Preparation of Microprotoplasts for Partial Genome Transfer via Microprotoplast Fusion in Liliaceous Ornamental Plants

Abstract

We aimed to produce intergeneric hybrid plants with only one or a few alien chromosomes via microprotoplast fusion for genetic improvement and chromosome studies in Liliaceous ornamental plants. In order to apply this technique, it is essential to establish an efficient system for mass-preparation of microprotoplasts. We have established 2 different systems for isolating microprotoplasts, one from partially synchronized cell suspension cultures of Hemerocallis hybrida and the other from developing microspores of Lilium longiflorum. Here, the induction of micronucleated cells, isolation of microprotoplasts, and enrichment of smaller microprotoplasts containing one or a few chromosomes are described for both systems.

An Effective Procedure for In Vitro Culture ofEleusine coracana(L.) and Its Application

“…As a rule the cells with 3-4 micronuclei were observed after such treatment of finger millet cells. It is corresponding to the previously obtained data where CIPC in this concentration was successfully used to obtain microprotoplasts from developing microspores of lily species [38]. The system described here can be used for the transfer of one or a few chromosomes via microprotoplast fusion from finger millet to different economically important grasses or cereals.…”

“…Recently, an alternative asymmetric somatic hybridization method using microprotoplasts (microprotoplast fusion) has been developed [18]. Since microprotoplasts contain only one or a few intact chromosomes, a limited number of chromosomes can be transferred via microprotoplast fusion, resulting in the production of chromosome addition lines with even a single and specific intact chromosome between sexually incompatible species [18,38]. In order to apply this technique for improvement of monocot plant species, we established an efficient system for mass-preparation of microprotoplasts of E. coracana.…”

Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium

“…Within ornamentals, good results were obtained in Liliaceae (Saito and Nakano 2002;Horita et al 2003). Asymmetric protoplast fusion efficiently transmits diverse features such as disease resistance (Jarl et al 1999;Kinoshita and Mori 2001;Collonnier et al 2003), fatty acid composition and salt tolerance (Waara and Glimelius 1995).…”

Antimitotic Drugs For Microprotoplast-Mediated Chromosome Transfer In Plant Genomics, Cell Engineering And Breeding

“…and L. speciosum Thunb. 67 In this case, CIPC was used at a concentration of 10 µM, but the treatment time of microsporocytes was three to four days and its micronucleation properties turned out to be better than that of APM or propyzamide.…”

“…To date, sequential experiments helped establish that because of their high affinity for plant tubulin, dinitroanilines, phosphoroamides, propyzamide, and phenyl carbamate can effectively block metaphase and induce massive micronucleation in suspension-cultured cell 63,65,67,[70][71][72][73] and in developing microspores 66,67 of various plant species. Together with the development of effective fusion procedures for microprotoplasts, 74 these approaches made it possible for the subsequent development of the microprotoplast-mediated chromosome transfer technique, whose first adaptation to plants was the production of hydrids containing one potato chromosome and an entire set of tobacco (Nicotiana tabacum L.) 74,75 or tomato (Lycopersicon esculentum P. Mill./Solanum lycopersicum L.) chromosomes, 74 or containing one chromosome of N. plumbaginifolia and an entire set of tomato chromosomes.…”