Plant genetic transformation and molecular markers By Ashwani Kumar Pointer Publishers Jaipur India pp 288 ISBN 978-81-7132-613-6 The last three decades have seen momentous developments in plant transformation technologies; such that a large number of transgenic crop plants have now been released for commercial production. Advances in the technology have been due to development of range of Agrobacterium-mediated and direct DNA delivery techniques, along with appropriate tissue culture techniques for regenerating whole plants from plant cells or tissues in a large number of species. Now a days, Agrobacterium-mediated transformation, Virus-mediated, chemical mediated microinjection, elctroporation, particle-bombardment and other transformation methods are routinely used. Researches in plant transformation are mostly going on not much on the introduction of DNA into plant cells, but rather more on the problems associated with stable integration and reliable expression of the DNA after integration. Various transformation technologies used for various crop plants are explained in chapter contributed by Professor Anil Kumar from Indore. Rapid progress in nanotechnology has enabled its applications in the diverse fields that encompass medicine, engineering and plant biotechnology. Attempts were made to produce nanoparticles in the plants. One approach is to use plants as a production host for therapeutic proteins, which would be formulated with the nanoparticles and delivered through the oral route for rapid and timely absorption by the mucosal membranes. In second approach, nanoparticles would be employed to deliver the gene of interest; instead of conventional techniques of Agrobacterium mediated transformation or gene bombardment for a more precise mechanism of the foreign gene integration into the host plant system. The problem in the production of recombinant proteins up to a desirable level using nanotechnology is of considerable interest. The uses of nanotechnology in biotechnology are explained in detail by Bapat et al. second chapter. Agrobacterium tumefaciens, a Gram negative bacterium that allows the introduction of genetic information into cells. This has revolutionized the field, improving cultivars by expressing traits that would not be possible to obtain by classical methods. The process of transformation by Agrobacterium requires the presence of several genetic Elements. Details are provided by SOLÍS-RAMOS et al in 3rd chapter. Chloroplasts are photosynthetic organelles containing a part of plant genome usually called plastome. Introduction of foreign genes into plastids and creation of transplastomic plants has some advantages over classic nuclear transformation, methods but chloroplast gene expression machinery is diverse and complicated. A review is illustrated with the examples of successful application of this knowledge in introduction and expression of foreign genes in chloroplasts ( Lysenko chapter 4th). Clonal propagation of elite genotypes, which has the advantage of capturing all the genetic superiority without involving any gene segregation, combined with genetic transformation is a particularly attractive approach, as both can confer new or enhanced traits for forest tree species. European chestnut (Castanea sativa Mill.), English oak (Quercus robur L.) and European beech (Fagus sylvatica L.), belonging to the Fagaceae family, are of great ecological, environmental and economical interest, and are often dominant in northern temperate forests. In the review, we summarize the state of the art of both organogenic and embryogenic processes for these three species, and their use for tree improvement through genetic transformation approaches (BALLESTER and VIEITEZ, chapter 5th ) Sugarcane is an important sugar crop throughout the world. Although a tropical crop, sugarcane’s growth and productivity is markedly affected by ambient temperature fluctuations. Efforts have been undertaken to understand the effect of growth limiting low and high temperatures on sugarcane at whole plant and molecular levels. Various strategies have been proposed to improve crops for tolerance to these stresses (WAHID et al chapter 6th) Grapevine (Vitis sp.) is one of the most important crops for humankind from an economic and cultural point of view. In this chapter 7th (TORRE et al) , the state-of-art on genetic transformation of grapevine will be examined, including the target tissue used for transformation, the design of expression cassettes as well as the main transformation methods (Agrobacterium vs. biolistic) used in both genetic improvement and functional genomics approaches. Somatic Embryogenesis is useful for plant propagation and transformation. Unfortunately, efficient protocols are still lacking for producing large amounts of well formed embryos in many economically important species. A great deal of efforts has recently been concentrated on the identification of molecular markers related to embryogenic competence, that could be used to select valid genotypes and tissues. Somatic Embryogenesis Receptorlike Kinase (SERK) genes have first been identified as markers of embryogenic competence. SERK genes seem to be ubiquitous in plants and are generally present as a small gene family. Some connections between the different pathways mediated by SERK proteins are probable, as suggested by the antagonistic regulation of the BR-dependent growth pathway and the cell death pathway in Arabidopsis, or the pleiotropic role of Oryza sativa SERK1 in SE and in the resistance to a fungal disease(MAILLOT et al, chapter 8th) Genetic analysis of plants is a basic requirement for studies in genetics and plant breeding. Marker, be it morphological, biochemical or molecular marker, play a key role in genetic studies. Genetic markers have evolved over time from morphological markers through isozyme markers to the most advanced DNA-based markers. The DNA-based molecular markers have very rapidly and efficiently complemented the classical strategies for genetic analysis. With the increasing availability of information on genome sequences for different plant species, the sequence specific molecular markers (e.g. SNP, TRAP) are becoming more important and useful over the random DNA markers (e.g. RAPD, DAF). In this chapter (KUMAR chapter 9th ) an overview of DNA-based molecular markers has been presented with the underlying principle and methodology employed. These molecular techniques, in particular the applications of molecular markers, have been used to scrutinize DNA sequence variation(s) in and among the crop species and create new sources of genetic variation by introducing new and favorable traits from landraces and related crop species. Markers can abet selection for target alleles that are not easily assayed in individual plants, minimize linkage drag around the target gene, and reduce the number of generations required to recover a very high percentage of the recurrent parent genetic background. An overview of the current stage of these developments, examines some of the special issues that have been used in applying molecular techniques to molecular breeding of various crops (Yadav chapter 10th) . A remarkable increase in total grain production was observed between 1960 and 1980, but only a marginal increase was realized during 1980-2000. Much of the early increase in grain production resulted from increase in area under cultivation, irrigation, chemical fertilizers and improved cultivars. Yield plateau has already been observed in several crops, therefore, most of the productivity gains in future will have to be achieved through better natural resource management and crop genetic improvement. Chapter 11th ( Kumar ) provides an overview of the status and availability of genomic resources for crop improvement. Morphogenesis and development are key processes for multi-cellular organisms; regulation of this process requires nuclear accessibility of several genes and specific transcription factors that modulate this process. In Arabidopsis homeotic genes have been found as key controllers which involved the transcription factor WUSCHEL and fourteen other transcription factors that have conserved regions with WUSCHEL know as WUSCHEL HOMEOBOX family of genes. These factors are involved in the control of meristem cells as well as the first steps of cell differentiation. The impressive conservation of nucleotide sequences of the homeoboxes has fostered the efficient and rapid isolation of homeobox-containing genes from diverse sources (SOSA et al chapter 12th ) . Thus the book provides present day state of art knowledge about genetic transformation technologies and use of molecular markers for crop improvement and increasing crop productivity. This book is aimed towards the students, researchers and teachers in the field of plant biotechnology in Universities, Agricultural and horticultural Institutions, Engineering Institutes and industries dealing with plant biotechnology. Ashwani Kumar . Biodata of Professor Ashwani Kumar Born 1946 (Bandikui) Rajasthan, B.Sc. Agra University, M.Sc.(Botany) University of Rajasthan, awarded gold medal for standing first in order of merit, Ph.D. (1971) on ‘Studies on growth physiology and biochemistry of some edible plant tissues in culture’ under supervision of Professor H.C.Arya and post doctoral with Professor Dr. K-H.Neumann, Justus Liebig Universitat, Giessen, Germany with award of Alexander von Humboldt Fellowship. Professor Ashwani Kumar was also selected through Indian Administrative Services examination 1972 into Indian Police Service which he declined due to his interest in teaching and research as better option This decision was also influenced by the advice of his father Professor Swami Dayal Tewari who studied Masters of Science (Botany) at Allahabad University. He was appointed Assistant Professor of Botany in 1986, Associate Professor of Botany in 1986 and Professor of Botany in 1996 at the University of Rajasthan, Jaipur. Throughout his carreerr he held many important administrative and research positions like Head Department of Botany and Co-ordinator SAP(1995-1998), Director, Life Sciences, 2001-2004, Director Central Library and Information Science (2000-2005), Member Senate, Research Board, Convener Board of Studies Botany and Biotechnology, VC nominee for the Academic Council of MDS University, Ajmer and VC nominee for governing body of Subodh PG College Jaipur. Member , Governing body Academic Staff College, University of Rajasthan Jaipur. Member Board of studies and Academic Council of several Universities, Advisory committee on Biotechnology, Govt of Rajasthan, Medicinal plant board of state Government, member selection committees of Rajasthan and Himachal Public Service Commissions. He along with members of COC introduced Integrated Biotechnology 5 year course in Rajasthan which has also been adopted by several other Universities. He has also edited 4 Books on plant biotechnology and tissue culture including one along with Professor Dr Neumann published by Springer, Germany. In recognition of his research contributions he was awarded Alexender von Humboldt Fellowship in 1977-1979, British Council Visitorship, U.K.(1986) Visiting Professorship at Tomyama Medical and Pharmaceutical University, Japan (1999-2000) INSA-DFG visiting Professorship at Germany, 1997 and DAAD award 1999. He had excellent knowledge of German language and has diploma in German besides certificate in French. He has made important contributions to the understanding of development of photosynthetic apparatus in cultured plant tissues at light reaction and photosynthetic carbon fixation. He has also carried out research projects granted by UGC., USDA-ICAR., MNES, CSIR, DST and DBT and FACT. He has been invited by various Universities including University of California and Chicago, USA, University of Laval, Canada, Wageningen University , Netherlands, and visited UK , Japan, France, Netherlands, Denmark, Sweden, Spain and Portugal for scientific interactions. He has been co-chair and chair at several International Conferences including International Botanical Conference, Berlin and several World European Union conferences held in Europe. He has published around 180 papers in National and International journals and is member of editorial board of IBS several journals including Current Trends in Biotechnology and Pharmacy, He has also been awarded fellowships by different societies including FABAP -Fellow Association of Biotechnology and Pharmacology. FBS, (Fellow of Botanical Society), FPSI (Fellow of Phytopathological Society), FISMPP Indian Society of Mycology and Plant Pathology, FMA-Fellow Mendelian Association. He has guided research to 35 students to Ph.D. in various branches of botany including plant pathology, plant tissue culture and morphogenesis, plant biotechnology, plant physiology, pharmacognostisc and ethnobotany and renewable sources of energy. Details of his work can be seen at the blog