eChapters

1. Introduction

Cucurbits are group of plants belongs to family Cucurbitaceae which are mostly creeping or crawling in nature and distributed in tropical and subtropical zone of the globe. The family cucurbitaceae has around 120 genera and more than 800 species (Swarup, 2016). It holds the highest rank amongst all other plant families considering number and percentage of species used for human consumption either as fruit of the tender leaves and stems. Cucurbits have diverse daily use such as salad (cucumber and long melon); dessert (watermelon and muskmelon), curry or boil (bottle gourd, bitter gourd, sponge gourd, ridge gourd, summer squash, squash melon, pumpkin etc.); candy (wax gourd); pickles (pickling cucumber, pointed gourd); jam (pumpkin); utensil (bottle gourd), musical instruments (bottle gourd). From plant breeding point of view, the crops are cross pollinated and entomophilies.

For the first time, the Quality Planting Material (QPM) production from aerial stem node cutting and its comprehensive growth parameters are being reported in Valeriana jatamansi (Jones) (family: Valerianaceae), a highly endangered medicinal plant and a natural habitat in North Eastern Himalayan Region in the high altitude ranging from 1000 to 8000m Above Sea Level (ASL). The present communication identified a novel avenue exploring the unused plant part, aerial node cutting for producing QPM in a continuous and exponential rate without scarifying the rhizome and root, enriched with active ingredients like Valerenic acid and Valepotriates. This method also assures 100% plant emergence and more than 91±5.56% plant survival in the different field conditions without requiring hardening process as an additional merit. This first-time report for QPM production of an endangered medicinal plant, Valeriana jatamansi through aerial node cutting without following the hormone supplemented MS based tissue culture, a high cost process required sophisticated instruments and aseptic environment. This present process will have potentiality to explore commercially for mass clonal-propagation even at farmer level.

This study deals with the ethnomedicinal prospects of common weeds thriving in the wide array of terraced paddy fields used for rhizipisciculture by the Apatanis, a unique tribe indigenous to the remote Ziro valley in the Eastern Himalayas. Surveys were conducted in agricultural fields surrounding three villages in order to record the paddy field flora present on the partition bunds. Elderly villagers were also interviewed with the help of an interpreter to document their traditional knowledge about the medicinal uses of these weeds. A total number of 36 species of indigenous medicinal plants were recorded along with the local name, the plant parts used, the modes of preparation and administration of traditional herbal remedies and the illnesses treated. This easily accessible medicinal plant resource, ameliorating a wide range of common ailments, was found to be a reliable means for managing the primary healthcare of this indigenous agrarian community of the Eastern Himalayas.

Ethnobotany deals with different aspects of man-plant relationship with special emphasis on ethnic community of a region, nay country or any geographical territory. This perhaps gives us a new dimension, orientation and objectives to make it complete with vivid information on people and their resource use pattern including science background behind scientific use. The new form of ethno-botany is now reformed and re-oriented as beta-ethnobotany. Earlier taxonomists, anthropologists and social workers have made their work on a wide field of man-plant relationship which was very loose in the sense that no specification or special focus was there which has its origin so called alpha-ethnobotany. Beta-ethnobotany deals with objective based oriented studies, divine, dynamic and quantitative ethno-botany with special emphasis to assessment of credibility of folk claims, ethno pharmacology, ethno-pharmacotherapy and ethno-medicine. Presently scientists estimated that out of over 5000 plants have been used in ethno-medicine much less than 1000 are utilized in herbal drug industry in different indigenous systems of medicine like ayurveda, homoeopathy, unani and siddha. A wide variation of use and conservation strategies on that medicine or the raw sources of these materials are available in the lower tract of Chotanagpur plateau in red lateritic part of West Bengal so called the ‘ rarh’ region. Tribal people like santal, kol, sabar, kheria, oraon, munda etc. inhabiting the rarh region use the medicine as ethno-medicine since time immemorial and conserve the plants and similar plant propagules in their home gardens and nearby sal (Shorea robusta) dominated forests for future generation. Wide application and conservation strategies of a large number of ethno-medicine and ethno-medicinal plants have been presented here with special emphasis on landscape conservation to make the ecosystem pristine rather than degraded in near future.

1. Introduction

Valeriana jatamansi Jones is considered to be highly endangered medicinal plant found mainly in northern part of Himalayan region. Although it is also found in all temperate regions of world except in Australia (Jain, 1968; Polunin and Stainton, 1987; Bennet, 1987). It is grown in Himachal Pradesh, northern part of U.P., Sikkim and newly introduced in West Bengal Darjeeling region by us. It is enlisted in the endangered category by IUCN (Kaul and Honda, 2000) and it is enlisted as an endangered species in the national medicinal plant board, New Delhi (www.nmbp.nic.in). The plant was reported to be found in fragmented populations in different places of Kashmir Himalayan hilly region like Dara, Sonamarg, Harwan, Gulmarg, Ferozpur, Pahelgum, Yusmarg, Shajnar (Naqashi and Dar, 1982-86). It is found to be in the margin of extinction from the world though it is widespread in the temperate regions of whole world except Australia (Nayar and Sastri, 1998). Indiscriminate collection of rhizomes from the plants made potential threat of its existence in some previously founded localities (Wyatt, 1981; Nayar and Sastri, 1998; Airi et al, 2000; Nautiyal et al, 2003, CAMP, 2003).

Dioscorea L. commonly known as yam is the largest genus in the family Dioscoreaceae with 602 species distributed in the tropical and subtropical regions. It is an important food crop next to cereals and grains due to high yield storage of carbohydrates which form an important place in the dietary habits of small and marginal farmers especially in the food security of tribal population. The tubers are rich source of various secondary metabolites like Diosgenin, a commercially important bioactive sapogenin used as a precursor in the manufacturing of sex hormones, oral contraceptives and other pharmaceutically important steroidal drugs. Despite of its huge diversity only few species are recognized to be useful for medicinal and edible purposes. About 50 species of Dioscorea are distributed in India. The North East India, one of the hotspot of biodiversity is particularly rich in tropical root and tuber crops. Dioscorea species are in luxuriant growth in wild habitat in this region. The present study mainly addressed to explore the ethno medicinal uses, phytochemical analysis and also focused on safe conservation of sixteen species of Dioscorea found in this region.

1. Introduction

The North east India (located between 87Ú32’E to 97Ú52’E latitude and 21Ú34’N to 29Ú50’ N latitude) forms a distinct part of the Indo-Burma hotspot which rank the 6th among 25 biodiversity hotspot of the world is climatically, ecologically and ethnically very diverse. The region is largely dominated by tribals such as Naga, Khasi, Mizo, Miri, Adi, Aka, Apatani, Maripa, Mushard, Garo, Tagin Naga and Singhpho who are sociocultural different. The North east region, besides having large floral biodiversity, is yet not fully explored. The forest resources available in this area are most often ill managed due to lack of awareness and anthropogenic activities. Various wild roots and tuber crops are consumed by the indigenous group of North East India which have very less scientific documentation although. As wild tuber crops available in North East India, Dioscorea species play a prime role in providing food and medicinal requirements for the indigenous communities of this region. Hence the present paper focuses on the ethnobotanical importance and qualitative screening of few important phytochemicals present in the various species of Dioscorea, their conservation strategies and sustainable utilization in North East India.

1. Introduction

Jackfruit (Artocarpus heterophyllus) is one of the most significant trees in tropical and subtropical homegardens and perhaps the most widespread and useful tree in the important genus Artocarpus. Jackfruit belongs to the family Moraceae. The word “jackfruit” comes from Portuguese jaca, which is derived from the Malayalam language term, Chakka. Jack fruit native to India and seen abundant in Western Ghats (Wangchu et al.,2013). Besides India, jackfruit is commonly grown in home gardens of tropical and sub-tropical countries especially Sri Lanka; Bangladesh, Burma, Philippines, Indonesia, Thailand, Malaysia and Brazil. In India, it widely distributed in the states of Assam, West Bengal, Uttar Pradesh, Maharashtra, Kerala, Tamil Nadu and Karnataka and considered to be the “Poor mans food” (Prakash et al., 2009). In Malayalam, jack fruit is called as “Chakka” while the ancient Indian language Sanskrit refers as Atibruhatphala (Baliga et al., 2011). The morphology of the tree varies with 10-30 m tall; with long tap root and dense crown producing the largest tree born fruit in the world. The fruit weight up to 50 kg, but average weigh is considered to be 10 kg, while only 30-35% of the bulb is edible. Jack fruit is considered as national fruit in Bangladesh and highly appreciated in India due to cheap and availability in summer seasons were food is scarce (Muralidharan et al., 1997). It is a monoecious tree and both male and female inflorescences are found on the same tree. The fertilization is by cross-pollination and the propagation is mostly through seeds. The complete fruit development process takes about three to seven months from the pollination, varying in different countries. Jackfruit consists of lower fleshy edible region (bulb), middle fused region (syncarp) and out spiney region (spike). When ripe the fruit get fleshy, outer spines widened and flesh get soft and yellow. Except the thorny outer bark and axis are not edible (Baliga et al., 2011). The jackfruits were classified based on their phonotypical and organoleptic characteristics with variation in bulb colour as well as shape, size, odour, flake size, flake colour and period of maturity. Two types of ecotypes are recognized, one with soft and spongy flakes or bulbs while other with firm carpels.

1. Introduction

The interest in the uses of medicinal plants has grown considerably from the mid of 20^th century with the introduction of phytochemical analysis of natural products from various plants parts (Matos, 1996). Most studies survey specific local communities to examine patterns of use of plant part for treating various aliments along with the influence of economic and social variables on plant use (Albuquerque, 2006). Medicinal plants are the base for the development of new drug, which is the basic mode of survival of human and livestock from many diseases to maintain their healthcare (Uddin et al., 2014). Number of studies has been carried out globally on ethnomedicinal uses of plants among various indigenous communities (Alexiades and Sheldon, 1996; Basualdo et al., 1995; Martin, 1995). Particularly from the region Karbi Anglong, Assam (Teron and Borthakur, 2012, 2013; Terangpiet al., 2014; Teronpi et al., 2015) have also conducted many notable work on ethnomedicinal. While in all these studies qualitative approaches have not been adapted to document ethnobotanical information. The quantitative approaches on ethnobotany have grown steadily in the last two decades. Today researchers have developed and applied quantitative methods to the various ethnobotanical data to test different hypotheses about the relation between plant species and humans (Reyes-García et al., 2006). Quantitative ethnobotanical indices were applied to determine how important these plants are to ethnic and indigenous cultures whether, as food, veterinary medicine, human medicine or economic value from various non-timber forest products (Pieroni, 2001; Upadhyay et al., 2011; Reyes-García et al., 2006). Therefore, the aim of this study was to document the various uses of the medicinal plant Geophila repens (L.) I.M. Johnst by the Karbi tribe and to apply quantitative methods to determine the specific importance of these plants for treating particular ailment. G. repens is a very rare plant with high medicinal potential (Roy et al., 2011) and is global distribution is reported from India (Assam, Meghalaya, Andaman Islands and Western Peninsula); Bangladesh; Bhutan; Sri Lanka; Nepal; Malaysia; South China; Fiji; Tropical America; and Africa.

1. Introduction

Pummelo is considered as largest fruit among all the Citrus species from the Rutaceae family. It is also known as shaddock, papanus or chakotra. Although C. grandis (L.) Osbeck is more frequently used, however C. maxima (Burm.) Merr. is considered as correct under the International Code of Botanical Nomenclature (Singh and Navneet, 2017). It is considered as one of the four true species (natural, non-hybrid) of Citrus. The ltypical large size, thick rind is one of the causes for slow acceptance of pummelo in international market. However, it is presently considered as important fruit due to presence of various bio-active compounds related with human health and nutrition. The word ‘pummelo’ was originally derived from the Dutch word ‘pompelmoes’. The fruit is also named as ‘Shaddock’ after a Captain Shaddock who was believed to have introduced the fruit in Barbados on an East India Company ship on its way to England in the 17^th century. It is known by different names by different communities residing in India such as chakotra (Hindi), batabi lebu or jambura (Bengali), robab tenga (Assamese), nobab (Manipuri), ser trawk (Mizo), soh myngor or soh bah (Khasi), rabab tasing (Adi), papanas (Gujarati), chakota (Kannada), kambili naranga (Malayalam), pamparapanasa (Telugu), metukki or pampalimaku (Tamil) and bampara (Marathi) [Anon, 2019]. Pummelo can be distinguished from grapefruit by its larger size fruit, thicker albedo and the bunch bearing habit of grapefruit can also be considered for distinguishing it from pummelo (Scora et al., 1982). The crop is believed to have originated from South and Southeast Asia and it is now cultivated in many tropical and semi tropical countries. The top ten world producers of pummelo and grapefruit are USA, China, Mexio, South Africa, India, Argentina, Turkey, Cuba, Brazil and Tunisia (FAOSTAT, 2010). It is also cultivated in in Bangladesh, Cambodia, Chile, Philippines, Vietnam, Sri Lanka and Nepal. There is no report of its commercial cultivation in India till date; rather it is mainly grown in homestate gardens across the country including UP, Punjab, Maharashtra, Tamil Nadu and Karnataka. However, maximum genetic diversity in pummelo germplasm is found in North Eastern Region, West Bengal and Bihar (Singh et al., 2015).

While reviewing the status of education and research on medicinal plants in Bangladesh, it was found that there are 27 government and private educational institutions currently engaged in offering Bachelor or Diploma degree on Unani and Ayurvedic systems and training of traditional medicine practitioners in Bangladesh. Several other public and private universities have included different aspects of medicinal plants in their syllabus with other degrees at undergraduate and post graduate level. At present, there are 277 Unani, 201 Ayurvedic and 32 herbal drug manufacturers in Bangladesh with 6498 Unani, 4058 Ayurvedic and 537 herbal products registered by the Directorate General of Drug Administration. About 500 species of plants are identified which possess medicinal properties and are distributed throughout the country. There is no distinctive research organizations at present for conducting planned research solely on medicinal plants; though, the Bangladesh Forest Research Institute have some research programme on medicinal plants. Hamdard University- a private university is offering degree on Unani and Ayurvedic systems and conducting some research including drug manufacturing. Bangladesh Agricultural Research Institute with other public and private universities are conducting some hectic research on medicinal plants including some high yielding variety development. The Government of Bangladesh (GoB) have taken some initiatives for development of Unani and Ayurvedic medicine sector like making ‘Bangladesh Unani and Ayurvedic Practitioners Ordinance, 1983’, ‘National Drug Policy 2016’, establishment of ‘Board of Unani and Ayurvedic Systems of Medicines (BUASM)’, etc. But proper implementation of the ordnance, strengthening the activities of BUASM and taking organized research programme on medicinal plants are very much essential for having skilled professionals, quality drugs and research.

1. Introduction

There are a large number of indigenous rice varieties in India, which are still grown by the tribal people and small farmers of the remote areas where the modern agricultural practices, sufficient foods as well as healthcare systems are a dream. Nature has provided them some alternative ways. They have different indigenous rice varieties with its nutritional and medicinal values. Until recently, rice was considered only a starchy food and a source of carbohydrates and some amount of protein. Rice protein, though small in amount, is of high nutritional value (Chaudhary and Tran, 2001). It has extensive curative properties known from the ancient days. In fact, specific rice varieties with medicinal properties were cultivated and used in the treatment of some ailments in different counties of South East Asia.

During recent times, both at national level and international levels, the land races are being preserved in the gene banks. The scientists said that they were also checking these land races for some such characteristics which were not available in the existing varieties so as to incorporate the same in the present varieties (Sultan and Subba Rao, 2013). Traditional varieties are experiencing an increasing trend among consumers, due to their incredible health benefits for example the traditional varieties possess higher amount of glutamic acid, fiber and vitamins. Some people also credit traditional varieties with other health benefits, such as giving sensations of cooling in the body, improves vocal clarity, eyesight, fertility and mitigating rashes (Caius, 1999).

1. Introduction

Medicinal plants are used to be the valuable sources of new drugs (Nalawade et al., 2003; Chen et al., 2010;Chacko et al., 2010; Hamilton 2004). In Europe, about 1300 plants were used as medicinal plant, of which 90% are collected from wild resources. In United States, about 118 out of 150 prescribed drugs made up of natural sources (Balunas and Kinghorn 2005)

Moreover in developing countries, about 80 % people are totally reliant on herbal drugs and over 25 % of prescribed medicines derivative from wild plant species (Hamilton 2004). With the increasing demand for herbal drugs, natural health products, and secondary metabolites of medicinal plants, the use of medicinal plants is growing rapidly throughout the world (Nalawade et al., 2003). Between 50,000 and 80,000 flowering plant species are using for medicinal purposes worldwide according to the International Union for Conservation of Nature and the World Wildlife Fund (Bentley 2010). However, 20 % of the wild resources have already been exhausted with the increasing human population (Ross, 2005). Worldwide loss of species and habitat destruction has increased the extinction rate of medicinal plants, especially in China (Nalawade et al., 2003; Heywood and Iriondo, 2003), India (Heywood and Iriondo, 2003; Hamilton, 2008), Nepal (Hamilton, 2008), Kenya (Hamilton, 2008), Uganda (Zerabruk and Yirga, 2012) and Tanzania (Zerabruk and Yirga, 2012). China and India used highest numbers of medicinal plants i.e. 11,146 and 7500 species respectively (Rafieian-Kopaei, 2013). So, it is now necessary to find out new medicinally important plants or to utilize the well known medicinal plant sustainably. There are many plants in India that contains different medicinal and nutritional components, which can be exploited much more efficiently to establish a new era of medical science.

1. Introduction

The Aquilaria malaccensis (Agarwood) is an Appendix II species of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES, 1994). The species is also red listed under International Union for Conservation of Nature and Natural Resources (IUCN) and is classified as ‘Vulnerable’ globally but ‘Critically Endangered’ in India (Oldfield et al., 1998; IUCN, 2009) and almost ‘Extinct in wild’ in Assam (Anon., 2003; Saikia and Khan, 2012a). The species due to its multipurpose uses is overexploited which is threatening its existence from the wild (Lee and Mohamed, 2016; Talucder et al.,2016). There are 21 reported Aquilaria species, of which 13 are fragrant resin producers, while the rest are yet to be studied (Lee and Mohamed, 2016; Talucder et al.,2016). Eleven of the domesticated species are A. malaccensis, A. crassna, A. subintegra, A. hirta, A.rostrata, A. beccariana, A. filaria, A. khasiana, A.microcarpa, A. grandiflora and A. sinensis (Zuhaidi, 2016). Species reported from Asia viz., A. apiculata, A. baillonii, A. banaense, A. beccariana, A. brachyantha, A. cumingiana, A. filaria, A. hirta, A. khasiana, A. malaccensis, A. microcarpa, A. rostrata, A. sinensis, A. subintegra and A. crassna are known to produce essential oils (Sulaiman et al., 2015). Only three species i.e. Aquilaria khasiana, A. macrophylla and A. malaccensis has centre of origin in India with A. macrophylla endemic to Nicobar Islands (Anon., 2003; Giri, 2003) and A. khasiana endemic to Khasi Hills of Meghalaya and surroundings (Kanjilal et al., 1982), while A. malaccensis is native to evergreen rain forests in the foothills of northeastern India (Chakrabarty et al., 1994; Saikia and Khan, 2012b) and West Bengal (Barden et al., 2000).

1. Introduction

The global population is very cautious about gaining weight and diabetics. Calorie less and sugar free substances are gaining momentum in food sector. The miracle plant stevia is very fit in this scenario as it is both zero calorie and sugar free and one more additional benefit that it is completely natural. Stevia rebaudiana Bert. belonging to the family asteraceae has been used in many countries for thousands of years as a source of natural sweetener eg., in Paraguay and Brazil Central America, Thailand, Korea and China (Megeji et al., 2005, Sivaram and Mukundan, 2003; Jain et al., 2009). It is also known as sweet herb, sweet leaf, honey leaf, candy leaf, honey yerba (Carakostas et al., 2008) and meeti tulsi. It possesses an extensive root system and brittle stems producing small, elliptic leaves. In natural conditions S. rebaudiana grows in a form of a shrub and reaches even 1 m of height. Leaves are the major economic part. They are sessile and oppositely arranged, lanceolate to oblancoelate in shape, and are serrated above the middle. Trichome structures on the leaf surface are of two different sizes, one large and one small (Shaffert and Chetobar 1994). The white florets are, borne in small corymbs of 2–6 in number. Corymbs are arranged in panicles. Dr. Moises Santiago Bertoni discovered this plant in 1888 at Paraguay. In 1905, the plant was scientifically named as S. rebaudiana after a Paraguayan chemist Dr. Rebaudi. (Gupta et al., 2013). The active components of Stevia i.e., steviol glycosides (SGs) includes rebaudioside A (Reb A), rebaudioside C and stevioside (ST) that are 250-300 times sweeter than sugar (Chalapathi & Thimmegowda 1997). Stevia is a short-day plant and its flowering is induced when days are shorter. Long photoperiods stimulates leaf growth and steviol glycoside content. The plant can survive only mild winters of its native regions. It cannot survive in waterlogged areas. Stevia extracts can be used in a wide variety of food and beverage applications. Individual usages and use levels may vary depending on the country. They act synergistically when used in combination with other sweeteners. The higher market demand favors the cultivation and commercialization of this particular species.

1. Introduction

Man (Homo sapiens) since his appearance on the sylvan Blue Planet started his search for healthcare and beautycare, besides food and shelter. Eventually, he also became conscious about his mind and attempted at ‘mindcare’. He employed leaves, flowers, fruits, etc. or some plant products altering his mind to remain aloof from his heavy worldly affairs, stress or even incurable diseases. He also endeavoured to use plants for his amusement. He exerted more pressure on local biodiversity, modified his study and observations on biodiversity which resulted into ‘cultural biodiversity’. He intensified his method of ‘trial and errors’ to search new sources or biodynamic chemicals. In recent times, he is emphasizing value-addition of the products in hand for his welfare. After food, medicine was his next priority to sustain life in primitive societies. Naturally, his study and research laid emphasis more on food and medicine in comparison to mind-altering bioresources. Even the modern man also paid more attention on the former rather that the latter. These trends have perforce led in partial negligence of research for psychoactive plants in his ambience. The present author, therefore, extended efforts to prepare an inventory of the mind-altering vis-à-vis psychoactive plants in the state of Maharashtra.

1. Introduction

Plant is probably the greatest gift of nature. Plants provide us with food, cloth, house, papers and many others. Some of the plants have medicinal values and are known as ‘medicinal plants’. Our ancestors used these plants not only for protection from diseases but also for the treatment of several ailments. China has the highest number of the medicinal plants used (about 11,500 species) followed by India (about 7500 species). People of other countries also use medicinal plants in varying amount (Rafieian-Kopaei, 2013). Tradition is still going on. It is reported that even today 80% of the world population relies on medicinal plants in their medication (Hamilton, 2004). Few plants secrete essential oils full of aroma and are called ‘aromatic plants’. These plants are being utilized in industry for the production of essence and many other products. Medicinal and aromatic plants (MAPs), therefore, are in continuous use from past to present. In future also there will be tremendous scope of utilization of MAPs in drug discovery, aromatherapy, as cosmeceuticals, nutraceuticals as well as in animal husbandry. Present review will focus utilization of Medicinal and Aromatic Plants (MAPs) in past and present and its possible prospect in future.

1. Introduction

Vedic system of medicine, also called Hindu medicine, Aryan median, Indian system of medicine and in modern era as science of life (Ayurveda). It traces origin from mythical, through a semi-mythical, to historical beginning. Clues on uses of medicine are although obtainable since Indus civilization, it was propounded more in the Vedic period and also in post Vedic era. It is claimed to be the oldest medicinal system and heritage of India. The ancient Indians were first to cultivate astronomical science, mathematics, chemistry, grammar, lexicography, architecture and Dhanur-veda. Another compartment of human life is medicine, in which Indians never lagged behind. Vedic medicine is not born within four walls of a laboratory. It is nurtured in nature, human minds and hearts. The Vedic wisdom is engendered by trial and errors, experimentation, natural observations and timely criticism by the learned exponents. Obviously, there is no denial to the fact that the torch of learning was handed down to the other civilizations by the ancient Indians. Vedic medicine is not confined to the boundary of Indian nation but percolated in other cultures. Careful and critical examination of such a great heritage is a task of not of ordinary men. Still I took interest to review it, although almost impossible to justify it within the limits of few pages and time. The title of my paper amply suggests that Vedic medicine is a mine of information but I have tried to put on record it, in a nutshell. To tap down information in historical perspective, relevant literature has been borrowed and consulted to pinpoint the events, concepts wisdom of ancient Indians. These literary sources are enlisted under references.

1.Which one is the example of abiotic stress?

a.Drought

b.Salinity

c.Flood

d.All of these

2.Which one is the example of osmoticum?

a.NaCl

b.PEG

c.H₂O

Objective: To quantify aflatoxin content in given sample by Indirect competitive ELIS

Background information:

Enzyme linked immuno sorbent assay (ELISA) is a common biochemical technique that uses the principle of solid-phase enzyme immunoassay (EIA) for detection of substances mainly antigens in a liquid or wet sample.

ELISA is a very popular technique that is used for diagnostic purpose in health sector, plant protection and also for quality test in food and other industries. This technique applies the principle of antigen-antibody reaction on a solid surface. The attached antibody is linked to certain enzymes and in the end of reaction by addition of substrate that performs enzyme-substrate reaction and makes product that has some signal mostly color change and based on intensity of color the qualitative as well as quantitative estimation of antigens can be done using ELISA reader coupled with spectrophotometer.

In ELISA technique one antibody should be specific to certain antigen. For reaction of ELISA the ELISA plates are used that is also known as microtiter plate made-up of polystyrene. The sample to be tested for ELISA contain unknown amount of antigen to be detected, this sample containing antigen is immobiliz d on a solid support like polystyrene microtiter plate in two different ways; first is non-specific where antigens adsorbed to the surface and second is specific where the antigens of sample are captured by specific antibody to the antigen, like used in sandwich ELIS Once the antigens are immobilized non-specifically or specifically the detection antibody should be added that makes precise antigen-antibody complex. The detection antibody may be either covalently linked to certain enzyme or may be detected by another secondary antibody linked to bioconjugated enzyme. During ELISA steps several proteins or antibodies may non-specifically bound that can be removed by washing with mild detergent solution in between of each ELISA step. For washing of ELISA plates, ELISA plate washer may be used that mechanically wash the ELISA plates. At the end of ELISA the final wash is given and substrate is added that produces visible signal in the form of color change that indicates the presence of antigens in the sample and further quantification may be done using ELISA reader coupled with spectrophotometer.

Objective: To synthesize cDNA from isolated plant total RNA.

Background information:

Complementary DNA (cDNA) is double-stranded DNA made from template of single stranded RNA, [e.g., mRNA or microRNA] by reverse transcriptase enzyme. Mostly cDNA is used for cloning of eukaryotic genes in the prokaryotes. For the expression of a specific protein in a cell where the expression of that protein is not found (i.e., heterologous expression), the cDNA will be transferred that codes for the protein to the recipient cells. Retroviruses (e.g. HIV-1, HIV-2, simian immunodeficiency virus, etc.) also produce cDNA naturally and integrate in host's genome and further creates provirus (Croy, 1998). Major use of cDNA is in gene cloning or as gene probes or in the creation of a cDNA library. When scientists transfer a gene from one cell into another cell for the expression of new genetic material as a protein in the recipient cell, the cDNA will be added to the recipient (rather than the entire gene), because the DNA for an entire gene may have certain DNA that may not code for the protein or that interrupts the coding sequences of the protein (e.g., introns). Partial sequences of cDNAs are often obtained as expressed sequence tags.