Optimize 2,4-D concentration and callus induction time enhance callus proliferation and plant regeneration of three rice genotypes

Carsono, Nono; Juwendah, Endah; Liberty, Liberty; Sari, Santika; Damayanti, Farida; Rachmadi, Meddy

doi:10.13057/biodiv/d220702

Cited by 15 publications

(10 citation statements)

References 33 publications

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“…The formation of calli is very important to regenerate adventitious buds. Calluses of different ages showed different growth capacities and regeneration abilities ( Cheng et al., 2009 ; Carsono et al., 2021 ), which was also shown in our study. The highest frequency of callus and adventitious bud induction of E. urophylla × E. grandis DH32-29 was obtained when leaves were used to induce calluses for 15 days.…”

Section: Discussionsupporting

confidence: 88%

Agrobacterium-mediated genetic transformation of the most widely cultivated superior clone Eucalyptus urophylla × Eucalyptus grandis DH32-29 in Southern China

et al. 2022

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Eucalyptus, as an economically important species for wood and paper industries, still remains a challenge to genetic improvement by transgenic technology owing to the deficiency of a highly efficient and stable genetic transformation system, especially in cultivated superior clones. Eucalyptus urophylla × Eucalyptus grandis clone DH32-29 is most widely planted in southern China, but it is relatively recalcitrant to adventitious bud regeneration, which blocks the establishment of a genetic transformation system. Here, an efficient adventitious bud regeneration and transformation system of Eucalyptus was established using E. urophylla × E. grandis DH32-29 as material. The in vitro leaves from microshoots that were subcultured for 20–25 days were immersed into liquid Woody Plant Medium supplemented with 0.02 mg·L−1 of α-naphthaleneacetic acid (NAA) and 0.24 mg·L−1 of forchlorfenuron (callus-inducing medium (CIM)). After 15 days, explants were transferred to a medium containing 0.10 mg·L−1 of NAA and 0.50 mg·L−1 of 6-benzyladenine (shoot-inducing medium (SIM)) for adventitious bud induction. The highest regeneration efficiency of adventitious buds was 76.5%. Therefore, an Agrobacterium tumefaciens-mediated genetic transformation system was optimized. The leaves were precultured for 7 days and infected for 30 min with A. tumefaciens strain EHA105 grown to a bacterial density of 0.3 (OD600). After 72 h of cocultivation in the dark, leaves were transferred to CIM supplemented with 100 mg·L−1 of cefotaxime (Cef), 100 mg·L−1 of timentin, and 15 mg·L−1 of kanamycin (Kan) for 15 days to induce calluses. Then, the explants were transferred to SIM supplemented with the same concentration of antibiotics, and the fresh medium was replaced every 15 days until resistant adventitious buds appeared. After induction of roots in root-inducing medium supplemented with 200 mg·L−1 of Cef and 75 mg·L−1 of Kan, completely transgenic plants were obtained. With the use of the aforementioned method, the transformation frequency can reach 1.9%. This provides a powerful approach for genetic improvement of E. urophylla × E. grandis DH32-29 and gene function analysis in Eucalyptus.

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Section: Discussionsupporting

confidence: 88%

Agrobacterium-mediated genetic transformation of the most widely cultivated superior clone Eucalyptus urophylla × Eucalyptus grandis DH32-29 in Southern China

et al. 2022

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“…Auxins cause embryogenic and organogenic differentiation, cytodifferentiation, and cell division in tissue culture [ 24 ]. Among PGRs, the auxin 2,4-D (dichlorophenoxyacetic acid) is well-known for helping to accelerate the proliferation and expansion of embryogenic calli [ 25 ]. Many modifications in both type and concentration of PGRs have been evaluated.…”

Section: Introductionmentioning

confidence: 99%

Salinity Stress Response of Rice (Oryza sativa L. cv. Luem Pua) Calli and Seedlings

2022

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Soil salinity limits plant growth and production. This research investigated a suitable medium for callus induction and plantlet regeneration in the Luem Pua rice cultivar. The effect of salt stress on seedling growth was determined using in vitro culture and soil conditions. An efficient protocol for callus induction has been developed by culture sterilized seeds on the Murashige and Skoog (MS, 1962) medium containing 0.5 mg/l benzyladenine (BA) with 1 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) that resulted in a 100% callus induction. Plantlet regeneration percentage of 49% was recorded on the MS medium containing 4 mg/l BA with 0.5 mg/l 1-naphthaleneacetic acid (NAA) after 4 weeks. For salt stress investigation, the calli were treated on an induction medium containing various concentrations of NaCl (0, 50, 100, 150, and 200 mM), while two-week-old rice seedlings were planted in soil and treated with the same concentration of NaCl for 4 weeks. In vitro culture revealed that callus survival percentage decreased when NaCl concentration increased, similar to soil culture. Seedling growth under salinity treatment also decreased when NaCl concentration increased, while other physiological parameters such as total chlorophyll, chlorophyll a, chlorophyll b, green intensity, and chlorophyll fluorescence under light conditions increased under salinity stress. These changes define the growth and physiological salinity tolerance characteristics of Luem Pua rice calli and seedlings. They can be utilized as a baseline for demand-driven in vitro rice propagation, providing useful information that can be combined with other agronomic features in rice development or breeding programs to improve the flexibility of abiotic stress-tolerant cultivars.

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“…The medium contain 1 mg/L 2,4-D + 1 mg/L BAP (ACM1) generates the greatest calli weight (2.086 grams). The involvement of 2,4-D is to promote the mechanism of morphogenesis of calli, calli initiation, and to regulate the genetic structure of the plant tissues [16,17]. In this treatment, the use of endogenous or exogenous auxin and cytokinin promotes regeneration [18,19,20].…”

Section: Characteristics Of Calli Formationmentioning

confidence: 99%

The effect of 2,4-D, thidiazuron and BAP on calli induction of arabica coffee (Coffea arabica L.)

Arimarsetiowati

Putra

Suwastono

et al. 2023

IOP Conf. Ser.: Earth Environ. Sci.

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Four continents and numerous developing contries benefit economically from the annual income and jobs that coffee provides. The only polyploid species found in the genus Coffea is called Coffea arabica, and it belongs to the Rubiaceae family. In order to commercially propagate plants or to learn more about key concepts in somatic embryo induction and development, somatic embryogenesis is used. The purpose of this experiment was to compare how calli induction was impacted by several plant growth regulators in the Arabica coffee. Leaf tissues were cultivated on ½ MS medium with varying concentrations of 2,4-D and thidiazuron (1.0, 2.0, 3.0 mg/L) together with 1.0 mg/L BAP. Each experiment used a minimum of seven replicated abd was repeated three times using a completely random design. The varied grades of calli’s morphological examination was identified. Up to eight weeks, the proportion of calli development was measured every two weeks. Nearly sixty percent of calli formed in the medium of 1 mg/L 2,4D and 1 mg/L BAP, which had the greatest calli formation percentage. The maximum calli weight (2.086 grams) is achieved in medium of 1 mg/L 2,4-D + 1 mg/L BAP.

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Optimize 2,4-D concentration and callus induction time enhance callus proliferation and plant regeneration of three rice genotypes

Cited by 15 publications

References 33 publications

Agrobacterium-mediated genetic transformation of the most widely cultivated superior clone Eucalyptus urophylla × Eucalyptus grandis DH32-29 in Southern China

Agrobacterium-mediated genetic transformation of the most widely cultivated superior clone Eucalyptus urophylla × Eucalyptus grandis DH32-29 in Southern China

Salinity Stress Response of Rice (Oryza sativa L. cv. Luem Pua) Calli and Seedlings

The effect of 2,4-D, thidiazuron and BAP on calli induction of arabica coffee (Coffea arabica L.)

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