eChapters

Jam is prepared by boiling the fruit pulp with required quantity of sugar to thick consistency, firm enough to hold fruit tissues in position. It should contain not less than 68.5 per cent soluble solids. Jam may be made from a single fruit (apple, strawberry, banana, pineapple etc.) or from combination of two or more fruits. Jam contains 0.5-0.6% acid and invert sugar should not be more than 40%.

Purpose-The term ‘Grafting’ includes all forms of grafting and budding. It is the art of joining the part of together in such a manner that they will unite and continue their growth as one plant. A grafted plant consist of two parts a rootstock, also known as understock or stock, and a scion. The part of the graft combination which is to become the upper portion or top and performs the function of flowers production and fruits is known as scion and the part which is lower portion or root which performs the function of fixing itself in the soil, absorbs moisture and nutrient from the soil is known as rootstock.

When the scion part is a small piece of bark containing only one bud the operation is termed as budding, but when the scion part is a branch containing more than one bud, the operation is termed as grafting.

ABSTRACT

Bunium persicum (Boiss.) Fedtsch., (Family: Apiaceae) is an important plant of temperate regions and its seeds are used as carminative substance in various food recipes. The seeds of the species act as a good source of economy to the hilly and mountain people. As the species is restricted in its habitat, therefore, it requires mass cultivation practices. Many attempts are already made to cultivate species near to its natural habitat. In recent years some efforts have been done to domesticate this plant in certain areas of India like Shong area in Kinnaur district of Himachal Pradesh, India, but the cultivation remained very limited. Therefore, in addition to its collection from the wild sources the species needs more scientific intervention in agriculture sector. This species has typical Palaeo-Mediterranean distribution. In India the species is distributed in wild only in North West Himalaya (J&K, HP and UK) which makes its ocurrence rarity in the country. The species restricted its distribution between 1850-3100 meters above mean sea level. Phylogenetically the species is near to Carum carvi L., therefore, C. carvi is the potential adulterant of B. persicum.

ABSTRACT

Green revolution provided sufficient amount of foods but the way resources were utilized causes the degradation of natural resources in various aspect. The adverse effect on the ecological system can be visualized in the form of various diseases in human and loss of biodiversity. The population burst is the root cause of imbalanced ecological system because agricultural land decreases drastically. Therefore we must aim to develop and improve the agri- economic growth of farmers by introducing elite variety of Mucuna pruriens (L.) DC. M. pruriens, a member of Fabaceae family, is an adaptogen and known for its tonifying, strengthening and all around beneficial properties from every part of the plant. M. pruriens has range of medicinal applications, because of unusual amino-acid L-DOPA (L3, 4dihydroxyphenylalanine) a precursor of neurotransmitter dopamine (Anti Parkinson’s) and involved in various secondary metabolite pathways. It acts like aphrodisiac, anti-snake venom, anti-depressant, antidiabetic, anti-microbial. It has a nutritional quality comparable to soyabeans. M. pruriens exhibits a tolerance for various kinds of environmental stresses such as drought, low soil fertility, and high soil acidity. M. pruriens suppresses weeds through indeterminate growth and large leaves reduce the amount of light for weeds. It shows allelopathy and effective in lowering nematodes. M. pruriens is used as forage, fallow, green manure prevents the soil erosion. It has a longer time span to be used in rotations for management of various pathogens.

ABSTRACT

Phosphorus is a macro nutrient, acts as a limiting factor in most ecosystems. Phosphorus immobilization is a major issue in most agricultural soils of India. Phosphate Solubilizing Bacteria (PSB) help in the solubilization of insoluble mineral phosphates by the production of organic acids. The aim of this research was to isolate, identify and characterize phosphate solubilizing bacteria from different agricultural crop soils of river banks of Yamuna, Delhi. Experiments were established in the laboratory to isolate phosphate solubilizing bacteria from various soil samples. Biochemical tests were performed to identify and characterize phosphate solubilizing bacteria. The phosphate solubilization efficiency of bacteria was measured. Organic acid production by different PSB for solubilization of insoluble phosphates were estimated. PSB from the soils of four agricultural crops such as Fennel, Spinach, Coriander and Mustard were isolated, identified and their ability to solubilize mineral phosphates by the production of organic acids were studied. Different PSB isolated were characterized as Citrobacter sp., Pseudomonas sp., Staphylococcus sp. and Bacillus sp. The PSB varied in phosphate solubilizing efficiency and production of organic acids. Pseudomonas sp. produced the maximum amount of organic acids and hence was the most efficient PSB. These PSB can be used as biofertilizers instead of chemical fertilizers and lead to sustainable agriculture.

1. Introduction

Ornamentals include plants which have either attractive foliage or flowers. They are used for various purposes like bedding plants in the garden or as cut-flowers and have huge commercial importance in terms of nursery business. New cultivars of ornamentals, which have been developed by various plant improvement techniques (hybridization, mutation breeding and colchiploidy) are always an added attraction to the plant lovers and a highly prized commodity for commercialization.

New cultivars of ornamentals usually inherit characters of both the parents singly or in combination and produce an entirely new trait. The progenies, thus, express a range of phenotypic characters. One has to screen these plants and identify elite plant(s) having desired phenotypic characters.

Second part of this process is characterization of the selected plants. Various characters of these plants are studied as per laid down procedures, which include vegetative, floral, seed and rhizome characters. The recorded observations are then compared with the parental characters to establish their distinctiveness. Subsequently, these plants are subjected to field trials for studying their uniformity and stability of the desired phenotypic characters.

New cultivars of ornamentals have enough potential for commercialization and good source of earning on the part of breeders who may be farmers, nurseryman, and researchers. However, for claiming right over the new cultivars developed, it is now mandatory to get those new varieties registered with appropriate agencies. The registration authorities ask for every morphological details as well as parental history. Therefore, characterization is a must for the newly developed cultivars/varieties.

The term rare amino acid is misleading in some respects. It refers to all those amino acids that are not incorporated into proteins. We already got to know some of them like ornithine or citrulline that are rather common intermediates of the basic metabolism. Roughly 220 different structures are known, most of which occur in plant cells in a free state though glutamate-, oxalate- or acetyl- derivatives can also sometimes be found.

The phenomenon and behavior of supercritical fluid (SCF) has been the subject of research right from 1800's and suggestion of supercritical fluid chromatography (SFC) came in to exixtence in 1958. Supercritical fluid chromatography (SFC) was first developed in 1960, and that time, it was considered as a science fiction chromatography and a revolutionary separation technique. But its reputation has slowly ebbed over the years and it is now moving forward by leaps and bounds as a stable analytical method with many advantages over the existing chromatographic methods. It is an unit operation that exploits the high dissolving power of fluids at temperature and pressurre above their critical values for extracting analytes from sample matrices.The first commercial packed column of SFC was made available in 1981 and the first commercial capillary column SFC instrument was introduced in 1985. Several refrences are useful for understanding the super critical fluid extraction and super critical fluid chromatography

Organic meat food products are choice products for health conscious people. These are the products produced mainly in most natural way without using fertilizers, pesticides and growth regulators in their feed and fodders. In addition, meat animals are also reared without growth hormones, antibiotics drugs etc. and grazing of the animals is a prerequisite in organic farming system.

The term organic farming was first time used by Lord Northbourne. The term organic farming was derived from his concept of “the farm as organism”. It is a holistic, ecologically balanced approach to farming. In 1991, the European Commission formulated the first government system to regulate organic labelling. In India, standards for organic agriculture were announced in May 2001 and the National Programme on Organic Production (NPOP) is administered under the Ministry of Commerce. The concept of organic meat food products has been emphasized now a day due to animal welfare point of view, consumer’s conscious ness, less residue level of chemicals, antibiotics and drugs, Bovine spongiform encephalopathy (BSE), Foot and mouth disease (FMD), genetically modified food etc.

Organic meat food products as a whole are dependent on the life stages of the livestock like its origin, feeding, housing, health, breeding, transportation and slaughter. The other factors accounting for organic foods are its production, harvesting, preservation and processing. Inspite of low growth rate of organic red meat globally, growth in dairy, egg and poultry is increasing day by day. At present 1.2 million hectares (0.6%) of Indian land is producing organic products (Willer and Kelcher, 2011).

Soil temperature is an important environmental parameter for plant growth and development. It has direct effects on microbial growth and development, organic matter decay, seed germination, root development, and water and nutrient absorption by roots. In general, the higher the temperature the faster these processes occur. Variation in soil temperature is much more pronounced because of varying characteristics and composition of soil.

The soil gains heat by absorption of solar radiation and longwave counter radiation from the atmosphere, downward transfer of sensible heat and horizontal transfer of heat from the surroundings. On the other hand, soil loses heat by longwave radiation to the atmosphere, upward transfer of sensible heat to air when the soil surface is hotter than air and with evaporation and horizontal transfer of heat from the column to surroundings.

Atmospheric pressure is the force per unit area exerted on a surface by the weight of air above that surface in the atmosphere of earth. The gas molecules of the atmosphere are pulled towards the earth by the force of gravity and hence exert a force on earth surface. In most circumstances atmospheric pressure is closely approximated by the hydrostatic pressure caused by the weight of air above the measurement point. Atmospheric pressure influences other weather elements in a significant way. Small changes in air pressure may not be felt, but they are important for bringing a change in weather. Atmospheric pressure can be calculated as the summed weight of all gas molecules between a horizontal plane and top of the atmosphere and divided by the area of the plane. Low pressure areas have less atmospheric mass above their location, whereas high pressure areas have more atmospheric mass above their location.

Measurement of atmospheric pressure

The standard unit of pressure in the SI system is the Pascal which is the name given to one Newton per square metre (N/m²). For meteorological purposes, standard sea-level pressure is 101325 pascal (Pa). To simplify this large number, the millibar unit (mb) has been adopted, which is equal to 100 pascals. Units like kiloPascal (kPa) or hectoPascal (hPa) are also used to measure pressure. 1 kiloPascal is equal to 10 millibars and 1 hPa is 1 millibar. The standard sea-level pressure is represented as 1013.2 mb or hPa.

Water is the only substance that exists in the atmosphere in all the three phases, i.e., solid, liquid and gas. The moisture on the earth’s surface is in a constant phase change. The moisture content of the atmosphere is about 0.035% of all fresh water. The atmosphere receives its supply of moisture from the earth’s surface through evaporation from oceans, lakes, rivers, damp soil, etc. and through transpiration from vegetation. Water vapour is the gaseous state of water and is invisible.

The amount of water vapour present in the atmosphere is called atmospheric moisture or humidity. It is a measure of the amount of water vapour dissolved in the air, not including any liquid water or ice falling through the air. While humidity itself is a climate variable, it also interacts strongly with other climate variables. The humidity is affected by winds and by rainfall. Moisture content is determined by the local rate of evaporation, air temperature and the horizontal transport of moisture. Humidity indicates the likelihood of precipitation, dew or fog. It also plays a major role in the heat budget and the weather phenomenon of the atmosphere.

Water vapour changes to the liquid water through condensation process. Condensation is the direct cause of the various forms of precipitation. It occurs as a result of changes in air volume, temperature, pressure or humidity. Water vapour can also be converted directly into the solid form through a process called sublimation. The latent heat expended in evaporation is liberated in the process of condensation. Four mechanisms may lead to condensation.

Flowers, the aesthetic creations of God over the earth and symbolize beauty, purity, peace and love. Flowers have been associated with progress and prosperity from ancient times. They have long been associated with each civilization and culture in the world, and are integral part of traditional, religious ceremonies and rituals and no function, from birth till death, are complete without the use of flowers. They play a cardinal role in human behaviour and culture, bringing tranquility and peace of mind. Sensory elements of flowers like fragrance, colour, texture, softness, freshness, etc. influence human psyche in different manner.

From a symbol of love, flowers have transformed into an industry generating both income as well as employment. In the flower industry, significant changes are occurring in the competitive relationships worldwide due to increasing habit of “saying it with flowers” to celebrate major life events, expressing emotions in ways that words are deficient. Estimate of the annual consumption of commercially grown flower vary by source and range from 40-60 billion USD. Global exports over the last few years have grown by more than 10 per cent annually. Flower growing and its value addition viz., essential oils, natural dye extraction, dry flowers, floral ornaments, bouquets and artistic garlands, etc. is associated with variety of employment generation activities. Besides the traditional centres of production, new production centres are also developing.

In the beginning, human kind selected some of the useful wild flowers and brought them to gardens for decoration; mainly used for their floral and other values. The rose, which is an high value flower today, has long history, perhaps older than man. Carnation was grown in Greece during 300 B. C. Orchids were known for their attractive flowers and scent in China about 2500 years ago as mentioned by Confucius. Chrysanthemum is the another oldest flower grown in China as early as 500 B. C. At the time of Confucius, only a wild single-flowered type was known in chrysanthemum. Later on many new varieties arose and in Japan, the flower reached a high stage of development and popularity. From Japan, the chrysanthemum spread to Europe. Now some of the best known modern varieties have been developed in Europe, America, and Australia. Lotus, an aquatic ornamental flower, is native to India and is associated with religion, art and culture of Indian civilization. Eloquent references to the aesthetics of these ornaments are available from ancient literature and sculptures. Most of the garden flowers were introduced into Europe for cultivation from their wild habitats between 16^th and 19^th centuries.

Growing of flowers in gardens was patronized by the rulers themselves earlier. Commercial production of flowers gathered momentum globally, efforts to breed new varieties of flower crops intensified in the 70s, and 80s primarily due to their high commercial value. With the advent of WTO followed by the Plant Breeder’s Rights, the commercial breeding efforts got intensified further. Domestication, introduction, selection, hybridization, mutation, and polyploidy breeding have played a major role in the evolution of the present day ornamental plants.

4.1 Brief historical account

Flowers have long been associated with each civilization and culture in the world. Numerous cultures have incorporated flowers into their everyday lives expressions of beauty and art. Flowers are used to celebrate major life events and to decorate graves, expressing emotions in ways that words are deficient.

In the beginning, human kind selected some of the useful wild flowers and brought them into gardens for domestication. They were used for their floral and other values. According to Theophrastus (372-287 BC), carnation was grown in Greece during 300 BC. It was used for medicinal purpose in France. Romans prepared dye from calendula flowers. During early days, Bells of Ireland (Moluccella laevis) was used as a vegetable, but now after much improvement, it is used as a cut flower. The rose, which is an important flower today, has long history, perhaps older than man. Orchids were known for their attractive appearance and fragrance in China about 2500 years ago as mentioned by Confucius. Man’s interest in cultivating and growing orchids developed during the 19^th century in Europe. Chrysanthemum is the one of the oldest flower grown in China as early as 500 BC. Most of garden flowers were introduced into Europe from their wild habitats between 16^th and 19^th century. Ornamental annuals constitute an amazing group of plants widely grown in the parks and gardens in different seasons. The enchanting beauty of colour, shades, form and fragrance have made them an obvious choice for every garden. No other plant can influence the general landscape in such an incredible manner by riot of colours.

One of the important studies in the crop improvement programme is the discriminant function analysis. This is a method of exercising selection in plants, which show the extent of which character is genetically related to yield (Goulden, 1959). The technique of discriminant function was developed by Fisher (1936). But, the application of this method for plant selection was first made by Smith (1936) in wheat. He developed an index design for the selection of plant lines in wheat using the concept of discriminant function to derive a linear equation based on observable characteristics as the best available guide to the genetic value of each line.

Stephens (1942) reported about the interaction of yield components within the plant and within the population, which serves as an index for yielding ability. According to Hazel and Lush (1943), selection for a total score or net desirability is more efficient that selection for one trait at a time. The formulae presented by them for selection index give proper weight to each trait that are more efficient than selection for one trait at a time or for several traits with an independent culling level for each trait. According to Darlington and Mather (1949) discriminant function has been defined as a linear compound of series of varieties, obtained by giving the different varieties with individual coefficients which will minimize the difference between classes relative to variation within classes of objects of which the variates are measurements. Mather (1949) reported that these functions afford to the best available means of discriminating the classes and evaluating the genotype of a plant in terms of the observed values of various characters. Pundir and Raj (1971) reported that selection index based on three characters combination i.e., seeds per siliqua, 1000 seed weight and yield per plant was efficient in toria.

3.1 Introduction

There is now latest national thinking going on to conserve the known sources of energy such as firewood, cow dung, agricultural waste, kerosene, oil etc. i.e. by limiting their conventional uses and to propagate the use of non conventional energy especially in the rural area. The large scale indiscriminate use of non commercial fuels fully in the rural areas are not only providing to be inefficient but also inadequate to cope with the growing rural energy demand caused by growing population and growing consciousness among rural people for a better way of life. Planned input of energy are, therefore urgently require to protect the environment and to cope with raising rural energy demand. In order to assess the widening gap between the supply and demand of commercial energy and indiscriminate use of non commercial fuels in rural areas, there is a need for various types of statistical or quantified information to be collected and analyzed in an objective manner and presented suitability so as to serve as a sound basis for taking policy decision to make an optimum utilization of commercial energy in a balanced manner. This will require conducting of survey in areas to be covered.

The statistical data may be collected by complete enumeration or by a sample survey. The earlier method is very cumbersome, time consuming, cost ineffective and requires an intensive development of manpower, if domains of study is quite large. Complete enumeration and sample surveys presuppose the existence of a certain minimum of facilities such as funds, professional personnel for planning the survey methodology and supervision of field operations, sufficiently qualified enumerators, sampling frames, machine tabulation equipment, transport and communication facilities etc. These facilities or combinations there of do not always exist to the extent needed for a complete enumeration survey. As such, sample survey is preferred to complete enumeration for ethic type of studies giving due importance to sampling error.

In the integrated energy planning, there is an objective to find out the present consumption pattern of energy for different end uses, shortages being experienced and people’s reaction to using alternate sources of energy is also enumerated.

The survey methods determine – incident, distribution and the interrelation among sociological variable and psychological variable. The sociological variable s consists of age, education, income and attitude, behavior, motivation, belief and knowledge about the problem.

These three requirements are required to be followed in sequence for getting maximum benefits. However, integrated rural energy concept is a new area of work and not much experience is available in this field as compared to other rural development activities. It is difficult to give any guidelines for planning. One can only recommend approaches and not solutions to planning.

The rural area in developing countries are almost always rich in one or another sources of energy, which could be used separately or in combination with other sources in decentralized small scale units, or even on a household scale. Thus, the problem is one of good planning, management and a strong educational and promotions campaign, rather than the non availability of resources or the lack of technology. Further, after planning, prefect development of device and their effective operation is required.

The purpose of the Integrated Rural Energy Planning project should be clearly defined. It may design either for research and development or demonstration for Popularisation of system, education and training, promotion, health or combination of these.

It is usually better to start with a small size to gain experience and build up confidence before undertaking the large scale project. As defined earlier, one of primary requirement of Integrated Rural Energy Planning is to develop energy demand pattern, energy supply system and optimally match demand with supply.

Potato has special seed production problems as the quality characteristics of seed potatoes are influenced by a number of factors and important amongst them are the diseases and pests namely, viruses, fungi, bacteria and nematodes. Once the seed tuber is infected the pathogens, specially viruses, permeate through the plant system. The plant growth, therefore declines and the yield reduces progressively. It is therefore, important that seed stocks should not only be genetically pure but also should be in right physiological condition and disease free at the time of planting. Seed production technology developed for seed potato production, thus aims at producing disease free, genetically pure seed. There are now two independent channels of seed production for hills and plains.

Hill seed : The seed produced in hills (2500 metres above sea level) at suitable locations is called ‘Hill Seed’.

Plain seed : The seed produced in plains at suitable locations is called ‘plain seed’. Northern plains have emerged as an important source of potato seed production. The low aphid, plains seed is in right physiological condition at the planting time and yields higher than the traditional hill grown seed.

Once healthy seed potatoes are introduced into the system of growing them during low Aphid period accompanied by a systematic insecticide application, roguing and removal of haulms before the aphids attain critical number and the re growth is checked, the health standards for the seed crop could be maintained for a number of generations. This system of seed potato production has been designated as ‘Seed Plot Technique’.

Abstract

The anomalies observed in the reverine systems have been demonstrated to be the potential tool in deciphering the tectonically active zones. Such various drainage anomalies viz: radial drainages, anomalous compressed meanders, eyed drainage and the youthful floodplains observed in Annamali – Palani - Varushanad hill ranges of the Western Ghats of Tamil Nadu show that the N-S and NE-SW trending faults are tectonically active in the recent years.

Much has been talked about the mighty river systems and their migratory histories, since Purana and Vedic times. In Mahabharata the places where the Lord Krishna is said to have played with Gopis in the banks of River Yamuna are now forming flat lands many kilometers away from the present day Yamuna. In the same way, there is a mention in ancient literature about the ‘Indo-Brahm’ river which was originated from the Himalayas and flown through Brahmaputra valley, Indo-Gangetic plains, Rajasthan desert and met the Arabian Sea in Rann of Kutch. Such a mighty river is said to have fragmented into Brahmaputra, Ganges and the Indus rivers due to Tectonic activity in Himalayan regions. Similarly there are many evidences in the Tamil literature. For example in ‘Kalingathuparani’ it is mentioned that the river ‘Palar’ was flowing north of Kanchipuram but the same river is now flowing south of it.

All these facts show that the rivers have the tendency to frequently change their courses. The advent of modern Remote Sensing Technology has opened up new vistas in studying such migratory rhythms of the rivers, as such old and buried rivers exhibit spectacular ribbon like, linear, curvilinear and contorted features with typical darker tone in black and white images and reddish tone in false color composites (FCC) and other color coded images.

Abstract

The Remote Sensing and the followed up field based investigations show that the Tamil Nadu coast, India, is facing series of geoenvironmental problems due to tectonic movements and morphodynamic processes. Such problems pose a challenge to the engineering geologists. The paper discusses such challenging engineering geological problems and plausible remedial measures.

INTRODUCTION

Molecular tools generally refer to molecular genetic tools. Although the first use of these tools began more than four decades ago, whenLewontin & Hubby (1966) used allozymes to find out the amount of genetic variation present in wild populations of Drosophila, the lack of expertise and high cost of these tools have prevented their widespread use among ecologists in general and behavioural ecologists in particular. But recently the accessibility and costs of these techniques have come down a lot and with the right kind of collaboration, lack of expertise is no longer an issue.

As was mentioned above, the Lewontin & Hubby’s (1966) investigation was done using allozymes. Although still an option, user-friendly neutral nucleotide markers are more preferred by investigators nowadays. The discovery of the polymerase chain reaction (PCR) in 1983 by Kary Mullis was the single landmark in this field, if one has to mention one. The use of molecular markers has totally revolutionized the field of ecology. It has provided ecologists the tool to ask questions, which were hitherto impossible to tackle and also to answer questions with much more precision and accuracy than was possible till now. It has toppled long held beliefs, sometimes, romantic ones, by ecologists.

Introduction

Increasing production of cereal crops, oil seed crops, and horticultural crops is becoming more important than ever before owing to increasing population and changing food habits of the people. The modern tools of biotechnology are being exploited to the fullest throughout the world for developing transgenic crop plants with improved traits. Already about 20 countries in the world are growing transgenic crops, predominately with traits such herbicide tolerance and resistance to insect-pests. However, there is a constant fear of pollen flow from theses transgenics to the neighbouring non-transgenic fields or to their weedy or wild relatives. It has become a matter of great concern to all biotechnologists to develop alternative strategies to prevent the pollen flow.

The best alternative strategy available today to circumvent the nuclear- transformation associated bottlenecks in developing tenable crop transgenics is the chloroplast transformation technology. This technology allows introduction and insertion of foreign genes into the chloroplast genome, called plastome. Chloroplasts are the site for photosynthesis in plants mostly present in leaves. In general called plastids, they are also present in other parts of the plant. For example, chromoplasts in coloured leaves and fruits, amyloplasts in starch storing organs, leucoplats in roots, and etioplasts in dark-grown seedlings. The plastids arise from proplastids - the undifferentiated form of plastids present in most plant cells. Just like nucleus, these plastids (and also mitochondria) contain their own genetic material on the chloroplast genome.

ABSTRACT

Commercial propagation using tissue culture techniques started in seventies for orchids in USA. Since then, the science and technology of tissue culture have advanced particularly after the development of the most widely used medium by Murashighe and Skoog (1972). The science has led to the development of the fascinating area of gene transfer. Today, micropropagation protocols are available for almost all horticultural crops. But, there are certain grey areas where research need to be intensified to solve the problems confronting certain difficult to propagate species belonging to plantation and spice sectors. The major area that needs to be addressed is recalcitrance.

ABSTRACT

Microbial contamination is perceived as the most serious threat faced by plant tissue culture research laboratories and commercial production units. A culture is often viewed as contaminated when obvious microbial growth incited by fungi, yeast or bacteria is noticed on the culture medium. On the other hand, there can be situations when microbes may be present in a covert or latent form with serious side effects on the cultures. The present investigations were taken up with the objectives of elucidating the cause of poor growth, lack of regeneration response or modification of rooting observed with visibly clean cultures of different horticultural crops. Apparently clean cultures of grape, banana, watermelon, eggplant, gerbera and chrysanthemum of recent origin or after long-term in vitro culturing were used in this study. Indexing the medium of such cultures using fungal/ yeast and bacteriological media brought out the covert association of one or more bacterial types with varying proportions of incidence in such cultures. Indexing of tissue revealed bacterial presence in different parts of in vitro plantlets. In some instances, bacteria showed endophytic colonization while being not detectable in the tissue culture medium subsequent to a disinfection or antibiotic treatment. Such bacterial association appeared to be the cause of in vitro decline observed with long-term micropropagated cultures, lack of reproducibility of tissue culture protocols, failure of in vitro rooting etc. besides being a threat to in vitro gene banks and germplasm exchange.

ABSTRACT

Recognizing the need to apply plant tissue culture tools for generating planting materials of important spice crops especially vanilla, the Spices Board started Biotech Production Unit at its head quarters building at Kochi with an annual production potential of 8 lakhs plantlets. Complete tissue culture protocols were developed and scaled up for large scale production of vanilla and other spices viz. large cardamom, small cardamom and tree spices. The laboratory is equipped with all modern facilities and is established in an area of about 2000 sft. Department of Biotechnology, Govt. of India also participated in the venture by sanctioning a project entitled “Regional centre for large scale production of tissue culture plantlets of vanilla and other spices” with a financial out lay of Rs 95 lakhs equally shared by DBT, New Delhi and Spices Board, Cochin.

At present there is a stock of over 3.00 lakh vanilla cultures in the lab and already 1.25 lakh plant lets were sent for hardening. The Spices Board, in coll aboration with “Kudumbashree”, a poverty eradication mission of Govt. of Kerala, extend short term training in tissue culture techniques to undergraduates from ‘Below Poverty Line’ (BPL) families and engage them as long term trainees in the Biotech Production Unit. The Board also coordinates and extends support to Kudumbashree members for, establishing and maintaining hardening facilities for TC plantlets thus providing means for empowering women from BPL families.

ABSTRACT

Astudy was undertaken to evaluate the performance of tissue culture derived plants vis a vis open pollinated seedlings of cardamom (Elettaria cardamomum Maton). A total of 102.5 ha (units) covering 50 ha in Kerala, 37.5 ha in Karnataka and 15 ha in Tamil Nadu was selected for the present study out of which each demonstration plot (unit) had an area of one hectare in which 80% of the plants are tissue cultured and 20% are open pollinated. Elite cardamom clones were subjected to in vitro multiplication and their respective open pollinated seedlings were compared. The planting was initiated in 1989 and completed with 1991 in all the zones identified. Plant growth characters such as number of tillers, number of bearing tillers and panicles per clump were recorded and compared. The yield was recorded in 1993-94 and 94-95 seasons after its stabilisation. Results indicated that tissue culture plants performed better compared to open pollinated seedlings with regard to growth attributes and yield irrespective of the seasons and zones. In Kerala region tissue culture plants exhibited a yield increase of 24.3 and 36% over open pollinated seedlings for the 1989 and 1990 plantings respectively. Similar trend was observed in Tamil Nadu for 1989 and 1990 planting where the percentage increase in yield became 130 and 21 respectively. In Kerala, the yield performance of tissue culture plants and open pollinated seedlings was on par during 1990 planting but in the subsequent year (1991) tissue culture plants showed 24% increase in yield compared to their respective open pollinated ones. Further, the study enabled to identify cardamom clones suited to in various zones of the cardamom tract. Accordingly 14 clones with a yield potential of over 750 kg/ha under moderate management were identified and this indicated that cardamom clones are highly location specific in their performance.

ABSTRACT

Vanilla planifolia Andrews - the high value, exotic spice crop has recently gained wide acceptance in the southern and northeastern states of the country. Being an orchid, germination of naked embryo is limited and this favours genetic uniformity. An attempt was made to induce genetic variability in Vanilla through biotechnology tools for its exploitation in crop improvement programmes. In vitro embryo culture and mutagenesis were applied for the purpose. Plantlets were multiplied, hardened, planted out and evaluated for morphological, biochemical and molecular markers. Great interclonal variability was observed in the in vitro derived plantlets. The intraclonal variability was very low in tissue culture derived regenerants. Isozyme and RAPD assay confirmed genotypic variation. Six superior clones were identified and distributed to farmers for further evaluation.

ABSTRACT

Vanilla (Vanilla planifolia Andrews) is a high value spice crop having immense potential for cultivation in India as a source for natural vanillin. Though the crop was introduced way back in 1835, its cultivation gained momentum only in the recent past due to various reasons. Lack of sufficient good quality planting materials is one of the constraints for expanding the area under the crop. Conventionally vanilla is propagated by vine cuttings, but this method is inadequate to have large number of planting materials in a short span of time. So, micropropagation was attempted and proved to be highly successful. In the present study, performance of vanilla plants raised from tissue culture plantlets and vegetative cuttings was assessed on large-scale in planters’ field covering a total area of 111 ha in Kerala(68ha), Karnataka(28ha) and Tamilnadu(15ha) with the financial assistance of the Department of Biotechnology, New Delhi under the project entitled “Tissue culture Vanilla : Product Plan“. Disease free elite accessions of vanilla identified by the Indian Cardamom Research Institute (Spices Board), Myladumpara from its germplasm repository were used as mother plant source for tissue culture multiplication and also for raising the nursery for producing vegetative cuttings. The field planting for the demonstration plots was carried out during 1996-1999 on a cluster approach basis having 400 plants in each unit or plot of 0.25 ha. Accordingly a total of 444 evaluation plots were laid out under the project covering the aforesaid three states. Out of the 400 plants in a plot, 80% were nursery hardened grown up tissue culture plants and the balance( 20% )vegetative cuttings of comparable length. Observations on growth and yield attributes such as vine length, number of leaves per vine, internodal length, number of beans per vine, bean size, yield etc. revealed that performance of tissue culture plants is on par with that of conventional plants raised from vegetative cuttings of comparable length. It is also observed that if good management practices are adopted, tissue culture plants perform better at the full bearing stage of the plant. This proves that tissue culture plants of vanilla can be popularized as a cost effective and faster source of planting materials compared to conventional vegetative cuttings. As the demonstration plots were distributed under varied agroclimatic situations, the influence of different weather parameters on yield was also ascertained. Further, the project could convince the farmers about the comparable performance of tissue culture plants and that is quite evident from the ever increasing request for tissue culture vanilla plants received by the Spices Board.

ABSTRACT

Vitex trifolia L. (Verbenaceae) is known for its medicinal property. Micropropagation can provide the opportunity to obtain large number of homogeneous plants and there are no reports of micropropagation of this species. So, we attempted for in vitro propagation of this species and described here is an efficient and rapid propagation method. This report describes in vitro shoot induction and plant regeneration from mature nodal explants of Vitex trifolia L. on Murashige and Skoog (MS) medium fortified with benzylaminopurine (BAP), kinetin (KN), thidiazuron (TDZ), adenine (ADE), and 2- isopentenyladenine (2 - iP) (0.25-10.0 mM). Multiple shoots differentiated directly without callus mediation within 3 weeks when explants were cultured on medium supplemented with cytokinins. The maximum number of shoots (9 shoots per explant) was developed on a medium supplemented with 5.0 mM BAP. Shoot cultures was established repeatedly by subculturing the original nodal explant on the same medium. Rooting of shoots was achieved on half strength MS medium supplemented with 0.5 mM naphthaleneacetic acid (NAA). Rooted plantlets were transferred to pots containing autoclaved soil and vermiculite mixture (1:1) that showed 90 % survival when transferred to outdoor. Plants transferred to the field have established themselves in the soil and are growing well. The regenerated plants did not show any immediately detectable phenotypic variation. Natural stands of Vitex trifolia are fast disappearing in India because of indiscriminate collection and over exploitation and the outlined procedure here offers a potential system for mass multiplication and conservation of Vitex trifolia.

ABSTRACT

Considering the difficulties faced during ex vitro establishment of orchid plants the present investigation was undertaken to elicit information on the influence of different sucrose levels in the in vitro rooting medium and by subjecting the plants to different levels of triazole at the time of planting out in order to overcome the mortality percentage at the time of ex vitro establishment. Among the growth parameters observed viz. crop growth rate, net assimilation rate, and relative growth rate was found to be high in plants treated with 40 g/l of sucrose concentration and also at the triazole level of 5 mg/l. The photosynthetic rates were observed to increase meanwhile the transpiration rate was declined in plants grown under the above treatment levels of sucrose and triazole. The maximum survival percentage was observed when the in vitro plants grown in rooting medium with 40g/l of sucrose which is attributed to the influence of sucrose concentration on morphological characters studied viz. plant height, number of leaves per shoot and number of roots per shoot. Triazole treatment of plantlets at the time of planting out helped in better survival percentage which is the whole sum effect of increase in number of leaves per shoot, number of roots per shoot of plants grown under the triazole level of 5 mg/l. Where as the height of the plant was inversely proportional to triazole concentration,the biometric aspects studied viz root length, total fresh and dry weight of the plants with the above mentioned treatments were found to be superior. The present study envisages the measures to be taken during in vitro propagation and ex vitro establishment of tissue cultured plants in order to over come the field mortality.

ABSTRACT

Mucuna pruriens (L.) DC. is one of the important medicinal plants that contains L DOPA (L-3,4 dihydroxy phenylalanine) which is the precussor for the neurotransmitter dopamine that plays an important role in curing Parkinson?s disease. To accommodate the huge demand for L DOPA, in vitro production of the drug using cell cultures is felt essential. Hence, an experiment was carried out with completely randomized design with seven treatments viz., T₀ - Control; T₁ - MS + 2 mg l^-1 NAA; T₂ - MS + 2 mg l^-1 IAA; T₃ - MS + 2 mg l^-1 2,4-D; T₄ - MS + 2 mg l^-1 Kinetin; T₅ - MS + 2 mg l^-1 BA; T₆ - MS + 1 mg l^-1 IAA + 1 mg l^-1 BA and 3 replications. Among the various explants investigated stem bits registered the least days (12.38) for callus induction. The stem bits responded well with better callusability (78.57 per cent), relative growth rate (2.57) and callus index (249.16). MS medium + 2 mg l^-1 2,4-D was found to be the best for inducing callus in various explants namely stem bits, leaf bits and root bits. L DOPA accumulation was more in cell suspension cultured in liquid MS medium with 4 per cent sucrose + 1 mg l^-1 IAA + 1 mg l^-1 BA and that was confirmed by means of HPLC.

ABSTRACT

Tamarind, an economically important multipurpose tropical tree, is a species of choice for sustainable development of wasteland due to its hardy nature and adaptability to various agroclimatic conditions. However, there are very few reports on in vitro regeneration of tamarind. In an attempt to induce caulogenic response, effect of different cytokinins on nodal explants from mature trees was tested. The study extended to seedling explants, revealed that 6-Benzylaminopurine (0.08 - 44.39μM) and Zeatin (0.09 - 9.12μM) induced multiple shoots with callusing at the base of explant. Kinetin (0.09 - 9.29μM) and 6-γ,γ- dimethylallylaminopurine (2iP) (0.09-9.84μM) remained ineffective for multiplication of shoots and callusing was noted at the cut end of explant in contact of medium. Thidiazuron (TDZ) (0.09- 9.08μM) failed to induce morphogenic response and triggered dedifferentiation in the basal cut end. This led to profuse callusing in the explant and restricted morphogenesis. To avoid wound in the explant, tamarind seeds were cultured in medium containing TDZ.

Seedlings were germinated in medium with or without Thidiazuron (4.54, 9.08, 13.12, 18.16μM). Differentiation of the apical meristem to shoot was restricted and proliferation of the cotyledonary node meristem was triggered. Large number of meristematic buds appeared in a radial pattern around the node. The meristematic activity subsequently spread towards the cotyledonary bridge and also in the proximal part of the hypocotyl. These buds differentiated to form shoot primordia and subsequently to shoots in medium devoid of growth regulator. Plants developed by micrografting of these shoots on seedling derived rootstocks, survived in soil.

ABSTRACT

It is essential to prospect wild Zingiber species for soft rot resistance, since the cultivated ginger is monotonously susceptible to this disease. We evaluated the genetic diversity of Zingiber species and their response to Pythium aphanidermatum, the causal agent of soft rot. Diversity of 21 cultivars and three wild accessions of Z. officinale, 84 accessions of five wild species: Z. zerumbet, Z. cernuum, Z. neesanum, Z. purpureum and Z. wightianum and one accession of Hedichium flavescens were examined using AFLP markers. No polymorphism was detected at more than 500 AFLP loci screened in ginger cultivars, which to our knowledge, is the most extreme case of genetic narrowness in a species. Analysis revealed high variability in Z. cernuum and relatively low variations in other species. Strong correspondence was evident between the genetic diversity of a species and its mode of propagation. Selected accessions of Zingiber species were inoculated with Pythium zoospores and the disease progression was scored. The response to Pythium varied from total susceptibility in Z. cernuum and Z. officinale to hardy behavior among Z. zerumbet accessions. The present study provided an estimate of genetic diversity of Zingiber species common in Kerala. Species relationships and population genetic structure were discussed. The monotypic nature of ginger cultivars may hinder the possibility of their identification based on their nucleic acid profiles. The present study identified Z. zerumbet as a potential source of soft rot resistance, which will trigger off the application of genomics tools to access this trait in order to genetically engineer soft rot resistance in ginger.

ABSTRACT

India is the largest producer of banana, contributing to 17.3 per cent of the global production. In Kerala, it is the leading fruit crop, being cultivated in an area of 99,412 ha with an annual production of 7,31,650 tonnes. Biodiversity in the cultivars of banana, which consist of triploids, is complex especially in Nendran. Nendran belonging to Musa AAB plantain subgroup is the leading banana cultivar of the Kerala. in Kerala, several landraces are available, cultivated in different parts, known in different local names. Twelve Nendran ecotypes of banana belonging to AAB genomic group were collected from Banana Research Station, Kannara, Banana Nursery Farm, Palode and Instructional Farm, College of Agriculture, Vellayani to ana lyse genetic divergence using RAPD marker and morphological traits. Out of the forty one decamer primers screened for RAPD analysis, 34 could produce amplification. Twenty five primers showed high level of polymorphism. Finally, six of the most promising primers viz., OPA-01, OPA-03, OPA-13, OPB-04, OPB-10 and OPB-12 were used for RAPD analysis. Statistical analysis for the morphological traits and RAPD marker were conducted using the software programme NTSYS pc version 2.02e. The matrix average taxonomical distances (similarity coefficient) among the ecotypes ranged from 0.59 to 1.00 in RAPD profiles and morphological profiles (dissimilarity coefficient) ranged from 2.82 to 11.21. In the dendrogram (RAPD profiles), Koonoor Ethan showed the maximum genetic divergence. Quintal banana and Zanzibar came under a single clusters at a distance of 0.79 and clustered together at a distance of 0.82. Both the ecotypes showed next higher genetic divergence among the Nendran ecotypes. In the case of morphological traits, the same trend could be observed. Koonoor Ethan showed the maximum genetic divergence at a distance of 11.21 and Quintal banana showed next higher genetic divergence at a distance of 9.11. Koonoor Ethan, Quintal banana and Zanzibar were distinct ecotypes among the Nendran ecotypes of Kerala

ABSTRACT

Random Amplified Polymorphic DNA (RAPD) markers were used for characterizing eleven banana (Musa AAB Plantain Subgroup) clones using 40 decamer primers. Eight primers, which produced reproducible banding patterns, yielded 42 scorable bands with an average of 5.25 bands per primer. The amplification products ranged in size from 400 to 1500 bp. A genetic similarity matrix was constructed using the Jaccard’s coeffecient method. The pair wise similarity coefficient values varied between 0.3333 and 0.9355. Based on the coefficients, distances between the clones were computed using SYSTAT software package. In the dendrogram constructed by the nearest neighbour (single- link) method (Krzanowski, 1988), all the eleven clones were found grouped under five clusters. Attu Nendran, Changanasseri Nendran, Changazhikodan, Kaliethan, Koonoor Ethan and Quintal Banana formed the largest cluster. Manjeri Nendran and Myndoli formed the second cluster. Padalamurian, Mysore Ethan and Zanzibar formed three separate clusters. Zanzibar, belonging to the Horn Plantain group was different from the rest of the clones.

ABSTRACT

Genetic relationships were estimated among twenty five ashgourd (Benincasa hispida (Thunb.) Cogn.) landraces of different types using PCR-based random amplified polymorphic DNA (RAPD) markers. The molecular survey of the ashgourd germplasm by RAPD using three random primers resulted in twenty scorable bands (average of 6.66 bands per primer) of which two were monomorphic and rest, 18 were polymorphic (90.0%). Pair-wise genetic similarities among the landraces determined using Jaccard’s coefficient ranged from 0.14 to 1.00. The dendrogram based on clustering matrix separated the landraces with morphologically distinct smooth textured fruits from waxy textured fruits and the former group showed less divergence than the latter. With in the group of waxy textured fruits, limited variation was detected among landraces with small and medium sized fruits. Further, morphologically similar landraces with large fruits form distinct clusters with in the major clusters. The study demonstrated the usefulness of the RAPD marker analysis in effectively assessing the genetic relationship present among the ashgourd landraces examined.

Introduction

Genetic markers have assisted in genomic characterization of various crops species (Rafalski and Tingey, 1993). Most of the genetic marker systems rely upon DNA sequence polymorphisms (Staub et al., 1996). Each system has unique advantages and disadvantages based on cost, experimental objectives and type of information desired. Regardless of its inherent methodology, the effective use of any marker system for genetic analysis requires reliability and consistency.

The random amplified polymorphic DNA i.e. RAPD (Williams et al., 1990) or arbitrarily primed polymerase chain reaction (Welsh and McClelland, 1990) marker identifies DNA sequence polymorphisms. In this technique, segments of genomic DNA are amplified in a polymerase chain reaction i.e. PCR (Mullis et al., 1986) with random primers followed by agarose gel electrophoresis. This is simple, relatively inexpensive and is amenable to high throughput investigation. RAPD markers are usually dominant and polymorphisms are observed as the presence or the absence of DNA bands of a particular size. RAPDs are common molecular approaches employed in plant breeding, genotypic differentiation, gene linkage analysis, assigning evolutionary and taxonomic affinities and relatedness and DNA fingerprinting analysis (Lin et al., 1996).

ABSTRACT

Owing to paucity of available genetic / molecular markers, onion breeding still relies heavily on phenotypic selection methods. DNA markers can provide plant breeders genetic information for indirect selection of plants. All the DNA based procedures require high quality plant genomic DNA. Abundance of polysaccharides and phenolics present in plant tissue often impede PCR amplification. The polysaccharide contaminants can also produce downstream problems by inhibiting many of the enzymes that are employed in subsequent analysis and cloning. RFLP based markers in Allium species pose certain problems due to relatively large size of the genome compared to most other herbaceous crops. As an alternative, RAPD would prove advantageous in genetic analysis of Allium. Here we present our results on optimization of DNA extraction in onion to get amplifiable quality of DNA for further RAPD analysis. Leaf samples were collected from the onion germplasm maintained at Indian Institute of Horticultural Research (IIHR), Bangalore. Dried leaf protocol was suitably modified to obtain fair quality of DNA. After varying each of the ingredients of the extraction buffer we arrived at the optimal extraction buffer, which resulted in good quality DNA. Quality was analyzed by subjecting the DNA to agarose gel electrophoresis and restriction digestion. Extracted genomic DNA was quantified with ‘Hoefer’s Dynaquant’ (Pharmacia, USA). The DNA quantity ranged between 160-2400mg/gm of dried powder. The standardized 25ml PCR mixture contained template DNA (25ng), 5 pmol of primer (Operon Tech. Inc., Alameda), 2mM MgCl₂, I unit of Taq polymerase (Bangalore Genei), with 10x buffer along with 200 mM each of dNTPs. Amplifications were performed in thermal cycler (MJ Research, PTC 100) for 45 cycles after an initial denaturation of 95°C for 3 minutes. In each cycle denaturation for one minute at 95°C, annealing for one minute at 35°C and extension for two minute at 72°C was programmed with final extension step at 72°C for 5 minutes. This fine tuned protocol resulted in clear, informative RAPD profile.

ABSTRACT

The present investigation was undertaken to standardize the DNA extraction methods, to optimize the PCR conditions and generate RAPD’s to study the genetic variability in chrysanthemum. Total genomic DNA was extracted from freshly emerged leaves of chrysanthemum cultivars (Flirt, Nagpur Red, Snow Ball) using different DNA extraction methods (CTAB method, SDS potassium acetate method, SDS miniprep method and urea phenol method). The highest amount of DNA (349μg/g of leaves) was extracted using CTAB method, while it was lowest (130 μg/g of leaves) in SDS miniprep method. The absorbance ratio was high for DNA extracted by CTAB method (1.82) followed by SDS- potassium acetate (1.65) and SDS miniprep (1.60) methods. For standardization of PCR ampl ification conditions, optimization was carried out to determine the optimal concentrations of template DNA, MgCl₂, Taq DNA polymerase and annealing temperature. In general, either no products or products with inconsistent bands were obtained when the concentration of template DNA was 25 ng or MgCl₂ was 1.0 mM with high annealing temperature of 45°C.

ABSTRACT

Though the genetic control of sex determination is being explored in several animal systems, no much information is available on the genetic basis for sex specificities in dioecious or sexually polymorphic plant species. Date palm is a unique member of the family Palmaceae which exhibits dioecy. Here we report the sex-specific variation in the genomic DNA of male and female date palms as revealed from an extensive study conducted with palms representing different date palm growing regions of the Kingdom of Saudi Arabia. The genomic DNA content in females was less than 60 per cent compared to those of the male genotypes. Polymerase chain reactions carried out to amplify different segments of the alcohol dehydrogenase (AdhA) gene from the date palm genomic DNA yielded two DNA fragments each in the female genotypes where as the same primers amplified a single fragment in male plants. Genomic Southern analysis of Eco R1 restriction digests showed RFLP between the male and female genotypes with the male plants showing a single hybridization signal of approximately 14 kb size and the females showing six additional bands of varying molecular weights. The results of our investigations reveal the potential of molecular biology in differentiating the sex in date palm at DNA level. Further research is needed to develop DNA markers that will help to unequivocally identify the sex of dioecious plants at the seedling stage.

ABSTRACT

Piper colubrinum is a wild relative of the cultivated species of black pepper, Piper nigrum. The enzyme hydroxy methyl glutaryl Co A reductase which catalyses mevalonate synthesis, is the key enzyme in isoprenoid biosynthesis.This is the rate limiting step in the pathway which leads to the formation of sesquiterpene phytoalecins, a group of plant defense metabolites. Piper colubrinum is known to be tolerant to the dreaded disease called foot rot, incited by Phytophthora capsici. The hmgr gene is reported to confer resistance to fungal pathogens. Hence we made an attempt to isolate gene encoding this enzyme. cDNA was synthesised by reverse transcription from total RNA of P.colubrinum using oligo dt primers. Specific degenerate primers designed based on the conserved boxes among various plant species were used for amplifying hmgr gene from the cDNA. The 700 bp amplicon was cloned in pGEMT vector and sequenced with T7 primer. Multiple sequence alignment using clustal W 1.8 revealed homology of the cDNA fragment with hmgr genes in several other plant species. Maximum similarity was observed among P. colubrinum, Solanum xanthocarpum and Nicotiana tabacum. The cDNA library of P. colubrinum constructed in lTriplEx2 when screened with radiolabelled hmgr amplicon, yielded positive signals. Attempts are being made to isolate full length gene from these clones, which could be later used for imparting biotic stress tolerance to cultivated crop species.

ABSTRACT

Basic understanding of plant cell proliferation and differentiation is essential for applying modern techniques of genetic transformation. The lack of understanding about the physiological and molecular mechanisms underlying in vitro morphogenesis, together with the non-availability of reliable morphogenetic markers, is one of the most serious constraints for application of gene transfer technology to plant improvement. Plant tissue culture has therefore remained an empirical science. In recent years, concerted efforts are being made to understand the molecular control of cell differentiation. In light of this, present work was carried out to study the role of CKI1 (cytokinin independent 1) gene, one of the genes implicated in shoot regeneration, by transforming the tomato cotyledons through Agrobacterium mediated transformation with two different gene constructs one with CKI 1 gene and another without CKI1 gene. In both the constructs, nptII gene was used as a selectable marker. Transformation of tomato cotyledon explants with CKI 1 gene resulted in higher regeneration of putative transformant s (8.0%) as compared to transformation of explants without CKI1 gene (2.1%). The results show that CKI1 gene has a role in increasing the regeneration frequency in tomato.

ABSTRACT

Astudy was undertaken with the main objective to standardise the Agrobacterium mediated genetic transformation technique in Dendrobium Sonia 17. Protocorms and protocorm like bodies (PLBs) of Dendrobium orchid were transformed by Agrobacterium tumefaciens strains LBA4404 and EHA105 harbouring the plasmids pCAMBIA2301, pCAMBIA1301 and PBI121. The PLBs were generated in vitro by culturing the shoot apices in half strength MS medium. The protocorms were obtained by culturing the seeds in vitro. The sensitivity of orchid tissues to various antibiotics viz., ampicilin, rifampicin, cefotaxime, carbenicillin, kanamycin and hygromycin was evaluated to use the antibiotics as selection agents. The sensitivity was studied in detail by scoring the tissues, based on the morphological changes as those remaining fully green, partially discoloured, bleached, completely turning brown and dead. The various requirements for transformation viz., the type and size of explant, infection time, wounding, co-cultivation and incubation period, concentration of acetosyringone required, elimination of bacteria, and the selection of transformants were optimised to establish a successful transformation system. The pre- cultured explants were mixed thoroughly by gentle swirling with the bacterial cells obtained by centrifuging the bacterial suspension, and also with the bacterial suspension diluted to 1/5 with fresh medium. Then the explants were cocultivated with the bacteria for 2 days at 28°C. The transformation was performed through two consecutive stages of cultivation, the first stage on half strength MS medium containing cefotaxime and subsequent stage on selection medium containing kanamycin. Initially recovered PLBs following co-cultivation, were allowed for further selection with kanamycin. New PLBs and leaves were formed from the transformed PLBs when they were regenerated under kanamycin selection. Successful transformants were obtained with a maximum transformation efficiency of 3.0 per cent with the addition of acetosyringone 100 mM. Out of the kanamycin resistant PLBs 69.23 per cent were GUS positives. The newly formed PLBs and leaves when subjected to histochemical GUS assay indicated successful transformation and it was detected as distinct blue spots on the PLBs after 24’ hours. The PLBs which were not co-cultivated with Agrobacterium were green in colour. The transformation were further confirmed by PCR analysis.

ABSTRACT

Cassava has developed importance mainly as an industrial crop due to its ability to store large quantities of starch (74% to 85% of dry weight) within secondary root structures. Cassava starch is exploited as a source of human food, feed for livestock and as raw material for the food processing and chemical industries. Research accomplishments during the last four decades helped India to have the top position among cassava growing countries with an average yield of 24 t ha-1. However several traits like starch quality, high protein , post harvest deterioration etc can be altered only through the development of transgenics. The present study summarizes the results of the experiments undertaken to optimize the protocol for the development of transgenic cassava through Agrobacterium mediated gene transfer. Friable Embryogenic Callus (FEC) was developed in the cassava var. TMS 60444 us ing Murashige and Skoog (MS) media with picloram (50μM). Experiments were conducted to study the efficiency of different Agrobacterium stains viz, AGL 8, GV3101, C58C1, LBA 4404 and EH105 in transforming FEC. The strains differed in their growth rate. The strains EH105 and AGL were slower in growth as compared to other strains and the protocol was modified accordingly. Among all the strains, EH105 resulted in maximum recovery of transgenic tissues. The paper also summarizes the present status of development of transgenic cassava along with future thrusts.

ABSTRACT

An efficient in vitro micro propagation protocol was developed for direct shoot produc tion of Contoloupe (Cucumis melo variety (NS Abhijit) using cotyledons as explant. The excised cotyledons were cultured on Murashige and Skoog’s medium (MS) contain- ing plant growth regulator, 6 Benzyl adenine with various concentrations for the study of shoot production. The best condition for the shoot production was with 1.2mg/l Benzyl adenine (BA) in MS medium. The shooting frequency was 65% and three shoots were obtained from each explant after 14 days of culture. The shoots (~1.5 cm length) were rooted most effectively on 0.1mg/l of α Naphthalene acetic acid (NAA) supplemented MS medium. We have also standardized the transformation protocol for GUS gene in the muskmelon variety Eden Gem to see test the feasibility of this variety for Agrobacterium mediated transformation. The transformed explants were confirmed through Indigogenic GUS assay.

ABSTRACT

Biofertilizers can supplement the chemical nutrients and helps in reducing the production expenditure in agriculture. Crop biofertilization with microbial consortia is an important strategy to improve the physical, chemical and biological conditions of the soil. The facultative methylotrophs are capable of fixing atmospheric nitrogen, besides it produces phytohormones viz; IAA, GA and cytokinin, vitamin B₁₂, polysaccharides, PHB, and siderophores. The essential nutrients are considered as the most important, among the factors limiting growth and yield of the tomato crop. The application of adequate amount of fertilizers is a pre-requisite for exploiting the genetic potential of tomato variety. Therefore, efficient and economic use of the fertilizer with microbial consortia would help in decreasing the input costs for raising bumper crop.

More number of studies carried out on the effect of facultative methylotrophs in different parts of the world and in India. The results confirmed that the facultative methylotrophs have a greater importance in increasing the growth and yield of various crop plants. Keeping this in view an experiment was conducted to find out the interaction effect of facultative methylotrophs with other bioinoculants viz., Azospirillum, Azotobacter, PSB and VAM (by seedling dip method followed by PPFM phyllosphere spray) on yield of tomato var Ruchi. The results showed that the treatment with above microbial consortia (50.71 t/ha) was found to increase the yield of the tomato crop by 62.73 percent over uninoculated control (18.90 t/ha) and individual treatments.

ABSTRACT

Phytophthora rot is the most destructive disease of black pepper nursery inflicting heavy crop losses. Considering the seriousness of the disease, the present study was undertaken to isolate and select the efficient antagonists from pepper nurseries raised in Government farms at Chelakkara, Pazhayannur, Mannuthy and Pananchery of Thrissur district. A quantitative estimation of the rhizosphere microflora from different nurseries revealed more abundance of soil bacteria followed by fungi and actinomycetes. Based on cultural characters of the rhizosphere microflora, 22 fungi, 20 bacteria and five actinomycetes were selected for further studies. Antagonistic action of soil microflora against Phytophthora capsici was studied by dual culture method in comparison with that of standard culture of T. harzianum. The study revealed that all fungal isolates and five bacterial isolates were antagonistic towards the pathogen with varying degrees. Among the fungal isolates, 13 isolates including T.harzianum recorded cent per cent inhibition of P.capsici. From among the fungal isolates that showed antagonistic reaction in dual culture, further selection of efficient ones was carried out based on the antagonistic index (AI). The isolate 22 F₂ (Trichoderma sp.) showed the maximum AI of 3000 followed by isolate 34 F₂ (Trichoderma sp.) and T. harzianum with an AI of 1500 each. The cultural and morphological characters of the efficient native isolates (22 F₂ and 34 F) were studied and were identified as Trichoderma longibrachiatum Rifai aggr. and Trichoderma viride Pers. ex S.F. Gray aggr. The three antagonists were found parasitic on P.capsici as evidenced by excessive coiling, penetration and disintegration of the hyphae and thereby these Trichoderma spp. used in the study is found to have effective parasitic effect on the pathogen in addition to some degree of antibiosis and these fungi could be utilised for the management of Phytophthora rot in black pepper nursery.

ABSTRACT

Studies on the impact of bio-fertilizers with levels of NPK on bulb yield, dry matter production, nutrient uptake and cost: benefit ratio in onion (cv. Bellary Red) were conducted in medium black soils during kharif and rabi seasons of 1999 and 2000 both under irrigated and rainfed conditions at the Agricultural Research Station, Hiriyur located under Central Dry Zone of Karnataka. The treatments were; T₁-Control, T₂-Azotobacter, T₃-Azotobacter+75% N+PK, T₄-Azotobacter +100%N+PK, T₅-Azospirillum, T₆-Azospirillum+75% N+PK, T₇-Azospirillum+100%N+PK, T₈-VAM, T₉-VAM 75% P+NK, T₁₀- VAM+100%P+NK, T₁₁ ? recommended NPK(125:50:125 kgha-1) and T₁₂-75% recommended NPK. The experiments were laid out in RBD in three replications.

The bio-fertilizers Azospirillum brasilence (500g ha-1) and Azotobacter vinelandii(500g ha-1) were applied to seedlings at transplanting separately by dipping the roots in the slurry for 15 minutes. The VAM fungi - Glomus mossae was applied to soil @ 1 kilogram per square meter area of nursery bed. The recommended dose of FYM @ 25 t ha-1 was applied to all the treatments as basal dose.

The results of the investigations revealed that the plants provided with Azospirillum + 100% N+PK (T₇) produced highest bulb yield (250.29;339.02;232.98 q/ha), highest total dry matter production (7053.96;8992.78;6287.03 kg/ha), maximum uptake of N, P a nd K ( 1 5 7 . 8 8 : 7 3 . 5 6 ; 7 0 . 4 6 N PK k g / ha ) , ( 2 0 6 . 8 6 : 9 6 . 2 5 : 9 1 . 1 0 N PK k g / ha ) , (131.81:50.25:71.24 NPK kg/ha) and also the maximum cost:benefit ratio of 4.00 (1:400), 5.42 (1:5.42), 3.94 (1:3.94) under irrigated condition during kharif and rabi seasons and under rainfed situation during kharif season respectively.

Thus, among all the treatments, application of Azospirillum + 100% N + PK (Azospirillum + 125:50:125 NPK kg/ha) was found to be most remunerative considering the nutrient uptake, bulb yield and total dry matter production and cost:benefit ratio, which gave the highest returns. This level can be followed for Bellary Red onion irrespective of seasons both under irrigated and rainfed conditions in the Central Dry Zone of Karnataka.