Sugarcane (NCo310) Transient Transformation Using uidA Reporter Gene

Matroodi, Soheila; Motallebi, Mostafa; Zamani, Mohammadreza; Mousavi, Amir; Davoodi, Daryoush; Jahromi, Zahra Moghaddassi

doi:10.5812/ijb.11203

Cited by 5 publications

(8 citation statements)

References 27 publications

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“…They showed that out of the various combinations of helium pressure and target distances used in their experiments on rice calli, maximum transient GUS activity was observed at 1,100 psi pressure and 9 cm target distance. High frequency of transient GUS expression at a helium pressure of 1,100 psi and a target distance of 9 cm was also found to be optimum in many other studies (Zhang et al, 1996;Chen et al, 1998;Jiang et al, 2000;Ramesh and Gupta, 2005;Matroodi et al, 2013).…”

Section: Discussionmentioning

confidence: 62%

OPTIMIZATION OF PARTICLE BOMBARDMENT CONDITIONS FOR RICE (Oryza sativa L.) TRANSFORMATION

Mukhtar¹

2018

PAKJAS

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Many factors which affect bombardment efficiency were studied and optimized to develop a simple and reproducible system for transformation of rice through particle-bombardment. Conditions were optimized by varying particle size, helium pressure, target distance and DNA/particle loads. Effect of pre-and post-bombardment osmotic treatment and effective concentration of hygromycin B for selection of transformed cells were also investigated. Both types of explants gave an equally good response in transient GUS expression, however, mature embryos were found to be more suitable giving higher selection efficiency. Gold particles of 1 µm size, helium pressure of 1,100 psi, target distance of 9 cm and a DNA to particle ratio of 3:1 produced the highest transient GUS expression in embryos. Osmotic treatment resulted in 2.7 to 2.9 fold increase in transient GUS expression. Hygromycin B at 50 mg L -1 was found to be optimum for effective selection of transformed cells. Transgenic plants were recovered at an average transformation efficiency of 1.5%. Analysis of putative transgenic plants revealed that 96% of the recovered plants were positive for both GUS and hpt genes. All the established plants were phenotypically normal and fertile.

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Section: Discussionmentioning

confidence: 62%

OPTIMIZATION OF PARTICLE BOMBARDMENT CONDITIONS FOR RICE (Oryza sativa L.) TRANSFORMATION

Mukhtar¹

2018

PAKJAS

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“…In the present research, the gold particles were used as microcarriers for their biologically inert behavior, uniformity in shape, size and non-toxic nature. Efficiency of gold particles in comparison to tungsten particles as microcarriers has also been reported (Matroodi et al, 2013). Gold particles of 0.6 μm size resulted in maximum transformation efficiency in contrast to 1μm particle size.…”

Section: Discussion Discussion Discussionmentioning

confidence: 73%

“…Though, reports on successful genetic manipulations are available; however, the success rate is low (Jackson et al, 2013). A number of biological and physiological parameter affects the biolistic transformation process (Matroodi et al, 2013). As seed production is not possible in most of the region due to the lack of suitable; therefore, the induction of embryogenic calli for transformation is demanded.…”

Section: Discussion Discussion Discussionmentioning

confidence: 99%

“…In contrast, biolistic transformation has high transformation efficiencies. Most of the published research on sugarcane transformation is based on biolistic transformation of sugarcane (Jackson et al, 2013;Matroodi et al, 2013). Altpeter and Oraby (2010) preferred biolistic technique because of its applicability to a large number of species.…”

Section: Introduction Introduction Introduction Introductionmentioning

confidence: 99%

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Biolistic transformation of sugarcane (Saccharum officinarum L.) with the Oryza sativa L. H+-PPase gene for improved salt stress tolerance

ULLAH

KHAN

JAN

et al. 2022

Not Bot Horti Agrobo

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Several biotic and abiotic stresses adversely affect sugarcane crop. Amongst the abiotic stresses, salinity causes substantial losses to sugarcane yield. Lack of conducive environment for sugarcane flowering and unstable genome are the main constraints in sugarcane crop improvement via conventional breeding. Therefore, genetic engineering of sugarcane using Oryza sativa L. H+-PPase gene (OVP1) to confer salt tolerance is the preferred alternative strategy. In the present research, tissue culture and biolistic transformation protocols for sugarcane were optimized. For callus induction, Dichlorophenoxyacetic acid (2,4-D) was most effective. Maximum callus induction (50.22%) was achieved on CM-3 augmented with 2.5 mgL-1 2,4-D. The induced calli was subjected to biolistic transformation. For biolistic transformation, the gold particles (0.6 and 1.0 µm) and plasmid DNA (100 and 50 ng) were used for 1X and 2X bombardment. Significant differences (P≤0.05) were observed amongst the gold particles and plasmid DNA concentrations. Maximum transformation frequencies were observed when calli were bombarded 2X with 0.6 µm gold particles coated with 100 ng of DNA (23.60%). Selection and regeneration of transgenic calli were performed on MS media augmented with BAP (2.5 mgL-), NAA (1 mgL-1) hygromycin (25 mgL-1) and 2% coconut water. Regenerated plants after root induction on MS media containing NAA (2 mgL-1) and 2% coconut water were transferred to loam and clay (1:1) and acclimatized to greenhouse condition. The transgenic plants were confirmed through PCR and RT-PCR.

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“…In our study, we obtained the maximum transformation efficiency of sugarcane calli with 8.5 µg of recombinant pGreenII0129 plasmid. Matroodi et al (2013) reported a higher concentration of plasmid as vital for enhanced transformation efficiencies; however, they used pUBI121 plasmid instead of pGreenII0129. Kim et al (2012) reported the impact of plasmid concentration on the biolistic transformation of sugarcane calli.…”

Section: Discussionmentioning

confidence: 99%

Transformation of the Osc3h52 Gene Promoter in Tissue Culture of Sugarcane

2020

THE JAPS

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Sugarcane growth is negatively affected by abiotic stresses. Genetic engineering of sugarcane with abiotic stressresponsive genes has been the method of choice to confer abiotic stress tolerance. In the present work, we optimized callus induction and transformation of the sugarcane variety CP-77-400 with the promoter region of the OsC3H52 gene to analyze its regulatory function under drought and salt stress. Calli were induced using different callus induction media (CIM). The promoter region of the OsC3H52 gene that belongs to the zinc finger protein family was cloned into two different expression vectors i.e. pBI221 and pGreenII0129, which were subsequently transformed to sugarcane calli through agrobacterium and biolistic transformation methods. Among various callus induction media, CIM3 (5 mg L-1 2,4-D + 10% coconut water) showed high callus induction (93%). Biolistic transformation using recombinant pGreenII0129 at 8.5µg/µl concentration resulted in higher efficiency of transformation (28%). While the agrobacterium mediated transformation using recombinant pBI221 plasmid gave 13% transformation efficiency. Amongst the various concentrations of acetosyringone used in agrobacterium mediated transformation, 100 µM showed higher efficiencies (13%) of transformation. Selection of the transformed calli was performed at 25 mg L-1 Hygromycin and 25 mg L-1 Hygromycin + 300 mg L-1 cefotaxime for 15 days, both for biolistic and agrobacterium mediated transformation, respectively. Transformed calli were then successfully confirmed through PCR and GUS expression. The regulatory role of the OsC3H52 promoter in transgenic calli was further evaluated under drought and salt stress. Quantitative real-time PCR analysis of the GUS reporter gene showed high expression in transgenic calli under drought and salt stress conditions.

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Sugarcane (NCo310) Transient Transformation Using uidA Reporter Gene

Cited by 5 publications

References 27 publications

OPTIMIZATION OF PARTICLE BOMBARDMENT CONDITIONS FOR RICE (Oryza sativa L.) TRANSFORMATION

OPTIMIZATION OF PARTICLE BOMBARDMENT CONDITIONS FOR RICE (Oryza sativa L.) TRANSFORMATION

Biolistic transformation of sugarcane (Saccharum officinarum L.) with the Oryza sativa L. H+-PPase gene for improved salt stress tolerance

Transformation of the Osc3h52 Gene Promoter in Tissue Culture of Sugarcane

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