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## Meet the editor

Dr. Kin-Ying To received a Ph.D. degree at the Institute of Botany, National Taiwan University, Taiwan. After postdoctoral training at Chang-Gung University (Taoyuan, Taiwan), Harvard University (Boston, USA), and Academia Sinica (Taipei, Taiwan), Dr. To joined the Agricultural Biotechnology Research Center, Academia Sinica, Taiwan, in 1999. Currently, Dr. To is working as an Associate Research Specialist. Dr. To has published

over 30 research articles and reviews, 3 book chapters, and also received 2 patents and several research grants. In addition, Dr. To has been serving as a reviewer for a range of journals and a lecturer in some courses. Research interests in Dr. To's laboratory include plant tissue culture and genetic engineering in valuable medicinal and crop plants, *Agrobacterium*-mediated transformation, molecular breeding and plant biotechnology.

Contents

Agrobacterium-Mediated Transformation

*Brassica oleracea* Transformation *by Penny Hundleby and Monika Chhetry*

RNA Interference: An Overview

in Higher Plants

Improvement

*and Wagh Yogesh Sahebrao*

*by Risha Amilia Pratiwi and Muhammad Imam Surya*

*by Devaiah Kambiranda, James Obuya and Janana Snowden*

Construction and Evaluation of Chloroplast Expression Vectors

*by Jitesh Kumar, Khushbu Jain, Priyanka Kumari, Auroshikha Mohanty,* 

Grapevine Improvement through Biotechnology

*by Po-Yen Chen, Yung-Ting Tsai and Kin-Ying To*

*Kumari Rajani, Ravi Ranjan Kumar and Tushar Ranjan*

RNAi Induced Gene Silencing Journey from Simple dsRNA to High-Throughput Intron Hairpin RNA Construct in Crop

*by Jadhav Pritam Ramesh, Ekatpure Sachin Chandrakant* 

*by Getachew Alamnie and Berhanu Andualem*

**Preface XI**

**Chapter 1 1**

**Chapter 2 15**

**Chapter 3 37**

**Chapter 4 47**

**Chapter 5 77**

**Chapter 6 99**

**Chapter 7 111**

Biosafety Issues of Unintended Horizontal Transfer of Recombinant DNA

## Contents


Preface

It is estimated that a 70-100% increase in cereal food supply is required to feed the predicted world population of 9.8 billion people by 2050. Unfortunately, climate change, decreasing environmental resources, and increasing human population are major factors limiting agricultural production worldwide. Climate change will aggravate the effects on crops of various stresses such as pathogen attack, extreme temperatures, drought, salinity and submergence with water, and is set to have a profound impact on food production. To breed crops with increased yield and resistance to biotic/abiotic stresses, it is necessary to effectively utilize the current methodologies and further develop new breeding technologies as soon as possible.

Genetic crossing, selection of natural or artificial mutations, and transgenics, are the major techniques for plant breeding. Among them, plant transformation has been widely adopted as the most effective method in understanding how the plant works, improving crop characteristics, and producing value compounds. Back to 1974, Marc Van Montagu and Jeff Schell at Ghent University (Belgium) were credited with discovering that the soil bacterium Agrobacterium uses a plasmid, now known as the Ti plasmid, to transfer its genetic material to a plant cell. Since then, Agrobacterium-mediated transformation has been rapidly developed and recognized as the most powerful method for genetic engineering and creation of transgenic plants. In 1987, the first field trial of a genetically modified (GM) potato was reported in the United Kingdom, and the transgenic potatoes had the GUS reporter gene. In 1994, Flavr Savr tomatoes (Calgene company) went on sale in the USA. The polygalacturonase gene which degrades pectin in cell walls and results in the softening of fruit, was knocked out in the transgenic tomato, so that the GM fruits remain firm for a longer time, and allowed them to develop a better flavor. This GM tomato was the first commercially available GM food. However, controversial debates regarding public concerns, risks, and biosafety issues have arisen since the first GM food became available on the market, and these controversies seem to continue. Nowadays, only a few important crops such as soybean, corn, canola, cotton, etc., have been successfully engineered for specific characteristics, and are

The first chapter of this book is an overview of Agrobacterium-mediated

transformation. This chapter extensively describes updated information related to Agrobacterium-mediated transformation, including the history of Agrobacterium, the natural process of Agrobacterium, comprehensive mechanism of gene transfer from Agrobacterium to host plant, and the general protocol of Agrobacteriummediated transformation in higher plants. In brief, this chapter provides fruitful information regarding Agrobacterium-mediated transformation, and will definitely be useful in the field of transformation. The second chapter extensively describes grapevine improvement through biotechnology. Grapes are one of the most widely cultivated fruit crops worldwide. Authors in this chapter mention the achievement of the sequencing of the multiple grape genomes and its applications to grape improvement. For grapevine breeding, several techniques including marker-assisted selection, RAPD markers, microsatellites, single-nucleotide polymorphism (SNP), genetic transformation, etc., have been clearly described. This information should

available on the market.
