High frequency plant regeneration from embryogenic callus of a popular indica rice (Oryza sativa L.)

Indica rice variety Ciherang is the most planted variety in Indonesia, but the micropropagation technique is restricted because it is known has low regeneration frequency and included as recalcitrant cultivar for tissue culture and tranformation activities. One of solution to resolve that problem is developing a technique of somatic embryogenesis in callus of ciherang rice cultivar. The aims of study were to determine medium composition for inducting embryogenic callus in Ciherang rice and to know the effectivity of rice regeneration using callus as explant. The methods were included induction of embryogenic callus in callus induction media (CIM) containing MS basal, Proline 600 mg/l, Casein Hidrolisat 300 mg/l, phytagel 2,5 g/l, BAP 0.25 mg/l, sukrosa 30 %, and 2,4-D in different concentration, from 2,4-D 0 mg/l as control (CIM 1), 2,4-D 2 mg/l (CIM 2), 2,4-D 3 mg/l (CIM 3), and 2,4-D 4 mg/l (CIM 4). It then be continued to regenerate the calli in RM 1 medium containing MS basal, NAA 0,2 mg/l, Kinetin 2 mg/l, Agarose 10 g/l, and sukrosa 30 %, pH 5,8 for six days in dark and RM2 medium containing MS basal , NAA 0,2 mg/l, Kinetin 2 mg/l, Agarose 8 g/l, sukrosa 30 %, pH 5,8 in light room. Results showed CIM 4 medium using 2,4-D 4 mg/l gave optimum result in calli induction with procentage 57,63% and CIM 3 using 2,4-D 3 mg/l gave optimum result in embryonic calli induction with procentage 53,63%. Result of embryonic calli gave regeneration frequency procentage is 9,6%. The total planlet obtained after five weeks old in regeneration medum were 11 planlets ready for aclimatization. Keywords: Somatic embryo, Indica rice (Oryza sativa L. var. Ciherang), Callus

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Effect of 2,4-D on Embryogenic Callus Induction of Malaysian indica Rice (Oryza sativa L.) Cultivars MR123 and MR127

Jurnal Teknologi ◽

10.11113/jt.v64.2048 ◽

2013 ◽

Vol 64 (2) ◽

Author(s):

Fauziah Illyas Ahmad ◽

Nur Shafiqoh Johan ◽

Alina Wagiran

Keyword(s):

Oryza Sativa ◽

Callus Induction ◽

Embryogenic Callus ◽

Staining Method ◽

Oryza Sativa L ◽

Indica Rice ◽

Simple Protocol ◽

Basal Media ◽

Embryogenic Callus Induction ◽

Mature Seeds

The aim of this study is to study the effect of various concentrations of 2,4-D on embryogenic callus induction of indica rice MR123 cultivar and MR127 cultivar using mature seeds. Optimal media for induction of callus of both cultivars was MS basal media supplemented with 30g/L sucrose, 500mg/L glutamine and 500mg/L proline supplemented with 2.5 mg/L 2,4-D. The highest percentage of callus induction was 70% and 76% for MR123 and MR127 respectively. Both cultivars produced an embryogenic callus from scutellum after a week in culture with white-yellowish in color. The viability tested using Evans blue show callus obtained was embryogenic. This simple protocol using staining method could be used for screening embryogenic callus before further experiments can be conducted.

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In vitro plant regeneration of some recalcitrant indica rice (Oryza sativa L.) varieties

Vegetos- An International Journal of Plant Research ◽

10.1007/s42535-021-00193-2 ◽

2021 ◽

Vol 34 (1) ◽

pp. 102-106

Author(s):

Ubaid Yaqoob ◽

Tanushri Kaul ◽

Irshad Ahmad Nawchoo

Keyword(s):

Oryza Sativa ◽

Plant Regeneration ◽

Oryza Sativa L ◽

Indica Rice ◽

In Vitro Plant Regeneration ◽

In Vitro Plant

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The effect of different antibiotics on the elimination of Agrobacterium and high frequency Agrobacterium-mediated transformation of indica rice (Oryza sativa L.)

Czech Journal of Genetics and Plant Breeding ◽

10.17221/77/2011-cjgpb ◽

2012 ◽

Vol 48 (No. 3) ◽

pp. 120-130 ◽

Cited By ~ 8

Author(s):

A.M. Priya ◽

S.K. Pandian ◽

R. Manikandan

Keyword(s):

Oryza Sativa ◽

High Frequency ◽

High Efficiency ◽

Oryza Sativa L ◽

Indica Rice ◽

Gus Expression ◽

Culture Conditions ◽

Embryogenic Calli ◽

Positive Effects ◽

Callus Proliferation

We report here the suitable explant with high efficiency of transformation and the positive effects of timentin over other antibiotics like carbenicillin and cefotaxime on the elimination of Agrobacterium tumefaciens during the genetic transformation of popular indica rice (Oryza sativa L.). The tissues assayed were embryogenic calli, embryos with endosperm contamination, intact seeds, leaf blades, leaf bases and coleoptiles. The frequency of transient β-glucuronidase (GUS) expression as revealed by histochemical assay was 90% for embryogenic calli, which was the highest among the explants used. On the basis of disc-diffusion assay, the maximum zone of inhibition (29 mm) at 250 mg/l was obtained for timentin. In tissue culture conditions the frequency of Agrobacterium recurrence after 20 days of infection was minimum (2.3%) at 200 mg/l of timentin. At 250 mg/l of timentin there was no Agrobacterium growth, besides, there were no negative effects on the callus growth unlike other antibiotics, hence it was selected as the optimum concentration for high frequency callus proliferation and regeneration.The effect of the parameters evaluated was determined by the callus proliferation during selection, reduction in browning, transient GUS expression and stable transformation efficiency (23.3%). The resultant plants were stable transformants as confirmed by a molecular analysis of the gus and hpt genes. The developed transformation protocol will be very helpful for the information on indica rice cultivars in general and on IR 64 in particular.

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