scholarly journals An efficient protocol for perennial ryegrass mesophyll protoplast isolation and transformation, and its application on interaction study between LpNOL and LpNYC1

Plant Methods ◽  
2017 ◽  
Vol 13 (1) ◽  
Author(s):  
Guohui Yu ◽  
Qiang Cheng ◽  
Zheni Xie ◽  
Bin Xu ◽  
Bingru Huang ◽  
...  
Keyword(s):  
Perennial Ryegrass ◽  
Mesophyll Protoplast ◽  
Protoplast Isolation ◽  
Interaction Study

scholarly journals Stochastic gene expression drives mesophyll protoplast regeneration

2021 ◽  
Vol 7 (33) ◽  
pp. eabg8466
Author(s):  
Mengxue Xu ◽  
Qingwei Du ◽  
Caihuan Tian ◽  
Ying Wang ◽  
Yuling Jiao
Keyword(s):  
Gene Expression ◽  
Low Frequency ◽  
Mesophyll Protoplast ◽  
Underlying Mechanism ◽  
Protoplast Isolation ◽  
Chromatin Accessibility ◽  
Cellular Level ◽  
Protoplast Regeneration ◽  
Expression Variability ◽  
Enhanced Expression

Cell pluripotency is fundamental to biology. It has long been known that differentiated somatic plant cells may reacquire pluripotency, but the underlying mechanism remains elusive. In many plant species, a single isolated mesophyll protoplast may regenerate into an entire plant, which is widely used in gene transformation. Here, we identified two transcription factors whose ectopic activation promotes protoplast regeneration. Furthermore, we found that their expression was induced by protoplast isolation but at a very low frequency. Using live imaging and single-cell transcriptomics, we show that isolating protoplasts induces enhanced expression variation at the genome level. Isolating protoplasts also leads to genome-wide increases in chromatin accessibility, which promotes stochastic activation of gene expression and enhances protoplast regeneration. We propose that transcriptome chaos with increased expression variability among cells creates a cellular-level evolutionary driver selecting for regenerating cells.

scholarly journals SOME FACTORS AFFECTING ISOLATION OF MESOPHYLL PROTOPLASTS FROM APPLE (MALUS DOMESTICA BORKH CV. FUJI)

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 693g-694
Author(s):  
Chang-Hoo Lee ◽  
N. I. Hyung ◽  
S. E. Kim
Keyword(s):  
Cell Division ◽  
Culture Media ◽  
Shoot Culture ◽  
Mesophyll Protoplast ◽  
Protoplast Isolation ◽  
Enzyme Treatment ◽  
Planting Density ◽  
Factors Affecting ◽  
Cell Structures ◽  
Viable Protoplast

Experiments were conducted to investigate the factors influencing mesophyll protoplast isolation in `Fuji' apple. Half an hour pretreatment in 0.6M mannitol gave the highest protoplast yield.The enzyme solution containing 2% Cellulase Onozuka R-10 and 0.5% macrozyme R-10 with CPW 0.6M mannitol at pH 5.5 was most effective for protoplast isolation from leaf. Effective incubation time for the enzyme treatment was found to be 15-20 hrs at 25°C in the dark. Use of 1.0-2.0% PVP and 0.5mM MES was essential for higher yield and viability of protoplast. Supplementation of BA and IBA to the shoot culture media gave the higher yield of viable protoplast. From these protoplast, new cell walls were regenerated and 4 cell structures developed from one protoplast by cell division in K8P medium supplemented with 3A and NAA. Planting density higher than 10 protoplasts/ml was required for cell division from protoplast in liquid or 0.5% agarose culture.

scholarly journals Optimization Conditions of Wheat Mesophyll Protoplast Isolation

2016 ◽  
Vol 07 (12) ◽  
pp. 850-858 ◽  
Author(s):  
Xingyue Jia ◽  
Xiaohui Zhang ◽  
Jingming Qu ◽  
Rong Han
Keyword(s):  
Mesophyll Protoplast ◽  
Protoplast Isolation

N transfer from white clover to perennial ryegrass, via exudation of nitrogenous compounds

Agronomie ◽  
2003 ◽  
Vol 23 (5-6) ◽  
pp. 503-510 ◽  
Author(s):  
Florence Paynel ◽  
Jean Bernard Cliquet
Keyword(s):  
Perennial Ryegrass ◽  
White Clover ◽  
N Transfer ◽  
Nitrogenous Compounds

USE OF CELL AND MOLECULAR GENETIC MANIPULATION TO IMPROVE PASTURE PLANTS

1988 ◽  
pp. 67-72
Author(s):  
D.W.R. White
Keyword(s):  
Cell Culture ◽  
Perennial Ryegrass ◽  
White Clover ◽  
Nutritional Quality ◽  
Rust Resistance ◽  
Herbicide Tolerance ◽  
Pest Resistance ◽  
Crown Rust ◽  
Wide Range ◽  
Pasture Plants

Cell culture and genetic engineering techniques can be used to develop improved pasture plants. To utilise these methods we have developed procedures for regenerating plants from tissue cultures of perennial ryegrass and white clover. In both, the plant genotype influences regeneration capacity. There was significant genetic variation among regenerated perennial ryegrass plants in a wide range of characteristics. Most of the regenerants were resIstant to crown rust and this trait was highly heritable. This rust resistance is being used to breed a new ryegrass cultivar. A system for introducing cloned genes into white clover is described. This capability is bemg used to incorporate genes with the potential to improve nutritional quality and pest resistance. Other possibilities for engineering genetic improvements in white clover, genes conferring herbicide tolerance and resistance to white clover mosaic virus, are briefly outlined. Keywords: Lolium perenne, Trifolium repens, cell culture, somaclonal variation, crown rust resistance, transformation, cloned genes, nutritional quality, proteinase inhibitors, Bt toxins, pest resistance, WCMV viral cross-protection, herbicide tolerance, Agrobacterium, Bacillus thuringenisis.

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