scholarly journals Temporal and Spatial Expression Analysis of Shoot-Regeneration Regulatory Genes during the Adventitious Shoot Formation in Hypocotyl and Cotyledon Explants of Tomato (CV. Micro-Tom)

2020 ◽  
Vol 21 (15) ◽  
pp. 5309 ◽  
Author(s):  
Myoung Hui Lee ◽  
Jiyoung Lee ◽  
Eun Yee Jie ◽  
Seung Hee Choi ◽  
Lingmin Jiang ◽  
...  

Enhancing the competence for plant regeneration in tissue culture studies is an important issue not only for efficient genetic transformation of commercial crops but also for the reproducibility of scientific reports. In this study, we investigated optimization of several tissue culture conditions including plant growth regulators, types and ages of explants, culture densities, and plant position in order to improve the competence of adventitious shoot formation of the tomato (Solanum lycopersicum cv. Micro-Tom). In addition, we examined the differential expression of D-type cyclin (CYCD3-1) and several shoot regeneration regulatory genes from hypocotyl and cotyledon explants of tomato during shoot organogenesis. A treatment of 1 mg L−1 Zeatin and 0.1 mg L−1 Indole-3-acetic acid (IAA) in Murashige and Skoog (MS) medium containing 3% sucrose was optimal for adventitious shoot formation from hypocotyl and cotyledon explants. The younger explants exhibited more shoot formation regardless of explant types. Additionally, those closest to the shoot apical meristem produced more shoots compared to the other regions in the hypocotyl and the cotyledon explants. Gene expression of CYCD3-1, SHOOT MERISTEMLESS (STM), and cytokinin dependent WUSCHEL (WUS) was significantly higher in younger explants than in older ones. Furthermore, an increase in CYCD3-1, STM, and WUS expression was evident at the distal part of hypocotyls and the proximal part of cotyledons compared to other regions. These differential gene expression profiles exhibited good agreement with the results of shoot formation obtained from diverse explants of tomato. These results suggest that temporal and spatial gene expression of shoot regeneration regulatory genes plays an important role in enhancing the competence and the reproducibility of adventitious shoot formation from tomato explants.

HortScience ◽  
1994 ◽  
Vol 29 (12) ◽  
pp. 1536-1538 ◽  
Author(s):  
P. Gercheva ◽  
R.H. Zimmerman ◽  
L.D. Owens ◽  
C. Berry ◽  
F.A. Hammerschlag

Shoot regeneration from apple (Malus domestica Borkh.) leaf explants following particle bombardment at various acceleration pressures was studied. Basal leaf segments of micropropagated `Royal Gala' apple were bombarded with 1 μm gold particles, accelerated using helium pressures of 4.5, 6.2, 7.6, 9.3, or 13.8 MPa (650–2000 psi), and cultured on shoot regeneration medium consisting of N6 salts supplemented with 10 μM TDZ for 5, 10, or 20 days in darkness. Bombarded and control explants exhibited 63% to 100% shoot regeneration. With a 5-day dark period, average shoot production per explant ranged from 6.1 to 14; bombardments of 4.5 and 6.2 MPa significantly increased shoot production over the controls. With a 10-day dark period, average shoot production per explant ranged from 9.1 to 22 following bombardment at 9.3 and 6.2 MPa, respectively. Following bombardment at 6.2 MPa, 75% of the explants produced more than 20 regenerants per explant. With a 20-day dark period, average shoot production per explant ranged from 8.9 to 19 following bombardment at 13.8 MPa and following no bombardment, respectively. Shoot production per explant was significantly less than the controls following bombardments ranging from 6.2 to 13.8 MPa. Shoot production was highest per explant with particle bombardment at 6.2 MPa followed by incubation in darkness for 10 days. Chemical name used: thidiazuron (TDZ).


HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 749-750 ◽  
Author(s):  
Michael E. Compton

Organic competence of different explant sizes and locations on watermelon seedlings was determined by calculating the percentage of cotyledon explants that produced adventitious shoots. About 52% (214/412) of explants prepared from the proximal region of cotyledons formed shoots, whereas only ≈6% (24/411) of distal explants did so. Shoot formation was limited to the proximal end of basal explants but was not restricted to any specific region on distal ones. The percentage of explants that produced harvestable shoots was greater from basal halves than basal quarters in `Sweet Gem', `Crimson Sweet', and `Minilee', but explant size did not affect adventitious shoot regeneration of `Yellow Doll', resulting in significant interaction between cultivar and explant size. This study indicates that cultivars that respond poorly to in vitro procedures may have fewer cells competent for shoot regeneration, requiring special care during explant preparation.


Biologia ◽  
2011 ◽  
Vol 66 (3) ◽  
Author(s):  
Natalija Burbulis ◽  
Aušra Blinstrubienė ◽  
Ramunė Kuprienė

AbstractBreeding linseed (Linum usitatissimum L.) using haploid techniques allows breeders to develop new cultivars in a shorter time period. Many research groups successfully created new linseed genotypes through anther culture; however ovary culture has been the subject of only a few earlier studies. In the present study, the effect of genotype and growth regulators combination on callus induction and shoots regeneration in ovary culture of nine commercially important linseed cultivars was investigated. Ovaries were cultured on modified MS medium supplemented with three different combinations of plant growth regulators. Variable callogenic responses were expressed by all of the genotypes tested on different induction media. The results suggested that specific combination of growth regulators for callus induction must be designed for each genotype. Shoot regeneration from ovary derived callus is a critical phase of the whole gynogenetic process. Differences in adventitious shoot formation frequency among genotypes were demonstrated and four responsive genotypes have been selected. Ovary derived callus from cultivar ‘Mikael’ manifested the highest adventitious shoot formation frequency with a high number of shoots per explant. The optimum ratio of growth regulators for shoot regeneration was shown to depend on the genotype. Cultivars ‘Linola’, ‘Mikael’ and ‘Szaphir’ showed the highest shoot regeneration frequency when callus had originated on induction medium supplemented with 2 mg L−1 BAP and 2 mg L−1 NAA, while combination of 1 mg L−1 BAP and 2 mg L−1 IAA promoted shoot formation in ovary-derived callus of ‘Barbara’. The highest rate of shoots per explant has been obtained in second subculture.


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 571d-571
Author(s):  
Jim Hruskoci ◽  
Paul E. Read

In an effort to increase somaclonal variation in blueberry, a protocol was developed to regenerate viable shoots from internode segments. The explant consisted of the last-formed, fully developed internode taken from 3 different genotypes of Vaccinium grown in vitro. Explants were cultured 6 weeks on Zimmerman's Z-2 medium supplemented with 2iP, zeatin, thidiazuron, kinetin, or BA at concentrations of 5, 25, 50, and 100 uM. Explant response to the treatments varied and included: no response, callus growth only, callus growth and subsequent shoot formation originating from the callus mass, and adventitious shoot formation directly from the internode segment without an intervening callus. Greatest shoot regeneration (20-25 shoots/explant) was obtained on medium supplemented with zeatin at 5, 25, and 50 uM, however treatment response was not consistant across all genotypes. Regenerated shoots could be readily sub-cultured, rooted in soil mix and will be evaluated for somaclonal variation.


2016 ◽  
Vol 11 (3) ◽  
pp. 373
Author(s):  
Ira Djajanegara

Coconut water and rotten banana fruits are commonly found in traditional markets as organic wastes. One way to overcome the problems caused by these organic wastes is to convert these unuseful matter into an important and economically useful matter by using them as components of tissue culture media. One important commodity that is usually propagated by tissue culture is Phalaenopsis orchid type 229 (Phalaenopsis amabilis). Therefore, it would be more benefit to substitute the expensive chemicals with organic wastes such as coconut water and banana puree.In this experiment, addition of coconut water and banana pure to the minimum media containing commercial fertilizer red Polyhyponex, sucrose and commercial agar did not show any inhibition of Phalaenopsis orchid plantlet growth. This probably caused by sufficient macro and micro nutrients provided by those organic matter and Polyhyponexfertilizer. Moreover, addition of 100 mL/L of coconut water and 100 mg/L banana puree gave the optimum leaf and adventitious shoot formation. On the other hand, addition of 150 mL/L coconut water gave the optimum height and root formation. In this case,growing Phalaenopsis orchid plantlet should be done in 2 subculture period. The first subculture is to obtain maximum amount of leaf and shhot formation while the second subculture is to obtain optimum height and root formation.Key words : Waste, coconut water, banana puree, tissue culture, Phalaenopsis orchi


2014 ◽  
Vol 62 (1-2) ◽  
pp. 11-15 ◽  
Author(s):  
Anna Stojakowska

A method of micropropagation of <i>Urginea maritima</i> (L.) Baker s.str. (<i>Liliaceae</i>) by adventitious shoot formation was developed. Bulb scales and leaf fragments were used as primary and secondary explants, respectively. The most favourable for shoot regeneration were media: MS supplemented with BAP or kinetin (bulb scales) and medium C containing NAA along with BAP (leaf explants). No difficulties in rooting and adapting of plants to greenhouse conditions were observed.


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