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Aonla is said to be indigenous to tropical south-eastern Asia, particularly in central and southern India (Firminger, 1947). The genus comprises of about 350 species. It is also called Indian gooseberry, amla, nelli, myrobalan, amlaki, amali, ambala, amlika, sirikai, amalakamu in different regional languages in India. Aonla is one of the richest sources of natural ascorbic acid which is protected by high polyphenols content during storage and processing. Aonla is indigenous to central and southern India. It is found it Sri Lanka, Malaysia, China and Malacca. In India, it is widely cultivated in Uttar Pradesh, Tamil Nadu. Madhya Pradesh, Rajasthan, Gujarat, Chhatisgarh and Jharkhand.

Cultivars/Varieties: Some of the well-known improved varieties are Banarasi, Chakaiya, Krishna (NA-5), Kanchan (NA-4), Hathi Jhul, Neelam (NA-7), Amrit (NA-6), Balwant, Anand-1, 2 & 3.

Uses

The fruit is widely used in ayurvedic preparation due to its pharmacological properties. The fruits are made into preserve (murabba), sauce, candy, dried chips, etc., Fruits are pickled and made into, jelly, toffee, etc., bark and leaves are used for tanning and wood is used for preparing hookah.

Fruits have been one of the earliest foods of mankind. Ancient Indian texts like Charak Samhita and Sushruta Samhita, religious books and puranas are replete with mention of various kinds of fruits. The mango is believed to have been grown for more than 4000 years. Mention of apple, the forbidden fruit, in the Holy Bible is well known. The date-palm is recorded as the fruit crop grown by mankind since 7000 BC. The first book in the world on fruit culture is said to have been the one written on litchi in 1056 AD. In the earlier period of human civilization, fruits served both as food and medicine. Fruits, nuts and tubers were the staples for early wanderers and gatherers. The settled agriculture made mankind more dependent on cereals, pulses and oilseeds for staples and fruits, vegetables and nuts became more a medicine than food.

Many evolutionary and anthropological changes occurred in course of time with changing food habits. At every stage of major change, food habits determined the life expectancy. After completing a full cycle or helix, time has returned to focus on fruits and vegetables again as food, from a stand point of health and wellbeing. From being trees of wild and forests in earlier period, fruit crops became confined to palaces and orchards of Kings in medieval period. In modern Era, it came to stay as commercial commodities dictating the terms of trade. Due to its growing commercial importance the governments across the globe focused to expand the area under fruit crops and cross border trade in several fruits and vegetables. Horticulture research started prioritizing the traits in fruits that determine the economics of trade. In line with these developments, Indian government assigned great significance to horticultural crops since 8^th five-year plan period, and the allocation for horticultural research was increased to `110.2 crores from `31.96 crores in the previous plan. During the 10^th five-year plan period in 2005-06, the National Horticulture Mission was started. As the net outcome of these efforts, the area under fruit crops increased from 2.87 million ha in 1991-92 to 6.48 million ha in 2016-17. The production increased from 28.63 million tonnes to 92.85 million tonnes and productivity from 9.96 tonnes/ha to 14.33 tonnes/ha during the same period (Anon, 2017).

Out of the total value of output of agricultural crops, fruits and vegetables contribute the lion’s share of 23%. The major fruit crops producing states are Andhra Pradesh, Maharashtra, Uttar Pradesh, Gujarat, Madhya Pradesh, Karnataka, Tamil Nadu, Bihar, West Bengal and Chattisgarh. The major fruit crops grown are banana, mango, citrus fruits (sweet orange, mandarin, lime & lemons), peach, guava, papaya, pomegranate, sapota, apple, pineapple, ber, litchi, avocado, etc. Several minor crops like jackfruit, aonla, bael, jamun, custard apple, fig, karonda, phalsa, carambola, etc., are also grown. Some exotic fruit crops like dragon fruit, kiwi, durian, Malayan apple, rambutan, etc., are also getting imported either due to its extra ordinary taste or exotic flavor.

Food, Water and Air are three essential elements for existence and sustenance of life. Primarily food is manufactured by the plants by trapping the energy of Sun in presence of carbondioxide and water. Foods can be broadly classified into energy foods and protective foods. While energy is required for all bodily functions, protective foods confer a degree of resistance against diseases, corrects some disorder and prevents occurrence of certain conditions that occur due to deficiency of certain elements and phytochemicals. The balanced diets are said to be those which have an appropriate amount of carbohydrates, proteins and fats. All these three components play certain essential role in the process of metabolism. Deficiency or insufficiency of these major food components may lead to energy deficient disease conditions and are considered responsible for malnutrition world wide. Energy deficiency can threaten the existence of life. Food grains, tubers and roots are major sources of carbohydrates, and proteins. Sufficient intake of carbohydrates, proteins and fats alone does not ensure health. Health and wellbeing are the two important aspects of life, which makes it worth living. For being healthy, besides energy, body requires minerals and vitamins which are present mainly in fruits and vegetables. The role phytochemicals play in cleansing the body and fight against diseases, was understood quite recently. Therefore, for a healthy life we need to consume a balanced diet comprising of appropriate amounts of carbohydrates, proteins, fats, fruits, vegetables and nuts. While the former three can be consumed based on the energy requirement of an individual, one should consume at least 180 and 300 grams of fruits and vegetables respectively everyday.

India is the second most populous country in the world with a population of 1.3 billion and is expected to reach 2 billion by 2050. Though the country transformed itself from a food deficit nation to surplus donor in agricultural commodities in a short span of 70 years, it also encountered several stumbling block along the way. In post independence era, the governments of the day adopted a multi-pronged strategy of population control, development of irrigational infrastructure, high yielding varieties of crops and agricultural development schemes that resulted in green revolution, white revolution, rainbow revolution. etc., which saddled the country in a comfortable position as far as food production is concerned. In the process, what went unnoticed was the hidden hunger resulting in malnourished population with different kinds of ailments. ‘Hidden hunger’ manifested due to an imbalanced diet, lacking in micronutrients–vitamins, minerals and other essential diet components. Serious among them, which plague the country even today are iron deficiency causing anaemia, Vitamin A deficiency (VAD) causinig blindness in children and increased risk of other illnesses and death. Anaemia and calcium deficiency among women and stuntend growth in children are the other diet related diseases. About 10% of population are also suffering due to over feeding or mal-noursihment. Economic burden and loss of human productivity due to lifestyle diseases is taking a toll in India during the last couple of decades.

At present, the world can very broadly be divided into two main categories. (i) Industrialized countries where food is plentiful and the diet is varied, and the most prevalent diet-related disorders are cardiovascular disease (CVD), cancer, obesity, diabetes and osteoporosis. (ii) Developing countries, where food is scarce and malnutrition is a major problem or where traditional foods and cultural practices are being replaced by western-style diets resulting in lifestyle diseases/disorder. India falls in the second category. Diversified food habit is one of the direct impacts of increasing disposable income in India. Global epidemiological studies with reference to food habits have shown that increase in lifestyle diseases are directly related to reduced consumption of fruits and vegetables and increased consumption of animal food (meat and animal fat).

Downy mildew has been reported on a wide range of ornamentals, especially those in home gardens, but only in a few instances the diseases have been sufficiently severe and widespread .Generally the diseases have been of most concern in a few ornamentals grown as crops either out doors or under glass or in nurseries, where many seedlings are raised in close proximity, often under humid conditions which favour this disease. Downy mildew having importance on ornamentals and flowering plants are:

11.1. Downy Mildews of Anemone (Windflower)

Two downy mildews are reported on cultivated anemones. The one is caused by Plasmopara pygmaea while the other is caused by Peronospora sp.
Pathogen .1. : Plasmopara pygmaea
It was first recorded in Britain on decaen anemones near Liskeard, Cornwall, in september 1917

Symptoms
It causes blackening of the foliage of the planting. The sporangiophores emerged from stomata at the periphery of the blackened zone. The fine white mildew covers the under surface of the leaves. Plants become discolored with aborted stamens. Oospores are found in the discolored tissues.

10.1. Downy mildew of Plantago psyllium

Downy mildew is the major and serious disease of Plantago particularly P. ovata. It causes enormous quantitative and qualitative loss and often becomes a limiting factor in the successful cultivation of the crop (Desai and Desai, 1969). Usually the disease appears at the time of spike initiation.

Pathogen: Peronospora alta and P. plantaginis

Geographical distribution
Plantago ovata under commercial cultivation has been reported to suffer from downy mildew disease. Downy mildew occurs on ripe seeds of Spanish, French and European plantago. The downy mildew is a native of Mediterranean region especially Southern Europe, North Africa and West Pakistan.
The plantago commercially cultivated in Gujarat and to a limited extent in Haryana and Rajasthan provinces in India is observed to be infected with downy mildew.

9.1. Downy mildew of Opium poppy
Pathogen: Peronospora arborescens (Berk) de Bary

Geographical distribution and losses
This is one of the serious and widespread diseases of opium poppy. The disease was observed epiphytotic in Argentina, Germany, Iran, Hungary, Austria and India. The disease was first recorded in India in 1918 and with the introduction of susceptible varieties, the disease rapidly became serious due to favourable environment in its cultivation area in India. Downy mildew now occurs in most regions in India (Rajasthan, Madhya Padesh and Uttar Pradesh). Downy mildew caused 20-30% loss of yield of opium latex. The secondary infection of downy mildew was found to reduce 11-22% latex.

Symptoms
The infection spread upwards from lower leaves. The entire leaf surface is covered by downy mildew coating, which is composed of sporangiophores and sporangia of the pathogen (Fig 9.1). The stem, branches and even capsules are attacked resulting in the premature death of the plants. The infection start from the moment the seed germinate and persist during the whole period of growth (Yossi fovitch 1929). Khristov (1943) reported that the pathogen often cause abnormal development in opium poppy. The pathogen also causes hypertrophy and curvature of stem and flower stalks.

8.1. Downy mildew of cruciferous vegetables

Cruciferous vegetables are important Kharif vegetable crops, which are grown both for table and seed purpose. This vegetable group constitutes crops like cauliflower, cabbage, radish, turnip, broccoli, brussel’s sprouts; knol-khol and rutabaga .The vegetable crops affected by downy mildews pathogen Peronospora parasitica are members of the genus Brassica and the downy mildew diseases on these crops are widespread in those regions of the world, which have cool and wet climate.

Geographical distribution

The downy mildew disease on cruciferous crops has worldwide distribution. The disease was first recorded in 1883 in USA on Brassica spp. (Farlow, 1883). In India, this disease was first reported by Butler in 1918. Subsequently various authors reported this disease from many countries of the world (Channon, 1981) particularly Angola (Serafim and Serafim, 1968), Argentine (Lindquist, 1939), Australia (Samuel, 1925; Anon., 1955), Austria (Glaeser, 1970), Bermuda (Waters-ton, 1940), Brazil (Grillo, 1937), Britain (Moore, 1959), Brunei (Herb. IMI), Canada (Jones, 1944; Downey and Bolton, 1961), Chile (Mujica and Vergara, 1960), China (Porter, 1926; Pai, 1957), Columbia (Orjuela, 1965), Costa Rica (McGuire and Crandall, 1967), Cuba (Fernandez Rosenada, 1973), Cyprus (Herb.IMI), Czechoslovakia (Rydl, 1968), Denmark (Gram and Rostrup, 1924), Dominica (Anon., 1972a), Egypt (Elarosi and Assawah, 1959), Ethiopia (Herb. IMI), Fiji (Anon, 1969), Finland (Herb. IMI), France (Darpoux, 1945), Germany (Neumann, 1955), Greece (Herb.IMI), Guatemala (Muller, 1950), Haiti (Anon., 1972a), Hong-Kong (Johnston, 1963), India (Thind, 1942), Iran (Ershad, 1977), Iraq (Herb.IMI), Israel (Peleg, 1953), Italy (Ciferri, 1961), Jamaica (Leather, 1967), Japan (Hiura and Kanegae, 1934), Kampuchea (Soonthronpoct, 1969), Kenya (Anon., 1957), Korea (Anon., 1972b), Libya (Herb.IMI), Malawi (Peregrine and Siddiqi, 1972), Malaysia (Mcintosh, 1951), Malta (Herb.IMI), Mauritius (Orian, 1951), Mexico (Rodriguez, 1972), Morocco (Herb. IMI) , Mozambique (Decarvalho, 1948), Nepal (Bhatt, 1966), Netherlands (Thung, 1926b), New Zealand

7.1 Downy mildew of grape
Pathogen: Plasmopara viticola

History and Epidemics
In France downy mildew is called as “le mildiou” while in Germany it is called as “falscher Meltau”. Downy mildew is undoubtedly of American origin and was probably indigenous on the wild grapes of the eastern United States, from which it spread to the cultivated vineyards when these were established by the colonists. It was not known to occur outside America until 1870, although it was first described from this country as early as 1834. In the early part of the last century various horticultural writers referred to the destructive action of the mildew on cultivated grapes.
Its history in Europe dates back to the time when French vine-yards were suffering from the grape Phylloxera, an aphid insect pest that was also introduced into Europe from America. It was found that rootstocks of the American species were more resistant to this aphid than was the European vinifera grape, and therefore consequently around 1870 importations of the American Vines were made into Europe, and somewhere around 1875 the downy mildew fungus evidently was also introduced with these importation and became established, and by 1878 it was prevalent enough to cause alarm. Since the European vinifera grapes were far more susceptible to this disease than were the American species, it soon became a real menace to grape culture throughout the humid sections of Europe.

6.1. Downy mildew of Sugarcane

Pathogen: Sugarcane downy mildew can be caused by four species:

    1)    Peronosclerospora sacchari
    2)    P. miscanthi
    3)    P. spontanea and
    4)    P. philippinensis

Major Host
Hosts in the Saccharum genus include: S. officinarum, S. robustum, S. spontaneum and S. edule.

Minor Host
Other hosts include Zea mays (maize), teosinte (Euchlaena mexicana), Sorghum halapense (Johnson grass), S. sudanense (Sudan grass), Tripsacum dactyloides (Gama grass), Sorghum bicolor (broom corn). Other studies suggest there are 18 species that are systemically infected in the sub-family Panicoideae; these include species of Andropogon, Bothriochloa, Eulalia, Schizachyrium and Sorghum (Bonde & Peterson 1981)

Geographic Distribution
The disease is restricted to the Pacific, South Asia and South East Asian regions viz Fiji, India, Indonesia, Japan, Papua New Guinea, Philippines, Taiwan and Thailand (Suma & Magarey 2000).

5.1 Downy Mildew of hemp

Pathogen: Pseudoperonospora cannabina

Geographical distribution
 
It’s spread has been limited to Asia and Europe.

Symptoms

Downy mildew begins as yellow leaf spots of irregular size and angular shape, limited by leaf veins. Opposite the spots, on the underside of leaves, the fungus emerges from stomata to sporulate. Mycelial growth on the underside of leaves is best seen in early morning when dew turns the mycelium a lustrous violet-grey colour (Fig 5.1). Lesions enlarge quickly and affected leaves become contorted. Leaves soon become necrotic and fall off. Whole plants and entire fields may follow this pattern.

4.1. Downy mildew of sunflower

Pathogen: Plasmopara halstedii (Fort.) Berl.& de Tom.

Hosts

Over 100 host species from a wide range of genera in the family Asteraceae have been reported susceptible to this pathogen, including wild and cultivated species of Helianthus, e.g. sunflowers, which is the principal economic host. Wild Asteraceae hosts (e.g. species of the genera Helianthus, Artemisia, Xanthium, etc.) may also occur widely in the corresponding areas but their potential as reservoirs of the pest is not yet known (Virányi, 1984). For lists of hosts see Leppik (1966) and Novotel’nova (1966).

Geographical distribution

EPPO region: Present wherever sunflowers are grown, in Albania, Austria, Bulgaria, Czech Republic, Egypt, Estonia, France, Germany, Hungary, Italy, Moldova, Morocco, Poland (unconfirmed), Romania, Slovakia, Spain, Switzerland, Turkey, Russia (European, Siberia), Ukraine and Yugoslavia.
Asia: Azerbaijan, China, Georgia, India, Iran, Iraq, Israel, Japan, Kazakhstan, Pakistan, Russia (Siberia), Turkey. 
Africa: Egypt, Ethiopia, Kenya, Morocco, Zimbabwe, Uganda.

3.1. Downy mildew of peas
It is an important disease of pea crop particularly in those areas where cool and wet climate is prevalent during its cultivation. The disease during favourable weather conditions causes considerable yield losses.
Pathogen: Peronospora viciae (formerly P. pisi)
Host range
Vigna faba, Vigna sativa. Pisum arvensis, P.sativum, Lathyrus sativus, Vicia hirsuta, Vicia sativa, Vicia villosa (Dixon, 1981).
Geographical distribution
Reports of the pathogen are from different countries on different crop host.
On Pea- Berkeley in England first discovered the disease in 1846 (Chupp and Sherf, 1960) and since then it has been found in all parts of the world where the crop is grown intensively. The occurrence of the disease has been reported from Argentina, Australia, Canada, Chile, Denmark, France, Germany, Kenya, Netherland, New Zealand, Rhodesia, Spain, Tasmania, USA, Ukrain, Italy, Poland and Australia (Dixon,1981). In India it is prevalent in Indogangetic plains including Uttar Pradesh and Punjab (Bains and Dhaliwal, 1986).

2.1. Downy mildew of Sorghum
Pathogen: Peronosclerospora sorghi (W. Weston & Uppal) C.G. Shaw, (1978)
Host plants
Major hosts: Sorghum bicolor (sorghum), Sorghum caffrorum, Sorghum Sudanese (sudan grass), Zea diploperennis, Zea mays. 
Minor hosts: Andropogon sorghi, Panicum trypheron, Pennisetum glaucum (pearl millet), Sorghum halepense (Johnson grass), Zea mexicana (teosinte) 
Geographic Distribution
It is found worldwide in tropical and subtropical regions. Sorghum downy mildew is widely distributed in following countries.
Asia: Bangladesh, China, India, Iran, Israel, Japan, Nepal, Pakistan, Philippines, Thailand and Yemen (CMI, 1988; Jeger et al., 1998).In India though the losses are reported from the states of U.P. and Madhya Pradesh, the disease is very common and destructive in Karnataka, Tamil Nadu, Andhra Pradesh, and Maharashtra

Downy mildew is refered to a condition of plant disease where the fungal growth and infection appears on the dorsal side of the leaf surface while the corresponding ventral side of the leaf shows faint yellowish color. It is also refered to the fungus which causes this disease.The word mildew in Latin language means mil + dew i.e honey like dews.The fungal growth on plant surface resembles to the milky dews in the beginging which turn to golden brownish honey like dews in later stage. Subsequently the affected areas turn grayish brown and dry. Besides the infection of leaves, the fungus also infects the fruits. The downy mildew infection on fruits causes the deformities on the affected fruits. The infection of the downy mildew pathogen on some of the cereals like pearlmillet causes the symptoms like green ear disease .

Downy mildew pathogens can cause major damage in the nursery, in greenhouse, in field crops and in landscape in the form of leaf spots, blights, and distortions with mildew growth. Downy mildews are primarily foliage blights with mildew growth of the fungus on affected portion. The fungus attack and spread rapidly in young, tender green leaf, twig, and fruit tissues. The fungal development are severe when a film of water is present on the plant tissues and the relative humidity in the air is high during cool or warm, but not hot, periods. Downy mildews can cause severe losses in short periods of time. Downy mildews often cause rapid and severe losses of young crop plants still in the seedbed or in the field. They often destroy from 40 to 90% of the young plants or young shoots in the field, causing heavy or total losses of crop yields. The severity of loss depends on the prolonged presence of wet, cool weather during which the downy mildews fungus sporulate profusely, cause numerous new infections, and spread into and rapidly kill young succulent tissues.

1.    Remove the growth media
2.    Wash 2 times with warm PBS (w/o Ca and Mg)
3.    Add 5ml of trypsin /EDTA 0.35 % (Invitrogen), place at 370C in a CO2 incubator for 4-5 min
4.    Wait until cells are detached/lifted and make sure by examining on microscope
5.    Add equal volume (5ml-1:1) of growth medium as appropriate and mix with 5 strokes
6.    Transfer into 15 ml centrifuge tube
7.    Centrifuge at room temperature for 4 min in 980 rpm
8.    Remove supernatant complete but carefully without touching the cell aliquot
9.    Re-suspend in 10ml DPBS (with Ca and Mg)-5 strokes
10.    Immediately after mixing transfer 100µl of cell suspension into Casy vial, containing 10ml Casyton

1.    Collect the bone marrow from iliac crest of rabbit, dog, goat and sheep; sternum of pigs by 16-18 G bone marrow biopsy needle under sedation/anesthesia and maintain strict asepsis protocol
2.    To aspirate bone marrow, negative pressure should create in the syringe by pulling back the plunger. Syringe should be first flushed with heparin before bone marrow collection
3.    Collect approximately 2.5-5 ml of bone marrow from each animal
4.    Mix the bone marrow sample with equal volume of low DMEM having 10% FBS and 1 ml of PS and then layer on 5 ml of histopaque/Ficol-Paque Plus in a falcon tube
5.    Centrifuge at 3000 rpm for 30 min
6.    Collect stem cells from the interface with the help of a micropipette
7.    Cells wash thrice with DPBS 
8.    Centrifuged at 2000 rpm for 5 min

1.    Collect fat tissues from euthanized animals into a Falcon tube containing DPBS + 1% PS ( for transferring lab to lab)
2.    Wash 3 times with DPBS + 1% PS
3.    Inside a laminar cabinet, transfer the fat to a Petridis (use an sterile spatula to collect the fat)
4.    Cut the fat into small (mm) pieces using scalpel and forceps or scissors
5.    Transfer the fat into a Falcon tube (eg into a 50 ml Falcon tube you can add upto 10 ml fat tissues)
6.    Wash the fat by adding approx 40 ml DPBS; Centrifugation: 430g, 10 min (without break)
7.    Transfer the fat into a fresh 50 ml Falcon tube (with the help of sterile spatula); Repeat steps 5 to 7 for 2-3 times

Collect the femur and tibia and clean them very well from blood, tendons, muscle, skin and other tissues (using scalpel)

Cut off epi-and metaphysis from the long bones with electric/manual bone saw
Transfer the open bones to a falcon tubes containing cell culture medium/DPBS without FBS but containing 1% PS (Penicillin-streptomycin)
Transfer the bones to a Petri dish containing 5 ml medium without serum (FBS) and without antibiotic (40-50 mm Petri dish)
Remove the pteri dish to the laminar cabinet
Collect the bone marrow by flushing out to a 50 ml falcon tube (use syringes and needles to flush out the bone marrow; flush out the bone marrow with complete (10% FBS, 1% PS, 90% DMEM-LG) culture medium
Centrifugation: 300rpm, 8 min (in the meanwhile prepare 5 ml of Ficoll into a 15ml falcon tube)
Aspire the medium

Bone is the only tissue in the body able to heal without microscopical scarring. On the other hand fracture healing is a slow process causing long immobilization periods with consequently high costs for fracture healing giving rise to delayed healing, fibrous healing or non-healing with subsequent problems for both, the patient and the orthopedic surgeon. Fracture healing is complex phenomenon which involves numerous cells, regulators of cell function and biochemical interactions in the repair process. The healing potential of bone, in a fracture model is influenced by a variety of biochemical, biomechanical, cellular, hormonal and pathological mechanisms. A continuous occurring state of bone deposition, resorption and remodeling facilitates the healing process.

Biodegradable and bioinert ceramic materials such as hydroxyapatite (HA), tri­calcium phosphate (TCP), aluminum-calcium-phosphate (ALCAP) ceramics provides scaffold to support the attachment and migration of newly formed bone cells into the osseous defect and also help in formation of a vascular network through the newly formed bone. Bioceramic also act as a carrier to deliver stem cells for osteogenesis.

Specific bone-inductive proteins/growth factors can induce bone formation and healing in vivo. These morphogens are therefore, to be an ideal alternative to autogenous bone grafts. These findings initiating intensive research into bone regeneration orchestrated by putative soluble signals and lead to the discovery and identification of an entirely novel family of protein initiators collectively called the bone morphogenic protein (BMPs), which belong to transforming growth factors-ß (TGF- ß) super family. These growth factors might possess a biological activity that is site and tissue specific in the induction of bone formation. Different growth factors have been demonstrated to possess osteoinductive and fracture healing properties.