scholarly journals Histology of Microspores of Chile Apple Capsicum pubescens R and P. and in vitro Culture

2021 ◽  
Vol 2 (7) ◽  
pp. 01-06
Author(s):  
J. L. Rodríguez-de la O ◽  
F. Pérez-Pérez ◽  
M. Pérez-Grajales

In plant biotechnology, in vitro culture of gametic or sexual cells, microspores or pollen grains, has been described as a successful tool to accelerate genetic improvement, obtaining haploid, homozygotic plants or pure lines in a short time. In chile apple, Capsicum pubescens R and P. Anthers were sown in vitro, and their cytological analysis, locating the meiotic division stage of microspores or pollen grains. Flower buds with diameters from 2.5 to 4.4 mm were pre-incubated at 4°C, in ascorbic and citric acid at 100 and 150 mg-L-1 for 24 h. Five semisolid culture media (A1, A2, A3, A4 and A5) were used, with Murashige and Skoog (1962) salts (MS), modifying iron and vitamin chelates, sucrose, and L-cysteine, 2,4-dichlorophenoxyacetic acid (2,4-D) and Kinetin (Kin). Anthers, in vitro, were plated, in light and dark, for 70 days. Two differentiation media (R1 and R2) were evaluated with 100% MS salts, glycine, kinetin and myo-inositol. The anthers seeded, coincided with the first mitosis of the microspore, the anthers, formed callus in the media (A1) 100 % EDTA-Fe, 0.40 mg-L-1 thiamine, 3 % sucrose) and (A3) 100 % EDTA-Fe, 0.40 mg-L-1 thiamine, 3 % sucrose, 0. 3 mg-L-1 of 2,4-D, and differentiated pro-embryonic structures in (A3) and (A5) 200 % EDTA-Fe, 0.4 mg-L-1 thiamine, 50 mg-L-1 pyridoxine, folic acid, riboflavin and niacin, 0.3 mg-L-1 2,4-D plus 0.3 mg-L-1 Kinetin, as well as roots in (A1). Light influenced the formation of pro-embryos and roots, in the dark callus. The media (R1) and (R2) favored the formation of pro-embryos.

2016 ◽  
Vol 6 (2) ◽  
pp. 91
Author(s):  
Yati Supriati

<p>Micropropagation Efficiency of Banana cv Kepok<br />Amorang through Modifications of Culture Media and<br />Incubation Temperature. Yati Supriati. The budless<br />banana cv Kepok Amorang is potentially commercialized<br />due to its sweet taste and does not have flower bud, hence<br />reduced the potential of being infected by the blood disease<br />pathogen. Enhancement of banana industry needs continuous<br />supplies of large number banana seedlings. In vitro<br />culture enable the production of seedlings in a large scale,<br />uniform, quick. The research aims: (1) to formulate an<br />efficient medium for in vitro multiplication of cv Kepok<br />Amorang shoot, (2) to identify efficient growth environment<br />for in vitro culture of cv Kepok Amorang, and (3) to formulate<br />an efficient culture medium for roots inductions of cv<br />Kepok Amorang. The plant material used was in vitro culture<br />of Kepok cv Amorang, 2 cm in height without leaf and root.<br />The media formulation for shoot multiplication were full<br />strength, half strength, one fourth strength MS media,<br />supplemented with either 1, 3, or 5 ppm IBA. On optimization<br />step, the media tested were MS, Knop, Knop and<br />Heller, Hyponex N, Growmore N, and Rosasol N containing<br />of 1 ppm BA. The explants were incubated in culture room<br />with 8, 12, and 16 hours photoperiod with temperatures 30oC<br />(non air conditioned) and 25oC (air conditioned). The root<br />induction trial was done using MS, Knop, Knop and Heller,<br />Hyponex N, Growmore N, and Rosasol N media containing<br />of 1 ppm and 3 ppm IBA. The results showed that the best<br />medium formula for shoot multiplication was ¼ MS + 1 ppm<br />IBA. The best incubation condition was 16 hours photoperiods<br />at 30oC. The best media for root induction was<br />Hyponex 2 g/l + 1 ppm IBA. This culture method reduced<br />cost by Rp 261.7 per plantlet through efficiency of media<br />formulation and electricity use.</p>


2016 ◽  
Vol 19 (2) ◽  
pp. 168
Author(s):  
Toni Herawan ◽  
Mohammad Na'iem ◽  
Sapto Indrioko ◽  
Ari Indrianto

Sandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood.


Genome ◽  
1988 ◽  
Vol 30 (4) ◽  
pp. 511-518 ◽  
Author(s):  
C. Doré ◽  
Y. Cauderon ◽  
M. C. Chueca

In vitro culture of immature inflorescences of F1 hybrid plants originating from the cross between the common wheat cultivar Roazon and two inbred lines of rye was carried out with 2,4-dichlorophenoxyacetic acid in the medium. After 3 or 8 weeks of culture as undifferentiated callus, 62 plants could be regenerated. Of these, 47 reached the adult stage; 46 of which were analysed for chromosome counts and chromosome pairing. For 44 of these, chromosome counts showed high stability and phenotypes were similar to those of the mother plants. In vitro culture of immature inflorescences could therefore be considered as a possible vegetative multiplication method to obtain numerous copies from one individual. The three other plants exhibited variations in phenotype and chromosome number: one was an amphiploid, and a chromosome was lost in each of the other two. All meristematic cells and pollen mother cells were involved in these numerical changes for each plant. The amphiploid plant (2n = 56) was nonchimeric and all the spikes were highly self-fertile. It could be an efficient method for chromosome doubling. This underscores the usefulness of inflorescence tissue culture for overcoming the sterility barrier in interspecific hybrids.Key words: wheat × rye hybrids, tissue culture, amphiploidization, aneuploidy, meiotic behaviour.


2019 ◽  
Vol 27 (2) ◽  
pp. 91-98
Author(s):  
Jaber Panahandeh ◽  
Nasrin Farhadi

AbstractHaploid induction using in vitro cultures of unpollinated flowers has been recognized as an important tool to produce homozygous plants for genetic studies and breeding programs. In this study the potential of gynogenic haploid induction in four ecotypes of Allium hirtifolium under different combinations of benzylaminopurine (BAP) with 2,4-dichlorophenoxyacetic acid (2,4-D), or α-naphthaleneacetic acid (NAA) was investigated. Unpollinated flower buds were excised from an umbel 5 to 3 days before anthesis, and cultured onto B5 medium containing 7.5% sucrose and 2 mg·dm−3 BAP with auxin. The experiments revealed that NAA increased the percentage of gynogenesis induction and number of gynogenic embryos per flower in all ecotypes. Somatic organogenesis from basal callus or other floral parts was most effective on the media containing 2,4-D. Plants obtained by gynogenesis were haploid in 70–77% and plants from somatic tissue were mostly diploid.


HortScience ◽  
1996 ◽  
Vol 31 (7) ◽  
pp. 1225-1228 ◽  
Author(s):  
Rida A. Shibli ◽  
M.A.L. Smith

Ohelo (V. pahalae Skottsb.) and bilberry (V. myrtillus L.) shoots were regenerated via direct organogenesis from whole leaves and leaf sections and also from hypocotyl explants of bilberry. Explants preincubated for 1 to 2 weeks in darkness yielded ≈75% regeneration frequencies and the highest number of regenerating shoots/explant on TDZ-supplemented media (0.9 to 2.7 μm). When 2iP or zeatin were substituted as the cytokinin source, frequencies of regeneration and shoot productivity were significantly lower. Explants held under constant illumination (no dark pretreatment) had significantly lower regeneration frequencies in all tested cytokinin-supplemented media. 2,4-D stimulated callus formation, but did not support regeneration from vegetative explants. Cells from callus and suspension cultures did not exhibit regeneration in any of the media that supported organogenesis from leaves. Regenerants were successfully micropropagated, although callus formation caused by zeatin and high 2iP levels interfered with shoot proliferation. Zeatin induced hyperhydricity in shoots from both species, but more severely in ohelo. Ex vitro rooting after treatment with 4.9 μm IBA or 5.4 μm NAA was 95% and 60% successful for bilberry and ohelo, respectively, and plants were readily acclimatized after an interval in a fog chamber. Bilberry microshoots also rooted in vitro in the absence of growth regulator treatment. Chemical names used: 1H-indole-3-butanoic acid (IBA); N-(3-methyl-2-butenyl)-1-H-purine-6-amine (2iP); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA); thidiazuron=1-phenyl-3-(1,2,3-thiadiazio-5-yl)urea (TDZ); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).


Author(s):  
RAVIKUMAR S ◽  
KRISHNA V ◽  
AJITH S

Objective: In the present study, a new method for the production of anticancerous compound podophyllotoxin (PTOX) was developed for Bridelia scandens Wild. by feeding coniferyl alcohol. Methods: The production of anticancerous compound PTOX through leaf explant derived calli of B. scandens. Murashige and Skoog (MS) medium fortified with 0.5 mg/l 6-Benzylaminopurine (BAP) and 0.5 mg/l 2,4-D (2,4-Dichlorophenoxyacetic acid) induced luxuriant mass of callus growth. Suspension culture was initiated by sterile MS media fortified with 0.1–1.0 mg/l BAP and 0.1–1.0 mg/l 2,4-D. and growth product was analyzed by the high-pressure liquid chromatography method. Results: Phytochemical analysis of the B. scandens leaf and leaf calli showed the presence of PTOX at the concentrations of 0.69 and 1.81, respectively. The callus cell suspension was established with the same callogenic media also it is augmented with 10–70 mg/l of coniferyl alcohol to elicit the biosynthesis of PTOX. Successive cultures of the calli suspension yielded stable production of PTOX of 3.91 mg/g dry cell weight at 50 mg/l coniferyl alcohol in the media. The biosynthesis of PTOX was ideal when plant cells were cultivated in the dark with an agitation speed of 100 rpm. Conclusion: The growth and production of PTOX were found to be better with glucose than with sucrose as the medium carbon source. The harvesting of the secondary metabolite from the in vitro grown leaf calli of B. scandens is a better way to stop the exploitation of medicinal plants.


Author(s):  
◽  
Chrismawan Ardianto ◽  
Junaidi Khotib ◽  
Djoko Agus Purwanto ◽  
Wirdhatul Muslihatin ◽  
...  

AbstractBackgroundCatechin is one of the secondary metabolites in Camellia sinensis L. that is alternatively produced through in vitro cultures. The in vitro culture product is possibly improved by optimizing the culture medium with the addition of growth regulators and precursors. The purpose of this study was to confirm the success of the secondary catechin metabolite production through the in vitro culture of C. sinensis L in a relatively short time.MethodsThe secondary catechin metabolite product is obtained in about 40 days. The study was conducted by (1) leaf cutting for inoculation in Murashige and Skoog media with 1 μg/mL of 2,4-dichlorophenoxyacetic acid growth regulator; (2) the inoculation of callus multiplication on the same medium as a partially modified inoculation media condition with the addition of 1 μg/mL of 6-benzylaminopurine (BAP) and 2 μg/mL of 2,4-dichlorophenoxyacetic acid at concentration; (3) callus multiplication developed on a new medium containing phenylalanine precursors (300 μg/mL); (4) testing growth by harvesting the callus and weighing the wet weight of its biomass and (5) identification of the callus qualitatively and quantitatively by using high-performance liquid chromatography (HPLC).ResultsThe level of secondary catechin metabolite produced was 2.54 μg/mL and 12.13 μg/mL in solid and suspension media, respectively.ConclusionsIt is concluded that the method is effective and efficient in producing catechin product from C. sinensis L.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 460e-460 ◽  
Author(s):  
Marisa F. de Oliveira ◽  
Gerson R. de L. Fortes ◽  
João B. da Silva

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.


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