Selection and characterization of a new switchgrass (Panicum virgatum L.) line with high somatic embryogenic capacity for genetic transformation

Xu, Bin; Huang, Linkai; Shen, Zhengxing; Welbaum, Gregory E.; Zhang, Xunzhong; Zhao, Bingyu

doi:10.1016/j.scienta.2011.05.016

Cited by 27 publications

(33 citation statements)

References 27 publications

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“…Xu et al (2011) identified the switchgrass genotype HR8, a line derived from the cv. Alamo that has extremely high tissue culture capacity [33]. In this study, SE calli were induced from mature seeds of HR8.…”

Section: Discussionmentioning

confidence: 99%

“…Mature seeds were harvested and stored at 4 C before use. The switchgrass genotype HR8 [33], derived from cv. Alamo, which has an exceptional tissue culture capacity as well as a high germination rate, was used to induce somatic embryogenic (SE) calli in this study.…”

Section: Plant Materialsmentioning

confidence: 99%

“…Switchgrass tissue culture was performed as described previously [33]. In brief, mature switchgrass seeds were dehulled in 60% sulfuric acid for 30 min, and then sterilized with 50% bleach for 45 min at room temperature.…”

Section: Plant Tissue Culturementioning

confidence: 99%

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Production of Autopolyploid Lowland Switchgrass Lines Through In Vitro Chromosome Doubling

et al. 2013

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Switchgrass is considered one of the most promising energy crops. However, breeding of elite switchgrass cultivars is required to meet the challenges of large scale and sustainable biomass production. As a native perennial adapted to North America, switchgrass has lowland and upland ecotypes, where most lowland ecotypes are tetraploid (2n=4x=36), and most upland ecotypes are predominantly octoploid (2n=8x=72). Hybridization between lowland and upland switchgrass plants could identify new cultivars with heterosis. However, crossing between tetraploid and octoploid switchgrass is rare in nature. Therefore, in order to break down the cross incompatibility barrier between tetraploid lowland and octoploid upland switchgrass lines, we developed autooctoploid switchgrass lines from the lowland tetraploid cv. Alamo. In this study, colchicine was used in liquid and solid mediums to chemically-induce chromosome doubling in embryogenic calli derived from cv. Alamo. Thirteen autooctoploid switchgrass lines were regenerated from seedlings and identified using flow cytometry. The autooctoploid switchgrass plants exhibit increased stomata aperture and stem size in comparison with the tetraploid cv.Alamo. The most autooctoploid plants were regenerated from switchgrass calli that were treated 2 with 0.04% colchicine in liquid medium for 13 days. One autooctoploid switchgrass line, VT8-1, was successfully crossed to the octoploid upland cv. Blackwell. The autooctoploid and the derived inter-ecotype hybrids were confirmed by chromosome in-situ hybridization and molecular marker analysis. Therefore, the results of this study show that an autooctoploid, generated by chemically-induced chromosome doubling of lowland cv. Alamo, is cross compatible with upland octoploid switchgrass cultivars. The outcome of this study may have significant applications in switchgrass hybrid breeding.

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Section: Discussionmentioning

confidence: 99%

Section: Plant Materialsmentioning

confidence: 99%

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Production of Autopolyploid Lowland Switchgrass Lines Through In Vitro Chromosome Doubling

et al. 2013

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“…Therefore, Odjakova & Conger (1999) and Burris et al (2009) used specific single genotypes Alamo 2702 and Alamo 2, respectively, in their experiments, and Alexandrova et al (1996b) developed a micropropagation procedure with node culture for the multiplication of such selected genotypes. Interestingly, the selected line HR8, having high somatic embryogenic capacity, was recently bred using recurrent tissue culture selection to allow easy induction of embryogenic callus from mature seeds (Xu et al, 2011).…”

Section: Switchgrass Regeneration Systemsmentioning

confidence: 99%

“…Interestingly, this system also employed Agrobacterium strain EHA105, suggesting that it may be highly compatible with 'Performer'. Switchgrass transformation using a EHA105 with the switchgrass selected line HR8, which has high somatic embryogenic capacity (Xu et al, 2011) (see section 3.1), may be a promising avenue for future research.…”

Section: Switchgrass Transformation Systemsmentioning

confidence: 99%

Molecular Breeding of Grasses by Transgenic Approaches for Biofuel Production

Takahashi¹,

Takamizo²

2012

Transgenic Plants - Advances and Limitations

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Inside out: high-efficiency plant regeneration and Agrobacterium-mediated transformation of upland and lowland switchgrass cultivars

et al. 2015

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Selection of pre-embryogenic callus from a core structure from mature seed-derived callus is the key for high-efficiency plant regeneration and transformation of switchgrass different cultivars. Switchgrass (Panicum virgatum L.) has been identified as a dedicated biofuel crop. For its trait improvement through biotechnological approaches, we have developed a highly efficient plant regeneration and genetic transformation protocol for both lowland and upland cultivars. We identified and separated a pre-embryogenic "core" structure from the seed-derived callus, which often leads to development of highly regenerative type II calluses. From the type II callus, plant regeneration rate of lowland cultivars Alamo and Performer reaches 95%, and upland cultivars Blackwell and Dacotah, 50 and 76%, respectively. The type II callus was also amenable for Agrobacterium-mediated transformation. Transformation efficiency of 72.8% was achieved for lowland cultivar Alamo, and 8.0% for upland cultivar Dacotah. PCR, Southern blot and GUS staining assays were performed to verify the transgenic events. High regenerative callus lines could be established in 3 months, and transgenic plants could be obtained in 2 months after Agrobacterium infection. To our knowledge, this is the first report on successful plant regeneration and recovery of transgenic plants from upland switchgrass cultivars by Agrobacterium-mediated transformation. The method presented here could be helpful in breaking through the bottleneck of regeneration and transformation of lowland and upland switchgrass cultivars and probably other recalcitrant grass crops.

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Selection and characterization of a new switchgrass (Panicum virgatum L.) line with high somatic embryogenic capacity for genetic transformation

Cited by 27 publications

References 27 publications

Production of Autopolyploid Lowland Switchgrass Lines Through In Vitro Chromosome Doubling

Production of Autopolyploid Lowland Switchgrass Lines Through In Vitro Chromosome Doubling

Molecular Breeding of Grasses by Transgenic Approaches for Biofuel Production

Inside out: high-efficiency plant regeneration and Agrobacterium-mediated transformation of upland and lowland switchgrass cultivars

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