Tag Archives: regeneration

xxx L.Q. Cao, P.D. Nguyen, X.H. Pham, V.H. La, X.V. Nguyen, T.D. Nguyen
Comparison of in vitro regeneration and transformation efficiency among five vietnamese indica rice cultivars
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Comparison of in vitro regeneration and transformation efficiency among five vietnamese indica rice cultivars

L.Q. Cao¹†, P.D. Nguyen¹, X.H. Pham¹, V.H. La², X.V. Nguyen², T.D. Nguyen²*

¹Department of Molecular Pathology, Institute of Agricultural Genetics, Vietnam Academy of Agricultural Sciences, Hanoi 11956, Vietnam
²Institute of Biotechnology and Food technology, Thai Nguyen University of Agriculture and Forestry, Thai Nguyen 24153, Vietnam
*Correspondence: dungnt@tuaf.edu.vn

Abstract:

This study optimized the in vitro regeneration and transformation processes for five popular indica rice cultivars in Vietnam named as BC15, Bac Thom 7 (BT7), Khang Dan 18 (KD18), OM5451, and TBR225. The highest callus induction rates (83-93.67%) were achieved on a medium containing 2.5 mgL-l 2,4-D, supplemented with 500 mg L-l each of L-proline, L-glutamine, and casein. Higher or lower concentrations of 2,4-D resulted in reduced callus induction. Among tested cultivars, BT7 exhibited the highest callus induction rate of 93.67%. Shoot regeneration was optimized on a medium with a combination of 0.5 mg L-l kinetin, 2.0 mg L-l BAP, and 0.5 mg L-l NAA, resulting in regeneration rates ranging from 80.23% to 90.77% across cultivars, with BT7 exhibiting the highest regeneration rate (90.77%). Hygromycin at 20 mg L-l was optimal for selecting transgenic calli, with BT7 showing the highest transformation efficiency opdetermined to be optimal for selecting transgenic calli, with BT7 achieving the highest transformation efficiency (44% hygromycin-resistant callus and 29% GUS expression). PCR analysis confirmed a high proportion (87.5%) of transgenic plants in the BT7 cultivar. These findings indicate that the BT7 is the most suitable cultivar for genetic transformation and gene editing applications.

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468–477 D. Lazdiņa, I. Bebre, K. Dūmiņš, I. Skranda, A. Lazdins, J. Jansons and S. Celma
Wood ash – green energy production side product as fertilizer for vigorous forest plantations
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Wood ash – green energy production side product as fertilizer for vigorous forest plantations

D. Lazdiņa*, I. Bebre, K. Dūmiņš, I. Skranda, A. Lazdins, J. Jansons and S. Celma

Latvian State Forest Research Institute ‘Silava’, Riga str. 111, LV-2169 Salaspils, Latvia
*Correspondence: dagnija.lazdina@silava.lv

Abstract:

Notable amounts of wood ash containing plant macro and micronutrient elements in balanced proportions are produced in Latvia. If bioenergy production source product is plant material, and facilities are operating well, then ‘side product’ fermentation residues or wood ashes should not contain elements in toxic concentrations. Wood ash contains P and K which are lacking in acidic organic soils and could work as fertiliser as well as a long term liming agent, besides that, all micronutrient elements necessary for physiological processes are present in wood ash. Wood ash could also be used as ‘revitalization agents’ – fertilisers to improve the growth of plantation forests. The aim of this research is to find and describe the positive effect of wood ash fertilisers on Norway spruce (Picea abies) and other economically valuable tree species. Research results show positive wood ash application effect on tree growth and vitality within the first 4 years when used for recultivation and revitalization purposes. Recycling of wood ash (0.5–3 t ha- 1 before planting) for fertilisation of and Picea abies forest plantations are a sustainable and effective solution for the improvement of tree growth as well as an environmentally safe method of utilization of bioenergy production residues.

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563-575 M. A. Zaidi, M. Narayanan, R. Sardana, I. Taga,, S. Postel, R. Johns,M. McNulty, Y. Mottiar, J. Mao, E. Loit, I. Altosaar,
Optimizing tissue culture media for efficient transformation of different indica rice genotypes
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Optimizing tissue culture media for efficient transformation of different indica rice genotypes

M. A. Zaidi¹, M. Narayanan¹⋅², R. Sardana¹, I. Taga¹,³, S. Postel¹, R. Johns¹,M. McNulty¹, Y. Mottiar¹, J. Mao¹, E. Loit¹, I. Altosaar¹,*

¹ Agricultural Biotechnology Laboratories, Department of Biochemistry, Microbiology andImmunology, University of Ottawa, 451 Smyth Rd. Ottawa, ON., K1H 8M5, Canada
² Present address: Department of Agricultural Entomology, Agricultural College and ResearchInstitute, Tamil Nadu Agricultural University, Madurai-625 104, India
³ Clinical Biochemistry and Nutrition Unit, Biochemistry Department, Faculty of Sciences, P OBox 24157, University of Douala, Douala, Cameroon
* Corresponding author: altosaar@uottawa.ca Tel: +1(613) 562-5846, Fax: +1 (613) 562-5440

Abstract:

This article is dedicated to the rye breeder Count Friedrich Georg Magnus von Berg, celebratingthe 160th year of his birth. An efficient system was established for high frequency embryogenesis and regeneration of indica rice, Oryza sativa L. cv. MDU 5. Basic media, carbohydrate sources and concentrations, agar concentrations, amino acids, cytokinins and auxins were evaluated in callus induction and regeneration media to establish the most efficient regeneration protocol for MDU 5 indica rice. The optimized callus induction and regeneration media consisted of the basic MS salt mixtures and vitamin solutions supplemented with 4% maltose, 1g/L casein hydrolysate and 50 mg/L tryptophan, solidified with 1% agar. The callus induction medium was supplemented with 2,4−D 2 mg/L, kinetin 0.5 mg/L, indole acetic acid 1 mg/L, and 6−benzyl aminopurine 0.5 mg/L whereas the media for regeneration phase comprised kinetin 2 mg/L, indole acetic acid 1 mg/L and 6−benzyl aminopurine 2 mg/L. The media optimized for MDU 5 was analyzed for response of 73 other indica genotypes. Of these indica varieties 64 genotypes yielded 98.5% callus induction within eight days, 59% embryogenic calli formation, initiation of multiple green buds within eight days and a regeneration rate of 90%. The embryogenic calli derived were used for transformation with Agrobacterium tumifaciens (LBA 4404 or EHA 101 strains). Two binary vectors (pKHG4 and pIG121Hm) containing hph and GUS genes were used in these transformation studies. Fifty-one genotypes responded to the optimized media by producing hygromycin−resistant calli. The -histochemical test for ß−glucuronidase activity was positive from 32 genotypes with the transformation efficiencies ranging between 7.0% and 8.3%.

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