eChapters

Reproductive development determines flowering, pollination, fruit set, fruit growth, maturation, and ultimately horticultural productivity. This chapter introduces the physiological processes controlling flowering and reproductive development and explains the influence of environmental factors and plant hormones. Major plant growth regulators such as auxins, gibberellins, cytokinins, ethylene, and abscisic acid are considered in relation to horticultural applications. PGRs can influence rooting, flowering, fruit set, fruit size, thinning, ripening, dormancy, and senescence when used appropriately. The chapter emphasizes crop-specific concentration, timing, method of application, and environmental conditions, highlighting the importance of responsible PGR use for improving yield, quality, and marketability.

Gametogenesis

Male Gametogenesis: Male gametogenesis is the formation of male gametes and it occurs in the anthers. The end products of meiosis in higher plants are four microspores. Two mitotic divisions occur in microspores. The first division produces a tube nucleus and a generative nucleus. The second mitotic division occurs only in the generative nucleus resulting into production of two sperm cells (Fig. 3.1). The microspore is haploid (n) and develops into pollen grain in the anther, and the pollen grain further develops into a pollen tube on the stigma and in the style. The pollen tube, when it reaches the embryo sac is the fully developed male gametophyte and is trinucleate, two sperm cells and one tube nucleus.

A tissue culture laboratory can be placed in any geographical area. The internal controlled environmental conditions permit isolation with a minimum of external influence. To decide where to build the tissue culture laboratory, the following factors must be taken into consideration:

The response to selection (R) or selection advance or genetic gain is the difference between the mean phenotypic value of the offspring of the selected parents and the whole of the parental generation before selection. The response to selection is simply the change of population mean between generations following selection. Similarly, the selection differential (S) is the mean phenotypic value of the individuals selected as parents expressed as a deviation from the population mean (i.e., from the mean phenotypic value of all the individuals in the parental generation before selection). Response to selection is related to heritability by the following equation:

Introduction

Restriction enzymes, recognizes specific base sequences in double-helical DNA and cleaves at specific places, both strands of a duplex containing the recognized sequences. To biochemists, these exquisitely precise scalpels are marvelous gifts of nature. They are indispensable for analyzing chromosome structure, sequencing very long DNA molecules, isolating genes, and creating new DNA molecules that can be cloned. Werner Arber and Hamilton Smith discovered restriction enzymes, and Daniel Nathans pioneered their use in the late 1960s.

Principle

Since the entire DNA sequence of the bacteriophage is known, we know exactly where a restriction enzyme will cleave the viral DNA and the size of the DNA fragments generated. This restriction enzyme digest will create a pattern of specific DNA fragments that will serves as a “DNA ladder”.

Principle

Restriction digestion is one of the most common reactions performed in molecular biology. Restriction endonucleases (RE) are also known as restriction enzymes, which recognize short, specific DNA sequences. There are three types of RE: Type-I, Type-II and Type-III. Type II RE, most commonly used for DNA analysis and genetic engineering, cleave double-stranded DNA (dsDNA) at specific sites within or adjacent to their recognition sequences. All Type II RE recognises a unique nucleotide sequence at which it cuts a DNA molecule and nowhere else. The recognition sequence is often a six base pair palindromic sequence (the top DNA strand from 5' to 3' is the same as the bottom DNA strand from 5' to 3'), but others recognize four or even eight base pair sequences. Many RE will not cut DNA that is methylated on one or both strands of the recognition site, although some require substrate methylation.

REs can also differ in the way they cut the DNA molecule. Some enzymes cut in the middle of the recognition sequence, resulting in a flush or “blunt end”. Other enzymes cleave in a staggered fashion, resulting in DNA products that have short single-stranded overhangs (usually two or four nucleotides) at each end. These are often called “cohesive ends”, as these single-stranded overhangs could potentially come together again through complementary base-pairing, as shown below:

Restriction Enzymes

Restriction enzymes are enzymes isolated from bacteria that recognize specific sequences in DNA and then cut the DNA to produce fragments, called restriction fragments. Restriction enzymes play a very important role in the construction of recombinant DNA molecules, as is done in gene cloning experiments. Another application of restriction enzymes is to map the locations of restriction sites in DNA.

Restriction endonucleases, often called restriction enzymes, cut DNA within the molecule by hydrolyzing the phosphodiester bonds between the nucleotides. Restriction enzymes were discovered in bacteria and there are now more than 1200 known restriction enzyme types. These enzymes are named using a simple system. EcoRI, for example, was isolated from E. coli and was the first enzyme isolated from a particular strain, hence the designation of I. HaeIII was isolated from Haemophilus aegyptius. HindIII was isolated from Haemophilus influenzae, and was the third enzyme discovered in a particular strain.

Usually, restriction enzymes only cut the DNA at or near a very specific nucleotide sequence known as a recognition site. This “restrictive” nature of these enzymes allows molecular biologists to pin-point exactly where the DNA is to be cut. In many cases, the recognition site of a restriction enzyme consists of a palindromic sequence where the order of the bases is read the same on each side of the helix. The two sides of the helix are antiparallel and the recognition sequence reads from 5’ to 3’ on either side of the helix. For example the recognition site for EcoRI is as follows;

Introduction

The primary tools used by the molecular biologists in manipulating DNA are restriction enzymes and other DNA/RNA modifying enzymes. Over the years, the number and uses of these enzymes have increased as new molecules have been discovered. Restriction endonucleases are bacterial enzymes that cleave double-stranded DNA. They are present in bacteria presumably to destroy DNA from foreign sources. By cleaving the foreign DNA at specific sites. The host bacterial DNA os protected from cleavage because specific recognition sites are modified, usually by methylation form methylase at one of the bases in the site, making the site no longer a substrate for RE cleavage. Host bacteria used to propagate cloned DNA in the laboratory arte usually mutant in the host restriction genes, thus their intracellular enzyme activities will not destroy the foreign recombinant sequences. The endonuclaase with its accompanying methylase is called a restriction modification (R-M) system.

At least four different kinds of R-M systems exist characterized on the basis of the subunit composition, cofactor requirements and type of DNA cleavage. Most characterized enzymes belong to type II class, together with type IIS class. They comprise the commercially available restriction enzymes used for DNA analysis and manipulation. Type I and Type III enzymes are relatively uncommon and a few additional enzymes fit none of the classes.

Among more than 3000 different enzymes isolated from bacterial strains many share common specificities. Restriction enzymes that recognize identical sequences have been called as isoschizomers.

The genetic information which makes up the genes of higher plants is stored in DNA sequences of the nuclear chromosomes and the organelle genome. There is such an enormous amount of DNA in higher plant cell, that no two organisms are likely to be identical in DNA base sequence. 

In 1985, Sir Alec Jeffreys developed restriction fragment length polymorphism (RFLP), which quickly became the standard technique for DNA testing throughout the 1980s. RFLP provided the world with the first form of genetic testing based on DNA, the body’s genetic material.

Definition : RFLP analysis is a difference in homologus DNA sequences that can be detected by the presence of fragments of different lengths after digestion of the samples in question with specific restriction endonucleases (RE).

A restriction fragment length polymorphism , or RFLP, (commonly pronounced “rif lip”), is a variation in the DNA sequence of a genome that can be detected by breaking the DNA into pieces with restriction enzymes and analyzing the size of the resulting fragments by gel electrophoresis. It is the sequence that makes DNA from different sources different, and RFLP analysis is a technique that can identify some differences in sequence (when they occur in a restriction site). Though DNA sequencing techniques can characterize DNA very thoroughly, RFLP analysis was developed first and was cheap enough to see wide application. Analysis of RFLP variation was an important tool in genome mapping, localization of genetic disease genes, determination of risk for a disease, genetic fingerprinting, and paternity testing.

Retrospection

India is a unique country in the world with many religions and cultural diversity. Indian culture has been continuing through the ages because of its adjustability with others and openness. Unity in diversity is one of the major characteristics of Indian culture. It is a diverse nation where people of many religions live together peacefully. Religion in India is characterized by the diversity of religious beliefs and practices. In Indian history, religion has played an important role in the cultural set up of the country.

Hinduism is the main Indian religion and considered as a universal order, a way of life which is followed by the people. The origin for Hinduism is very old having no exact date of its inception. This is a belief and practices followed by people through the ages and considered as a process of humanity. The earliest known sacred text of Hinduism is ‘Vedas’ which dates back to at least 3000 BCE. However, it is believed that these texts are of divine origin and timeless. Hinduism was more philosophically and theoretically described in Upanishads, written roughly between 800 and 400 BCE. The Mahabharata and Bhagavad-Gita are other best-known book and widely cited on Hinduism. The Ramayana is the other most important epic book on Hinduism which teaches about life, principles, duty, justice etc. through the life-story of Rama.

Dasharatha was a great Hindu king of the Kosala kingdom and Ayodhya was the capital from where He ruled. He was the son of King Aja of Kosala and Queen Indumati of Vidarbha. Dasharatha had three primary queens namely - Kausalya, Sumitra, and Kaikeyi. Unfortunately, the King had no sons to continue his dynasty after him. Despaired Dasharatha summoned Rishyashringa, a forest rishi for a ritual to fulfill his desire of have sons. By that way, the King had four sons. Queen Kaushalya gave birth Rama and similarly, Kaikeyi gave birth Bharata and Sumitra gave birth two sons, namely - Lakshmana and Shatrughna. Lakshmana was very much attached and devoted to Rama while Shatrughna was devoted to Bharata.

What are Retrotransposons?

The presence of mobile elements, capable of moving from one region of the genome to another, has been known and thoroughly studied since the 1950’s. These elements are capable of exciding and inserting themselves from one region of the chromatin to another, through their peculiar terminal repeat regions. This process is mediated by enzymes encoded by the transposon itself.

Retrotransposons are a class of mobile elements, widespread in plant genomes, where they account for a significant part of noncoding DNA. Barley, for example, contains thousands of copies of the Bare-1 element, dispersed throughout the euchromatic regions of chromosomes and exhibiting sequence identity higher than 95% to each other.

Unlike other classes of mobile elements, retrotransposons insert into the genome via an RNA intermediate, synthesized by their own reverse transcriptase, without the loss of the parental copy. Unique sequence properties show that retrotransposons of the Ty1-copia group are close to retroviruses, and their mode of integration in the host genome is also very similar.

The number and exact point of insertion of retrotransposons may differ from one individual to another and this can be exploited to characterize individuals, varieties or species, depending on how recent the transpositional event is. Furthermore, insertion of a retrotransposon is an irreversible event, a characteristic that can be helpful when tracing an ancestry.

Introduction
It has been universally acknowledged that breeding for hybrids is the most effective conventional plant breeding technique. When two genetically distinct parent lines typically inbred lines are crossed, the resultant progeny is referred as a hybrid. It has been demonstrated that hybrids are larger, more productive, and hardier than their parental lineages. When we talk about hybrids, it is also imperative to understand two other related jargons: heterosis and hybrid vigour. The word “heterosis” was first coined in 1914 by Gottingen to refer the hybridization process in which two homozygous individuals (inbreds) are crossed to produce a hybrid. Darwin first used the phrase “hybrid vigour” in 1876 to denote the superiority of the F1 offspring produced by heterosis (i.e., the hybrids show superior performance with increased characteristics when compared with the two parental lines).
In general, the goal of plant breeding is to create better crop varieties by utilizing genetic variation found in the germplasm of the given plant species. Traditionally, two genetically different accessions of the given crop were selected and crossed to produce a hybrid. The genotype of this hybrid is the outcome of the fusion of the male and female gametes, those produced a zygote, from which the progeny eventually emerged. It would be appropriate to recollect the point that a plant’s life cycle consists of a gametophytic generation that produces gametes. Therefore, the genotypes of the gametophytes reflect the genetic diversity of the gametes.
Gametophytes are derived from spores, which are created via sporophytic reproduction during the course of the plant’s life cycle. Meiosis is a specialized kind of cell division that produces spores from differentiated cells that are located in the reproductive organs of plants. Two important processes (viz., recombination and chromosomal segregation) occur during meiosis and this results into independent assortment and creation of novel combinations of the genetic components of a diploid genome into a haploid genome of the gametophytes. Consequently, the genotype of a progeny is made up of the genotypes of the male and the female gametes that merged to create a new sporophyte. Therefore, meiosis can be viewed as a crucial stage in the life cycle of a plant and is responsible for generating genetic diversity. To generate desired plants with novel traits, this diversity or variability is frequently employed. In many cases, a hybrid plant outperforms a homozygous (parental) plant because it combines the best traits of the two parents. However, the creation of such hybrids is quite difficult. In order to evaluate the combining ability of hybrids, multiple putative parental lines are first produced at homozygous conditions by employing several generations of inbreeding and selection. The best pairs are then kept, and the corresponding parental lines produce a vigorous hybrid. Although hybrids were originally generated in maize during the 1930s, they are now employed in a wide variety of economically important plant species because they significantly surpass their parental inbreds. The main reason quoted for the wide-spread application of hybridization in breeding programs is because of the reason that hybrid seeds are easier and faster to growing homogenous plants

ABSTRACT

This paper provides an overview on existing IP protection and registration systems of India and adds information culled from publicly available sources pertaining to modernization efforts of Government of India towards establishment of foolproof IP protection in accordance with international treaties, conventions and bilateral and multilateral agreements.

Rice (Oryza sativa L., 2n = 2x = 24, monocot, family Poaceae formerly called as Gramineae, grass family, self-pollinated crop) is one of the Four Big Crops (rice, wheat, corn and soybean) globally and is the world’s most important food crop after wheat and corn. In crop year 2018, there was around 167 million hectares of paddy (rough, un-milled rice) cultivated area worldwide with production of 782 million tons resulting into a productivity of 4.68 tons/ha. The top ten countries in rice production during this period globally were China, India, Indonesia, Bangladesh, Vietnam, Thailand, Myanmar, Philippines, Brazil and Pakistan. Chinese production was 212 million tones followed by India with 172 million tons (FAOSTAT). Indian production of 172 million tons of rough rice can be converted into milled rice by multiplying a conversion factor of 0.67 and this way it comes to 115 million tons.

I. INTRODUCTION

Over the last few decades, protection of intellectual property rights (IPRs) has become a global issue. The existing international intellectual property rights regime has been governed by the Trade- Related Aspects of Intellectual Property Rights (TRIPS) Agreement.1 Inspite of maturity of intellectual property laws in post TRIPS era it still desiderate to established coherence with human rights laws particularly right to health and maintain a distance with the socio-economic dimension of intellectual property protection. On the other hand the international human rights instruments from the very beginning appear to be considerable to make certain links with intellectual property rights.2 The issue that is now debated that the adoption of the TRIPS Agreement provisions have serious impact on the realization of certain basic human rights such as the right to health because of non availability and affordability of much needed medicines for the poorer populations of the world particularly in majority of developing countries. Meanwhile TRIPS flexibilities as confirmed by the Doha Declaration on Public Health were adopted in 2001 by the global community to address medicines as public health issue.3 Keeping in view the above developments the problem of right to health and the relationship between medical patents and access to medicine in the case of the HIV/ AIDS and T.B epidemics is complex challenge in the context of the 2030 Agenda for Sustainable Development Goals in 2015 adopted by Member States of the United Nations in Sep, 2015. One of the aims of the agenda is to ensure healthy lives and promote the well-being of people of ages.4 In such a situation it is high time to revisit the existing intellectual property regime in the broader context to protect everyone who is likely to be negatively affected by intellectual property standards imposed by developed nations. Under this backdrop the present paper wishes to examine the various kinds of potential impacts of intellectual property regime on the realization of access to medicine as part of human right to health as universal entitlements amongst the most disadvantaged and marginalized peoples and communities in the broader context of sustainable development goals.

11.1 Principles and Framework for Risk Assessment
11.1.1 Introduction
Risk assessment is an indispensable scientific tool in evaluating the safety and potential impacts of genetically modified (GM) crops on human health, animal welfare, and the environment. It provides a structured, evidence-based framework for identifying hazards, characterizing risks, and proposing mitigation strategies before the approval and commercialization of GMOs. In the context of biotechnology and regulatory biosafety, risk assessment ensures that GM crops meet both national and international standards for biosafety and sustainability.
11.1.2 Definition of Risk Assessment
Risk assessment is defined as the systematic process of identifying, evaluating, and characterizing the potential adverse effects arising from exposure to a GMO or its products, under specified conditions of use.
Risk = Hazard × Exposure
Where hazard is the potential to cause harm, and exposure is the likelihood of encountering the hazard.
11.1.3 Objectives of Risk Assessment in GM Crops
• To evaluate the safety of GM crops for human and animal consumption.
• To assess the potential environmental impacts, including effects on biodiversity, gene flow, and ecosystem interactions.
• To ensure that regulatory decisions are science-based, transparent, and precautionary.

Introduction 
In healthcare settings, risk management is not a novel idea. There is a wealth of research on risk control in the supply of medications, in-vitro diagnostics production, pharmaceutical manufacture, health information protection, & healthcare process management. Risk management is an important part of patient safety programs in developed countries. An important aspect of quality improvement in laboratories is risk management, which requires an evaluation of every testing method. All the working laboratories must perform this repeated preventive means at least once a year. The safety of laboratory workers, the caliber of results, and environmental preservation all depend on risk management in good laboratory practices (GLP). These are a few GLP risk management techniques. GLP- The Concept In accordance with excellent laboratory practice, risk management should be implemented by evaluating every laboratory procedure to guarantee the safety of both patients and staff. When a GLP facility implements a risk-based inspection procedure, risk assessments will be documented. QA inspection programs will frequently cover widespread operations, but the frequency & extent of the inspection may not take into account or represent the risk associated with each action. By putting quality control procedures into practice, one can learn about the shortcomings of a particular activity and use this information to focus the quality assurance program. According to the OECD Principles of GLP, or “the principles,” there are three different kinds of quality assurance inspections: process-, study-, and facilitybased. Risk Management The process usually begins with the creation of management committee for risk, which consists of important Laboratory staff members and a few ad hoc people chosen by the primary members. All technical laboratory procedures (pre-, examination-, & post-examination) will undergo a process flow analysis

Principle

RNA is negatively charged at neutral pH and when an electric field is applied, it migrates towards the anode. RNA retains much of its secondary structure during electrophoresis unless it is first denatured. The addition of formaldehyde to the agarose gel maintains the RNA in its linear (denatured) form. The following protocol is based on Formaldehyde Agarose Gel Method.