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Apple (Malus x domestica Borkh.) is a premium fruit of the world. It is a typical temperate fruit and main fruit crop of J&K State. The area under this fruit in J&K State is 1,62,971 hectares with an annual production of 17,26,834 metric tones. The acreage under this fruit crop in Jammu Division is 18,146 hectares and the annual production is 38,421 metric tones. In Jammu Division, apple is being grown in upper areas of Doda, Kishtwar, Ramban, Poonch, Udhampur, Reasi, Kathua and Rajouri districts.

Climate and soil

Apple is a typical temperate fruit crop requiring 1000-1600 hours of rest period below 7^oC. The average temperature during growing season should be 21⁰C to 24⁰C. Sunshine is essential for colour development of the fruit. In general, a cool climate with low winter temperature and little rainfall in summer is most suitable for apple cultivation.

Apple is grown on a wide range of soils. However, a well drained slightly acidic (about 6.5 -6.7 pH) loam soil with good depth (more than 45 cm) is ideal for apple cultivation. Apple can’t tolerate wet field, and water logging may cause collar rot and death of trees.

Pecan (Carya illinoensis) is a highly nutritive and energy giving nut fruit. It has nutty flavor and superb tree characteristics. Area under pecannut in J&K is 414 hectares with annual production of 86 metric tonnes.

Climate and Soil

Pecan need warm temperate climate and extremes of both temperate and sub-tropical climates are undesirable. It needs adequate chilling hours ranging from 400-600 hours below 7.2^oC for different cultivars. Mean monthly temperature ranging from 24.0 to 29.5⁰C, with little diurnal variation during growing period is best for growth and fruiting. High humidity prevents pollination, increases the incidence of diseases and premature splitting of nuts.

Although pecan nuts are grown in varied soil conditions, but sandy loam soils (pH 5.0-8.0) which are deep, fertile and are rich in organic matter are best for its growth and fruiting. Symbiotic mycorrhiza and fibrous roots permeate the top 1 meter of soil.

Phalsa (Grewia subinaequalis) is one of the hardiest, drought resistant and quick growing fruit crop. Different parts of the plant are used for curing variety of ailments. The ripe phalsa fruits are consumed fresh, in desserts or processed into refreshing fruit and soft drinks enjoyed during hot summers.

Climate and Soil

Phalsa prefers dry conditions and can therefore, withstand drought. It flourishes well under variable climatic conditions, and requires protection from the freezing cold temperature. Also, adequate sunlight and warm or hot temperatures are required for fruit ripening, development of appropriate fruit colour and good eating quality. Kandi areas are, therefore, quite suitable for its cultivation. It can also be grown under irrigated conditions. Phalsa plant grows vigorously under wide range of soils including fine sand, clay or even limestone. However, best results are obtained in well-drained, loamy soils.

Ber (Zizyphus mauritiana Lamk.) is one of the ancient and common fruits, indigenous to India. It is growing in wild, semi wild and cultivated forms all over the country in sub-tropical and tropical areas. Area under ber in J&K is 5,376 hectare with annual production of 10,779 metric tonnes.

Climate and Soil

Ber plant is hardy and can grow successfully even under poor soil and unfavourable climatic conditions, where most other fruit trees fail to grow. It prefers a hot and dry climate for its successful cultivation, but needs adequate water during its first fruiting season. It can be grown up to an elevation of 1000 meters above sea level. Excessive atmospheric humidity seems to be a limiting factor in its successful fruiting.

The ber tree develops a deep tap root system with in a short period of its growth and as such adopts itself to wide variety of soils. However, deep sandyloam soils appear to be better for its plantations. It is well known in its ability to thrive under adverse conditions of salinity, drought and water logging.

Guava (Psidium guajava), the apple of tropics, is one of the most common fruits in India. It is the fourth most important fruit in area and production after mango, banana and citrus. Being hardy, it gives assured crop even with little care, and is good for rainfed areas also. This fruit is popularly being grown in sub-tropical regions of Jammu on 2480 ha with annual production of 9153 MT.

Climate and Soil

Guava is successfully grown under tropical and sub-tropical climate. In areas with distinct winter season, the yield tends to increase and quality improves. It can be grown upto an altitude of 1515 m above mean sea level. Older plants can tolerate drought. High temperature at the time of fruit development can cause fruit drop.

Guava trees are very hardy and can thrive on all types of soils, but are sensitive to water logging. The best soils are deep, loamy and well drained.

The litchi (Litchi chinensis) is a delicious juicy fruit of excellent quality. It is liked very much as table fruit. Presently, litchi occupies 940 ha area in Jammu region with an annual production of 2315 MT.

Climate and Soil

Litchi being a sub-tropical fruit thrives best under warm humid sub-tropical climate. However, frost in winter and hot winds in summer limits its growth and production. Hot winds results in cracking of fruits before reaching maturity while excessive rainfall at the time of flowering interfere with pollination. The prolonged dry spell after fruit set reduces the crop considerably.

Litchi flourishes well on deep, well drained loamy soil, rich in organic matter and free from hard substrata.

Citrus (Citrus spp.) is one of the most important fruits grown in sub-tropical and foot hill regions of Jammu province (Kathua, Hiranagar, Samba, Jammu, Akhnoor, parts of Basohli, Nowshera, Ramnagar, Ramban and Udhampur) upto an elevation of 800 m amsl. The most important commercial citrus fruits in Jammu region are kinnow mandarin, sweet orange, grapefruit, lemon and lime. The area under citrus fruits in Jammu division of state is 14,542 ha with an annual production of 34,191 metric tones.

Climate and soil

Citrus plants being sub-tropical cannot with stand extended cold period. Temperature below 3^oC and above 40^oC are injurious to the citrus. Hot winds and excessive heat during flowering and fruit set cause fruit drop and sun burning. These are the light loving crops.

Citrus fruits thrive well in deep, loose, well aerated soils with the pH of 5.5 to 6.2 devoid of any hard pan layers of calcium carbonate in the rooting zone up to 150 cm. High water table and poor drainage system are quite unsuitable for citrus fruits which cause drying of roots and nutritional imbalance, respectively.

Mango (Mangifera indica L.) is one of the choicest and most ancient fruits known to mankind. It occupies a pre-eminent place amongst the subtropical fruits grown in Jammu region and occupies an area of 12,739 ha with annual yield of 24,150 metric tons. It is extensively grown in Jammu district and also in the sub-tropical areas of Udhampur, Reasi, Kathua and Rajouri districts.

Climate and Soil

Mango thrives well up to 600 m above mean sea level provided locality is frost free and there is no high humidity or rains during flowering. The favourable temperature is 24⁰ C to 27⁰ C, however, it can tolerate temperature as high as 48⁰ C provided that trees are getting regular irrigation.

Mango has been found to grow on a wide range of soils. However, deep and well-drained sandy loam soils are most suitable for its cultivation. Heavy black cotton, saline and alkaline soils should be avoided. The ideal range of soil pH for mango cultivation is 5.5 to 7.5.

Jammu division with three agro-climatic zones viz., sub-tropical, intermediate (low hills) and temperate (Fig. 1) has vast potential for horticultural industry by way of improving the productivity of existing orchards and using vast tracts of underutilized wastelands, kandi areas of sub-tropical zone and hilly areas of intermediate and temperate zones for fruit production. For this, there is need for plantation of suitable varieties, budded/grafted on proper rootstock and following of improved orchard management practices viz., training and pruning, nutrient and water management and pest and disease control. At present, fruits occupy an area of 118805.00 ha with the production of 274006.70 MT and productivity of 2.31 MT/ha in Jammu division.

Sub-tropical zone

It comprises both irrigated and unirrigated areas and goes upto an attitude of 500 m above mean sea level. This zone is characterized by hot summers and relatively dry but pronounced winters. The land is suitable for cultivation of mango, citrus crops (sweet orange, mandarin particularly kinnow, lime, lemon), guava, litchi, ber, papaya and other minor fruits. Some low chill varieties of temperate fruits like peach, plum, pear and strawberry have been introduced at Advanced Centre for Horticulture Research (ACHR) of SKUAST-J, Udheywalla and peach cultivars Shan-e-Punjab, Flordasun and Flordared and pear cultivars Patharnakh and LeConte have shown good yield and quality crop. Besides, strawberry cultivars Chandler, Gorella, Fern and Belrubi have also shown promising yield with proper management practices. For rainfed areas, hardy fruits like aonla, phalsa, ber, guava, lasoda, karonda and custard apple are more suitable with aonla, phalsa and ber being commercially more viable.

North East India is a hub of ethnomedicinal diversity. Nearly 8,000 flowering plants are distributed and approximately 200 ethnic tribes and subtribes reside in the region. The cross-cultural diversity of the ethnic tribes and subtribes has enriched the ethnomedicinal practice in the region. Due to inaccessible terrains the ethnomedicinal practices are mostly confined to this region than the rest part of the country.More than 175 dialects are practiced in NE India. The ethnomedicinal documentation of the region in universally accepted languages was started in early 21^st century. The ethnomedicinal documentation requires multidisciplinary aspects viz., correct identification of plant species, updated nomenclature, geographical area of occurrence, IUCN status, conservational approach, vernacular names of the species, formulations, documentation of traditional knowledge informants, medicine men, specific indications of treatments, mode of preparation, mode of consumption etc, Optimum and sustainable utilization of ethomedicinal practice holds a good future for economic and societal development of the region. An insight covering various factors like challenges, scopes, legislative modalities of ethnomedicinal practices are discussed here along with a compiled list of commonly used ethnomedicinal species of the region.

Several ethnic communities inhabiting in the biodiversity-rich state of Meghalaya are primarily dependent upon the forest for their livelihood needs like food, vegetables, materials for construction of houses, firewood, medicine for curing numerous ailments and many other purposes. They have acquired vast knowledge about the wealth of plant and their utilisation. The present study was aimed to document such less-known wild edible plants used by Khasi people of the state. During the current investigation, a total of 94 species of such importance belonging to 80 genera under 47 families were documented. Among these, 46 are herbs followed by 29 shrubs, 12 trees and 02 are creepers. Based on the availability and commercial potential, 37 species have been sold in local markets for income generation. Of which 17 species are sold as fruit, 11 as leaves, six as root, rhizome and tuber, five species as a stem, two as inflorescence and one species sold as fronds. All the plants are arranged alphabetically in the tabular form, followed by families, vernacular name(s), plant part(s) used, habit (s), distribution and methodology. It has been observed that the knowledge of less known wild edible plants is significantly declining day by day. These plants are known for their importance in as food nutritional supplement and its medicinal value; if the information is not collected and documented, it may not last soon.

The review deals with 151 species of plants covering 138 genera and 66 families used by the tribes for curing women diseases involving 172 practices. Fabaceae is the dominant family with 13 species followed by Caesalpiniaceae (9 spp) and others curing a variety of ailments. The useful plants be subjected to phytochemical and pharmacological tests to determine the effective constituents and characteristic biological activity.

1. Introduction

Primitive man gets much enthusiasm towards green plants and started to examine the property of using plants by trial and error and obtained different beneficial properties. India is one of the 12 megadiversity countries and comprises of two hot spots. The World Health Organization (WHO) has compiled a list of 20,000 medicinal plants used in different parts of the globe. In India the scheduled tribe population is 104,545,716 constituting 8.6% of country total population of 1,210,726,932 (Census 2011) belonging to over 550 tribal communities and 227 ethnic groups. They inhabit about 5000 forested villages and occupying about 15% of country’s geographical area.

1. Introduction

Chirata (Swertia chirayita) belongs to the family Gentiniaceae, a potential medicinal plant used as bitter tonic in treatment of fever and for curing various skin diseases, is a perennial herb and distributed throughout the eastern Himalayan region of India. The plant is a native of temperate Himalayas, found at an altitude of 1200-3000 m (4000-10000 ft), from Kashmir to Bhutan and in the Khasi hills at 1200-1500 m (4000 to 5000 ft). It can be grown in sub-temperate regions between 1500 and 2100 m altitudes. It is used in traditional medicine to treat numerous ailments such as liver disorders, malaria, and diabetes and are reported to have a wide spectrum of pharmacological properties. Its medicinal usage is well-documented in Indian pharmaceutical codex, the British, and the American pharmacopeia’s and in different traditional medicine such as the Ayurveda, Unani, Siddha, Horopathy and other conventional medical systems. This ethno medicinal herb is known mostly for its bitter taste caused by the presence of different bioactive compounds that are directly associated with welfare of human health. The increasing high usage of Swertia chirayita, mostly whole plant, as well as the illegal overharvesting combined with habitat destruction resulted in a drastic reduction of its populations and has brought this plant to the verge of extinction. The increasing national and international demand for Swertia chirayita has led to unscrupulous collection from the wild and adulteration of supplies. The bitterness, antihelmintic, hypoglycemic and antipyretic properties are attributed to amarogentin (most bitter compound isolated till date) 1, swerchirin, swertiamarin and other active principles of the herb. Due to the high demand, this plant is being cultivated in many places of Darjeeling district both for domestic use within India and for exporting purpose. During our roving survey (2011-2014), it was observed that this high value medicinal plant facing a number of diseases (like seedling blight, leaf spot, Leaf blight etc.), ultimately, cultivation of Swertia chirayita is getting hampered. The quality of plant is affected due to the occurrences of diseases as whole plants part being used as medicinal purpose and also affects the marketing scenario; the demand of good quality chirata is increasing at a rate of 10% annually. To harvest the benefit of megabiodiversity zone of eastern Himalaya needs urgent attention to combat the upcoming epidemic of diseases of chirata. Here, we are trying to throw some light related to present status of diseases of chirata and standard management practices.

1. Introduction

Over the centuries, the use of medicinal herbs has become an important part of daily life despite the progress in modern medical and pharmaceuticals research. It is estimated that 40% of the world populations depends directly on plant based medicine. India is one of the world’s 12 biodiversity centres. Herbal industry in India uses about 8000 medicinal plants. Indian export of raw drugs has steadily grown at 26% to Rs.165 crores in 1994-’95 from Rs.130 crores in 1991-’92 and this is likely to touch US $5 trillion by 2050. Thus, India has a great role to play, as supplier of herbal products not only to meet the domestic needs, but also to take advantage of the tremendous export potential.

Biotic stress related problems are important bottlenecks for the successful commercial cultivation of any crop. Medicinal plants are not the exception and are vulnerable to be attacked by several pests and pathogens resulting crop loss in terms of both quantity as well as quality. Therefore, the “health of these healthy plants” should be concerned. Without proper identification and prior knowledge of the pests and diseases, the management will be a waste of time and money and can lead to further plant losses.

1. Introduction

Intellectual property is a term referring to creations of the intellect for which a monopoly is assigned to designated owners by law. While intellectual property law has evolved over centuries, it was not until the 19^th century that the term Intellectual Property began to be used, and not until the late 20^th century that it became commonplace in the majority of the world.

Agriculture (including horticulture) and forestry are the two most important sectors in the land use system of India covering know 46.1% (141.2 m ha) and 22.2% (69.02 m ha), respectively of total geographic area of the country. Intellectual Property Right (IPR) was of limited significance in cultivation and improvement of agricultural, horticultural and forest crops and aggregation of these three sections comprises the Plant Genetic Resources (PGRs). IPRs were not applied to plants in earlier days. PGRs are the foundation for the development of food and nutritionally secured nation. Food is the foremost important to sustain life and livelihood followed by shelter and cloth. Thus, the slogan of 3Fs may be modified as, “Food, Frame (shelter) and Fibre (cloth)” instead of Food, Fodder and Fire-wood. All these 3Fs we get from PGRs.

1. Introduction

Medicinal plants are well known for their secondary metabolites which are alternatively called as active principles responsible for curing of diseases and disorders. India is a biodiversity rich country belongs within 12 mega diversity zones of the world and possesses four biodiversity hot spots, 8 phyto-geographic regions and 10 vegetation zones (Khandekar and Srivastava, 2014). This subcontinent holds 47000 different species of plants out of which 8000 plants are medicinal in nature (Hamilton, 2004, Owolabi et al., 2007). High rate of endemism and 426 habitats for specific species are the unique resource of the country (BSI, 2009; Khandekar and Srivastava, 2014). Still majority of the herbal raw materials are wildcrafted and hence habitat destruction and wildcrafting pose a serious problem to the natural population particularly where roots are the targeted plant part as the percentage of root yielding medicinal plants are higher in the subcontinent. Since these plants are purely industrial and not having alternative uses the commercial production is not flourishing at a higher pace. Still the production of medicinal and aromatic plants in India has reached 1031000 tonnes from 634000 hectares.

In the post-CBD regime with global developments in aspects of Plant Genetic Resources and Intellectual Property Rights, the outlook towards these lesser exploited species has been changed to bioprospecting from collection and documentation.Here lies the importance of value addition in medicinal plants. Value addition means a change of state of the produce which will give a manifold return than the additional investment. Value-added products can be obtained through processing of raw herb or crude drug which will improve its look and also increase its quality, longevity, efficacy and obviously the return.

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.

Objective: To check the integrity of isolated RNA on agarose gel.

Background information:

The most common method used to assess the integrity of total RNA is to run RNA sample on agarose gel (denaturing) stained with ethidium bromide (EtBr). While native (non-denaturing) gels can be used, the results can be difficult to interpret. The secondary structure of RNA alters its migration pattern in nativegels so that it will not migrate according to its true size. Nondenaturing conditions also result in bands that are not as sharp, and even multiple bands representing different structures of a single RNA species.

Intact total RNA run on a denaturing gel will have sharp, clear 28S and 18S rRNA bands (eukaryotic samples). The 28S rRNA band should be approximately twice as intense as the 18S rRNA band. This 2:1 ratio (28S:18S) is a good indication that the RNA is completely intact. Partially degraded RNA will have a smeared appearance, will lack the sharp rRNA bands, or will not exhibit the 2:1 ratio of high quality RNA. Completely degraded RNA will appear as a smear (having very low mw). Inclusion of RNA size markers on the gel will determine the band size or smears size and will also serve as a good control for ensuring proper gel run. Note: Poly selected samples will not contain strong rRNA bands and will appear as a smear from approximately 6 kb to 0.5 kb (resulting from the population of mRNAs, and depending on exposure times and conditions), with the area between 1.5 and 2 kb being the most intense (this smear is sometimes apparent in total RNA samples as well).

Objective: To isolate RNA from plants using TRIzol reagent and its quantification by spectrophotometer.

Background information

RNA (Ribonucleic acid) is a polymeric substance present in living cells and many viruses, consisting of a long single-stranded chain of phosphate and ribose units with the nitrogen bases adenine, guanine, cytosine, and uracil, which are bonded to the ribose sugar. RNA is necessary in all the steps of protein synthesis in all living cells and carries the genetic information for many viruses.

The isolation of RNA with high quality is a crucial step required to perform various molecular biology experiment. TRIzol Reagent is a ready-to-use reagent used for RNA isolation from cells and tissues (Chomczynski, 1993).TRIzol works by maintaining RNA integrity during tissue homogenization, while at the same time disrupting and breaking down cells and cell components. Addition of chloroform, after the centrifugation, separates the solution into aqueous and organic phases. Aqueous phase contains RNA.

After transferring the aqueous phase, RNA can be recovered by precipitation with isopropyl alcohol. DNA and proteins can be recovered by sequential separation after the removal of aqueous phase. Ethanol precipitation needs DNA from the interphase, and an additional precipitation with isopropyl alcohol requires proteins from the organic phase. Total RNA extracted by TRIzol Reagent is free from the contamination of protein and DNA The isolated RNA can be further used in Northern blot analysis, in vitro translation, poly (A) selection, RNase protection assay and molecular cloning.

Objective:To transfer DNA on nylon membrane by Southern blotting.

Background information

A Southern blot is a method used in molecular biology to detect a specific DNA sequence in DNA samples. Southern blotting combines transfer of electrophoresis-separated DNA fragments to a filter membrane and subsequent fragment detection by probe hybridization.

The British biologist Edwin Southern (Southern, 1975) discovered this technique the technique is known by his name. Southern blotting technique uses digested genomic DNA with restriction enzymes. Southern blotting technique may be used to identify the gene copies number in a genome. A probe that hybridizes only to a single DNA segment that is uncut with restriction enzyme will produce a single band on a Southern blot, whereas multiple bands will likely be observed when the probe hybridizes to several highly similar sequences (e.g., may be due to sequence duplication).

In the southern blotting technique a short DNA probe is used that matches based on complementary DNA sequence fragment on filter membrane. It proves that the DNA fragment on membrane contain specific sequence for which DNA probe is prepared. When DNA is transferred on the membrane from gel it permits binding of radioactive/Biotin labeled hybridization probe to the restricted DNA. Southern blotting is also important for autoradiography.

Southern blotting is an important molecular biology technique that may be used for homology based cloning, screening a DNA library, screening of cloned DNA fragments and to identify methylated sites in genes.

Objective: To amplify DNA using RAPD primers and study on agarose gel.

Background information:

RAPD markers are decamer (10 nucleotide length) DNA fragments from PCR amplification of random segments of genomic DNA with single primer of arbitrary nucleotide sequence and which are able to differentiate between genetically distinct individuals. RAPD is applied to analyze the genetic diversity by random primers. Unlike traditional PCR analysis, RAPD does not require any prior DNA sequence information of the target organism: the identical 10-mer primers will or will not amplify a segment of DNA, depending on positions complementary with primers' sequence. For example, no fragment is produced if primers annealed too far apart or 3' ends of the primers are not facing each other. Therefore, if a mutation has occurred in the templ that was earlier complementary with primer, a PCR product will not be produced, resulting in a different pattern of amplified DNA segments on the gel. RAPD is a dominant marker means it cannot distinguish between heterozygous; single copy and homozygous; two copies. Rarely RAPD markers may be Co-dominant where different fragment length of DNA is amplified from the single locus. PCR is a molecular technique used for amplification of DNA, a complete enzyme based chemical reaction, therefore, the concentration and quality of template DNA, concentrations of PCR components, and the PCR cycling conditions may greatly influence the outcome. Thus, the RAPD technique is notoriously laboratory dependent and needs carefully developed laboratory protocols to be reproducible. Mismatches between the primer and the template may result in the total absence of PCR product or decreased amplified product. Thus, the RAPD results can be difficult to interpret.

Objective: To check quality of genomic DNA on agarose gel electrophoresis.

Background information

Agarose gel electrophoresis is a common method of gel based electrophoresis technique used in biochemistry, molecular biology, genetics and clinical chemistry for separation of Protein and nucleic acids from a mixture of macromolecules. Agarose is a major component of agar that is used as a matrix in gel baseelectrophoresis. The nucleic acids (DNA and RNA) are separated based on length of fragments while, proteins are separated based on size and/or charge (Kryndushkin, et al. 2003). For separation of bio-molecules the electric field is applied that leads to movement of charged particles via agarose gel matrix and bio-molecul s are separated based on difference in size in the agarose gel matrix (Sambrook and Russel, 2001).

Gel casting with agarose is easy and has very few charged groups, hence found most suitable for nucleic acid separation in most of the biochemistry, molecular biology and biotechnology laboratories. Separated DNA/RNA ma be seen and recorded with stain under UV light. Further DNA fragments can be eluted from agarose gel with an ease for further down applications. The percentage of agarose gel used is in the range of 0.7 - 2. Lower percent of agarose gel is used for long DNA fragments while, higher percent of agarose gel is used for short DNA fragments with suitable running buffer.

Objective: To isolate DNA from plant sample by CTAB method and quantification by spectrophotometer.

Background information:

DNA isolation from different plant tissues viz., leaf, stem, root, flower and seed is a challenging task due to hindrance of variety of variety of biochemical compounds present and further problem increases with diverge plant species. In case of animal tissues the diversity in cellular bio-molecules in different species is much lower compared to plants. Two major phyto bio-molecules that have wide variation between species and pose serious problems in DNA isolation are: polyphenols and polysaccharides. These phyto bio-molecules if contaminated in DNA then may hinder further down applications like PCR, restriction digestion, sequencing, etc.

Several methods have been developed and reported that can remove polyphenols and polysaccharides efficiently during DNA isolation from plants. For this purpose two different chemicals are widely used,the first one is CTAB (cetyl trimethyl ammonium bromide) that is a cationic detergent and helps in removal of polysaccharides and the second one is polyvinylpyrrolidone that helps in removal of polyphenols. Due to above properties the CTAB based DNA extraction buffer is very common for plant DNA extraction.

Significance of different types of chemicals found in CTAB based plant DNA extraction method is explained as under:

Objective: To study peroxidase isozyme variation in rice seedlings by electrophoresis.

Background information

For electrophoresis and PAGE please see practical no. 4. For peroxidase please see practical no. 7.(Page 17-22)

Isozymes (isoenzymes or more generally as multiple forms of enzymes) are the enzymes that are differing in the sequence of amino acids but they work on same substrate or in other words catalyze same biochemical reaction. The isozymes have difference in K_M values and regulatory properties. Isozymes are very useful for proper tuning of metabolic activities and to meet the specific need of particular tissue or developmental stage, e.g. lactate dehydrogenase (LDH). Isozymes are coded by homologous genes that have diversity during evolutionary process. Although, strictly speaking, allozymes represent enzymes from different alleles of the same gene, and isozymes represent enzymes from different genes that process or catalyse the same reaction, the two words are usually used interchangeably.

Objective: To extract and perform enzyme assay of Superoxide dismutase from plant.

Background information:

Superoxide dismutase (SOD, EC 1.15.1.1) is the enzyme that alternately catalyzes the dismutation (or partitioning) of the superoxide (O₂^-) radical into either ordinary molecular oxygen (O₂) or hydrogen peroxide (H₂O₂). Superoxide is produced as a by-product of oxygen metabolism and, if not regulated, may cause cell damage (Hayyan, et al. 2016). Hydrogen peroxide is also responsible for cell damage and may be degraded by other enzymes like catalase. Thus, SOD is an important antioxidant defense in nearly all living cells exposed to oxygen. One exception is Lactobacillus plantarum and related lactobacilli, which use a different mechanism to prevent damage from reactive O₂^-.

Objective: To extract and perform enzyme assay of guaiacol peroxidase from plant.

Background information:

Peroxidases are a large family of enzymes that typically catalyze a reaction of the form:

Objective: To extract and perform enzyme assay of catalase from plant.

Background information

Catalase is a commonly distributed enzyme and present in approximately all organisms exposed to oxygen (such as bacteria, plants and animals). Catalysis of H₂O₂ into H₂O and O₂ is done by catalase enzyme (Goodsell, 2004).

Objective: To extract and perform enzyme assay of polyphenol oxidase from plant.

Background information:

Polyphenol oxidase (PPO or monophenol monooxygenase or polyphenol oxidase I, chloroplastic) is a tetramer that contains four atoms of copper per molecule, and binding sites for two aromatic compounds and oxygen (www.worthington-biochem.com). The enzyme catalyses the o-hydroxylation of monophenol molecules in which the benzene ring contains a single hydroxyl substituent to o-diphenols (phenol molecules containing two hydroxyl substituents). It can also further catalyse the oxidation of o-diphenols to produce o-quinones.

PPO causes the rapid polymerization of o-quinones to produce black, brown or red pigments (polyphenols) that cause fruit browning. The amino acid tyrosine contains a single phenolic ring that may be oxidised by the action of PPOs to form o-quinone. Hence, PPOs are some time referred as tyrosinases (Mayer, 2006).

Jolley et al. (1974) referred to PPO as 4 electron-transferring phenol oxidase and oxygen. It is responsible for browning reactions throughout the phylogenetic scale.

Objective: To separate protein fragments by using SDS-PAGE method.

Background information

Electrophoresis is the motion of dispersed particles relative to a fluid under the effect of limited uniform field of electricity (Dukhin, and Derjaguin, 1974). Ferdin and Frederic Reuss (Reuss, 1809) first time observed such phenomena of electrokinetic nature in 1807. They found that the clay particles move in the water by giving constant supply of current. It happened due to interface of charged particles surface and the surrounding media. The technique is used widely in chemistry, biochemistry and molecular biology to separate different molecules based on difference in charge, binding affinity and size. Electrophoresis may be of two types viz., Anaphoresis and Cataphoresis. Anaphoresis is the electrophoresis of anions (-ve charged ions) while, Cataphoresis is electrophoresis of cations (+ve charged ions). Electrophoresis is a major analytical technique in biochemistry that is used for separation of macromolecules like protein, enzymes and nucleic acids. Electrophoresis is a technique used in laboratories in order to separate macromolecules based on size.

PAGE is abbreviated for Polyacrylamide gel electrophoresis that describes a technique widely used in biochemistry, forensics, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility. Mobility is a function of the length, conformation and charge of the molecule.

Objective: To estimate total protein in the plant sample by Folin-Lowry's method.

Background information:

Protein assay can be done to determine the level of protein in solution by the Folin-Lowry biochemical method. The concentration of protein leads to change in color of the solution and based on different color in different protein concentration solution the change in color may be digitized by colorimeter or spectrophotometer. This technique was developed by Oliver H. Lowry in the year 1940s. The tyrosine and trytophan amino acid residues in a protein sample produces purple or blue color in the absorption maxima at 660 nm by using Folin-Ciocalteau (having phosphate and sodium tungstate molybdate) chemical reagent. The color intensity of protein sample depends on the concentration of above aromatic amino acids (tyrosine and tryptophan), hence every sample will vary for color. For protein estimation the BSA (Bovine serum albumin) is widely used as standard protein molecule due to its cost effectiveness, purity and easy availability. This method of protein estimation is very sensitive and can detect up to 10 μg/ml. This is a relative method of protein estimation and may be interfere by salts, Tris buffer, nonionic & cationic detergents, EDTA, lipids and carbohydrates. For reproducible assay the incubation time, pH (9-10.5) is critical in this method (Lowry, et al. 1951).

Objective: To identify salt tolerant genotypes by Glycine Betaine (GB) method in laboratory.

Background information:

Salt or salinity is a biggest danger that causes reduction in crop productivity worldwide. Salinity reduces the germination percentage, plant vigour and ultimately loss in final crop yield. There are different physiological parameters that are affected by high level of salinity viz., oxidative stress, physical alteration of bio-membrane, nutritional imbalance, loss of cell division & cell expansion, water stress, genotoxicity and ion toxicity. Together, these effects reduce plant growth, development and survival. During the onset and development of salt stress within a plant, all the major processes such as photosynthesis, protein synthesis and energy and lipid metabolism are affected. During initial exposure to salinity, plants experience water stress, which in turn reduces leaf expansion (Petronia, et al. 2011).

The water-soluble compounds which, are low in molecular weight, accumulate at certain concentration known as “compatible solutes” or “osmolytes” is the common strategy adopted by many organisms to combat the abiotic stresses. Some common compatible solutes are betaines, sugars (mannitol, sorbitol and trehalose), polyols, polyamines and amino acid (proline). Their accumulation is favored under water-deficit or salt stress as they provide stress tolerance to cell without interfering cellular machinery. The tolerant or sensitive species show difference in the stress tolerance level based on levels of accumulation of these compounds during abiotic stress condition. Accumulation of osmolytes like glycinebetaine (GB) in cells is known to protect organisms against abiotic stresses via osmoregulation or osmoprotection. Other roles of GB like cellular macromolecule protection and ROS detoxification have been suggested as mechanisms responsible for abiotic stress tolerance. In addition, GB also influences expression of several endogenous genes in plants.