Embryogenic Callus Induction and Plant Regeneration in Triticum tauschii, the Diploid D-Genome Donor for Bread Wheat (Triticum aestivum)

1996 ◽  
Vol 44 (4) ◽  
pp. 489 ◽  
Author(s):  
S Afsharsterle ◽  
ECK Pang ◽  
JS Brown ◽  
JF Kollmorgen

Immature embryos of seven accessions of Triticum tauschii (Coss.) Schmal. were used to produce embryogenic callus suitable for initiation of suspension cultures. Several modifications of Murashige and Skoog basal medium (MS) were evaluated for callus induction from scutellar tissues of embryos. Nodular, embryogenic calli were induced from all accessions when MS medium was supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and a mixture of L-glutamine, L-asparagine and L-proline. Early differentiation of these embryogenic calli was overcome by substituting Dicamba for 2,4-D. Addition of 575 mg L-1 of L-proline gave a rapid increase in the production of nodular embryogenic callus in most of the accessions. Using this protocol, the embryogenic capacity of this type of callus was maintained for more than a year following further modification of the MS medium. A clear genotype dependency as well as media effects on the production of callus were observed.

2003 ◽  
Vol 55 (3-4) ◽  
pp. 77-80 ◽  
Author(s):  
Aneta Bijelovic ◽  
Marko Sabovljevic

Callus induction of moss species Aloina aloides (Schultz) Kindb. was obtained on Murashige and Skoog (MS) medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) or with 1.0 mg/L 2,4-D and 1.0 mg/L kinetin (KIN) or with 0.2 mg/L indole-3-butyric acid (IBA) and 2.0 mg/L 6-benzylaminopurine (BAP) or with 7.5 g/L of sucrose or with 15 g/L of sucrose or hormone - free and sugar free MS basal medium. The callus can be maintained for a long period of time without bud formation subcultured on the above media, at 16 h day/8 h night, 25 ? 2?C, 60-70% air humidity and irradiance of 50 ?mol m-2s-1. To obtain plant regeneration pieces, calli were transferred onto MS media supplemented with different concentrations of auxins and cytokinins (1.0 mg/L 2,4-D and 2 mg/L KIN; 0.2 mg/L IBA and 2 mg/L KIN; or 0.2 mg/L IAA and 2 mg/L BAP). In these media after subculturing, callus enlarges and turns to gametophytes with buds. Except for a smaller size, the plants obtained on the callus did not differ morphoanatomically from the shoots in the nature.


Aquilaria malaccensis Lam. and Aquilaria subintegra Ding Hou belong to the family of Thymelaeaceae which is commonly known as gaharu or agarwood. It is a commercially important tree and identified as a potential aromatic plant. The overwhelming responses in the lodging sector reduce gaharu species in the forest. Mass propagation through plant tissue culture technology will substitute this problem. The present study was conducted to investigate the embryogenic callus induction between these two species. The most optimum sterilization method for both species was sodium hypochlorite 5.0% which gave the highest percentage of aseptic culture (95%) with the absence of tissue browning. The leaves of both species were cultured on Murashige and Skoog, (1962) (MS) media supplemented with combination of various concentrations of 6-benzylaminopurine (BAP) (0.5, 1.0, 2.0 and 2.5 mg/L) and 2,4-dichlorophenoxyacetic acid (2, 4-D) (0.5, 1.0, 1.5 and 2.0 mg/L) and kept under dark condition. The explants produced embryogenic, white and compact callus at the end cut of the explants after two weeks of culture in all treatments. The highest frequency of embryogenic callus formation was observed in explants cultured on 2.0 mg/L BAP and 0.5 mg/L 2,4-D for both species. From the present study, the optimum sterilization technique and embryogenic callus induction for A. malaccensis Lam. and A. subintegra were established.


1989 ◽  
Vol 19 (10) ◽  
pp. 1303-1308 ◽  
Author(s):  
D. T. Webb ◽  
F. Webster ◽  
B. S. Flinn ◽  
D. R. Roberts ◽  
D. D. Ellis

Zygotic embryos of Piceaglauca (Moench) Voss from five half-sib seed families and P. engelmanii Parry from one half-sib family, collected on July 13 and 27 and August 24, were cultured in the presence of 2,4-dichlorophenoxyacetic acid, N6-benzyladenine, and sucrose ranging from 0.5 to 4% for the induction of embryogenic callus and the production of stable embryogenic callus lines. Embryogenic callus was induced from all three collections with all seedlots. The July 13 collection was two to four times more embryogenic than the later collections. Embryogenic callus was induced at all sucrose levels, but 4% sucrose was clearly inferior, whereas 1% was best overall. Factors that favored the induction of embryogenic callus also favored the production of stable embryogenic callus lines. There was a 40% decline in callus line establishment compared with embryogenic callus induction and some seed lots failed to yield embryogenic lines from the two later collections. The formation of caulogenic callus was promoted at 3 and 4% sucrose. Embryos from the August 24 collection were more caulogenic than those from the earlier collections.


2019 ◽  
Vol 7 (4.14) ◽  
pp. 107
Author(s):  
D S M Soder ◽  
D N A A Khalid ◽  
A Saleh ◽  
F Pardi ◽  
N J Sidik

Barringtonia racemosa is mangroves type of plant which had been extensively utilized in conventional practices for relieving ailments of pain and inflammation. Many studies have been done on ethnobotanical profiles, pharmacological activities and chemical compounds in Barringtonia racemosa. However, there is a limited study on callogenesis of this plant particularly from different maturity stage of fruits. The present study is to identify the callogenesis of Barringtonia racemosa from endosperm explants of immature and mature fruits in MS medium supplemented with different concentrations of hormones 2,4-Dichlorophenoxyacetic acid (2,4-D) (0, 0.5, 1.0, 1.5 and 2.0 mg/L) and Kinetin (KIN) (0, 0.5, 1.0, 1.5 and 2.0 mg/L). The optimum hormone combination was found in callus grown on endosperm of immature fruits in MS medium supplemented with 1.5 mg/L 2,4-D and 1.0 mg/L KIN. It was also found that the callus in this treatment grew profusely with highest fresh weight (0.513 ± 0.022 g), 100% callus induction and friable callus texture. The callus fresh weight on endosperm explants was higher in immature fruits compared to mature fruits for all the hormone combinations. Therefore, callogenesis were found more efficient from endosperm explant of immature fruits in Barringtonia racemosa species.   


HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 788E-789
Author(s):  
Hae Young Na* ◽  
Dong Jin Shin ◽  
Changhoo Chun

Pimpinella brachicarpa (Chamnamul in Korean) is an indigenous plant that grows in Korean mountain areas. It has not been cultivated yet but is gathered to use as a vegetable. Its difficulty of propagation by seeds is one of the major reasons not to be cultivated as a horticultural crop despite its demand. As a promising propagation method for the Chamnamul, we have developed a micropropagation system using somatic embryogenesis. In the present study, induction of embryogenic callus of the Chamnamul affected by part of explants (leaf and stem) and concentration (0, 0.1, 0.5, 1.0, 1.5, and 2.0 mg·L-1) of growth regulators (2.4-D, IAA, IBA, and NAA) was investigated to find the best conditions for embryogenic callus induction. A full strength of MS medium was used for a 50-day culture for all the treatments. The embryogenic callus was firm and light yellow in color and was distinct from the non-embryogenic callus that was friable and semitransparent. More embryogenic callus was induced in the treatments that the stem was used as an explant comparing with the treatments that the leaf was used. The 2.4-D treatments resulted in the better induction of embryogenic callus than other growth regulator treatments, and 1.5 mg·L-1 was the most effective among all the 2,4-D concentration treatments. Addition of 0.1 mg·L-1 BA to 2.4-D treatments retarded the induction of embryogenic callus of the Chamnamul, while the promotion of induction and multiplication of embryogenic callus was reported in many plant species by adding BA with low concentration to an auxin-base medium. The better induction was found in the treatments of darkness and dim lighting (10 μmol·m-2·s-1 of PPF) than in treatments of the higher PPF.


2017 ◽  
Vol 1 (2) ◽  
Author(s):  
Nining Intan Toharah ◽  
Dwi Soelistya Dyah Jekti ◽  
Lalu Zulkifli

This study aims to determine the concentration of growth regulators BAP and   2,4-D which have the highest effect in stimulating the formation of callus melon plants (Cucumis melo) Mai 119 variety. Completely randomized design (CRD) was used in this research. Media used on callus induction was MS medium with addition of several concentration of BAP  (0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L) and 2,4-D (0 mg/L, 1 mg/L, 2 mg/L, 3 mg/L) either alone or in a combination of both. Parameters measured were the time appearing of callus, callus diameter, callus texture, and callus color. Anova followed by Tukey's test was used to the analyse of time appearing of callus. Data of callus diameter was analyzed using Kruskal Wallis test followed by Mann-Whitney test. In the analysis of parameter related to the callus texture and callus color, descriptive test were used. The results showed that there were differences in the effect of growth regulators on the callus formation. The fastest callus induction and the largest diameter of callus were obtained on media with concentration of 2 mg/L BAP and 3 mg/L BAP.Keywords: BAP (benzyl amino purine), 2,4-D (2,4-dichlorophenoxyacetic acid), callus induction, melon (Cucumis melo) varieties Mai 119


2015 ◽  
Vol 10 (5) ◽  
pp. 1934578X1501000 ◽  
Author(s):  
Aya Inoue ◽  
Shinjiro Ogita ◽  
Shinpei Tsuchiya ◽  
Reiko Minagawa ◽  
Hamako Sasamoto

Callus induction, maintenance and protoplast cultures were achieved from immature seeds of a woody leguminous mangrove, Caesalpinia crista. Axenic cultures were possible during 1.5 months of pod storage in 0.1% benzalkonium chloride solution. Callus induction was achieved using 1 mL liquid medium in a 10 mL flat-bottomed culture tube. Protoplasts were isolated using Cellulase R10, Hemicellulase, and Driselase 20 in 0.6 M mannitol solution and sub-culturable calluses were obtained in 50 μL liquid medium using a 96-microplate method. The optimal hormonal concentration was 10 μM each of 2,4-dichlorophenoxyacetic acid and benzyladenine in liquid Murashige and Skoog's basal medium for both callus induction and maintenance, and protoplast cultures. Similarities and differences in amino acid profiles and culture conditions are discussed among woody mangrove species and non-mangrove leguminous species. Caesalpinia crista cultures were unique as they secreted a large amount of amino acids, including proline, into the liquid culture medium.


2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.


1999 ◽  
Vol 47 (4) ◽  
pp. 611
Author(s):  
Shoukat Afshar-Sterle ◽  
James F. Kollmorgen ◽  
Geoffrey B. Fincher

Immature embryos of 10 accessions of Triticum tauschii, the D genome donor of Triticum aestivum, were used to produce embryogenic callus for the initiation of suspension cultures. For the long-term maintenance of embryogenicity of these suspensions, it was necessary to use different media for the initiation, establishment and maintenance phases. The initiation phase required media supplemented with L-proline, L-asparagine and L-glutamine, together with Dicamba at 12 mg L −1 and maltose. In the establishment phase, it was essential to reduce the concentration of Dicamba to 6 mg L −1 for the rapid production of fine suspension cultures. Finally, the long-term maintenance of a capacity for regeneration depended on the inclusion of 1.1 mg L −1 2,4-dichlorophenoxyacetic acid and 30 g L −1 sucrose in the medium. By the use of these procedures, long-term embryogenic fine suspension cultures were established from two accessions, while non-embryogenic fine suspension cultures were produced from five accessions. Over 90% of plants regenerated from fine suspensions of 1-year-old embryogenic cultures were fertile, and embryogenic suspension cultures retained their regeneration capacity for more than 3 years.


1970 ◽  
Vol 3 (2) ◽  
pp. 40-45
Author(s):  
M.F. Mohamad Bukhori ◽  
Norzulaani Khalid ◽  
Ch'ng Lou Ven

To explore the potential for embryogenic callus induction protocol through cell suspension culture forhigh frequency plant regeneration of Maspine pineapple (Ananas comosus L.), eight different culturemedia formulation were evaluated for their effects on the induction of somatic embryos from suckerexplants. Explants were cultured on MS medium supplemented with various media concentration(NAA, Dicamba and BAP, Picloram, Kinetin and NAA, 2,4-D, TDZ, and TDZ and BAP).Embryogenic callus induction percentage, color and texture of the callus were assessed after fivemonths of culture. The optimum medium for the proliferation of in vitro shoots from sucker explantswas MS medium supplemented with 3 mg/L BAP. Meanwhile, the optimum medium for the inductionof fastest and high percentage of embryogenic callus growth from in vitro leaf-based was MS mediumsupplemented with Picloram. Results of mean comparison showed that 3 mg/L Picloram were moreeffective on explants than 10 mg/L. Results of the double staining method proved that somaticembryogenesis occurred in MS supplemented with 3 mg/L Picloram. Under microscopic observations,the globular-stage of the embryos were revealed in callus cells which is relatively suitable forsuspension cells inoculums, indicating that the tested PGR were significantly effective for somaticembryogenesis formation in this species. Most embryogenic callus from sucker explants wasyellowish-mucilaginous-wet-friable. The developed protocol potentially leads to the production ofembryogenic callus from sucker explants and plant regeneration through somatic embryogenesis.


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