scholarly journals Regeneration of Genetically Stable Plants from in Vitro Vitrified Leaves of Different Carnation Cultivars

Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 950 ◽  
Author(s):  
Ho Thi Minh Thu ◽  
Aung Htay Naing ◽  
Hui Yeong Jeong ◽  
Chang Kil Kim

This study was conducted to investigate the efficacy of shoot regeneration from different leaf types (normal leaves and vitrified leaves) from three different carnation cultivars ‘Kumbuyl’, ‘Denev’, and ‘Jinju’ using different combinations of 3-indole butyric acid (IBA) and thidiazuron (TDZ) concentrations. The shoot tips cultured on Murashige and Skoog (MS) basal media (Type 1 media) produced normal leaves, while those cultured-on media supplemented with plant growth regulators and/or vitamin (Type 2 media and Type 3 media) produced vitrified leaves for all cultivars. Culture of normal leaf segments on MS medium containing different combinations of IBA and TDZ concentrations induced callus in all treatments; however, the callus was unable to induce shoots and finally became necrotic. In contrast, no callus induction was observed in the control (hormone-free treatment). When vitrified leaf segments underwent the same treatments, shoots were induced from the vitrified leaves (derived from Type 2 media) but were unhealthy and gradually died, whereas those induced from Type 3 media were vitrified and healthy. The optimal combination for the best shoot regeneration and number of shoots per explants varied depending on the genotypes used. The vitrified shoots induced from the leaves of Type 3 media transformed into normal shoots and survived well under greenhouse conditions. According to the results of random amplified polymorphic DNA (RAPD) analysis, the banding patterns of twelve primers that were detected in vitrified leaf-induced normalized shoots were identical to those of normal in vitro grown plants, indicating that no genetic variation had occurred during the procedure. Taken together, this study indicates that vitrified leaves can be used for shoot regeneration of recalcitrant carnation cultivars, regardless of the genotypes and types of vitrified leaves. However, as the number of shoots per explants was still low, further investigation is warranted to obtain a more efficient shoot regeneration protocol for genetic transformation of the cultivars.

2009 ◽  
Vol 36 (No. 4) ◽  
pp. 140-146 ◽  
Author(s):  
J.K. Kanwar ◽  
S. Kumar

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in <I>Dianthus caryophyllus</I> L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.


2017 ◽  
Vol 14 (2) ◽  
pp. 24-31 ◽  
Author(s):  
S S Riva ◽  
A Islam ◽  
M E Hoque

An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration.  It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium.  For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.The Agriculturists 2016; 14(2) 24-31


1984 ◽  
Vol 62 (9) ◽  
pp. 853-858 ◽  
Author(s):  
Erwin Regoeczi ◽  
Paul A. Chindemi ◽  
Maria T. Debanne

125I-labeled asialotransferrin types 1 and 2 were administered in small doses to rats. The protein still in the plasma after 1–12 h was partially repurified and electrophoresed at pH 8.1, together with a transferrin standard that is composed of all six forms of the protein with respect to sialic acid content. The electrophoretic mobility of both asialotransferrins increased with time, type 2 being affected sooner than type 1. The changed mobility was due to increased electronegativity that was fully reversible by treatment of the samples with neuraminidase, thus identifying the underlying cause as partial resialylation. Asialotransferrin incubated in vitro with serum, plasma, or whole blood for 16 h exhibited no change in electrophoretic mobility. In conjunction with an earlier study on asialotransferrin type 3, it was found that the apparent speeds of resialylation of the three asialotransferrins were in the same order as their affinities for the asialoglycoprotein-binding hepatic lectin. This suggests the involvement of an endo- rather than of an ecto-transferase. Transfer of 59Fe from asialotransferrins to the liver was used to monitor the frequency of hepatocyte–asialotransferrin interactions. Iron deposition in the liver took place much more rapidly than the appearance of detectable quantities of partially resialylated asialotransferrin molecules in the circulation. It is concluded that each asialotransferrin molecule probably undergoes several passages through the hepatocyte before its glycans become modified.


2021 ◽  
Vol 5 (3) ◽  
pp. 575-584
Author(s):  
Omar Mardenli ◽  
Hadi Awad Hassooni ◽  
Mahdi Saleh Mohammad Al-Kerwi

In the current study, the hypothesis of the effects of luteinizing hormone (LH) and follicular fluid (FF) derived from follicles of varying size on in vitro embryo production of the Shami goat breed were tested. The caprine follicular fluid (cFF) was obtained from healthy female’s ovaries by aspiration method and classified into two main classes (follicles with a diameter of ≤ 2mm and ≥3mm). The resulting cFF was added to the culture medium TCM-199 through six Treatments (A, B and C with a source of follicle size of ≤ 2mm; D, E and F with a source of size of ≥3mm). LH was added only to four of the previous Treatments with the levels of 50 µg ml-1 (B and E) and100 µg ml-1 (C and F). Results of the study showed that the oocytes incubated in Treatment F achieved a clear superiority (p=0.001) in the rates of maturation (87.0%), fertilization (80.0%) and cleavage (82.3%). The oocytes incubated in the same Treatment (F) continued to outperform (p= 0.006) by achieving the best rates across cleavage stages at 2-16 cell (16%; the lower value of arrest) and blastocyst (42%). Significant differences (P=0.03) were observed among the rates of Type 1embryos (the highest rate: 45.3%; Treatment F) and Type 3 embryos (the highest rate: 45.1%; Treatment A). No significant differences were observed in the rates of morula and Type 2 embryos. It is advised to add 15% of the cFF derived from follicles with a diameter of ≥3mm and 100 µg of LH ml-1 in the maturation media to obtain higher rates of maturation and cleavage of goat oocytes.  


HortScience ◽  
2006 ◽  
Vol 41 (1) ◽  
pp. 193-201 ◽  
Author(s):  
Ana Carolina Espinosa ◽  
Paula M. Pijut ◽  
Charles H. Michler

A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 μm 6-benzylaminopurine (BA), 0.49 μm indole-3-butyric acid (IBA), and 0.29 μm gibberellic acid (GA3). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 μm thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 μm naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 μm BA in combination with 0, 0.54, 1.07, or 5.37 μm NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 μm TDZ plus 0.54 μm NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 μm TDZ plus 1.07 μm NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.2) was obtained with 2.5 μm IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 μm IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage.


2021 ◽  
Vol 51 ◽  
Author(s):  
Tecla dos Santos Silva ◽  
Rosembrando Sosthenes Leite Carvalho Filho ◽  
Priscila Tavares Fonseca ◽  
José Raniere Ferreira de Santana

ABSTRACT Myracrodruon urundeuva Fr. All. is a tree threatened with extinction, which has wood and medicinal potential. This study aimed to analyze the in vitro shoot regeneration in M. urundeuva, in order to increase the species multiplication. Two experiments were conducted: 1) concentrations of 6-benzylaminopurine (BAP) (0.0, 2.0, 4.0, 8.0 and 16.0 µM), in association with naphthaleneacetic acid (NAA) (0.0, 1.5 and 3.0 µM), in explants (cotyledon, hypocotyl and cotyledonary node); 2) concentrations of meta-topolin (mT) (0.0, 2.0, 4.0, 8.0, 16.0 and 32.0 µM) in explants (biaxillary, medial uniaxillary and apical basal nodal segment). The percentage of explants responsive to shoot regeneration, percentage of callus explants, number of shoots and shoot length were evaluated. In the first experiment, the shoot regeneration occurred only in explants of the cotyledonary node and hypocotyl type, with the highest responsiveness percentage (76.67 %) and number of shoots (1.97 and 1.63) obtained for the cotyledonary node in the presence of 3.0 µM of NAA in association with 2.0 (1.97 shoots/explant) and 4.0 µM (1.63 shoots/explant) of mT. In the second experiment, the resolution of the obtained quadratic equation indicates that the use of basal explant with 24.59 µM of mT added to the culture medium leads to the highest number of shoots (1.86). However, despite the mT having increased the mean number of shoots, all treatments containing this cytokinin showed callus formation. As a conclusion, it is possible to regenerate shoots in M. urundeuva from the cotyledonary node using BAP in association with NAA.


2010 ◽  
Vol 59 (5) ◽  
pp. 511-520 ◽  
Author(s):  
Thomas S. Murray ◽  
Michel Ledizet ◽  
Barbara I. Kazmierczak

Pseudomonas aeruginosa is an opportunistic Gram-negative pathogen capable of acutely infecting or persistently colonizing susceptible hosts. P. aeruginosa colonizes surfaces in vitro by either biofilm formation or swarming motility. The choice of behaviour is influenced by the physical properties of the surface and specific nutrient availability, and subject to regulatory networks that also govern type 2 and type 3 protein secretion. Biofilm formation by clinical isolates has been well-studied. However, the swarming behaviour of human isolates has not been extensively analysed. We collected isolates from 237 hospitalized patients without cystic fibrosis and analysed motility and secretion phenotypes of each isolate. We found biofilm formation and swarming to be negatively associated, while swarming was positively associated with the secretion of both proteases and type 3 exoenzymes. Most isolates were capable of type 3 secretion and biofilm formation, even though these traits are considered to favour distinct modes of pathogenesis. Our data demonstrate that while clinical isolates display diverse motility, biofilm and secretion phenotypes, many of the predicted relationships between swarming motility and other phenotypes observed in laboratory strains also hold true for bacteria isolated from human patients.


2016 ◽  
Vol 24 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Jelili Opabode ◽  
Oluyemisi Akinyemiju

Abstract An efficient and reproducible micropropagation protocol of Solanecio biafrae (Oliv. & Hiern) C. Jeffrey has been developed from nodal stem segments. Shoot development was obtained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) alone and in combination with zeatin and 1-naphthaleneacetic acid (NAA). Elongated shoots were rooted in the presence of zeatin or 3-indole-butyric acid (IBA) alone or in combinations. The highest number of explants forming shoots (100%) as well as the highest number of shoots per explant (3.4) and the longest shoots (22 mm) were recorded on medium containing 4.0 mg·dm−3 BAP, 2.0 mg·dm−3 NAA, and 1.0 mg·dm−3 zeatin. About 76% of shoots formed roots on half-strength MS medium free of plant growth regulators. The best root formation (approximately 88%) was recorded on the medium containing 1.0-1.5 mg·dm−3 IBA. The micropropagated shoots with well-developed roots were efficiently acclimatized under greenhouse conditions. The random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) amplification products were monomorphic in micropropagated plants and similar to those of mother plant showing their genetic uniformity. This is the first report of micropropagation of S. biafrae, which will facilitate in vitro mass propagation, conservation, and germplasm exchange of this endangered African vegetable.


2019 ◽  
Vol 41 (2) ◽  
Author(s):  
Leonardo Soriano ◽  
Eveline Carla da Rocha Tavano ◽  
Marcelo Favaretto Correa ◽  
Ricardo Harakava ◽  
Beatriz Madalena Januzzi Mendes ◽  
...  

Abstract The in vitro organogenesis of Fremont (Citrus clementina x ), Citrus reticulataThomas (Citrus reticulata), and Nules (Citrus clementina) mandarins was evaluated aiming to optimize a regeneration protocol that could be applied in genetic transformation. The use of epicotyl-derived explants resulted in higher explant responsiveness and number of shoots developed per explant when compared with the use of internodal-derived explants. The highest efficiency in shoot regeneration was observed in the presence of 1 mg L-1 of BAP, regardless of the explant type and cultivar. The in vitro organogenesis protocol produced transgenic plants from three mandarin cultivars expressing attA gene under the control of phloem-specific promoters.


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