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Ebook Plant biotechnology: Principles and applications - Part 2

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Ebook Plant biotechnology: Principles and applications - Part 2

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 ts is distributed into the nucleus, plastids, and mitochondria. Plastid has a central role of carrying out photosynthesis in plant cells. Plastid tran

sformation is an advantage to nuclear gene transformation due to higher expression of transgenes, absence of gene silencing and position effect, and t Ebook Plant biotechnology: Principles and applications - Part 2

ransgene containment by maternal inheritance, i.e.. plastid gene inheritance via seed not by pollen prevents transmission of foreign DNA to wild relat

Ebook Plant biotechnology: Principles and applications - Part 2

ives. Thus, plastid transformation is a viable alternative to conventional nuclear transformation. Many genes encoding for industrially important prot

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 ese advances, it remains a challenge to achieve plastid transformation in non-green tissues and recalcitrant crops regenerating via somatic embryos. I

n this chapter, we have summarized the basic requirements of plastid genetic engineering and discuss the current status and futuristic potential of pl Ebook Plant biotechnology: Principles and applications - Part 2

astid transformation.7.1IntroductionGenetic material in plants is divided into three organelles of the nucleus, mitochondria. and plastid. The plastid

Ebook Plant biotechnology: Principles and applications - Part 2

when present in green form in plant is called as chloroplast. which carries its own genome and expresses heritable traits (Ruf et al. 2001). Chloropl

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 AbdinDepartment of Biotechnology. Jamia Hamdard. New Delhi 110062. Indiap. SoniCTPD, Department of Biotechnology, Jamia Haindard, New Delhi 110062, In

dias. Kumar (El)International Centre for Genetic Engineering and Biotechnology.Anina Asaf Ali Marg. 110 067 New Delhi. Indiae-mail: skrhode@icgeb.res. Ebook Plant biotechnology: Principles and applications - Part 2

in© Springer Nature Singapore Pte Ltd. 2017M.z. Abdin et al. (eds.). Plant Biotechnology: Principles and Applications, nni IO IM7/O7V nil in ÌOĂI s 11

Ebook Plant biotechnology: Principles and applications - Part 2

91192M.z. Abdin et al.in 1986 by two Japanese research teams. Since then, over hundreds of chloroplast DNAs from various plant species have been seque

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 lastids per cell and the number of ptDNA per plastid vary species to species. For example, an Arabidopsis thaliana leaf cell contains about 120 chloro

plast organelles and harbors over 2000 copies of the 154 Kb size plastid genomes per cell (Zoschke et al. 2007). whereas Nicotiana tabacum leaf cell c Ebook Plant biotechnology: Principles and applications - Part 2

ontains about 10-100 chloroplast organelles per cell and harbors over 10.000 copies of ptDNA per cell (Shaver et al. 2006). The photosynthetic center

Ebook Plant biotechnology: Principles and applications - Part 2

of the plant cells and eukaryotic algae provides the primary source of the world’s food (Wang et al. 2009). Other important activities that occur in p

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 Daniell 2007).Transformation of the plastid genome was first accomplished in Chlantydonionas reinhardtii, a unicellular alga (Boynton et al. 1988). f

ollowed by plastid transformation in /V. tabacum, a multicellular flowering plant (Svab et al. 1990; Daniell et al. 2004). Plastid transformation sinc Ebook Plant biotechnology: Principles and applications - Part 2

e has been extended to Porphyridium, a unicellular red algal species (Lapidot et al. 2002). and the mosses Physcomitrella patens (Sugiura and Sugita.

Ebook Plant biotechnology: Principles and applications - Part 2

2004) and Marchantia polymorpha (Chiyoda et al. 2007). In higher plants, plastid transformation is reproducibly performed in ;V. tabac II m (Svab and

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 t el al. 2005: Kanamoto et al. 2006). potato (Nunzia 2011). and cabbage (Liu et al. 2007: Tseng et al. 2014). Monocols as a group are still recalcitra

nt to plastid transformation. It is assumed that in the next few years, there may be surge in commercial applications using this environmental-friendl Ebook Plant biotechnology: Principles and applications - Part 2

y technology due to several advantages over conventional nuclear transformation, like gene containment and higher expression levels of foreign protein

Ebook Plant biotechnology: Principles and applications - Part 2

s, the feasibility of expressing multiple proteins from polycistronic mRNAs. and gene containment through the lack of pollen transmission (Kiltiwongwa

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 enetically stable chloroplast transgenic also know n as transplastomic plant, all plastid genome copies should be uniformly transformed with foreign g

ene.7.2Tools and Elements for Chloroplast EngineeringRuhlman et al. (2010) emphasized the role of endogenous regulatory elements and flanking sequence Ebook Plant biotechnology: Principles and applications - Part 2

s for an efficient expression of transgenes in chloroplasts of different plant species.7 Plastoine Engineering: Basics Principles and Applications1937

Ebook Plant biotechnology: Principles and applications - Part 2

.2.1 PromotersAn efficient gene expression level in plastid is determined by the promoter. It contains the sequences which are required for RNA polyme

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 ne, the first requirement is a strong promoter to ensure high levels of inRNA. Chloroplast-specific promoters are essential to ensure an efficient acc

umulation of foreign protein into chloroplasts in algae and plants (Gao et al. 2012; Sharma and Sharma 2009).Plastid transcription is regulated by the Ebook Plant biotechnology: Principles and applications - Part 2

combined actions of two RNA polymerases recognizing different promoters, a T7-like single-subunit nuclear-encoded polymerase (NEP) and a bacterium-li

Ebook Plant biotechnology: Principles and applications - Part 2

ke a20p' plastid-encoded polymerase (PEP). Transcription in undifferentiated plastids and in non-green tissues is primarily regulated by the NEP. The

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 Many plastid promoters contain both the PEP and NEP transcription start sites (Allison et al. 1996; Hajdukiewicz et al. 1997).The 16S ribosomal RNA pr

omoter (Prrn) like psbA and alpA gene promoters are commonly used for chloroplast transformation. These promoters drive the high level of recombinant Ebook Plant biotechnology: Principles and applications - Part 2

protein expression in plastid transformation. Prr/Ỉ contains both PEP and NEP transcription start sites, whereas P/>.vM contains only a PEP transcript

Ebook Plant biotechnology: Principles and applications - Part 2

ion start site (Allison el al. 1996).7.2.25'UTRsThe 5' ƯTR is important for translation initiation and plays a critical role in determining the transl

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 al. 1994). Many reports have revealed that translational efficiency is a ratelimiting step for chloroplast gene expression (Eberhard et al. 2002). Th

us, 5' UTRs of plastid mRNAs are key elements for translational regulation (Nickelsen 2003). and many chloroplast genes are regulated at the posttrans Ebook Plant biotechnology: Principles and applications - Part 2

criptional level (Barkan 2011). However, the nature of these internal enhancer sequences has not been studied well (Klein et al. 1994).The most common

Ebook Plant biotechnology: Principles and applications - Part 2

ly used 5' UTRs are those of the plastid psbA gene. rbcL. and the bacteriophage T7 gene 10. It has been incorporated into many chloroplast transformat

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 2003; Venkatesh and Park 2012).194M.z. Abdin el al.7.2.33'UTRsThe 3' ƯTR plays an important role in gene expression, and it contains the message for

transcript polyadenylation that directly affects mRNA stability (Chan and Yu 1998). Plastid 3' UTRs, cloned downstream of the slop codon, contain a ha Ebook Plant biotechnology: Principles and applications - Part 2

irpinloop structure that facilitates RNA maturation and processing and prevents degradation of the RNA by ribonucleases (Stern cl al. 2010). Valkov cl

Ebook Plant biotechnology: Principles and applications - Part 2

al. (2011) reported the roles of alternative 5' UTR and 3' UTRs on transcript stability and translatability of plastid genes in transplaslomic potato

Chapter 7Plastome Engineering: Basics Principles and ApplicationsMalik Zainul Abdin, Priyanka Soni. and Shashi KumarAbstract Genetic material in plant

Ebook Plant biotechnology: Principles and applications - Part 2 ficiency of transgcncs expression in chloroplasts (Monde et al. 2000). 3' UTRs like rpsìồ, rbc!.. psbA. and rp!32 3' UTRs are being commonly used in c

hloroplast transformation system. The most commonly used 3' UTR is VpsbA (Gao et al. 2012: Kittiwongwattana et al. 2007). Ebook Plant biotechnology: Principles and applications - Part 2

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