scholarly journals In vitro induction and identification of polyploid Neolamarckia cadamba plants by colchicine treatment

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12399
Author(s):  
Wee Hiang Eng ◽  
Wei Seng Ho ◽  
Kwong Hung Ling
Keyword(s):  
Crop Improvement ◽  
Colchicine Treatment ◽  
Chromosome Count ◽  
Nodal Segments ◽  
Tetraploid Cell ◽  
Breeding Programs ◽  
Spad Value ◽  
Stomata Size ◽  
In Vitro Induction

Polyploidization has played a crucial role in plant breeding and crop improvement. However, studies on the polyploidization of tropical tree species are still very scarce in this region. This paper described the in vitro induction and identification of polyploid plants of Neolamarckia cadamba by colchicine treatment. N. cadamba belongs to the Rubiaceae family is a natural tetraploid plant with 44 chromosomes (2n = 4x = 44). Nodal segments were treated with colchicine (0.1%, 0.3% and 0.5%) for 24 h and 48 h before transferring to shoot regeneration medium. Flow cytometry (FCM) and chromosome count were employed to determine the ploidy level and chromosome number of the regenerants, respectively. Of 180 colchicine-treated nodal segments, 39, 14 and 22 were tetraploids, mixoploids and octoploids, respectively. The highest percentage of polyploidization (20% octoploids; 6.7% mixoploids) was observed after treated with 0.3% colchicine for 48 h. The DNA content of tetraploid (4C) and octoploid (8C) was 2.59 ± 0.09 pg and 5.35 ± 0.24 pg, respectively. Mixoploid plants are made up of mixed tetraploid and octoploid cells. Chromosome count confirmed that tetraploid cell has 44 chromosomes and colchicine-induced octoploid cell has 88 chromosomes. Both octoploids and mixoploids grew slower than tetraploids under in vitro conditions. Morphological characterizations showed that mixoploid and octoploid leaves had thicker leaf blades, thicker midrib, bigger stomata size, lower stomata density, higher SPAD value and smaller pith layer than tetraploids. This indicates that polyploidization has changed and resulted in traits that are predicted to increase photosynthetic capacity of N. cadamba. These novel polyploid plants could be valuable resources for advanced N. cadamba breeding programs to produce improved clones for planted forest development.

scholarly journals Types and concentrations of cytokinins in the micropropagation of Anthurium maricense

2018 ◽  
Vol 12 (2) ◽  
pp. 117
Author(s):  
Cecília Moreira Serafim ◽  
Arlene Santisteban Campos ◽  
Priscila Bezerra Dos Santos Melo ◽  
Ana Cecília Ribeiro de Castro ◽  
Ana Cristina Portugal Pinto de Carvalho
Keyword(s):  
Light Intensity ◽  
Culture Medium ◽  
Culture Media ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Viable Option ◽  
Growth Room ◽  
In Vitro Induction

Faced with the demand for plants potted for their foliage, Anthurium maricense is seen as a viable option. However, most of the studies on obtaining micropropagated plantlets are for A. andraeanum, with nothing yet reported for A. maricense. The aim of this study therefore, was to evaluate the effect of four cytokinins in different concentrations, on the in vitro induction of shoots from nodal segments of A. maricense. The experimental design was completely randomised in a 4 x 4 factorial scheme, with four cytokinins (BAP, ZEA, CIN and 2iP) and 4 concentrations (0, 2.22, 4.44 and 6.66 μM), for a total of 16 treatments, with 6 replications of five test tubes, and using one nodal segment. Cultures were kept in a growth room at 25 ± 2°C, a photoperiod of 16 h and a light intensity of 30 μmolm-2 s-1 for 60 days. After this period, the number of shoots formed per node was evaluated. The addition of a cytokinin to the culture medium was determinant for the in vitro regeneration of shoots in A. maricense. The greatest estimated number of shoot formations in A. maricense were obtained in the culture media containing ZEA (3.87) and BAP (3.30), both at concentration of 6.66 μM.

In vitro induction and identification of autotetraploid of Bletilla striata (Thunb.) Reichb.f. by colchicine treatment

2017 ◽  
Vol 132 (3) ◽  
pp. 425-432 ◽  
Author(s):  
Han Pan-pan ◽  
Liu Wei-xu ◽  
Liang Hui-hui ◽  
Xiang zeng-xu
Keyword(s):  
Colchicine Treatment ◽  
Bletilla Striata ◽  
In Vitro Induction

In vitro induction of polyploidy by colchicine treatment of shoots and preliminary characterisation of induced polyploids in two Miscanthus species

2010 ◽  
Vol 32 (2) ◽  
pp. 88-96 ◽  
Author(s):  
K. Głowacka ◽  
S. Jeżowski ◽  
Z. Kaczmarek
Keyword(s):  
Colchicine Treatment ◽  
In Vitro Induction

scholarly journals A new and rapid micropropagation protocol for Eucalyptus grandis Hill ex Maiden

2020 ◽  
Vol 29 (1) ◽  
pp. eSC04
Author(s):  
Anabel-Viviana Di-Gaudio ◽  
Esteban Tubert ◽  
Leandro-Ezequiel Laino ◽  
Jose-María Chaín ◽  
Sandra-Irene Pitta-Alvarez ◽  
...  
Keyword(s):  
Woody Plants ◽  
Eucalyptus Grandis ◽  
Nodal Segments ◽  
Ex Vitro ◽  
Breeding Programs ◽  
Hydroponic System ◽  
Rate Of Success ◽  
Production Dynamics ◽  
Intermediate Rate

Aim of the study: We developed a faster micropropagation protocol specifically designed for Eucalyptus grandis. Eucalyptus breeding programs use micropropagation protocols to obtain high quality cloned seedlings, but current protocols are excessively time consuming.Area of the study: The protocol has been developed in Argentina, but it can be applied in anywhere.Materials and methods: We used nodal segments as initial explants to obtain micropropagated shoots, which were then simultaneously rooted ex vitro and acclimated in a hydroponic system. Nodal segments were cultured in a MS medium supplemented with 1 mg l-1 6-benzylaminopurine, 30 g l-1 sucrose, 1 g l-1 active charcoal and 8 g l-1 agar and incubated for four weeks at 25 ± 2°C under 16 h day photoperiod. Then, micropropagated shoots were exposed 15 seconds to 5000 ppm indol-butyric acid prior to being transferred to a hydroponic system, allowing simultaneous ex vitro rooting and acclimatization.Main results: 73 ± 9% of nodal segments grew to generate 1.73 ± 1.03 shoots per explant (length: 0.76 ± 0.44 cm). After four weeks in hydroponic system, 46 ± 4 % of micropropagated shoots developed roots, which represents an acceptable and intermediate rate of success, compared to the reported in vitro rooting rates.Research highlights: Our protocol allowed to obtain micropropagated seedlings in a total timespan of 8 weeks. Our results show that, by utilizing a hydroponic system, traditional protocols to micropropagate Eucalyptus can be substantially enhanced, allowing for improved production dynamics and potentially resulting in better organized seedling manufacturing facilities.Keywords: Woody plants; silviculture; nursery seedlings; rooting methods; hydroponics; acclimatization.

scholarly journals In vitro plant propagation and crop improvement in Lisianthus (Lisianthus Russelianus Hook.)

2016 ◽  
Vol 73 (2) ◽  
pp. 168
Author(s):  
Rodica Pop ◽  
Maria Cantor ◽  
Erzsebet Buta ◽  
Iudita Csete
Keyword(s):  
Crop Improvement ◽  
Ornamental Plants ◽  
Basal Medium ◽  
International Market ◽  
Nodal Segments ◽  
F1 Hybrids ◽  
Vase Life ◽  
Eustoma Grandiflorum ◽  
Planting Material

Romania assists at the present time to an increase of production crops for ornamental plants and as a consequence an increased demand of planting material. Thus, improvements of the current multiplication methods are sought after. Lisianthus russelianus Hook. (Eustoma grandiflorum Grise.) is a relatively new floral crop to the international market, known for beautiful flowers of various colors and for having a long vase life. This study focused on the development of an efficient protocol for rapid regeneration of this species following known basic and applied aspects of lisianthus biotechnology but exploring new potentials. In the course of experiments conducted, for in vitro multiplication there were used nodal segments (1.5 cm) with axillary buds from three F1 hybrids ‘Echo Lavender’, ‘Flamenco White’, ‘Mirage Pastel Pink’ that were inoculated on MS basal medium supplemented with 0.50 mg 1-1 TDZ, 1.0 mg 1-1 BAP and 0.50 mg 1-1 AIA. The results show that the medium with BAP was most effective for obtaining the highest shoots number compared to medium containing TDZ. For rooting induction, two different concentrations of auxin IBA 0.5 mg 1-1 and 1.5 mg 1-1 were used simultaneously on MS basal medium. The highest roots number occurred when using 1.5 mg 1-1 IBA. Both the number of shoots and rooting regeneration were dependent on the cultivar. The highest shoots number was achieved for ’Mirage Pastel Pink’ hybrid (6.91) on the medium containing 1.0 mg 1-1 BAP and 0.50 mg 1-1 IAA.

scholarly journals In vitro Induction of Banana Autotetraploids by Colchicine Treatment of Micropropagated Diploids

1992 ◽  
Vol 40 (6) ◽  
pp. 887 ◽  
Author(s):  
SD Hamill ◽  
MK Smith ◽  
WA Dodd
Keyword(s):  
In Vitro Selection ◽  
Morphological Characteristics ◽  
Colchicine Treatment ◽  
Shoot Tips ◽  
Cell Permeability ◽  
Root Tip ◽  
Optimum Treatment ◽  
Morphological Criteria ◽  
In Vitro Induction

Alternative breeding strategies, based on colchicine-induced autotetraploids, have been proposed as a means of introducing disease resistance into banana breeding programs. This paper describes techniques for the in vitro induction of banana autotetraploids by the use of colchicine on cultured explants. The technique can be readily applied and large numbers of autotetraploids produced. The optimum treatment involved immersing shoot tips in a 0-5% w/v colchicine solution for 2 h under aseptic conditions. Dimethyl sulfoxide (DMSO) was applied with the colchicine treatments to increase cell permeability and so absorption of colchicine, resulting in the optimum treatment unchanged at 0-5% colchicine, but including the addition of 2% v/v DMSO. Of the shoot tips treated over 30% were induced to the autotetraploid level. Methods for in vitro selection of induced tetraploids from treated diploid plantlets were also developed. Tetraploid plants were more robust with thicker pseudostems, roots and broader leaves than diploids and they could be selected on these morphological characteristics. Mean stomatal lengths of diploid banana plants growing in vitro were significantly smaller (16-0 mum) than the tetraploids (26.9 mum) and were used as a more reliable indicator of ploidy than morphological criteria alone. A root tip squash technique using carbol fuchsin was developed for positive confirmation of ploidy change by chromosome counts- Although chimerism and reversion to the diploid form occurred, it was not considered a problem because of the large number of autotetraploids induced. Stable autotetraploids were recovered and established in the field and were characterised by their large, drooping leaves and thick pseudostems. They have retained these characteristics for more than 3 years in the field.

In vitro induction of polyploidy by colchicine treatment in herbaceous peony (Paeonia lactiflora Pall.)

2017 ◽  
pp. 265-278 ◽  
Author(s):  
D.X. Wei ◽  
Z.J. Tang ◽  
J.A. Teixeira da Silva ◽  
X.N. Yu
Keyword(s):  
Colchicine Treatment ◽  
Paeonia Lactiflora ◽  
Herbaceous Peony ◽  
In Vitro Induction

scholarly journals Effect of different culture tube caps and concentrations of activated charcoal and sucrose on in vitro growth and budding induction of Annona glabra L.

2011 ◽  
Vol 35 (5) ◽  
pp. 916-923 ◽  
Author(s):  
José Raniere Ferreira de Santana ◽  
Renato Paiva ◽  
Ana Valéria de Souza ◽  
Lenaldo Muniz de Oliveira
Keyword(s):  
Activated Charcoal ◽  
Dry Weight ◽  
Test Tube ◽  
Leaf Abscission ◽  
Nodal Segments ◽  
Bud Induction ◽  
Pvc Film ◽  
Annona Glabra ◽  
In Vitro Induction

The present work evaluated the effects of different types of culture flask seals and varying concentrations of sucrose and activated charcoal on the in vitro induction and growth of buds of Annona glabra L.; an edible fruit-producing species popularly known as "araticum". Nodal segments obtained from A. glabra plants maintained in green houses were surface sterilized and inoculated into a WPM culture medium solidified with 7 g L-1 of agar and supplemented with sucrose (0.00; 29.21; 58.63 and 116.84 mM), activated charcoal (0.0 and 2.0 g L-1), and 250 mg L-1 benomyl. In addition to the varying concentrations of sucrose and activated charcoal, we evaluated the efficiency of two types of test tube seals: PVC film, and cotton plugs. All possible combinations of caps and nutrient media were tested with 4 repetitions with 5 tubes each, evaluating the number of buds, the percentage of explant responses, the number of expanded leaves per bud, the length of the largest leaves, leaf abscission, and the length and dry weight of the buds. The type of seal influenced organogenesis in nodal segments of A. glabra, and no bud induction was observed in the absence of sucrose. The largest number of expanded leaves were obtained when 58.42 mM of sucrose was used in tubes sealed with cotton plugs, and leaf abscission was halved in the presence of activated charcoal. The greatest bud length and dry weight were obtained in tubes sealed with cotton plugs and in the presence of activated charcoal.

scholarly journals In vitro Induction and Characterization of Tetraploid Lychnis senno Siebold et Zucc.

HortScience ◽  
2006 ◽  
Vol 41 (3) ◽  
pp. 759-761 ◽  
Author(s):  
Li-ping Chen ◽  
Yan-ju Wang ◽  
Man Zhao
Keyword(s):  
Diploid Number ◽  
Leaf Size ◽  
Morphological Characterization ◽  
Nodal Segments ◽  
Flow Cytometry Analysis ◽  
Stem Height ◽  
Solid Media ◽  
In Vitro Induction

In this study, in vitro induction of tetraploid Lychnis senno Siebold et Zucc. and its cytological and morphological characterization were conducted. For polyploid induction, nodal segments with axillary buds from in vitro grown plants were kept for 3 days in MS (Murashige and Skoog, 1962) liquid or solid media added with a series of concentrations of colchicine. Out of total 588 recovered plants, 15 tetraploids and 6 mixoploids determined by flow cytometry analysis were obtained. The tetraploid contained 48 chromosomes, twice the normal diploid number of 24, as observed under light microscope. The tetraploid plants exhibited much larger but less stomata than diploid plants. Moreover, significant differences in stem height and leaf size between the diploid and tetraploid plants were noted. The tetraploid plants were more compact than diploids.

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