scholarly journals Biological Activity of the tzs Gene of Nopaline Agrobacterium tumefaciens GV3101 in Plant Regeneration and Genetic Transformation

2013 ◽  
Vol 26 (11) ◽  
pp. 1359-1365 ◽  
Author(s):  
Zhao-Fen Han ◽  
David M. Hunter ◽  
Susan Sibbald ◽  
Ji-Shu Zhang ◽  
Lining Tian

Agrobacterium tumefaciens has been widely used in plant genetic transformation. Hormone-encoding genes residing in the T-DNA region have been removed, resulting in disarmed Agrobacterium strains that are used in various transformation experiments. Nopaline Agrobacterium strains, however, carry another hormone gene, trans-zeatin synthesizing (tzs), that codes for trans-zeatin in the virulence region of the tumor-inducing plasmids. We investigated the activity and function of the tzs gene of a nopaline Agrobacterium sp. strain GV3101 in plant in vitro regeneration. Leaf explants of tobacco and Nicotiana benthamiana co-cultured with strain GV3101 exhibited active shoot regeneration in media without added plant growth regulators. On medium without plant growth regulators, transgenic shoots were also induced from explants co-cultured with GV3101 containing a binary vector. Enzyme-linked immunosorbent assay showed that cell-free extracts of Agrobacterium sp. strain GV3101 culture contained the trans-zeatin at 860 ng/liter. Polymerase chain reaction using tzs-specific primers showed that the tzs gene was present in strain GV3101 but not in other Agrobacterium strains. The study showed that the tzs gene in GV3101 was actively expressed, and that trans-zeatin produced in the Agrobacterium strain can promote plant shoot regeneration.

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 512F-513
Author(s):  
Hak Tae Lim ◽  
Y.S. You ◽  
E.J. Park

In order to regenerate explants of Brassica campestris ssp. pekinensis, known to be one of the most difficult crops to regenerate via organogenesis, three different explants, cotyledon, hypocotyl, and leaf, were cultured on MS basal medium supplemented with several plant growth regulators. In the medium containing NAA at 0.5 mg/L and BAP at 3.0 mg/L, the shoot regeneration, when hypocotyl was used as explant, was found to be quite effective. In the case of cotyledon, the most suitable combination of plant growth regulators was NAA at 1.0 mg/L and BAP at 3.0 mg/L. Treatment of AgNO3 (1.0 mg/L) for shoot regeneration gave positive results in general. Zeatin at 2.0 mg/L was very effective in shoot induction of leaf explant, especially when combined with BAP at 2.0 mg/L, NAA at 1.0 mg/L, and AgNO3 at 0.5 mg/L. A system to produce transgenic plants in Brassica spp. has also been developed using hypocotyl and cotyledonary-petiole segments and shoot-tips. An explants from 4-day-old seedlings were inoculated with an Agrobacterium tumefaciens strain containing a disarmed tumor-inducing plasmid pTiT37-SE carrying a chimaeric bacterial gene encoding hygromycin and kanamycin resistance, along with other genes of interests. The explants were co-cultured for 2 to 6 days before transfer to hygromycin and kanamycin selection media. Shoots regenerated directly from the explants in 1 to 4 weeks and were excised, transferred to shoot elongation medium, rooted in root induction medium, and planted in soil. Genetic transformation was confirmed by kanamycin or hygromycin resistance, GUS activities, and Southern blotting.


2016 ◽  
Vol 26 (1) ◽  
pp. 85-96 ◽  
Author(s):  
Mohamed R Rady ◽  
El Sayed ◽  
M Mervat ◽  
Mohamed El Sayed ◽  
Usama I Aly

Leaf explants were cultured to evaluate the effect of different auxins and cytokinins and its concentrations; MS salts on micropropagation of the promising biodiesel Jatropha curcas plant under Egyptian conditions. Results showed that shoot initiated on 0.5 mg/l BA + 0.25 mg/l IBA. Multiplication and elongation were found to be the best using 0.5 mg/l BA in combination with 0.05 mg/l IBA. The multiple shoots were cultured on MS or half strength of MS supplemented with different concentrations of IAA and IBA for rooting phase. Half strength of MS containing 1.0 mg/l IAA was the best for rooting of micropropagated shoots. The rooted plantlets were acclimated in sand : peat?moss mixture (1 : 1) successfully.Plant Tissue Cult. & Biotech. 26(1): 85-96, 2016 (June)


2014 ◽  
Vol 5 (2) ◽  
pp. 85 ◽  
Author(s):  
Ejiroghene Felix Lawyer ◽  
Z. O. Jamaleddine ◽  
P. T. Lyam ◽  
I. T. Borokini ◽  
A. A. Adedeji ◽  
...  

Growth regulators especially auxins and cytokinins are critical for plant in-vitro regeneration. The effect of these plant growth regulators on in-vitro propagation of Saccharum officinarum L (Sugarcane) was investigated. In vitro response of two different varieties of sugarcane (NCS 005 and NCS 008) to Plant Growth Regulators was obtained in this study. Formation of buds was obtained on shoot apical meristem when cultured on MS (Murashige and Skoog) medium supplemented with 0.1mg/l BAP (6-Benzylaminopurine). After two weeks of initiation, regenerated meristem was inoculated into MS (Murashige and Skoog) fortified with different concentrations and combination of cytokinins. Shoot multiplication was optimal on 0.5mg/l BAP + 0.25 mg/l Kin(Kinetin) for NCS 005 variety while for NCS 008 variety, no significant (P≥0.05) difference was observed between 1.5mg/l BAP and 1.5mg/l BAP +0.5mg/l Kin. The best root induction for in vitro derived shoots was obtained on 1.0 mg/l NAA (Naphthalene acetic acid) and 2.0 mg/l IBA( Indole butyric acid) for both varieties of sugarcane within ten days of culture transfer. Successfully established plantlets showed excellent growth response when weaned under regulated green house conditions.


2019 ◽  
Vol 256 ◽  
pp. 108648 ◽  
Author(s):  
Vu Quoc Luan ◽  
Le Kim Cuong ◽  
Hoang Thanh Tung ◽  
Vu Thi Hien ◽  
Tran Hieu ◽  
...  

Author(s):  
Arash Mokhtari ◽  
H. Alizadeh ◽  
B. Yazdi Samadi ◽  
M. Omidi ◽  
M. Otroshy ◽  
...  

2014 ◽  
Vol 32 (2) ◽  
pp. 170-179 ◽  
Author(s):  
Hernando Criollo ◽  
Margarita Perea ◽  
Mariano Toribio ◽  
Johanna Muñoz

Lulo is a species of great importance to the fruticulture of Colombia, but has significant phytosanitary problems that require an aggressive breeding program oriented toward the production of genotypes with tolerance to phytopathogens. These programs need to establish highly efficient mass plant propagation protocols, such as somatic embryogenesis. This study focused on research on the somatic embryogenesis of lulo using kinetin, naphthalene acetic acid-NAA (Plant Growth Regulators, PGRs), and different sucrose concentrations in a MS medium. Two lulo varieties, Solanum quitoense var. septentrionale and S. quitoense var. quitoense, and two explant types (hypocotyl and cotyledon) were used, incubated in dark conditions at 25±2°C. The highest production percentage of the embryos was obtained when 50 mM of NAA were added to the medium with sucrose (50.0 and 263.1 mM) for the two explant types used. In lulo with spines, the highest percentage of embryonic structures (50%) was observed with cotyledonary leaf explants and 50 mM of NAA ; while in the spineless lulo, the embryonic structures were observed in the same type of explant with 50 mM of NAA + 263.1 mM of sucrose (32%).


2020 ◽  
pp. 34-43
Author(s):  
Fayza R. Al Gethami ◽  
Hameda El Sayed Ahmed El Sayed

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.


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