In vitro regeneration of Turkish dwarf chickling (Lathyrus cicera L) using immature zygotic embryo explant

Disinfected mature seed embryos of Picralima nitida, were cultured in MS medium supplemented with different concentrations and combinations of 2,4-D, BAP and NAA to determine an efficient protocol for in vitro propagation. Nine culture media made of combination of different components were used in a factorial design with three replications. Results showed up to 80 ± 4% disinfection rate with combination of triton x- 100 (0.2%) and sodium hypochlorite (30%). Embryo germination was highest on control medium. Rooting was higher (2±1 roots per embryo) after 4 weeks on control medium and on BAP supplemented medium at 0.8 μM while the longest root (1.5±0.5 cm) was observed on 2,4-D supplemented medium at 1.8 μM. Black soil was suitable for leaf formation (4 ± 2 leaves) and shoot elongation (2±1 cm) after 8 weeks in acclimatisation. These results show efficient disinfection, regeneration and acclimatisation of Picralima nitida. Plant Tissue Cult. & Biotech. 31(2): 143-151, 2021 (December)

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In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Local Varieties of Mungbean (Vigna radiata (L). Wilczek)

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v29i1.41981 ◽

2019 ◽

Vol 29 (1) ◽

pp. 81-97

Author(s):

Sujay Kumar Bhajan ◽

Setara Begum ◽

Mohammad Nurul Islam ◽

M Imdadul Hoque ◽

Rakha Hari Sarker

Keyword(s):

Genetic Transformation ◽

Vigna Radiata ◽

Zygotic Embryo ◽

In Vitro Regeneration ◽

Multiple Shoots ◽

Direct Organogenesis ◽

Pcr Analysis ◽

Root Induction ◽

Half Strength

An efficient Agrobacterium-mediated transformation compatible in vitro regeneration protocol was developed for two important varieties of mungbean (Vigna radiata (L.) Wilczek) cultivated in Bangladesh, namely Binamoog-5 and BARI Mung-6. Two different zygotic embryo derived explants, such as cotyledonary node (CN) and cotyledon attached decapitated embryo (CADE) were used for direct organogenesis of shoot. MS supplemented with 4.0 μM BAP was found to be the best for the development of highest number of multiple shoots from CADE in both the varieties of mungbean. While in case CN the best shoot formation was achieved on MS containing 4.0 μM BAP and 0.5 μM NAA in both varieties. Half strength of MS with 2.0 μM IBA was found to be most effective for producing healthy root from regenerated shoots. Following root induction, the in vitro raised plantlets were successfully transplanted to soil for their establishment. Considering overall responses, genetic transformation efficiency was found to be better with CADE explant using Agrobacterium tumefaciens strain LBA4404 harboring the binary plasmid pBI121 conferring GUS and nptII genes. Different factors influencing transformation was optimized during this study. Selection of transformed shoots was carried out by gradually increasing the concentration of kanamycin and such transformed shoots were eventually selected using 200 mg/l kanamycin. Stable expression of the GUS gene was detected in various parts of regenerated transformed plantlets. Transformed shoots were rooted on half strength MS containing 2.0 μM IBA and 100 mg/l ticarcillin. Rooted transformed plantlets were successfully transferred to soil. Stable integration of GUS and nptII genes in the putative transformed shoots was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 81-97, 2019 (June)

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The effect of auxins in inducing organogenesis or somatic embryogenesis in mature sunflower zygotic embryo derived apex

Notulae Botanicae Horti Agrobotanici Cluj-Napoca ◽

10.15835/nbha48111591 ◽

2020 ◽

Vol 48 (1) ◽

pp. 150-161

Author(s):

Adriana AURORI ◽

Imola MOLNAR ◽

Elena RAKOSY-TICAN

Keyword(s):

Zygotic Embryo ◽

In Vitro Regeneration ◽

Mature Stage ◽

Induction Medium ◽

Naphthaleneacetic Acid ◽

Axillary Shoots ◽

Mature Embryos ◽

Cytokinin Treatment ◽

2,4 Dichlorophenoxyacetic Acid

Induction of shoots or of somatic embryos is the key step for gaining the morphogenetic potential in sunflower (Helianthus annuus L.), species known as recalcitrant to in vitro regeneration. In the immature zygotic embryo derived tissues or in other juvenile tissues resulted from seedlings, the acquisition of the competence for regeneration can be achieved directly by cytokinin treatment or by preconditioning the explants on cytokinin containing medium. In this paper is presented a new type of explant for sunflower in vitro culture, consisting of the apex with primordial leaves, resulted from ungerminated mature zygotic embryo, in which a specific morphogenetic response was triggered by the exogenously applied auxins. Among the auxins tested, indole-3-acetic acid, indole-3-butyric acid and 1-naphthaleneacetic acid are inducers of an organogenetic response, apical/axillary shoots and adventitious buds being regenerated while 2,4-dichlorophenoxyacetic acid, 3,6-dichloro-2-methoxybenzoic acid and 4-amino-3,5,6-trichloropicolinic acid led to somatic embryo formation. Among the auxins tested only 4-amino-3,5,6-trichloropicolinic acid sustains the embryos development up to mature stage. A high amount of sucrose (120 g L-1) supplied during the auxin treatment promotes the maturation of the embryos directly on the induction medium for all tested auxins with embryogenic effect. These findings show that regardless of the type of morphogenetic response aimed in sunflower meristematic tissues resulted from mature embryos, the presence of auxins is mandatory.

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In vitro Regeneration and Over Expression of Pea DNA Helicase 45 (PDH45) Gene into the Local Cultivars of Chickpea (Cicer arietinum L.) through Agrobacteriummediated Genetic Transformation

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v28i1.37204 ◽

2018 ◽

Vol 28 (1) ◽

pp. 125-140

Author(s):

Nuram Mubina ◽

MI Hoque ◽

RH Sarker

Keyword(s):

Genetic Transformation ◽

Cicer Arietinum ◽

Zygotic Embryo ◽

In Vitro Regeneration ◽

Dna Helicase ◽

Salt Tolerant ◽

Agrobacterium Strain ◽

Cicer Arietinum L ◽

Gus Histochemical Assay

In vitro regeneration studies compatible to Agrobacterium-mediated genetic transformation were carried out using two different types of zygotic embryo derived explants namely, decapitated embryo (DE) and decapitated embryo with single cotyledon disc (DEC) from three varieties of chickpea (Cicer arietinum L.) such as BARI chhola-4, -5 and -9 cultivated in Bangladesh. The best responses towards in vitro shoot regeneration was obtained from decapitated embryo with DEC on MS containing 0.5 mg/l BAP, 0.5 mg/l Kn and 0.2 mg/l NAA. Healthy and effective roots from the regenerated shoots were developed on MS supplemented with 0.2 mg/l IBA. Genetic transformation was carried out with Agrobacterium strain LBA4404 containing the binary plasmid pCAMBIA1301- PDH45 to integrate salt tolerant PDH45 gene in locally grown varieties of chickpea. The transformed plantlets were successfully established in soil following adequate hardening. Integration of salt tolerant PDH45 gene within the genomic DNA was confirmed through GUS histochemical assay and PCR analysis.Plant Tissue Cult. & Biotech. 28(1): 125-140, 2018 (June)

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