scholarly journals In vitro selection for Fusarium oxysporum f. sp. conglutinans resistance in brassica vegetables

Author(s):  
Oushadee A. J. Abeyawardana ◽  
Martin Koudela

<p class="abstract"><strong>Background:</strong> Fusarium wilt is an issue of concern in economically and nutritionally important Brassica vegetable cultivation. Thus, it deserves measures against the adverse production impact caused by Fusarium wilt.</p><p class="abstract"><strong>Methods:</strong> In this study, development of resistance to <em>F. oxysporum</em> f. sp. <em>conglutinans</em> in six white head cabbage cultivars, by in vitro chemical mutagenesis and selection, through direct and indirect organogenesis was examined. 6 day and 10 day old hypocotyl, shoot tip and calli, from 6 day old hypocotyl explants, were subjected to chemical mutagenesis treatment (DMSO (4% v/v) + EMS (0.3% v/v) for two hours at 28±2°C) were incubated in MS shoot induction medium (MS+ NAA (0.2 mg/l), BAP (3 mg/l), GA3 (0.01 mg/L) and AgNO3 (0.5 mg/l)). Shoots developed from hypocotyl and shoot tip explants (in the MS shoot induction medium and then in MS + NAA (0.2 mg/l), BAP (3 mg/l), GA3 (0.01 mg/l) for shoot development) and calli following mutagenesis treatment were screened for Fusarium resistance subjecting to 15% and 20% <em>Fusarium</em> culture filtrate for 30 and 60 day selection periods for each strength respectively.  </p><p class="abstract"><strong>Results:</strong> Developed plantlets from all six cultivars tested, showed resistance to<em> </em>Fusarium culture filtrate in the in vitro conditions with different survival frequencies ranging between 12.5% to 84.0% from hypocotyl and 0.0% to 86.7% from shoot tip explants among cultivars indicating development of resistance to <em>Fusarium</em> by in vitro chemical mutagenesis.</p><p class="abstract"><strong>Conclusions:</strong> Direct organogenesis, and 10 day old hypocotyl and 6 day old shoot tips are potential explants for successful application of in vitro chemical mutagenesis for Fusarrium resistance development in in cabbage.</p><p class="abstract"> </p>

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.


1970 ◽  
Vol 20 (2) ◽  
pp. 179-184
Author(s):  
N.P. Anish ◽  
M.G. Rajesh ◽  
Jiby Elias ◽  
N. Jayan

Shoot tip explants from in vitro germinated seedlings of Solanum capsicoides All. inoculated on MS containing 2 mg/l BA produced maximum shoot induction response (26 shoots per explant). Rooting of the microshoots (19.4 roots per explant) was obtained better in half strength of MS supplemented with NAA (0.5 mg/l). Well rooted plantlets were successfully hardened with 80 per cent survival rate.   Key words: Solanum capsicoides, Propagation, Therapeutic agent   D.O.I. 10.3329/ptcb.v20i2.6912   Plant Tissue Cult. & Biotech. 20(2): 179-184, 2010 (December)


HortScience ◽  
2001 ◽  
Vol 36 (6) ◽  
pp. 1102-1106 ◽  
Author(s):  
V.R. Bommineni ◽  
H. Mathews ◽  
S.B. Samuel ◽  
M. Kramer ◽  
D.R. Wagner

Improved in vitro clonal propagation methods are valuable tools for nurseries and growers, and are essential for manipulation and improvement of tree fruit germplasm using the tools and techniques of biotechnology. We have developed a rapid shoot multiplication procedure for clonal propagation of apple, Malus ×domestica cv. Gale Gala and pear, Pyrus communis L. cv. Bartlett. Rapid clonal multiplication was achieved after the following series of steps: pre-conditioning of micropropagated shoots, sectioning pre-treated stems into thin slices, placing slices onto shoot induction medium and incubating directly under cool-white fluorescent lights or after a brief dark incubation. Multiple induction of shoots recovered from stem slice explants within three weeks of culture. A maximum of 37% of cultured apple stem slices, and 97% of pear stem slices, showed induction of shoots. More shoots were recovered on phytagel solidified shoot induction medium than on agar. Cultured stem slices of both apple and pear showed maximum recovery of shoots from shoot induction medium supplemented with thidiazuron (TDZ) compared to medium supplemented with BAP and kinetin. Under ideal conditions, pear stems generated four times the shoots as the same quantity or length of apple shoots. Micropropagated shoots were rooted and transferred to the greenhouse and field nursery for further evaluation. Chemical names used: N-phenyl-N′-1,2,3-thidiazol-5-ylurea (thidiazuron or TDZ); 6-benzylaminopurine (BAP).


2008 ◽  
Vol 21 (1) ◽  
pp. 43-48
Author(s):  
S. M. H. Kabir ◽  
M. S. Ali ◽  
M. K. Islam

The Experiment was conducted to establish an efficient plant regeneration protocol from hypocotyl sections of soybean. Callus initiation, shoot and root development were observed by using different concentrations and combinations of growth regulators. The best result for callus induction was observed in MS medium supplemented with 1.5 mg/l Kinetin and 2.0 mg/l NAA. The calli were transferred to shoot induction medium. The best shoot induction occurred in the medium containing 3.0 mg/l BAP and 0.5 mg/l NAA. The elongated shoots developed roots on MS medium supplemented with different IBA concentrations where 1.5 mg/l IBA was the best for root development. Plantlets with a well developed root system were transplanted in plastic container with a soil mixture of cowdung and fine sand. Plantlet survival rate was 70%. Through this experiment, a general suitable regeneration protocol from hypocotyls of soybean has been developed which can potentially be used for micropropagation and future transformation research in soybean.DOI: http://dx.doi.org/10.3329/bjpbg.v21i1.17049


2018 ◽  
Vol 44 (3) ◽  
pp. 459-463
Author(s):  
PK Roy

An efficient protocol was developed for in vitro mass propagation of Paulownia tomentosa (Thunb.) Steud. using shoot tip and leaf segment explants from field grown plant. Different concentrations and combinations of BAP, Kn, zeatin and NAA were used for multiple shoot regeneration. Among two types of explants, leaf segment produced the highest number of shoots per explant (12 ± 0.4) when they were cultured on MS supplemented with 3.0 mg/l Kn and 0.5 mg/l NAA. Addition of 10% CW to above mentioned medium increased the number of shoots (18) per culture. Shoot tip explants also produced multiple shoots in the same medium, but their performance was not good as leaf segment explants. For shoot elongation, 100 mg/l urea was more effective when added with best shoot induction medium. Shoots rooted well in halfstrength MS supplemented with 2.0 mg/l NAA, within 12 - 15 days. Regenerated plantlets were successfully acclimatized and established in poly bag containing a mixture of soil and compost in 2:1 ratio. About 90% plantlets survived under open field conditions.


2012 ◽  
Vol 43 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Roberson Dibax ◽  
Giovana Bomfim de Alcantara ◽  
Marília Pereira Machado ◽  
João Carlos Bespalhok Filho ◽  
Ricardo Augusto de Oliveira

The objectives of this study were to establish appropriate conditions for obtaining plant regeneration and acclimatization of the 'RB92579' and 'RB93509' sugarcane cultivars and to elucidate the shoots origin through histological analysis. For both cultivars, obtaining shoots showed better results with the culture of explants on a callus induction medium containing 2.0mg L-1 2,4-dichlorophenoxyacetic acid, followed by cultivation on a shoot induction medium containing 0.1mg L-1 kinetin and 0.2mg L-1 benzilaminopurine. The MS medium without growth regulators proved to be appropriate for elongation and rooting of shoots and the use of the composed substrate of vermiculite + MS salts was effective for acclimatization. Histological analysis revealed that the origin of the shoots in both cultivars occurred through indirect organogenesis.


2021 ◽  
Author(s):  
Jorge Fonseca Miguel

The response on callus and shoot formation under different light incubation conditions was evaluated in cucumber (Cucumis sativus L.). Four-day-old cotyledon explants from the inbred line 'Wisconsin 2843' and the commercial cultivars 'Marketer' and 'Negrito' were employed. A four-week culture was conducted on MS-derived shoot induction medium containing 0.5 mg L-1 IAA and 2.5 mg L-1 BAP, under an 8-h dark/ 16-h light regime, or by a one- or two-week dark pre-incubation followed by the same photoperiod. Significant differences were obtained for the regeneration of shoots in all cultivars. The response in both frequency and number of shoots under continuous photoperiod was at least 3-6 fold higher than with dark pre-incubation. The highest genotypes response was obtained by 'Negrito' and 'Marketer' with identical values. All explants formed callus, and in two of the three cultivars, the response on callus extension was not significantly affected by incubation conditions. The results clearly show that shoot induction under continuous photoperiod regime was beneficial for adventitious shoot regeneration in cucumber.


Biologia ◽  
2008 ◽  
Vol 63 (5) ◽  
Author(s):  
Suleyman Cenkci ◽  
Mustafa Kargioglu ◽  
Sergun Dayan ◽  
Muhsin Konuk

AbstractThis report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium supplemented with 0.5–20 µM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin (2–8 µM) alone or in combination with a low level (0.5 µM) of 2,4-D or NAA. The best regeneration of shoots from root explants was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were achieved from cotyledonary explants at 4 µM BA and 0.5 µM NAA. The presence of 0.5 µM or higher levels of 2,4-D in shoot induction medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction.


HortScience ◽  
2010 ◽  
Vol 45 (3) ◽  
pp. 453-456 ◽  
Author(s):  
Fatemeh Haddadi ◽  
Maheran Abd Aziz ◽  
Ghizan Saleh ◽  
Azmi Abd Rashid ◽  
Hossein Kamaladini

An efficient micropropagation system for strawberry cv. Camarosa was developed. Sterilized runner tips were cultured on hormone-free Murashige and Skoog (MS) medium with 3% sucrose, 1 mL·L−1 Plant Preservative Mixture, and solidified using 0.25% phytagel to produce in vitro stock plants. Shoot tips derived from the in vitro stock plants were cultured on MS media containing 0, 2, 4, and 8 μM thidiazuron (TDZ) and 0, 4, 9, 18, and 27 μM N6-benzylamino-purine (BAP) for shoot induction. Shoots produced on the best shoot induction medium were rooted on MS media containing 1, 2, 3, and 5 μM of either indole-3-butyric acid (IBA) or naphthaleneacetic acid (NAA). Results showed that MS medium with 2 μM TDZ and 4 μM BAP was optimum for shoot multiplication from the shoot tips. The most suitable medium for inducing the highest number of roots per explant, the highest percentage of explant with roots, and the highest mean root length were 1 μM NAA, 1 μM IBA, and hormone-free MS medium, respectively. Plantlets were transplanted into substrate consisting of perlite + vermiculite + cocopeat (2:1:2 v/v/v) resulting in 90% survival. After 1 month, plants were irrigated using Hoagland's solution and runners were produced after 3 months.


Agrociencia ◽  
2020 ◽  
Vol 54 (3) ◽  
pp. 387-399
Author(s):  
Soledad Mora-Vásquez ◽  
Silverio García-Lara ◽  
Edgardo J. Escalante-Vázquez ◽  
Guy A. Cardineau

Plant tissue culture provides an alternative approach to improve the quality of soybean (Glycine max (L.) Merrill) cultivars. This study was undertaken to analyze the susceptibility of Mexican soybean for direct shoot regeneration and to determine the critical factors that affect in vitro performance. Our hypothesis was that Mexican soybean is suitable for in vitro regeneration using a cotyledonary node as explant. The effects of the seed disinfection procedure, soaking pretreatment before germination, soybean variety, as well as the culture medium composition of the shoot induction medium, were evaluated by two split-plot statistical designs. According to the statistical analysis, the seed disinfection procedure, the soaking pretreatment before germination, and the soybean genotype were the factors that brought about a significant effect (p£0.01), while the hormones composition of the shoot induction medium did not have a significant effect. The best response for multiple shoot formation was observed using a chlorine gas seed disinfection method, 3% hydrogen peroxide soaking pretreatment, and Huasteca-100, Nainari and Suaqui soybean genotypes. A robust protocol was developed, and under these selected conditions, it is possible to obtain more than 10 shoots per explant. Well-developed plantlets were obtained after 60 d of in vitro culture.


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