Development and Applications of Transplastomic Plants; A Way Towards Eco-Friendly Agriculture

With distribution of genetic materials and advance molecular characteristics, the chloroplast is prokaryotic compartments within the eukaryotic plants that have turned into a crucial source for the genetic engineering and transplastomic plants are becoming more popular means of agricultural development with elevated crop yield. To address global agricultural problems, genetic modification of crop plants is a rapid and promising solution to adapt the environment-friendly and well-controlled farming system. The transplastomic plant with high accumulation of foreign proteins (up to 45-46% TSP) and stable transgene expression with gene containment can be a unique choice for the agricultural innovation of coming centuries. Although the transplastomic plants still facing encumber to ensure the full potential exploitation and expansion as an economical means, the removal of hardness and obstacles of this technology and commercialization can contribute for the sustainable development of future agriculture. In this book chapter, we intend to recapitulate the up to date development and achievement of transplastomic plant including gene transfer procedures in plastid genomes, regulable expression of plastid transgenes, plant trait improvement by foreign gene expression, biopharmaceuticals production, engineering of metabolic pathways in plant, study of transformation mediated RNA editing technologies, bio-safety issues and public concerns on transplastomic plants and overall beneficial aspects. We believe that the utilization of transplastomic plants will ensure an eco-friendly approach in agriculture with minimized hazards and public concerns. © Springer Nature Switzerland AG 2020

Sugar Cane Genome Numbers Assumption by Ribosomal DNA FISH Techniques

Conventional cytological method is limited for polyploidy plant genome study, especially sugar cane chromosomes that show unstable numbers of each cultivar. Molecular cytogenetic as fluorescent in situ hybridization (FISH) techniques were used in this study. A basic chromosome number of sugar cane was estimated with three information; 1) number of 18S rDNA sites, 2) number of 5S rDNA sites and 3) total number of chromosomes. 18S and 5S rDNA were located by FISH techniques, the number of hom (e) ologous sites were illustrated in range of 7 to 9 and 13 to 15 sites. 110 chromosome numbers were shown in tapetal cells of flower buds of sugar cane. The implications of these results can predict about 14 basic chromosomes numbers but 5S rDNA seem reliable indicate for basic chromosome number and 18S rDNA were discussed about nucleolar dominance phenomenon of the sugar cane “KPS 00-25” cultiva

Rescue of Deletion Mutants to Isolate Plastid Transformants in Higher Plants

Plastid transformation is an attractive alternative to nuclear transformation enabling manipulation of native plastid genes and the insertion of foreign genes into plastids for applications in agriculture and industrial biotechnology. Transformation is achieved using dominant positive selection markers that confer resistance to antibiotics. The very high copy number of plastid DNA means that a prolonged selection step is required to obtain a uniform population of transgenic plastid genomes. Repair of mutant plastid genes with the corresponding functional allele allows selection based on restoration of the wild type phenotype. The use of deletion rather than point mutants avoids spontaneous reversion back to wild type. Combining antibiotic resistance markers with native plastid genes speeds up the attainment of homoplasmy and allows early transfer of transplastomic lines to soil where antibiotic selection is replaced by selection for photoautotrophic growth. Here we describe our method using the wild type rbcL gene as a plastid transformation marker to restore pigmentation and photosynthesis to a pale green heterotrophic rbcL mutant.</p