Cell size: a key determinant of meristematic potential in plant protoplasts

Metabolic pathway reconstruction and gene edits for native natural product synthesis in single plant cells are considered to be less complicated when compared to the production of non-native metabolites.Being an efficient eukaryotic system,plants encompass suitable post-translational modifications.However,slow cell division rate and heterogeneous nature is an impediment for consistent product retrieval from plant cells.Plant cell synchrony can be attained in cultures developed in vitro.Isolated plant protoplasts capable of division,can potentially enhance the unimpaired yield of target bioactives,similar to microbes and unicellular eukaryotes.Evidence from yeast experiments suggests that‘critical cell size’and division rates for enhancement machinery,primarily depend on culture conditions and nutrient availability.The cell size control mechanisms in Arabidopsis shoot apical meristem is analogous to yeast notably,fission yeast.If protoplasts isolated from plants are subjected to cell size studies and cell cycle progression in culture,it will answer the underlying molecular mechanisms such as,unicellular to multicellular transition states,longevity,senescence,‘cell-size resetting’during organogenesis,and adaptation to external cues.

Plant regeneration of protoplasts isolated from suspension cells derived from leaf blale of Oryza sative

We used the leaf blade of rice (cultivars were Nonghu 6, Sugeng 2, Huyou 2 and Hanfeng) as initial material for protoplast culture, and a great number of regenerated plants were obtained. Rice seeds were sterilized and germinated. The immature leaves were cut into 3-5 mm pieces when the third or forth leaf appeared. Leaf pieces were inoculated on MS medium with 2,4-D 4 mg/1, NAA 2mg/1 and IAA Img/1. After 2 wk culture, calli were induced and subcultured once or twice for multiplication. 3-5 g calli were transferred to the modified MS liquid medium with 2,4-D 2 mg/1 and KT 0.5mg/1 for suspension culture. Embryogenic cell suspension was established after 2 mo culture. The effect of the growth period of suspension cells on the

Plant Regeneration from Protoplasts of Wide Compatible Rice

Rice selection 02428 and T984(Oryzasativa L.ssp.japonica)were germplasmresources with wide compatibility.Mature embryos of rice cultured on Lin-smaier and Skoog medium with 2.5 mg/l2,4-D,1.0 mg/l thiamine-HCL,3%(W/V)sucrose and 0.7%(W/V)agar,pH 5.8(LS2.5)were used for callus initiation.Cultures were

Plant regeneration from mesophyll protoplast of indica rice Qiugui’ai 11 (Oryza sativa L.)

In the recent decade,plant regeneration fromprotoplast has been obtained through embryo-genic cell suspension cultures of rice.Howev-er,not only the establishment of embryogeniccell suspension cultures of rice was difficult,but also the protoplasts became less and lessregenerable and the genetic change was gradu-ally accumulated during the prolonged culture.Since 1976(Deka.),extensive efforts have

Gene delivery into isolated Arabidopsis thaliana protoplasts and intact leaves using cationic, a-helical polypeptide

The application of gene delivery materials has been mainly focused on mammalian cells while rarely extended to plant engineering. Cationic polymers and lipids have been widely utilized to efficiently deliver DNA and siRNA into mammalian cells. However, their applica- tion in plant cells is limited due to the different membrane structures and the presence of plant cell walls. In this study, we developed the cationic, a-helical polypeptide that can effectively deliver DNA into both isolated Arabidopsis thaliana protoplasts and intact leaves. The PPABLG was able to condense DNA to form nanocomplexes, and they exhibited significantly improved transfection efficiencies compared with commercial transfection reagent Lipofec- tamine 2000 and classical cell penetrating peptides such as poly（L-lysine）, HIV-TAT, arginine9, and poly（L-arginine）. This study therefore widens the utilities of helical polypeptide as a unique category of gene delivery materials, and may find their promising applications toward plant gene delivery.