding programs through direct organogenesis. The study aimed to develop a tissue culture (shooting & rooting) protocol on two important Turkish broad bean cultivars Filiz99 and Eresen87 using MS medium containing 0.05, 0.15, 0.25, 0.35, 0.45, 0.55 mg l -1 TDZ using embryonic axis as explant. Maximum number of 5.33 and 3 shoots per explant were noted on cv. Filiz99 and Eresen87 on MS medium containing 0.15 mg l -1 TDZ. The developing shoots were rooted on MS medium containing 1 mg l -1 IAA after three weeks of culture. The rooted plants were transferred to pots containing peat under maintained under controlled greenhouse conditions for acclimatization The acclimatized plants bloomed and set seeds. Present results underscore importance of seed hydropriming before taking of explants to achieve high micropropagation on faba beans to overcome recalcitrance.

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Section: Discussionsupporting

confidence: 93%

Section: Discussioncontrasting

confidence: 86%

Establishment of A Novel Plant Micropropagation System From Mature Hydroprimed Seeds Of Two Turkish Broad Bean Cultivars

Turan

Khawar

2022

Türk Tarım Ve Doğa Bilimleri Dergisi

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“…Alfalfa is one of the significant forage plants which is commonly used in animal feed due to its high protein, carbohydrate, and vitamins in many regions. It is a well-known forage queen and crops domesticated in ancient times by humans (Nofouzi et al, 2019;Yazıcılar et al, 2021). Turkey has been considered the secondary center of genes for alfalfa crops.…”

Section: Introductionmentioning

confidence: 99%

Embryogenic Callus Differentiation in Short-Term Callus Derived from Leaf Explants of Alfalfa Cultivars

Yazıcılar

Chang

2022

natprobiotech

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Alfalfa is a forage crop that accounts for one of the best sources of protein and is commonly grown all over the world. In vitro callus and embryogenic induction of alfalfa have been investigated previously; however, most of the investigations were almost limited with callus formation. In this study, leaves of 5 Medicago sativa L. cultivars (Alsancak, Sazova, Plato Iside, and Bilensoy) have been used. Influences of culture medium contents and hormones applications on callus and embryogenic callus induction were determined to optimize in vitro culture mediums of alfalfa (0.0125 g kinetin 2,4 D 1 mg mL-1; 1 g kinetin 2,4 D 1 mg mL-1 ; 0.25 g kinetin 2,4 D 2 mg mL-1; 0.5 g kinetin 2,4 D 2 mg mL-1). Callus formation was detected at a rate of 74% in 5 different cultivars used in the experiment. The five alfalfa cultivars were classified into four categories in terms of embryogenic differentiation capacity. The tested alfalfa cultivars varied in their callus formation and embryogenic callus differentiation. Sazova, Alsancak, and Bilensoy were detected for better callus formation; similarly, the same cultivars responded with better embryogenic callus formation in the culture mediums including various hormone concentrations. The present study shows that our methods have beneficial impacts on the somatic embryo induced by alfalfa. However, it depends strongly on genotype, hormone concentrations and the other medium components.

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“…The success of genetic transfer efficiency with A. tumefaciens depends on, Agrobacterium strain used, bacteria concentration, antibiotic types and concentration used for in vitro selection, inoculation time, and temperature [2]. Besides, the type of the target plant, plant explants, hormone combinations used in in vitro regeneration, pH, etc., are among the factors affecting the recovery of transgenic plants [3]. Increasing the transformation efficiency by reducing the limiting factors in genetic transformation studies will significantly contribute to the success rate [4].…”

Section: Introductionmentioning

confidence: 99%

Gamma Radiation Effect on Agrobacterium tumefaciens-Mediated Gene Transfer in Potato (Solanum tuberosum L.)

Aycan¹,

Oğuz²,

Özgen³

et al. 2021

Solanum Tuberosum - A Promising Crop for Starvation Problem

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Potato (Solanum tuberosum L.) is one of the major crops of the world. Significant improvements can be achieved in terms of yield and quality by the determination of efficient transformation methods. On the other hand, low transformation frequency seriously limits the application of molecular techniques in obtaining transgenic crops. In the present study, the effect of gamma radiation on Agrobacterium tumefaciens-mediated transformation to the potato was firstly investigated. Sterile seedlings of potato cv. ‘Marabel’, which was grown on Gamborg’s B5 medium in Magenta vessels, were irradiated with different gamma radiation doses (0-control, 40, 80, 120 Gy 60Co). Stem parts having axillary meristems were excised from irradiated seedlings and inoculated by A. tumefaciens (GV2260), which harbors the binary plasmid p35S GUS-INT contains and GUS (β-glucuronidase) gene controlled by 35S promoter (CaMV) and nptII (neomycin phosphotransferase II) gene driven by NOS (nopaline synthase) promoter). Inoculated stem parts having axillary meristems explants were then directly transported to a selection medium containing duocid (500 mg l−1), and kanamycin (100 mg l−1), 4 mg l−1 gibberellic acid, 1 mg l−1 BAP and 0.1 mg l−1 NAA. The adult transgenic plants were detected by polymerase chain reaction (PCR) analysis. According to the number of transgenic plants determined by PCR analysis, results obtained from explants treated with 40 Gy gamma gave the best results compared to the control (0 Gy) application. The doses over 40 Gy were also found statistically significant compared to the control (0 Gy). It is expected that the protocol described in this study make the transformation studies easier by skipping the stages of ‘co-cultivation’, ‘culturing explants on selection medium’ and ‘recovery of transgenic shoots on selection medium’ not only for potato but also for other crop plants. This study was supported by a grant from the Scientific and Technological Research Council of Turkey (TUBİTAK) (Grant number 113O280 to Prof. Dr. Mustafa YILDIZ).

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Improvement of the in vitro regeneration and Agrobacterium-mediated genetic transformation of Medicago sativa L.

Cited by 8 publications

References 26 publications

Establishment of A Novel Plant Micropropagation System From Mature Hydroprimed Seeds Of Two Turkish Broad Bean Cultivars

Establishment of A Novel Plant Micropropagation System From Mature Hydroprimed Seeds Of Two Turkish Broad Bean Cultivars

Embryogenic Callus Differentiation in Short-Term Callus Derived from Leaf Explants of Alfalfa Cultivars

Gamma Radiation Effect on Agrobacterium tumefaciens-Mediated Gene Transfer in Potato (Solanum tuberosum L.)

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