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This book presents up-to-date information on various vector-less/direct (physical, chemical) and vector-mediated/indirect (Agrobacterium-mediated) plant transformation techniques. It summarizes various strategies that facilitate a gene from lower organism to be expressed in higher plants and also in silico designing of synthetic gene for higher expression. It also highlights the importance of strong promoters to drive the expression of transgene(s). This book encompasses the advantages and drawbacks of cisgenesis and transgenesis, their implications towards sustainable crop improvement, and their future prospects. The importance, limitations, challenges, recent developments, and future prospects of molecular pharming is also discussed.  The book concludes with a chapter that summarizes the major contribution of GM-crops towards global food security and economy, advances in genome editing for crop improvement, challenges and risk associated with the release of GM-crops, and the future of GM technology.

This book is meant for students and researchers in the field of life sciences, food science, and agriculture. 

Cisgenics and Transgenics: Strategies for Sustainable Crop Development and Food Security

Preface

Foreword

Abbreviations

Section 1:Plant transformation techniques

1. Direct DNA Transfer

2. Physical gene transfer methods

2.1 Electroporation

2.2 Particle bombardment/microprojectile

2.3. Macroinjection

2.4. Microinjection

2.5. Liposome-mediated transformation

2.6. Silicon carbide-mediated transformation

2.7. Ultrasound/sonication -mediated transformation

2.8. DNA transfer via pollen

3. Chemical gene transfer methods

3.1. PEG-mediated gene transfer

3.2. Calcium-phosphate co-precipitation

3.3. The polycation DMSO technique

3.4. DEAE dextran procedure

3.5.DNA imbibition by cell, tissues, embryos and seeds

4. Indirect DNA transfer

4.1. Agrobacterium-mediated transformation

4.1.1. Importance of Agrobacterium-mediated transformation

4.1.2. Transformation protocols :

4.1.2.1. Agrobacterium-mediated transformation of Arabidopsis

4.1.2.2. Agrobacterium-mediated transformation of rice

4.1.2.3. Agrobacterium-mediated transformation of chickpea

4.1.2.4. Agrobacterium-mediated transformation of tomato

4.1.2.5. Agrobacterium-mediated transformation of potato

4.1.2.6. Agrobacterium-mediated transformation of cotton

4.1.2.7. Agrobacterium-mediated transformation of stevia

4.1.2.8. Agrobcterium-mediated transformation of sugarbeet

4.1.2.9. Agrobcterium-mediated transformation of maize

4.1.2.10. Agrobcterium-mediated transformation of melon

4.1.2.11. Agrobcterium-mediated transformation of poplar

4.1.2.12. Agrobcterium-mediated transformation of sugarcane

4.1.2.13. Agrobcterium-mediated transformation of apple

4.1.2.14. Agrobcterium-mediated transformation of flax

4.1.2.15. Agrobcterium-mediated transformation of sweet pepper

4.1.2.16. Agrobcterium-mediated transformation of soybean

4.1.2.17. Agrobcterium-mediated transformation of  canola

4.1.2.18. Agrobcterium-mediated transformation of  alfalfa

4.1.2.19. Agrobcterium-mediated transformation of squash

4.1.2.20. Agrobcterium-mediated transformation of eggplant

Section 2: Strategies to enhance the expression of the transgene in plants

5. Enhancement of transgene expression in plants 

6. Designing of coding sequence of the gene

6.1. Avoiding sequence motifs and codons that direct mRNAdegradation

6.2. Incorporation of elements for high-level expression

6.2.1.Use of strong promoter(s)

6.2.2. Untranslated regions (UTR) and sequences

6.2.3. Translation initiation context (TIC)

7. Subcellular targeting of recombinant protein for accumulation and stability

Section 3: Cisgenics and crop improvement

8. Difference between cisgenics and transgenics

9. Limitations of cis-genesis

10. Cis-genesis and sustainable crop improvement

                                                

Section 4: Transgenics and crop improvement

11. Crop improvement through transgenic technology

12. Transgenics for herbicide resistance

12.1.The story of transgenic mustard

12.2 Transgenics for pest resistance

13. Bt-technology

13.1. The story of BT cotton

13.2. The story of BTbrinjal

14. Transgenics for disease resistance

14.1. Pathogenesis related proteins (PR proteins)

14.2. Ribosome inactivating proteins

14.3. Use of anti-microbial protein

14.4. Pathogen-derived resistance (PDR) for viral diseases

14.5. Non-pathogen-derived resistance (non-PDR) for viral diseases

15. Transgenics for stress resistance

15.1. Production of osmoprotectants in plants

15.2. Na+/H+ antiporters for improved salt tolerance

15.3. COR and heat-shock regulons

15.4. Expression of enzymes involved in scavenging ROS

15.5. Production of antioxidants

15.6. Transgenics for nutrient biofortication and yield

16.Engineering plant protein composition

17. Engineering plants for Vit A composition

18. Biofortified rice

19. Biofortified maize and cassava

20.Engineering plant mineral composition

21. Biofortified rice and wheat

22. Enhancement of photosynthesis for improved yield

Section 5:Molecular pharming

23. An introduction to molecular pharming

24. Molecular pharming of carbohydrates

25. Molecular pharming of lipids

26. Molecular pharming of proteins

Section 6: Future prospects of GM plants

27. The current state of transgenic crops

28. Ethical issues and risks associated with the transgenic crops

29. Advances in genome editing for crop improvement