In vitro regeneration from leaf-base segments in three genotypes of Urochloa spp.

2018 ◽  
Vol 69 (5) ◽  
pp. 527 ◽  
Author(s):  
Diliane Harumi Yaguinuma ◽  
Luciana Midori Takamori ◽  
Adriana Mendonça de Oliveira ◽  
Luiz Gonzaga Esteves Vieira ◽  
Alessandra Ferreira Ribas
Keyword(s):  
In Vitro Regeneration ◽  
Forage Grasses ◽  
Leaf Base ◽  
Breeding Programs ◽  
Seed Formation ◽  
Regenerated Plants ◽  
Tropical Forage ◽  
Half Strength ◽  
The Media

The key agricultural species of Urochloa P.Beauv. (signal grass), important as tropical forage grasses, are characterised by asexual seed formation (apomixis), and this presents a challenge for breeding programs. Biotechnological approaches could be an option to develop improved cultivars. We evaluated the regenerative potential from three commercial genotypes, U. brizantha cv. Marandu, U. decumbens cv. Basilisk and U. ruziziensis cv. Ruziziensis, by using leaf-base segments as explants. We tested two auxins (2,4-D and picloram) and one cytokinin (TDZ) at four concentrations (1, 2, 3 and 4 mg L–1). Seeds were scarified, peeled and disinfected before inoculation on half-strength MS media in the dark for 14 days. Leaf-base explants were sectioned in thin slices and inoculated into the media. We analysed the number of primary calluses, number of calluses with shoots clusters and the average of regenerated plants. The lowest concentration of auxins tested (1 mg L–1) yielded the highest number of regenerated plants for Marandú and Basilisk, whereas the optimum for Ruziziensis was 2 mg L–1. Medium with higher concentrations of TDZ (4 mg L–1) was required to produce high frequency of plants for all genotypes. Explants cultured on media with TDZ produced very few calluses. These results indicate that the auxins and cytokinin tested can induce plant regeneration from Urochloa leaf-base segments, and may be used to produce transgenic plants in genetic transformation studies.

scholarly journals In Vitro Regeneration of Phyllanthus amarus Schum. and Thonn.: An Important Medicinal Plant

Our Nature ◽  
1970 ◽  
Vol 7 (1) ◽  
pp. 110-115 ◽  
Author(s):  
A. Sen ◽  
M.M. Sharma ◽  
D. Grover ◽  
A. Batra
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Phyllanthus Amarus ◽  
Regenerated Plants ◽  
Half Strength ◽  
Important Medicinal Plant

An efficient in vitro plant regeneration protocol was developed for the medicinally potent plant species Phyllanthus amarus Schum. and Thonn. (Euphorbiaceae) using nodal segment as explant. Maximum multiplication of shoots (15.275±0.96) was achieved on Murashige and Skoog’s medium supplemented with BAP (0.5 mg/l) after 3-4 weeks of inoculation. The shoots were separated from cluster and subcultured for their elongation on the same medium. In vitro flowering was also observed on the elongated shoots after 3–4 weeks of sub culturing on the shoot elongation medium. In vitro rooting was obtained on half strength MS medium supplemented with IBA (0.5 mg/l).  Regenerated plants were successfully hardened and acclimatized, 80 % of plantlets survived well under natural conditions after transplantation.Key words: In vitro regeneration, multiple shoots, nodal segments, Phyllanthus amarusDOI: 10.3126/on.v7i1.2557Our Nature (2009) 7:110-115

scholarly journals In vitro: Influence of Various Concentrations of Plant Growth Regulators (BAP & NAA) and Sucrose on Regeneration of Chenopodium quinoa Willd. Plant

2020 ◽  
pp. 34-43
Author(s):  
Fayza R. Al Gethami ◽  
Hameda El Sayed Ahmed El Sayed
Keyword(s):  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
In Vitro Regeneration ◽  
Chenopodium Quinoa ◽  
Naphthalene Acetic Acid ◽  
Cotyledonary Nodes ◽  
Half Strength ◽  
The Media

In vitro: regeneration of Chenopodium quinoa Willd. was achieved from cotyledonary nodes explants. In this study, used 6-Benzylaminopurine (BAP) and α-Naphthalene Acetic Acid (NAA) of plant growth regulators with different concentrations individually as well as in combination and used different concentrations of sugar (sucrose) with different concentrations. For was rooting, used half strength (½MS), full-strength MS and ½ MS supplemented with 0.2 mg/l of NAA. The results mentioned, explant responding (%) to multiplication was about 73% for all BAP treatments compared with control and average numbers of shoot increased with increased BAP concentration except 5 mg/l of BAP. The highest explant responding (%) was in media supplemented BAP without NAA compared other treatments noted that the media with combination of BAP and NAA gives formation of callus in bases of the plantlets. Also, the result inducted the combinations between (BAP–NAA) was highly significantly (P≤ 0.001) and less effective on number of shoots where the highest number of shoot was 3.40 in media with 3 mg/l BAP compared other treatments. The highest of explant responding 93.33% was in media supplement with 10 g/l sucrose and (10 g/l sucrose + 3 mg/l BAP), but sucrose level for good greening and developed shoots (4 shoots) was in medium supplement with 10 g/l sucrose. The shoots rooted well on half-strength MS medium with 60% percentage of root. The rooted shoots were acclimatized and transferred to green house to follow their development.

EFFECTS OF DIFFERENT PHYSIOCHEMICAL FACTORS ON REGENERATION OF CYMBOPOGON POLYNEUROS STAPF VIA CALLUS CULTURE AND SUBSEQUENT CHROMOSOMAL STABILITY

1999 ◽  
Vol 47 (3) ◽  
pp. 195-198
Author(s):  
Anath Bandhu Das
Keyword(s):  
Callus Culture ◽  
High Frequency ◽  
Potassium Salt ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Naphthalene Acetic Acid ◽  
Leaf Base ◽  
Regenerated Plants ◽  
The Individual

In vitro regeneration of Cymbopogon polyneuros Stapf was obtained through callus culture using leaf base, node, and root as explants. Callus was induced from different explants with 2–5 mg/1 α-naphthalene acetic acid (NAA) and 1–2 mg/1 kinetin in Murashige and Skoog's (MS) basal medium. High frequency shoots were noticed from leaf-base callus supplemented with 3.5 mg/1 6-benzylaminopurine (BA), L-arginine, adenine, and a low level of NAA (0.2 mg/1). About 80–85 shoot buds were obtained from ca. 200 mg of callus per culture. The individual shoots produced root in the presence of 0.5–3 mg/1 indole 3-butyric acid or its potassium salt. Regenerated plants were cytologically and phenotypically stable. Regenerants were transplanted into soil and subsequently transferred to the field.

scholarly journals Plant Regeneration in Buckwheat (Fagopyrum esculentum Moench.) via Somatic Embryogenesis and Induction of Meristemoids in Abnormal Embryos

2019 ◽  
Vol 29 (1) ◽  
pp. 33-47 ◽  
Author(s):  
Ribha Saraswat ◽  
Mithilesh Kumar
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Hypocotyl Explants ◽  
Regenerated Plants ◽  
Half Strength ◽  
Fagopyrum Esculentum Moench ◽  
Full Utilization

An efficient in vitro regeneration protocol is reported for common buckwheat. A combination of 0.5 mg/l 2,4-D and 0.2 mg/l BAP with sucrose showed highest induction of somatic embryogenesis from cotyledon and hypocotyl explants. More than 35% of normal somatic embryos matured on MS. MS with 2% sucrose were found best for germination and conversion of somatic embryos to plantlets. In tissue culture, abnormal somatic embryos usually occur. In this report, abnormal embryos are also used to induce shoot organogenesis, adding to the number of final regenerants and ensuring full utilization of regenerative propagules. A treatment of 0.2 mg/l BAP induced meristemoids in 60% of underdeveloped embryos and a combination of 0.5 mg/l BAP and 0.5 mg/l AgNO3 led browning and senescence-free progression of shoot buds to well developed shoots, which were subsequently rooted in half strength MS containing 2% sucrose and 0.25 mg/l IBA. The regenerated plants survived acclimatization, flowered and set seeds. Plant Tissue Cult. & Biotech. 29(1): 33-47, 2019 (June)

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

scholarly journals Indirect in vitro Regeneration of Viola canescens Wall. ex, Roxb. by using Leaf Calli

2018 ◽  
Vol 28 (2) ◽  
pp. 215-222 ◽  
Author(s):  
Arun Kumar Khajuria ◽  
NS Bisht
Keyword(s):  
Plant Regeneration ◽  
Natural Population ◽  
Plant Tissue ◽  
In Vitro Regeneration ◽  
Soil Condition ◽  
Medicinal Herb ◽  
Growth Chamber ◽  
Half Strength ◽  
Number Of Shoots

An efficient indirect plant regeneration protocol was developed for Viola canescens, an important medicinal herb used in broad spectra of diseases in number of folk medicines since aeon. Excessive use of this plant without any rehabilitating measure has led to decline its natural population. Present investigation reports the use of zeatin to regenerate the plant from the callus on MS following its acclimatization on the soil condition. Calli of the plant responded positively to zeatin and maximum number of shoots 13.07 ± 2.01 were obtained when 9.12 μM concentration of zeatin was used. Regenerated shoots were subsequently rooted with IBA on MS and half strength MS and showed maximum number of roots 14.13 ± 1.64 after 60 days when medium was fortified with 4.92 μM IBA, followed by transferring them to soil condition, acclimatization of the plantlet was carried in growth chamber and then finally to the field for their survival where it showed 80% survival. Plant Tissue Cult. & Biotech. 28(2): 215-222, 2018 (December)

scholarly journals An efficient in vitro regeneration protocol to generate stable transgenic lines of Black gram (Vigna mungo)

2018 ◽  
Vol 78 (3) ◽  
Author(s):  
Bidyut Kumar Sarmah ◽  
Trishna Konwar ◽  
Borsha Borah ◽  
Arun Kumar Handique ◽  
Sumita Acharjee
Keyword(s):  
Transformation Efficiency ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Vigna Mungo ◽  
Ms Medium ◽  
Black Gram ◽  
Transgenic Lines ◽  
Half Strength ◽  
The Individual

An efficient and quick in vitro regeneration protocol was developed for black gram (Vigna mungo) using wounded embryonic axis with cotyledon as explant. Murashige and Skoog (MS) medium supplemented with 4.44 μM BAP and 2.32 μM Kinetin was found to be effective in producing maximum number (mean 7.80) of multiple shoots. The individual shoots elongated to 4.5 cm when MS medium was supplemented with 2.89 μM GA3 along with 0.44 μM BAP and 0.46 μM KIN. A novel in vitro rooting technique was also optimized for black gram using half-strength liquid MS medium supplemented with 1.34 μM NAA. The shoots in this medium produced the highest number (mean 7.50) of roots with root length of 6.02 cm. The plantlets were transferred to soil mixture and placed in greenhouse where more than 80% successfully grew to maturity. The same protocol was successfully used to generate transgenic black gram lines carrying Bt-Cry2Aa gene through Agrobacteriummediated transformation with a transformation efficiency of 0.42%. The rooted T0 plants grew to maturity and produced T1 seeds with the presence and expression of transgene in T1 plants. Thus, we have standardized an in vitro regeneration protocol suitable for generation of stable transgenic black gram plants.

scholarly journals Fibrous Carbohydrate Fractions and in Vitro True and Apparent Digestibility of 10 Tropical Forage Grasses

1969 ◽  
Vol 58 (3) ◽  
pp. 293-304
Author(s):  
J. Coward-Lord ◽  
J. A. Arroyo-Aguilú ◽  
O. García-Molinari
Keyword(s):  
Nutritive Value ◽  
Panicum Maximum ◽  
Apparent Digestibility ◽  
Forage Grasses ◽  
Mean Values ◽  
Brachiaria Ruziziensis ◽  
Tropical Forage ◽  
The Difference ◽  
Tropical Climates

Ten tropical forage grasses (Pangola, Digitaria decumbens; Congo, Brachiaria ruziziensis; Signal, Brachiaria brizantha; Buffel, Cenchrus ciliaris; Guinea, Panicum maximum; Jaragua, Hyparrhenia rufa; Giant Pangola, Digitaria valida; African Crab, Digitaria swazilandensis; Venezuelan Elephant, Pennisetum setosum; and Limpo, Hemarthria altissima) lightly fertilized with NH4NO3 at the rate of 350 kg/ha, were harvested by hand (machete) every 30 days up to 180 days of age, beginning August 20, 1970, at the College of Agricultural Sciences grass collection, University of Puerto Rico at Mayagüez. They were evaluated chemically for cell-wall constituents or neutraldetergent fiber (NDF), acid-detergent fiber (ADF), permanganate lignin (L), cellulose (C) and silica (Si), as outlined by Goering and Van Soest and for in vitro true digestibility (IVTD), utilizing the Goering and Van Soest modification of the Tilley and Terry procedure. Hemicellulose (H) was determined as the difference between NDF and ADF. Statistical analyses of variance were conducted. Highly significant differences in NDF, ADF, H, L, L/ADF, C, and Si contents were observed between species. IVTD and in vitro apparent digestibility (IVAD) also revealed highly significant differences between species. Guinea and Buffel grasses exhibited the highest mean values in NDF and ADF contents but the lowest in IVTD and IVAD, suggesting that, as total fiber and lignocellulose contents increase, digestibility decreases. Similar trends were observed in other species. Highest mean values were obtained in Guinea grass for C, in Jaragua grass for Si and in Limpo grass for L and H. As the grasses advanced in maturity from 30 to 180 days, NDF, ADF, L, L/ADF, C and Si revealed highly significant increases while IVTD and IVAD revealed highly significant decreases with HC remaining almost constant throughout. The largest increase in NDF, ADF, L and C contents and the largest decrease in IVTD and IVAD evaluations occurred between the 30- and 60-day intervals, probably due to the high environmental temperature and high transpiration causing early onset of lignification and silicification in tropical climates. Pangola grass underwent little change in nutritive value as compared to other species. Evidence is presented that fibrous fractions and IVTD evaluations of all species compared favorably among themselves and that all fibrous fractions (except H) and IVTD differed significantly with plant age.

scholarly journals In vitro regeneration of two high-yielding eggplant (Solanum melongena L.) varieties of Bangladesh

1970 ◽  
pp. 08-12
Author(s):  
Sabina Yesmin, Mst Muslima Khatun, Tanzena Tanny ◽  
Anica Tasnim Protity ◽  
Md Salimullah ◽  
Iftekhar Alam
Keyword(s):  
In Vitro Regeneration ◽  
Solanum Melongena ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Healthy Plant ◽  
Ms Medium ◽  
Best Response ◽  
Half Strength ◽  
Multiple Shoot Regeneration

An in vitro regeneration protocol was developed for two high-yielding eggplant varieties (Solanum melongena L.) namely BARI begun-4 and BARI begun-6. Multiple shoots were regenerated from cotyledonary explants through organogenesis with growth regulators of different combinations and concentrations.  The best response towards multiple shoot regeneration was achieved from cotyledon explants on MS media complemented with 1 mg/l BAP + 0.2 mg/l IAA in both the two varieties of eggplant. Elongation of shoots was achieved on hormone free MS medium. Regenerated shoots of both the varieties produced   active in vitro root system on half strength of MS medium supplemented with 0.2 mg/l IBA.  The in vitro grown plantlets were acclimatized in soil, grew up to maturity, flowered, fruited and produced seeds as normal healthy plant like the control.

scholarly journals A Semi-Automated SNP-Based Approach for Contaminant Identification in Biparental Polyploid Populations of Tropical Forage Grasses

2021 ◽  
Vol 12 ◽  
Author(s):  
Felipe Bitencourt Martins ◽  
Aline Costa Lima Moraes ◽  
Alexandre Hild Aono ◽  
Rebecca Caroline Ulbricht Ferreira ◽  
Lucimara Chiari ◽  
...  
Keyword(s):  
Principal Component ◽  
Genotyping By Sequencing ◽  
Snp Markers ◽  
Correct Identification ◽  
Forage Grasses ◽  
Mendelian Segregation ◽  
Breeding Programs ◽  
Tropical Forage ◽  
Genotypic Analysis ◽  
Shiny App

Artificial hybridization plays a fundamental role in plant breeding programs since it generates new genotypic combinations that can result in desirable phenotypes. Depending on the species and mode of reproduction, controlled crosses may be challenging, and contaminating individuals can be introduced accidentally. In this context, the identification of such contaminants is important to avoid compromising further selection cycles, as well as genetic and genomic studies. The main objective of this work was to propose an automated multivariate methodology for the detection and classification of putative contaminants, including apomictic clones (ACs), self-fertilized individuals, half-siblings (HSs), and full contaminants (FCs), in biparental polyploid progenies of tropical forage grasses. We established a pipeline to identify contaminants in genotyping-by-sequencing (GBS) data encoded as allele dosages of single nucleotide polymorphism (SNP) markers by integrating principal component analysis (PCA), genotypic analysis (GA) measures based on Mendelian segregation, and clustering analysis (CA). The combination of these methods allowed for the correct identification of all contaminants in all simulated progenies and the detection of putative contaminants in three real progenies of tropical forage grasses, providing an easy and promising methodology for the identification of contaminants in biparental progenies of tetraploid and hexaploid species. The proposed pipeline was made available through the polyCID Shiny app and can be easily coupled with traditional genetic approaches, such as linkage map construction, thereby increasing the efficiency of breeding programs.

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