eChapters

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.

10.1 INTRODUCTION

‘Tissue culture’ is commonly used as a blanket term to describe all types of plant cultures, viz., callus, cell, protoplast, anther, meristem, embryo and organ cultures (George, 1993). It relies on the phenomenon of cell totipotency. The different techniques of culturing plant tissues may offer certain advantages over traditional methods of propagation.

Growing plants in vitro in a controlled environment, with in-depth knowledge of the culture conditions and the nature of the plant material, ensures effective clonal propagation of genetically superior genotypes of economically important plants. Micropropagation has become an important part of the commercial propagation of many plants (George and Sherrington, 1983; and Zimmerman et al., 1986) because of its advantages as a multiplication system (Pierik, 1997 and Razdan, 2003). Several techniques for in vitro plant propagation have been devised, including the induction of axillary and adventitious shoots, the culture of isolated meristems and plant regeneration by organogenesis and/or somatic embryogenesis (Williams and Maheswaran, 1986; and Gautheret, 1985).

Tissue Culture is becoming as an alternative means to vegetative propagation of plants. In vitro growing plants are usually free from bacterial and fungal diseases. Virus eradication and maintenance of plants in virus free stage can also be rapidly achieved in cultures (Sharma, 2016).

Introduction

The horticulture sector has been a driving force in stimulating a healthy growth trend in Indian agriculture. This sector contributes around 28 per cent of the GDP from about 13.08 per cent of the area and 37 percent of the total exports of the agricultural commodities (Rather et al., 2013)  India is currently producing 257.2 million tonnes of horticulture produce from an area of 23 million ha. Over the last decade, the area under horticulture grew by about 3.8% per annum but production rose by 7.6% per annum. The higher growth rate in horticulture was brought about by improvement in productivity of horticulture crops, which increased by about 28% between 2001-02 and 2011-12. (Anonymous, 2012)

1. INTRODUCTION

Food processing involves the use of wide range of unit operations and technologies to transform the raw, bulky and perishable food into more shelf-stable, palatable and marketable food products. Food processing is one of the most effective solution to reduce the wastages, improves the quality and safety aspects of food products, increase the food availability and marketability. Fruits and vegetables are considered as an integral part of any dietary system as these are rich sources of vitamins, minerals and dietary fibres. India is the second largest producer of fruits and vegetables after Brazil and China which contributes 10 % of world fruit production and 14 % of world vegetable production. Fruits and vegetables are more prone to spoilage as compared to cereals due to their nature and composition. The spoilage can be occurr at the time of harvesting, handling transportation, storage, marketing and processing that results in lot of wastage in the form of seeds, pulps, peels etc. Efficient management of these wastes can help in preserving vital nutrients of fruits and vegetables, and bringing down the cost of production of processed foods, besides minimizing pollution hazards. In India, processing of fruits and vegetables is extremely low as compared to the developed countries.

Biotechnology can help in the bio-processing of fruits and vegetables for the production of novel products having better sensory and nutritional qualities, improved shelf life. Bio-processes involve the use of different micro-organisms and enzymes along with modern biotechnological techniques like genetic engineering.

9.1 INTRODUCTION

India is the second largest producer of fruit crops with annual production of 68.4 million metric tonnes from an area of 6.1million ha (Anon 2010). The national horticulture mission commenced in 2005 had fixed a target to double the horticultural production by 2011-12. According to the available production data of horticultural crops, it seems that we are very close to achieve our target. But, if the loss caused by biotic and abiotic stresses is accounted, a sound conclusion can be drawn that still there is a huge scope for productivity improvement in fruit crops.

Among biotic factors bacteria, fungi, nematodes, viruses and insect pests, diseases and other pests are widely recognized as the biggest threat to the sustainable production of fruit crops. Every fruit crop is infected with one or other pathogen/pest, which causes a significant reduction in yield. The disease alone both in field and storage accounts for 25-30 per cent losses (Rangaswami and Mahadevan 2002). Along with diseases, insect-pests also cause reduction in the yield. About 600 insect and mite pests have been reported in temperate fruits alone in India (Sharma and Singh 2006).

Beside biotic factors, abiotic factors (salt, drought, low and high temperature) also limit the potential yield of fruit crops. It is estimated that nearly 10 million ha is affected by salinity/alkalinity in India (Singh 2006). Moreover, fruit cultivars are also to be modified for yield and traits of consumers preference e.g. seedlessness in grapes and citrus.

Genetic improvement can be achieved by following the conventional breeding approaches, when the gene of interest is present in related cultivars/species and the crop is sexually crossable. But due to long juvenility and genetically prevalent problems like incompatibility, sterility in most of the fruit crops and apomixis in citrus, mango and apple, conventional techniques seem to have limited role in the improvement of fruit crops.

Various biotechnological approaches either alone or in combination with conventional breeding approaches can provide a direct solution for the genetic improvement of fruit crops.

Origin, History and Definition

The term biotechnology was coined by Karl Ereky, a Hungarain engineer, in 1919. The origin of biotechnology can be traced back to prehistoric time when armers and gardeners have been making use of hybridization and grafting among plants with a view to bring about an improvement in existing attributes. Using these approaches, several hundred varieties in ornamental plants were produced. Microorganisms were already used for processes like fermentation, formation of yoghurt and cheese from milk, vinegar from molasses, production of butanol and acetone from starch by Clostridium acetobutylicum or the production of antibiotics like penicillin from Penicillium notatum. Four defferent phases of biotechnological research were witnessed during the last century. In 1920, Chain Weizmann used Clostridium acetobutylicum to convert starch into butanol and acetone (during world war I, acetone was used as an essential component of explosives). Alexander Fleming in 1929 discovered penicillin from Penicillium notatum and wounded soldiers were given treatment using this antibiotic during world was II. In 1970s with the discovery of restriction enzymes, which led to the development of recombinant DNA, technology brought a revolution in the field of biology. The latest phase in the history of biotechnology is the study of structure and function of whole genome (DNA) and whole proteome (protein). This area of research is described as genomics and proteomics, respectively. Some more important activities in the field of biotechnology are given in Table1

In fact, biotechnology is the product of interaction between science of biology and technology. It is the technological exploitation and control of biological system. Biotechnology has wide range of its uses and accordingly Office of Technology Assessment described biotechnology as: (i) old biotechnology that includes fermentation, i.e. formation of vinegar from molasses and that of cheese and yoghurt from milk, production of antibiotics from certain fungi and process of baking and brewing and (ii) new biotechnology that includes the techniques of recombinant DNA and PCR polymerase chain reaction, cell culture and fusion and bioprocessing which became possible only through the researches in molecular biology. However, no efforts have been made to distinguish between old and new biotechnology. Keeping in view the importance of biotechnology, attempts have been made to define it. Following are some of the definitions of biotechnology, however, none of the definitions of biotechnology seems to be complete. The first definition seems to be comprehensive and wide.

11.14. Nanotechnology 11.14.1. Concept • Nanotechnology is science, engineering, and technology conducted at the nanoscale, which is about 1 to 100 nanometers. • Physicist Richard Feynman is considered as father of nanotechnology. The idea and concepts behind nanoscience and nanotechnology started with a talk entitled “There is Plenty of Room at the Bottom” by physicist Richard Feynman at an American Physical Society Meeting at the California Institute of Technology on December 29, 1959, long before the term nanotechnology was used. In his talk, Feynman described a process in which scientists would be able to manipulate and control individual atoms and molecules. Over a decade a later, in his explorations of ultra precision machining, Professor Norio Taniguchi coined the term nanotechnolog

It is difficult to define ‘stress’ precisely. In engineering and the physical sciences, it is usually defined as the force applied per unit area. But as far as in biology, stress it is usually described more loosely as any factor that disturbs normal functioning of an organism. Any environmental factor potentially unfavorable to plant is termed as stress.

The effect of stress on plant condition is called strain. Stress is the action, whereas strain is the reaction. Stress may be defined as external conditions that adversely affect growth, development and productivity. Stress may further classified as:

• Zero stress: This is the level of exposure of plants to environmental factors that leads to neither injury nor reduction in growth and yield of crops.
• Stress escape: Capacity of the plant to complete its life cycle during wet season before onset of stress.

2.1 Insect Pests

Insects are often a major obstacle to the successful production of cucurbits (York, 1992). They can severely reduce yield, injure or kill plants, spread diseases and adversely affect fruit quality. A number of insect pests, mites and nematodes have been found to infest cucurbitaceous vegetables but fortunately, in India, fruit flies and a few species of beetles are of economic importance; aphids and blister beetles though of regular occurrence, seldom cause severe damage.
 
Other insect pests reported to infest the cucurbitaceous vegetables are pumpkin caterpillar, Diaphania indica (Sounders), snake gourd semilooper, Plusia peponis Fab., white fly, Bemisia tabaci Genn., stink bugs, Apspongopus spp. and flea beetle, Phyllotreta spp. etc. These biotic pressures on the crops are increasing day by day due to round-the-year intensive cultivation of photo and thermo insensitive cultivars as well as indiscriminate use of toxic agro-chemicals in the present era of chemicalized agriculture.

Bitter gourd (Momordica charantia) is a widely grown vegetable variety in India. It is one of the important members of the family Cucurbitaceae. The seed production can be done throughout the year in tropical climate and during spring, summer and rainy season in subtropical climates. In Tamil Nadu September – October is suitable for seed production. In hills seed production can be done during summer season.

Bitter gourd (Momordica charantia L., 2n = 2x = 22, family Cucurbitaceae, monoecious, cross-pollinated but genetically behaving like self-pollinated crop), also known as balsam pear, bitter melon, bitter cucumber, and African cucumber is rich source of vitamin C and iron and credited with medicinal use in management of diabetes. Fruits are cooked with other vegetables, stuffed, stir-fried, or added in small quantities to beans and soups to provide a slightly bitter flavor. The bitterness of most cucurbits is mainly due to cucurbitacins. The bitterness of bitter gourd is due to the cucurbitacin-like alkaloid momordicine and triterpene glycosides (momordicoside K and L) as reviewed by Behera et al. (2010). Bitter gourd is grown on approximately 340,000 hectares across Asia, with India and China accounting for more than half the production. Estimated area of bitter gourd in India is about 80,000 ha with production reaching to 10,00,000 resulting into a productivity of 12 tons/ha. States accounting for major area of bitter gourd are Chhattisgarh, Telangana, Andhra Pradesh, Orissa, Madhya Pradesh, Uttar Pradesh, Bihar, Tamil Nadu, Haryana and Assam.

It is rich in iron and Vit –C and native of tropical Asia particularly eastern India and Southern China (Sheshadri 1986 Mini Raja et al., 1993).
 
Botany: Monoecious annual, staminate flowers small, yellow and borne on slender pedicel. Pistillate flowers solitary have small pedicel, flowers are yellow, leaves segmented pentamerous. Filaments 3, two are bilocular and one is unilocular. There are three short styles ended with three bilobed or divided stigmas.

Chromosome 2n=22: 6-centromere in middle, 3-submedian, 2-subterminal  (Vargheuse, 1973) 

Momordica dioica: 2n=28

Momordica cochincniensis: Sweet gourd of Assam

Germplasm

KAU-Vellanikara, IIHR-Bangalore, NDUAT-Faizabad, CSAUAT-VRS, Kalyanpur, BPUA&T-Pantnagar and IARI- New Delhi are the major sources of bitter gourd germplasm.

Bitter gourd (Momordica charantia L.) is one of the important commercial cucurbitaceous cash crops grown mainly by small holding farmers in Asia where it is cultivated on more than 340,000 ha annually (McCreight et al., 2013). Eastern Asia, possibly Eastern India, or Southern China is most likely the center of origin for bitter gourd (Walters and Decker-Walters 1988; Miniraj et al., 1993). It is also known as bitter melon, bitter cucumber, balsam pear, bitter apple, or bitter squash (Morton, 1967). Fruits are known for abundant source of vitamins and minerals. Delicious preparations are made from fruits after stuffing and frying. It is also consumed in the form of Juice, Tea and Pills. Bitter gourd fruit abounds in phytonutrients (Dhillon et al., 2017) and often used to manage type 2 diabetes as a folk medicine. Clinical studies confirmed that elevated fasting glucose levels in pre-diabetics can be reduced by supplementing the diet with bitter gourd fruits. (Amirthaveni et al., 2018; Krawinkel et al., 2018). Medicinal properties present in the bitter gourd fruits are used in the curing of diabetes, asthma, blood diseases and rheumatism.

The current seed market of bitter gourd in Asia values ≈ 16 million Euros. The bitter gourd seed market in India alone is 400 metric tonnes having value of 350-400 crores. 70 % area is under hybrids and 30 % is covered by open pollinated varieties.

Bitter gourd or Bitter melon, Momordica charantia L under the family Cucurbitaceae (Chromosome no. 2n = 2× = 22) is widely grown in India, Indonesia, Malaysia, Singapore, Thailand, China, Japan, East Africa, the Caribbean and South America for its edible fruit. Tender fruits are widely used as cooked vegetables in a number of ways. Bitter gourd is an excellent source of vitamin C, iron, magnesium, potassium, calcium and ß carotene. The fruits contain non-toxic glycoside momordicin, which gives it bitterness and also health benefits. It also contains an insulin-like compound called Polypeptide-p or p-insulin and steroidal compound “Charantin” having hypoglycemic property.
Origin, domestication and adaptation in India
Wild or semi-domesticated variants spread across Asia in prehistorical period, and it was probably domesticated in Southeast Asia. It has probably originated in eastern India or Indo-Burma area or southern China. Uncarbonized seed coat fragments have been tentatively identified from the spirit cave in northern Thailand however, there have been no archeological reports of bitter gourd remains in China (Marr et al, 2004).Population resequencing revealed the divergence between wild and South Asian cultivars about 6,000 years ago, followed by the separation of the Southeast Asian cultivars about 800 years ago, with the latter exhibiting more extreme trait divergence from wild progenitors and stronger signs of selection on fruit traits.

Anthracnose
Diagnostic Symptoms
• Pathogen attacks all aerial parts of plants and at any stage of their growth.
• Symptoms are circular, black, sunken spots with dark center and bright red orange margins on leaves and pods.
• In severe infections, the affected parts wither off.
• Seedlings get blighted due to infection soon after seed germination.
• Infected pods have discolored seeds (Fig.3).
Etiology
Scientific Classification
Kingdom : Fungi
Division : Eumycota
Sub- division : Deuteromycotina

Bleaching is essentially whitening or decolourizing. It is an important step in the processing of dry plants. It involves chemical processes that can change the ability of colour bodies to absorb light by changing their degree of saturation (Robson, 1967). This chemical action removes the discolouration occurring during the dehydration phase of preservation of plants. It allows the effective use of dyes for colouring. Bleaching agent is a material that lightens or whitens a substrate through chemical action. To reduce the processing costs, multistage bleaching is suggested. In this method, both oxidative and reductive bleaching chemicals are used.

Plant biodiversity, in particular, is arguably the single greatest resource that humankind has garnered from nature during our long cultural development. Nowhere is the value of biodiversity more evident than in our food supply. Roughly one third of all plant species have edible fruits, tubers, nuts, seeds, leaves, roots, or stems. The World Health Organization estimates that 3.5 billion people in developing countries rely on plant-based medicine for their primary health care, in which tuber crops also play a major role (Tuxill, 1999). There are 25 hot spots of biodiversity of which Western Ghats and Eastern Himalayan are the most important ones in India. About 7% of world flora and 6.5% of world fauna are in India. Indo-Burma region is one of the eight hottest hot spots of the world, and is the centre of origin of tuber crops like Dioscorea, Colocasia, Amorphophallus and Alocasia.

Onion is an economically significant bulb crop cultivated across diverse agroecological environments. Its cultivars vary considerably in bulb size, shape, colour, pungency, maturity, storage behaviour and photoperiod response. Understanding its vegetative and reproductive morphology, growth habit, physiology and taxonomic relationships is fundamental to crop improvement and germplasm conservation. This chapter examines the botanical features and taxonomic position of onion within the genus Allium. It explains relationships among cultivated types, landraces and allied species and discusses varietal classification according to climatic adaptation and consumer preference. Classical characterization and molecular taxonomy are considered important tools for developing productive, resilient and market-preferred onion cultivars.

Botany: Bottle gourds belonging to the family cucurbitaceae is an annual vine having white flowers and smooth, large, hard-shelled gourds. Brownish seeds are numerous in a whitish green pulp. Each seed is somewhat rectangular in shape with grooved notches near the attached end. Grown most often in warmer climates, this squash grows from 6 to 36 inches long and 3 to 12 inches in diameter.

Seeds leaves, flowers and young stems are all edible. Helpful in shedding extra calories. It contains higher concentrations of Dietary Fiber, Vitamin A, Vitamin C, Vitamin K, Vitamin B6, Folate, Potassium, Manganese, Protein, Vitamin E, Thiamin, Riboflavin, Pantothenic Acid, Calcium, Iron, Magnesium, Phosphorus and Selenium. A rich source of minerals and vitamins, bottle gourds contains many healing and medicinal properties. The cooked vegetable is not only easy to digest but also contains cooling, calming (or sedative), diuretic properties. It contains low calories also has iron, Vitamin C and B complex. Regular consumption of this vegetable provides relief to people suffering with digestive problems, diabetics and convalescents.

Bottle gourd (Lagenaria siceraria, 2n = 2x = 22, family Cucurbitaceae, monoecious, cross-pollinated but genetically behaving like self-pollinated crop), is grown primarily in India on large scale for consumption as cooked vegetables. Bottle gourd is considered to be one of the earliest species of plants to be domesticated by humans. Bottle gourds are regularly grown and consumed in various parts of the world, particularly Asia. Fruits of bottle gourds are harvested young and used as a vegetable. The mature, dried fruits have been used throughout the world for making storage jars and utensils and hence the name bottle gourd. Shoots and leaves are also cooked and eaten. Lately, bottle gourd juice is also becoming popular in India as a health drink. Other countries growing bottle gourd on limited scale are Sri Lanka, Indonesia, Malaysia, Thailand, Philippines, China, Hong Kong, Tropical Africa, Colombia and Brazil. After seed scooping, the hard rind is used as containers also. Seeds are used for oil extraction and oil is used as cooking oil.

During 2017-18, India produced 27 lakh tons (2.7 million tons) of bottle ground from 1.60 lakh ha (0.16 million ha) and this translated into a productivity of 17 tons/ ha (Horticulture Statistics at a Glance-2018, MAFW, 2018). Top six states producing bottle gourd on significant scale are Bihar, UP, Haryana, MP, Odisha, and Pujab, production being in the range of 135,000 to 650,000 tons.