eChapters

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. 

Developments in recombinant DNA technology have made it possible to transfer gene from any source across species barrier. In fact, transfer and expression of foreign genes in plant cells is now a routine procedure. Of the several methods tried so far, the system based on Ti plasmid of Aprobacterium tumefaciens appears to be most successful for transformation of plant cells.

Protoplasts are cells which have had their cell wall removed, usually by digestion with enzymes. Cellulase enzymes digest the cellulose in plant cell walls while pectinase enzymes break down the pectin holding cells together. Once the cell wall has been removed the resulting protoplast is spherical in shape. Digestion is usually carried out after incubation in an osmoticum (a solution of higher concentration than the cell contents which causes the cells to plasmolyse). This makes the cell walls easier to digest. Debris is filtered and/or centrifuged out of the suspension and the protoplasts are then centrifuged to form a pellet. On resuspension the protoplasts can be cultured on media which induce cell division and differentiation.

DNA can be sequenced in both double stranded and single stranded templates by cloning in plasmid or phage vectors. There are two sequencing methods.

M13 is a single stranded filamentous DNA phage. Vectors have been developed from M13 phase by Joachim Messing (1983), which are suitable for preparation of single stranded DNA required for sequencing by Sanger’s method. These are derivatives of M13 wild type (size 6.407 kb).

The polymerase chain reaction (PCR) is a very powerful new method used for in vitro to amplify a deoxynibonuclic specific region that lies between two regions of known DNA sequence single molecule to million folds in short time. This is achieved by an enzyme taq. DNA polymerase which is thermostable using this enzyme can synthesize microgram quantities of DNA of specified length and sequence from pictogram quantities of complex template in a few hours time.

Complementary DNA (cDNA) is a double stranded DNA which is an exact copy of the mRNA, i.e. it consists of group of molecules possessing the information contained in various mRNAs but with termini equivalent to or related to those produced by restriction endonucleases. The use of cDNA is an absolute prerequisite for the expression of eukaryotic proteins in bacteria.

Recombinant DNA is advancement in molecular technology which allows manipulation of genomes of higher organisms and dissection of their complex regulatory circuits. This new methodology is being used to understand the biochemistry of living organisms. The isolation of gene begins with the cloning of fragmented DNA of the organism into appropriate vector. 

Exchange of genetic material rarely occurs between unreleated organisms and even similar species may not exchange chromosomal genes. A certain degree of DNA sequence homology is a prerequisite for genetic recombination in vivo. However, recombination between DNA molecules in vitro is not subject to such taxonomic restrictions.

Nucleic acid hybridization requires denaturation i.e. the breakage of H-bridges between the complementary strands of the DNA either by heating at high temperatures or by treatment with denaturing chemicals like formamide. The restricted DNA, therefore, is denatured before transfer from the gel onto synthetic membrane since only single stranded DNA binds to nitrocellulose.

DNA fragment to be used as a probe should be purified by gel electrophoresis. The band can be located after staining with ethidium bromide and electroeluted.

The in vitro radioactive labeling of DNA fragment is accomplished by the incorporation of a radioactive nucleotide during strand synthesis with Klenow enzyme (large subunit of DNA polymerase-I).

Polysomes consist of two or more ribosome traversing a strand of mRNA, translating the nucleotide sequence into the corresponding amino acid sequence. Usually at any time, many ribosomes are translating an mRNA simultaneously forming a structure called a polyribosome or polysomes. One reason for isolating polyribosomes is to resolve them on sucrose gradients to determine the extent of ribosome loading. The second main reason is to provide a source of mRNA for in vitro translation.

RNA is transcriptional product of DNA. The total RNA includes three classes, rRNA, tRNA and mRNA. The ribonucleic acids exists as ribonucleo protein particles in intact cells. The ribonucleo protein complex is dissociated by SDS in to RNA and protein deproutinised by phenol and the free RNA left in the aqueous solution is precipitated in cold by adding alcohol. 

There are various methods available for isolation of high molecular weight genomic DNA from plants. The basic steps involved in all these methods are removal of cell wall, nuclear membrane, proteins, lipids, carbohydrates, cell debris, RNA without affecting the integrity of DNA. The two major factors that affect DNA integrity are action of endogenous nucleases and impact of mechanical shearing.

Enterobacteria phage ? is a bacterial virus, or bacteriophage, that infects the bacterial species Escherichia coli (E.coli). It was discovered by Esther Lederberg in 1950. The wild type of this virus has a temperate life cycle that allows it to either reside within the genome of its host through lysogeny or enter into a lytic phase, during which it kills and lyses the cell to produce offspring. Lambda strains, mutated at specific sites, are unable to lysogenize cells; instead, they grow and enter the lytic cycle after superinfecting an already lysogenized cell.

This is a procedure for transforming bacterial cells with circular DNA plasmids containing an origin of autonomous DNA replication and a selective marker (antibiotic resistance). Logarithmically growing cells are made permeable to DNA by incubation in CaCl2 and the DNA is taken up from the surrounding buffer.

Plasmid is a double stranded, self replicating, circular extra chromosomal DNA of bacterium. It is used in recombinant DNA experiments to clone genes from other organisms and make large quantities of their DNA. Plasmid can be transferred between same species or between different species. The sizes of plasmids range from 1-1000 kbps. Plasmids are part of mobilomes (total of all mobile genetic elements in a genome) like transposons or prophages and are associated with conjugation.

Plant biotechnology can be defined as the introduction of desirable traits into plants through plant tissue culture and genetic engineering techniques. The recombinant DNA technology is used to modify the expression of genes already present in the plants or to introduce new genes of other species with which the plant cannot be bred conventionally.

Non-Response

Non response occurs when respondents are not available or do not occur in the survey and refuse to respond by any means. This non response is mainly of two types either item non response or unit non response.

If we are conducting an online survey and mailing a questionnaire to various persons. But sometimes we get a reply for some questions and sometimes not i.e., in case of questions in which income is concerned. In some cases, the mail is ignored due to lack of time. So, if the whole questionnaire is ignored by any means, then it is known as non-response and if some of the questions are ignored due to their fussy nature such type of response is known as a unit non response. Such types of non-response problems are very common in surveys. Whenever we carry out a survey non response always exists because it is almost impossible to get every response from each individual.

The individual chosen for the sample is not ready to participate in the survey is known as non-response. This type of selection bias is unit/item non response.

In our previous chapters, we have discussed various sampling designs or techniques such as simple random sampling, stratified sampling, systematic sampling and cluster sampling recently we have discussed unequal probability sampling i.e., probability proportional to size sampling. In all these sampling designs the variable of interest (generally there is only one variable of interest) which we have considered and perform our estimation based on that single variable. There are some other variables which are related to our variable of interest and if we utilize those variables, we may gain some efficiency in our estimator i.e., if we take some correlated variables with our variable of interest or study variable by this, we can enhance our estimation. The variable which is correlated with the study variable is usually known auxiliary variable or supporting variable or supplementary variable or benchmark variable. This auxiliary variable is utilized in two stages either in the design stage or estimation stage.

Graunt (1662) was the first who used auxiliary information to estimate the population size of England. After this Laplace used auxiliary information for the estimation of the population in France.

Introduction

In some practical field situations, the results are described by some percentages or proportions. In agricultural field conditions, if any disease prevalence is to find then in the form of percentage or proportion, we can say how much disease rate is present i.e., 10 % or may be any other. So, in those situations, we need to describe our results in proportions. Similarly, in many fields such as medical sciences, social sciences and other allied sciences this type of results in proportions we need to describe.

Yet now we have discussed probability sampling techniques in which sampling units are assigned with equal probabilities. But sometimes in practice if want to draw a sample with unequal probabilities i.e., every sampling unit is assigned with varying probabilities and we draw a sample from these units with the help of equal probability methods, then the results are not as much efficient as compared to unequal probability sampling.

Simple random sampling is a procedure wherein the chance of selection of every unit in the required sample is equal. However, the use of simple random sampling is not advisable when the units differ substantially in size as the significance of unit size is ignored.

The basic idea of this type of probability technique is that chance of each unit of a population being selected in the sampling is proportional to its size. The units vary significantly in size and the variable under study is largely related to the size of the unit. Therefore, the probabilities fixed for each unit are proportional to their sizes.

Non-Probability sampling methods are also known as non-random sampling methods or deliberate sampling. These types of methods are reliant on the researcher’s ability to select members at random. These sampling methods are not fixed or pre-defined selection processes making it difficult for all elements of the population to have equal opportunities to be included in the sample.

Types of non-probability sampling techniques

1. Judgmental or Purposive sampling

This technique involves the purposive selection of units of samples based on the judgement of the researcher. The researcher selects the sample on the basis of his a priori knowledge and experience.

Multistage Sampling

Multistage sampling is a form of random sampling where samples are selected in a sequence of stages. Where each sample is drawn from the exclusive previously selected samples. This sampling is not done once but at multiple stages. It differs from cluster sampling in the sense that cluster uses all the subjects within a cluster whereas multistage selects a sample from within the selected group. Groups are randomly chosen from a population; subgroups from these groups are randomly chosen and then randomly chosen to be surveyed.

This is an extension of cluster sampling, which is carried out in multiple stages say two, three or four stages. In the first stage, the universe is divided into some clusters from which certain clusters are selected at random at the first stage samples. In the second stage, the selected first stage samples are again subdivided into some clusters from which again certain clusters are selected at random as the second stage samples. In the third stage, the selected second stage samples are again subdivided into some clusters from which certain clusters are again selected at random as the third stage samples.

Cluster Sampling (Meaning and Concept)

Cluster sampling is a special type of sampling which is used when we have to make a large survey such as a survey for the whole country and whole state etc. to know the privilege. Under this method, the total population is divided into some recognizable subdivisions known as clusters. Out of all the clusters, a given number of clusters are chosen at random. All the items covered by the selected clusters are included in the sample.

A sampling technique in which we divide the whole population into groups or subgroups in such a way that there is heterogeneity within each group and homogeneity among the groups and then a simple random sample of n groups is selected from a total of N groups of population and finally all the units of selected clusters from our sample.

Systematic Sampling (Meaning and Concept)

In this chapter, we are going to discuss the systematic random sampling technique. This is a random / probability / quantitative sampling method. In our previous chapter, we discussed the stratified random sampling technique. We have said that this technique is used when the population is heterogeneous and needs to be stratified into homogeneous groups. We have also discussed steps to be followed when using stratified random sampling as well as the strengths and limitations of stratified random sampling. Systematic sampling is a commonly employed technique if a complete and up-to-date list of the sampling units is available. The procedure involves choosing the very first object arbitrarily and the remaining members of the sample will be selected naturally according to any pre-decided sequence containing regular spacing of units. In this method, units of the population are numerically, geographically, and alphabetically arranged either in ascending or descending order (generally ascending order is preferred). When the ordering of the elements is related to the characteristic of interest,

Stratified Sampling (Meaning and Concept)

The word ‘stratify’ comes from the Latin word, meaning ‘to make layers’, thus in stratified sampling the whole population is considered to be classified into some mutually exclusive layers or subgroups or classes also called strata. The strata do not overlap and they constitute the whole population.

Why the population is considered to be divided into different strata (or layers or subgroups or classes)?

Simple Random Sampling (SRS)

Simple random sampling is probability sampling and refers to a technique of sampling where each and every unit of the population have an equal and independent opportunity of being selected and included in the sample during the sampling. In the SRS the selection of the sample depends on the chance factor and is also known as the method of probability sampling.

There are two ways in which SRS can be performed:

• SRSWR: Simple Random Sampling With Replacement
• SRSWOR: Simple Random Sampling Without Replacement

Sample Design

Sample design is a specific system for obtaining a sample from a given population. For the effective planning and sample survey, this sample design is the framework or road map that serves as the basis for the selection of the survey sample and affects many other important aspects of a survey as well. It is a definite plan for obtaining a sample from a given population. Sampling design refers to the procedure that the researcher would adopt in selecting items as the sample. Also, sampling design is a working plan that specifies the sample frame, sample size, sample selection and estimation method in detail. The blueprint for obtaining a sample from the sampling frame and this technique used in drawing a sample is known as sample design or sampling design. This whole process is done before the determination of data collection.

Basic thought of Sampling

The data is very important in every field for decision making whether it is medical sciences, engineering sciences, agricultural sciences and management sciences etc. for this we need to plan the correct data. If we don’t have correct data then based on that we cannot plan good decisions and it will provide misleading results. So, to achieve or plan the right data it is necessary to have the right technique which is to be used while collecting data and an appropriate method of data collection is preferred. From this, the two important questions arise (i) what are those methods required to obtain paramount data or sample and (ii) how best to use the data or sample to estimate the characteristic of the whole population i.e., what are those methods by which we can study our population parameters to draw a proper statistical inference.

‘Extension’ acts as a potential link between ‘research’ and ‘teaching’ which also facilitates transfer of technology. In this context new concepts, like Rapid Rural Appraisal (RRA) and Participatory Rural Appraisal (PRA) are becoming more meaningful now-a-days.

Rapid Rural Appraisal (RRA)
i) RRA can be defined as a systematic but semi-structured study, carried out in the field by a multi-disciplinary team over a short period ranging from three days to three weeks, based on information collected in advance from published and/or unpublished sources, direct observations and interviews as to generate working hypotheses for subsequent action
ii) RRA study aims at generating information on rural life and conditions that is relevant, timely, accurate and usable in

A. Circular letter
• Circular letter is a quick and cheap method of communication.
• It is a letter reproduced and sent to many people by the extension worker, to publicize an extension activity (like meeting, exhibits etc.) or to give information on farm and home problems.
• Circular letter could be used as a mail before organizing any campaign or extension work.
• It is sent from extension wing weekly or fortnightly to maintain regular contact with the farmers depending upon

Women account for 43% of the agricultural labor force in developing countries (FAO, 2011) and produce 50% of the global food production (FAO, 2011). Regional statistics reveal that in Latin America about, 20 percent, in Asia, about 30 to 60 percent, and in Sub-Saharan Africa, 50 to 75 percent of the agricultural labour force is supplied by women (FAO, 2013). Moreover, they invest 10 times more than their male counterpart in the family well being especially education and nutrition of children (Maertens and Verhofstadt, 2013). Despite being the primary contributors for the world food production, their roles are highly underestimated. Considerable gender bias exists in the agricultural sector, both in terms of quantities of assets, agricultural inputs and resources that women
control (Deere et al., 2013). Failure to recognize the roles, differences and inequities [between men and women] poses a serious threat to the effectiveness of the agricultural development agenda

1. Introduction
“Educational Technology is the application of scientific knowledge about learning and the conditions of learning to improve the effectiveness of teaching and training. In the absence of scientifically established principles, educational technology implements techniques of empirical testing to improve learning situation”.
Educational technology includes numerous types of media that deliver text, audio, images, animation, streaming video that includes technology applications and processes such as computer-based learning as well as local intranet/ extranet and web-based learning. Information and communication systems whether freestanding or based on either local networks or the internet in

Introduction
India with exceeding population of 1027.01 million with 741 million living in rural areas is on the path of development. The lack of access to and control over resources is said to be cause of poverty (Satapathy and  Mishra, 2011).
Women form half of our society. They have significant role in overall development. Rural development concept in our country attaches more importance to women empowerment. To combat poverty ‘self help promotion’ is of recent paradigm that has attracted attention to sections of the society.

Group Discussion
Group discussion is the method by which two or more persons meet, express or suggest their ideas for getting the solution to the felt problems by them.
Objectives of group Discussion
• To develop the democratic leadership among the members.
• To conduct the systematic group discussion to solve the problems.
• To give opportunity to each member to share experiences.
• To promote the habit of talking, thinking, group planning and group action.
• To identify the group and focus the public interest and mobilize them for group action.

The following is the plan of action to hold Animal Health Camps
1. Attempt will be made to hold at Gram Panchayat level.
2. Before 2-3 days monadi of camps must be do.
3. The camp will be held for 1 day. It will start by 7 AM and close at 2 PM.
4. The venue should be selected so that it is in the central location to get cases from 3-4 villages in order to provide facilities of clinical examination, pathological tests, etc.
5. The venue should be centrally located and easily accessible to the general public.

Awareness campaign should help educate your community on a specific subject by sharing basic information and providing examples. This will help community members have healthy conversations and increase awareness. This will begin the prevention process. Any awareness campaign should include following step;
1. Choose Your Topic Or Goals
2. Find Your Resources: Awareness campaigns take a lot of resources. You might have a set budget to implement your campaign. Money is not the only resource you will be using. Resources such as time, marketing and promotional material etc also needed for successful completion of campaign.
3. Segment Your Audience
4. Create a Schedule of Events, Programs, and Initiatives

Principle of construction of flash cards
• Flash cards are non-projected visual aid in which series of illustrated cards are flashed as presented in a proper sequence to explain complete story.
• The story must be familiar, with specific heading and suggestions at the last.
• Simple words and local expressions should be used in flash cards.
• Total number of flash cards must be 10 -12 which are easy to remember to farmers
• Card: Select white or light coloured thick paper.
• Flash card should be large enough for everyone to see at least 22 x 28 inches and simple line drawing or photographs or cartoons having plenty of colours.
• Content: Each set should have a title card. The story should be divided into a number of scenes,

Principle of construction of poster making
A poster is to be bold in design, simple to understand and attractive in colour. Poster is displayed in a public place with the purpose of creating awareness amongst the people. A poster is generally seen from a distance and the person glancing at to get first idea about it. Construction of poster making should be based on following components.
1. Pictures 2. Words 3. Space 4. Colors 5. Layout

a) Picture or illustration
1. Picture should be drawn in such a way that produce message clearly.

TV Talk
The talk delivered through TV i.e. through an electronic audio-visual medium which provides pictures with synchronized sound. The medium is cosmopolite in nature and can be used to develop instant mass awareness.

Important points while writing a programme for broadcasting
• Writing should be catchy, readable, conversational sound, understood and absorbed by the audience.
• Writing should be in simple, known language avoiding too much technicalities
• Interest of the listener should be sustained continuously
• Application of figures should be done judiciously
 

Radio script should be prepared in such a way that listener should feel that somebody is talking with him in a friendly manner and with informal style. For this purpose:
1. Collect the material from your experience.
2. Keep the purpose of the script and audience in mind.
3. Decide the method of presentation of information.
4. Considering the time available for the talk, the script should be prepared.
5. Generally we talk two words per seconds and 8 to 10 lines are required per minute having 5-6 words per sentences.
6. Time for talk is always limited hence select one idea at a time never include maximum technical information in the script.
7. Arrange the information in the sequences.
8. Include supporting and illustrated facts.
 

Overhead Projector
Principles of Over Head Projector (OHP)
A projector, which projects the image over the head of an instructor on the screen, is called OHP. It worked on the principle of indirect projection. With the use of lens of short focal length and two mirrors the picture is reflected on the screen. The drawing, diagram, letter (may be in colour or in black and white) etc. are made on transparent sheet and are put on the glass platform of the OHP, through which strong light is passed. The rays of light are converged by a lens and reflected by a mirror held at an angle of 45°.

“Audio-Visual aids are those instructional devices that may be used by a communicator in order to facilitate better understanding on the parts of learner by involving their many senses, particularly those relating to seeing and hearing”.
As a potential link between research institutions and livestock owners the extension agent must have good skills in effectively communicating with various stakeholders which include policy makers, researchers, input suppliers, market agents, credit institutions etc.

Identification of key communicators
For effective introduction of innovations and transfer of technology among farmers, effective communication is the prime need. An extension worker passes his information more effectively if he communicates with farmers/livestock owners through local leaders and key communicators.
Leader:  Is a person who effectively influences a group to cooperate in setting and achieving the goal.
Leadership: Leadership is a process by which an individual influence, guide and direct the thought, feelings and impressions in desired way.
Key communicators: Key communicator is a person who has maximum information about his society and vicinity area on different aspects.

Demonstrations
Demonstration means showing by doing. The basic principle of Demonstration is learning by seeing and doing. The concept of demonstration was given by Dr. Seeman A. Knapp. Farmers like to see how a new idea works, both purposes can be achieved by means of a farm demonstration. A good, practical demonstration is an invaluable method in extension work. The demonstration is a particularly powerful method to use with farmers who do not read easily. There are two principal types of demonstration used by extension agents - method demonstration and result demonstration.

The word data means facts or information, specifically collected by the researcher through observation or measurement from which meaningful conclusions are drawn. Data may be of qualitative or quantitative types:
Qualitative data are descriptive/qualitative in nature and can be statistically analyzed only after processing and after having them classified into some appropriate categories.
Quantitative data are expressed in the form of numerals and are amenable to statistical analysis readily.
Primary data are those which are collected afresh and for the first time, specifically for the designated study.

1.1. Origin and distribution
• South East Asia, with the greatest number of species found in Borneo, Java, Sumatra, and Malay Peninsula.
1.2. Varieties
• Neelum, Bangalora, Alphonso, Rumani, Banganapalli, Kalepad, Peter, PKM 1, PKM 2, Senthura, Jahangir, Mulgoa, Himayuddin, Paiyur 1, Mallika, Amrapali, Salem Bangalora, Arka Anmol, Arka Aruna, Arka Neelkiran, Arka Puneeth, Sindhu, Ratna and Arka Suprabath

1.1. Establishment of orchard
• A long term investment and deserves a very critical planning.
1.1.1. Location and site
• Well established fruit growing region, because one could get benefit of experience of other growers.
• Market should be close to the area with assured demand.
• Climate should be suitable to grow the chosen fruit crops; free from cyclones, frost, hailstorms and strong hot winds.
• Adequate and good quality water supply should be available round the year.

1. Propagation
• Multiplication of plants is termed as propagation.
• Aim to achieve increase in number and preserve essential characteristics of mother plant.
• Most of the horticultural crops are propagated through vegetative means.
• Two types of propagation
• Sexual propagation - through seeds which is formed by sexual reproduction
• Asexual propagation - through vegetative parts or organs of a plant.