eChapters

The traditional medicines are actually used in human for number of years and therefore its clinical existence comes as a presumption. However, for bringing more objectivity and also to confirm traditional claims, systematic clinical trials are necessary. In Ayurvedic medicine research, clinical experiences, observations or available data becomes a starting point. In conventional drug research it comes at the last. Thus, the drug discovery based on Ayurveda follows a ‘Reverse Pharmacology’ path546.

Nevertheless, all the critical Pharmacopoeial tests such as dissolution time, microbial, pesticide and heavy metals contamination etc. must be in accordance with global standards. It is important to ensure that all the Ayurvedic medicine manufacture is in accordance with Current Good Manufacturing Procedures for herbal products547,548. There have been concerns about quality standards and safety issues of herbal medicines549. The need for new regulations for botanical medicines has also been frequently stressed and some such regulations are coming in force in different parts of the world550,551.

Very few traditional systems have a fair level of documentations in place. Most of the little traditions, the flow of knowledge is mainly through word of mouth, however, the great traditions such as Ayurveda and Traditional Chinese Medicine do have a good level of documentation. For instance, Ayurveda means science of life in Sanskrit and aims at holistic management of health and disease. It remains one of the most ancient medical systems widely practiced in Indian subcontinent and has a sound philosophical, experiential and experimental basis541. Charak Samhita and Sushrut Samhita (100-500 B.C.) are main Ayurvedic classics, which describe of over 700 botanicals along with their classification, pharmacological and therapeutic properties542,543. A classic diagnostic text Madhav Nidana includes over 5000 signs and symptoms. During the course of evolution and years actual practice, the interpretation of such documents developed different schools of thoughts and resulted in wide variations in understandings. Similar situation also exists in other systems including TCM. These differences often raise issues related to authenticity and correctness of Traditional Knowledge (TK). Thus systematic documentation, interpretations and harmonization of concepts and practices remain a major challenge in most of the systems of TM. Many governments, professional and community organizations have undertaken such documentation and harmonization exercises. Projects like Traditional Medicine Knowledge Digital Library, AyuSoft, Triskandha Kosha and Medicinal Plants Database of FRLHT are attempting systematic documentation of Ayurveda in India544. Other databases such as The NAPRALERT (NAtural PRoducts ALERT) developed by Professor Norman Farnsworth contain bibliographic and factual data on natural products, including information on the pharmacology, biological activity, taxonomic distribution, ethno-medicine and chemistry of plant, microbial, and animal (including marine) extracts. Another project relates to the Global IP, Benefit Sharing and Traditional Medicine Database. It is being developed as a component of the Global Information Hub on Integrated Medicine from the work of the Commonwealth Working Group (CWG) on Traditional and Complementary Health Systems, now directed by the Malaysian Ministry of Health. This Information Hub is planning to create a network of legal centers, scholars and NGOs working together to develop a comprehensive legislative and policy review, information resource and exchange on issues pertaining to IP rights over traditional medical knowledge and the use of medicinal plants545. Other than such efforts, there is limited systematic documentation of TM knowledge and practices.

Genetics forms the basis of human life. All individuals have a physical and physiological self, determined by the interactions of genes and environment. Pharmacogenetics is the connotation used to study how heritability affects an individual’s response to drugs.494 The term ‘Pharmacogenomics’ represents systematic identification of all human genes, their products, interindividual variation and intraindividual variation in expression and function over time. It focuses on candidate genes and may include transcriptome and proteome information that affect drug metabolism, pharmacokinetics, and pharmacodynamics. It helps in predicting the patient’s response to a drug and to design new drugs495. Pharmacogenetics is study of single genes and the effects on drug response while pharmacogenomics is related to overall variation in genome. A gene is considered polymorphic when a mutation appears in more than 1% population, which may be present in the form of single nucleotide polymorphism (SNP), simple sequence length polymorphism (SSLP) and nucleotide addition or deletion. After the Human Genome Project, study in this field is accelerated dramatically. It was found that out of 1.4 million SNPs identified in the human genome, over 60,000 are present in the coding region496. These SNPs lead to miscoding of the various proteins that plays crucial role in the biological processes. If a point mutation is present in the gene coding region for various drug-metabolizing enzymes, drug target and drug transporters, it can lead to many non-therapeutic and unpredictable drug responses. In United States, over two million people suffer from severe adverse drug reactions per year and about 100,000 cases are life threatening497.

In the area of pharmaceutical discovery, screening methods continue to play a major role in drug discovery. The screening process is simply testing molecular entities (compounds/natural products) for activity to advance those that exhibit the desired activity. This process has evolved over time in both the testing methods and in the degree of testing. Originally, the screening was done entirely by dosing animals and studying their responses. Then came test tube experiments and now, testing is conducted either with cell lines or with purified proteins in miniaturized test tubes called microplate wells. Screening by microplate has allowed the degree of testing to increase from a very small selected set of compounds to libraries, which contain thousands of test substances. Technological improvements in many areas of science and information technology have led to the evolution of screening methods from time consuming, animal dosing studies to high throughput screening (HTS) methods that have the potential to generate very large datasets of information in a very short time. Today, developments in bioinformatics have opened up the possibility of virtual (in silico) screens for targets where there is sufficient knowledge of the target or of its ligand. High throughput screening has become a pivotal part of drug discovery for identifying potential lead molecules for specific disease targets. The success of screening is measured by several factors including the throughput and costs of the assays, but ultimately by how many of the HTS hits progress forward through the drug discovery pipeline. Miniaturization of assays by conversion from 96- to 384- to 1536- wells to well-less platforms, such as microarrays and the utilization of automated equipments are methodologies implemented to increase the throughput and reduce the cost of the screen.

The main purpose of herbal drug technology is to convert botanical materials into medicines with a focus on standardization and quality control with proper integration of modern scientific techniques and traditional knowledge. The use of chromatographic techniques and marker compounds to standardize botanical preparations has limitations because of their variable sources and chemical complexity. DNA-based molecular markers have utility in fields like taxonomy, physiology, embryology, genetics etc. DNA based techniques have widely been used for authentication of plant species of medicinal importance. Pharmacognosy mainly addresses the quality related issues using routine botanical and organoleptic parameters of crude drugs, and chemoprofiling assisted characterization with chromatographic and spectroscopic techniques. The new pharmacognosy includes all the aspects of drug development and discovery where the biotechnology driven applications play an important role. Current focus on chemotype-driven fingerprinting and related techniques require integration with genotype-driven molecular techniques so that an optimal characterization of botanical materials is possible. This review provides a brief account of various DNA based technologies that are useful in genotyping and quick identification of botanicals with suitable examples.

Review of Health Claims

Regulators of different countries have different procedures and views to review health claims of traditional medicine, herbal medicines, botanicals, functional foods, nutraceuticals, health products, diatory supplements and such other natural forms of materials intended to use for health promotion. However, if such products carry any therapeutic or health benefit claims then the regulators may provide general guidance and demand for sufficient evidence base in support of such claims. For instance, U. S. Food and Drug Administration, Center for Food Safety and Applied Nutrition (CFSAN) provides general guidance to review of health claims in functional foods and dietary supplements. Ethnopharmacological preparations involving herbal medicine and traditional medicines prepared from natural ingredients also will need similar process. The scientific review process for health claims of such products is comprehensive and mainly focuses on review of individual studies³²⁴

This guidance explains when a botanical drug may be marketed under an over-the-counter (OTC) drug monograph and when FDA approval of a new drug application (NDA) is needed. In addition, this document provides guidance to sponsors on submitting investigational new drug applications (INDs) for botanical drug products, including those botanical products (or botanicals) currently lawfully marketed as foods and dietary supplements in the United States. This guidance also discusses several areas in which, because of the unique nature of botanicals, FDA finds it appropriate to apply regulatory policies that differ from those applied to synthetic, semisynthetic, or otherwise highly purified or chemically modified drugs (including antibiotics). In particular, the guidance states that applicants may submit reduced documentation of preclinical safety and of chemistry, manufacturing, and controls (CMC) to support an IND for initial clinical studies of botanicals that have been legally marketed in the United States as dietary supplements or cosmetics without any known safety concerns.

The development of a new drug begins typically in the chemistry laboratory and is followed by a wide range of biological studies in vitro (cells/tissues) and in vivo (animals), which forecast what the agent would do in humans. The focus of these studies moves from evaluating the safety of a drug for use in humans to looking at its clinical relevance. Pharmacodynamics involves a set of biological studies both in vitro and and in vivo to get a better understanding of the mechanism of action of a drug. Preclinical studies mainly include safety and toxicity to ensure its safety for use in humans. Typically, preclinical in vitro, in vivo, and Phase I testing may not be necessary if preliminary data on safety are available. However, preclinical data or clinical observations are still necessary to justify a rationale for use and safety in initial clinical studies.

Three main screening approaches are generally used in drug development. Simple Screening evaluates substances that have a particular property by one or two tests. There is no need for a series of tests in which the interpretation of results of one test may depend on others. Blind Screening is used for a new plant extract where no adequate pharmacological information is available. Blind screening may give clues for potential activity and worthiness to probe further for pharmacological activities. Programmed screening involves a new plant extract or substance with known pharmacological effects and results in more precise information.

Typically herbal pharmaceutics may include any medicinal or functional food preparations such as phytopharmaceuticals, single or poly herbal formulations, and nutraceuticals made out of botanical materials. This includes two broad categories. First, there are many evidence based formulations and mixures that may or may not be based on traditional knowledge or practices. The second group includes all the traditional or ethnopharmacological preparations. Bringing herbal medications from either group to the market will require a pharmaceutics approach balanced with an understanding of the original context in which the treatment was used.

As an example, Ayurvedic medicines have been used safely for many years on a wide variety of patients. Medications that have not been altered from their original form have de facto gone through a type of safety testing. When a traditional medicine is prepared differently, then it is essential to validate the drug safety to be sure that the effects of the new procedure don’t cause harmful effects. In this same way, the value of animal testing to establish safety and toxicity is not as critical if the botanicals are used in traditional forms. Suitable animal models help in understanding the mechanism of action or pharmacodynamics of medicines. However, it is well known that no good animal models exist for many human diseases, such as asthma, diabetes, and rheumatism. The use of modern technology in making traditional preparations more contemporary and scientific is important but it should not compromise the original basis and principles described in tracditional medicine. The following examples highlight some of the concerns of this approach.

Despite its existence and continued use over many centuries, and its popularity and extensive use during the last decade, traditional medicine has not been officially recognized in most countries. Consequently, education, training and research in this area have not been accorded due attention and support. The quantity and quality of the safety and efficacy data on traditional medicine are far from sufficient to meet the criteria needed to support its use worldwide. The reasons for the lack of research data are due to not only to health care policies, but also to a lack of adequate or accepted research methodology for evaluating traditional medicine235.

The statement from General Guidelines for Methodologies on Research and Evaluation of Traditional Medicines (World Health Organization, 2000) highlights the need for consensuous in addressing the problems related to the quality of botanical drugs, which ultimately is a critical determinant of safety and efficacy of medicine. Further, the distribution and widespread sale of adulterated, misbranded, spurious botanical drug products and marked increase in misleading health claims of these products demands proper regulations on botanical medicine to protect public health236. Considering these two issues development of parameters for quality control standardization and regulatory norms for controlling the growing business of botanicals is a big task. Various regulatory authorities, research organizations and botanical drug manufacturers are constantly contributing to developing guiding principles addressing issues related to quality, safety and efficacy237. In this chapter we have highlighted problems associated with quality control of the botanicals and tried to guide a way towards a globally acceptable quality standard.

Safety is a primary concern regarding traditional and complementary therapies. There are two aspects of safety evaluations. First, to ensure that the right quality of material and apt possesses are used from sourcing to marketing; and second, there is no contamination, adulteration or spiking. Few studies have reported adulteration with steroids in some traditional Chinese dermatological preparations. In an analysis of Chinese herbal creams prescribed for dermatological conditions, Keane et al. found that eight of eleven creams analyzed contained steroids194. Spiking with corticostreoinds has been observed in some market preparations claimed to be useful in the treatment of arthritis and asthma195. Recently, Saper et al. from Harvard Medical School have reported heavy metal content of Ayurvedic herbal preparations and have recommended mandatory toxic heavy metal testing196. There have been few reports of heavy metal toxicity following traditional medicine use197,198. Such studies are important and needed, however they are more related to the quality control failures of the mass manufacturing activities. Often they are wrongly used to limit the use of TM. In reality, such a QC failure should not be considered as a general negative notion to create a bias against TM. We certainly need effective QC and regulation of herbal medicines, without limiting public access to these preparations and ensuring public interest and constituting restrictive trade practice.

Ethnopharmacology and Traditional medines use natural resources as medicines. They have developed the knowledge of this through the generations of traditions and experiences. Therefore the protection of this knowledge and protection of natural resources both remain critical. Bioprospecting involves sustainable, ethical and legal processes to access natural resources and use them judicially so that the business needs do not pose any threat to nature and to people.

One important consideration is assuring the preservation of traditional knowledge, particularly oral knowledge, within communities, for the potential benefit of others. Developing mechanisms to document traditional knowledge, such as national inventories, can be beneficial in identifying cases where conservation or cultivation is needed, based on an understanding of existing natural resources used for medicinal purposes. Decisions about access and ownership of information can also be made (including those relating to intellectual property rights, where appropriate) once methods have been found to collect and preserve traditional knowledge. On the other hand, the livelihood of traditional healers may depend on maintaining the secrecy of their healing methods - one argument for consulting with traditional healers as part of the process of planning and executing the collection, storage and testing of traditional medicines.

Extending some form of intellectual property protection to traditional knowledge, of which traditional medical knowledge is a significant part, is being actively debated in WIPO and other international forums. Existing intellectual property regimes can offer protection for purified compounds (produced by scientists and companies) isolated from a natural medicine. But traditional medicines, whose method of action may not be well understood, and which often consist of mixtures of different active substances, may not meet criteria for patentability. The principal impetus for international debate is a desire to compensate the holders of traditional knowledge appropriately for the use of their knowledge by others, or to prevent so-called “biopiracy” where knowledge or genetic materials are used without the consent of the holder. China and Kenya are examples of countries that have modified their laws in an effort to accommodate traditional medicine. It is not intended that the Commission should enter into this debate, except in so far as aspects of it may be relevant to its terms of reference, in particular stimulating innovation in traditional medicine with the purpose of expanding access to affordable medicines.

Contrary to “Law of Finite Biology” though the number of genetic codes will exhaust soon in an exponential way (presumably leading to solutions to every known genetic anomaly related health problem), the number of NCE (New Chemical Entity), their number being finite⁹⁹ too, will run out of fuel soon too, despite unraveling of genomic decoding. The bleak scenario in the pharmaceutical field is a testimony to the latter surmise. As many a wise scholar has pointed out, the era of single molecule drug seem to drawing curtains. However, unlike what seems common sense, the era of combinational drug therapies of MM paradigm probably will not foot the bill. Just as very high speed serial computing simulates parallel processing but does not make real time parallel processing, mixtures of prima facie complementary medicinal molecules will perhaps not make combinational recipes. The fundamentals of non-linear dynamical complex systems (NLDCS) paradigm demand so. The odds against success with such efforts are close to odds of success of creating life one bit at a time out of mixing C, H and O molecules in a jar would be. The number of genes might be finite, but data on their combinations (and their corresponding effects) and more so on their permutations (one has to only imagine the vastness of those possibilities) would, as of now or in near future, cause an information overload and comprehension failure¹⁰⁰ (limitation of human biological brain apparatus) pitting researchers and industry against a futility barrier. However, TM provides a silver lining to these bleak analyses. Analytical comprehension of TM recipes, regimens or techniques might be unachievable by contemporary science, empirical testing of their efficacy and safety being doable and therefore taken care of, TM can provide an outlet to the drug discovery impasse because the same natural forces that shaped multi-gene interaction into complex and dynamical mode have created some of, if not all, TM solutions.

The R&D thrust in the pharmaceutical sector is focused on development of new drugs, innovative/indigenous processes for known drugs and development of plant based drugs through investigation of leads from the traditional systems of medicine. In addition, many nutraceuticals are being consumed from unregulated markets for their perceived benefits in health care and improvement of quality of life.

Natural pharmaceuticals, nutraceuticals and cosmeceuticals are of great importance as a reservoir of chemical diversity aimed at new drug discovery and can be explored as potential antimicrobial, cardiovascular, immunosuppressive, and anticancer drugs. Around 80% of all such products are of plant origin; their sales exceeded $ 65 billion in 2003. Examples of plant products and derivatives used by the pharmaceutical industry include Paclitaxel, Vincristine, Vinblastine, Artemisinin, Camptothecin, Podophyllotoxin and such.

The nutraceutical marketplace in Europe is estimated to be $ 9 billion, while the US marketplace, estimated to be $10-12 billion in 2003, is expanding at a compounded rate of more than 20% per year. US Congress has fueled the rapid growth of nutraceuticals with the passage of the Dietary Supplement Health and Education Act (DSHEA) in 1994. Globally, there have been efforts to monitor quality and regulate the growing business of herbal drugs and traditional medicine.

Historically, the terms alternative, complementary or traditional medicine (TM) all referred to a genre of health care practices or services that got bound together as a class through the logic of reductio-ad-absurdum, or because of their “absence from the mainframe of” what has come to be known as Modern Medicine (MM). However, since Stephen Fulder¹ , the identities of TM have become more distinct, MM-TM blending more active and, the line separating the two more fuzzy. To date, the World Health Organization offers the most comprehensive definition of TM² .

WHO defines traditional medicine as including diverse health practices, approaches, knowledge and beliefs incorporating plant, animal, and/or mineral based medicines, spiritual therapies, manual techniques and exercises applied singularly or in combination to maintain well-being, as well as to treat, diagnose or prevent illness.

The phraseology MM/TM is syntactically incorrect because it is contended that medicine as such, modern or otherwise, evolves and progresses through traditions. The major difference between the various traditions of health care practices is that MM rode the crest of the high wave of reductionist-scientistic rigor of modernity³ to reach the shore of global acceptability while the others never crossed the threshold of geographical and cultural locality, remaining untested, neither proved nor disproved. Darwinian principle of survival of the fittest also endorses a truism that survival is a proof of fitness. Given this principle, it makes sense that TM as we know of it today may be useful for promoting human health. Moreover, most TM considers human beings and their health in the context of their environment. This criteria, not only defines TM apart from MM but also provides for a reason as to why lately TM is emerging out of long hibernation. Developmentally, MM isolated “human” from its global environmental context as a static, stand-alone, complete, total and delimited system in itself. While this reductionist approach has a very high pay off for the first 90% of human health problems, it fails when dynamic non-linear complex systems elements kick in (I’m not exactly sure that you mean by this phrase). TM covers some parts of those hidden variables and thats the utility to modern mankind.

To fully explore the history of medical and drug development, it would be necessary to go back as early as 13,000 BCE, where drawings on the Lascaux caves in France suggest that humans were using plants for medicinal purposes. Since then, medical traditions have evolved and dissolved across societies all over the globe. Natural products including plants, animals and minerals have been the basis of treatment of human diseases. The last two centuries have seen a focus on scientific medicine, with a focus on drugs whose effects that have been proved by the scientific method. Yet given this setting, many of the major pharmaceutical corporations have renewed their strategies in favor of natural product drug discovery. Numerous drugs have entered the international pharmacopoeia via the study of ethnopharmacology and traditional medicine. At this point in history, it is possible for both traditional and western medicine to benefit from collaboration between the two disciplines. For traditional medicines, newer guidelines of standardization, manufacture and quality control and scientifically rigorous research on the scientific basis for traditional treatments will be required. In return, traditional medical traditions can offer a more holistic approach to drug design and myriad possible targets for scientific analysis. Powerful new technologies such as automated separation techniques, high-throughput screening and combinatorial chemistry are revolutionizing drug discovery. Traditional knowledge can serve as powerful search engine, which will greatly facilitate intentional, focused and safe natural product drug discovery and help to rediscover the drug discovery process. By looking at the historical trends in drug and medical developments, it is possible to understand how current drug development will benefit from this partnership.

3.1 Tomato

Tomato (Lycopersicon esculentum) is an important vegetable crop. Its 2n= 24.

Floral biology

Flowers are borne in small forked racemose cyme. Anthesis starts at 6.30 a.m. and continues up to 11.00 a.m. Anther dehiscence is longitudinal. It occurs 1-2 days after opening of corolla. The stamen sheds its pollen when the style grows up through anther tube, thus self-pollination occurs. When the stigma protrudes above the anther tube, chances of cross pollination through bees increases. The optimum temperature for pollination is around 21°C.

TROPICAL AND SUBTROPICAL FRUITS

2.1 Mango
 

Mango is considered to be an allopolyploid. The chromosome number is 2n =40. The polyembryonic seedlings of cv. Vellaikolumban, earlier reported to be 2n= 80 was later shown to be a diploid. In Canary Island, a putative tetraploid viz Gomera-1 has been identified.

Floral biology

Flower starts opening early in the morning from 4-7 a.m. and maximum flowers open between 9.30-10.30 a.m. and complete 11 a.m. Dehiscence of anthers takes place at 11.30 a.m. and it continues up to 3.45 p.m. The pollen grains are oval, or triangular or oblong. Stigma becomes receptive even 18 hours before flower opening. Mode of pollination is entomophily; nectar is present to attract the insects.

The flowering duration is usually of short i.e. 2 to 3 weeks. The mango inflorescence or panicle bears mainly two types of flowers - male and perfect. The number of flowers per panicle varies between 1000 to 6000 depending upon the variety and climatic factors. The percentage of perfect flowers varies between 0.74 per cent in Rumani, 16.41 to 55.7 per cent in Neelum and up to 69.8 per cent in Langra.

Horticultural crops include the fruit crops, vegetables, spices, plantation crops, medicinal plants, aromatic plants, commercial flower crops and ornamental plants.

Knowledge on the methods by which these reproduce is of primary importance from the standpoint of plant breeding. Besides, their annual or perennial nature also decides the methods of breeding to be adopted to get desired improvement.

Two divisions of methods of reproduction commonly recognized are:

1. Asexual i.e. vegetative parts of the plants are utilized to produce new individuals.

2. Sexual i.e. gametic fusion, the resultant zygote develops into seed which is utilized to produce new individual.

This group can be further placed in several sub divisions according to the mode of pollination. They are

a. Naturally self pollinated crop - (e.g. tomato, pepper, brinjal) wherein cross pollination occurs less than 4 % only.

b. Often cross pollinated crop- (e.g. chillies, bhendi) wherein though self pollination is more frequent, cross- pollination may occur so frequently that some methods of preventing cross pollination between varieties and strains of different genotypic constitution must be followed throughout the breeding programme.
Common breeding methods can be applied to the above two groups, however, the parents chosen for breeding under often cross pollinated crop need to be selfed for few generations to select desirable, homozygous lines.

Azadiractin (1%) was found effective for the control of Hadda beetle on Ashwagandha. Nineteen arthropods were recorded on isabgol. Among them, 13 were phytophagous species belonging to order Lepidopteran (09), Hemipteran (03) and Homopteran and six predatory species belonging to order Coleoptera (03), Neuroptera (01) and Odanata (02). The sequential occurrence of arthropods revealed the presence of aphid (Aphis gossypii) from second week of January to fourth week of February, whereas Lepidopteran (Helicoverpa armigera, Trichoplusia ni, Thysanoplusia oricalchae, Spliarictia sp. Hyposidra successari and Olene mendosa and true bugs (Graptostethus servus and Spilostethus pandurus) were observed associated during early crop growth stages (i.e. from second week of December). Infestation of aphid was uniformly severe, whereas that of Lepidopterans and true bugs were sporadic and less severe. However, presence of these pest arthropods caused significant reduction in seed yield.

Isabgol (Plantago ovata Forsk.)

The crop requires cool and dry climate during the growing season. Sowing of seeds at 4 kg/ha in 0.25-0.50 cm depth during November 20 to December 20 was recommended. In medium black cotton soil of Malwa region, second week of November was reported to be optimum time for sowing. Broadcasting of seeds followed by light sweeping with broom found to give uniform germination. A spacing of 30x 45 cm found to be most ideal to get higher seed yield in Isabgol under Madhya Pradesh situation. Response of chemical fertilisers was found low. However, a fertiliser dose of 25 kg/ha each of N and P₂O₅ as basal dose and 25 kg/ha N as top dressing at 30-42 DAS was recommended for commercial cultivation in Gujarat while 50 kg/ha N reported to increase the seed yield in Madsaur areas of Madhya Pradesh. At Anand three irrigations viz. first at the time of sowing and subsequently at 30 and 70 days after sowing (DAS) proved to be beneficial. However, four irrigations at sowing, 10,25, and 50 DAS at Mandsaur are recommended. Chemical weed control was found to be economical and a pre-sowing or pre-emergence application of Isoproturone (0.5 kg ai/ha) was recommended for weed control. One spray of Metalaxyl + two sprays of Mancozeb were found significantly superior in controlling downy mildew. In Gujarat, Pearl millet in Kharif and Isabgol in Rabi season are advocated as a suitable crop rotation. Groundnut-Isabgol rotation gave higher economic return followed by Soybean-Isabgol and Pigeon pea-Isabgol rotations in Maharashtra.

A high-yielding cultivar Vallabh Medha of mandukparni (Centella asiatica) was identified. Fresh herbage yield (12,331 kg/ha) and dry herbage yield (2,113 kg/ ha) of this variety were far more than the local variety (2,050 and 392 kg/ha respectively), besides higher active ingredients. Among mucuna selections, IIHR PS 15 with long duration, IIHR PS 6 with medium duration and IIHR PS 14 with short duration recorded significantly higher yield and high L-dopa yield/ plant. In Coleus, a promising hybrid, Hy 08-53, recorded significantly higher root yield (60.22g) and higher forskolin yield/plant (0.58 g). In ashwagandha, IIHR- WS 3 was found to be the best yielder (11.65 q/ha).

Introduction

Medicinal Aromatic Plants (MAPs) play a valuable and important role in economic, social, cultural and ecological aspects of local communities the world over.

According to World Health Organization (WHO) “a medicinal plant is plant which, in one or more of its organs, contains substance that can be used for therapeutic purpose, or which are precursors for chemo- pharmaceutical semi-synthesis”.

However, a simplest definition of the medicinal plant would be “Medicinal plants are those plants which are used in official and various traditional systems of medicines throughout the world”.

Other definition could be “Medicinal plants are plants that provide people with medicines - to prevent disease, maintain health or cure ailments”.

In Indian traditions, all the plants in this earth are considered as medicinal [Jivak in Astanga Hriday (Sutra: 9-10)].

Aromatic plants are a special class of plants used for their aroma and flavour. Many of them are exclusively used also for medicinal purposes in aromatherapy as well as in various systems of medicine.

Technique to collect fresh and hygienic inflorescence sap (Neera) from coconut

In the traditional method of collecting sap, the inflorescence to be tapped is connected with an earthen pot/bamboo receptacle for the sap collection. Spadix to be tapped is tied with plastic rope and a small portion is sliced off from the top. The spadix is gently stroked with a mallet in the morning and evening. After a period of 8 to 10 days, sufficient sap starts oozing out from the cut end. A type of clay soil, a gummy material or leaf extract is pasted on the surface and sides of the cut end. Since the sap is highly nutritious, both enzymatic and microbial fermentation takes place. The total sugar reduces progressively and it becomes more acidic and develops a harsh odour and becomes unpalatable. In order to prevent fermentation, lime is coated inside the pot. The above method is not hygienic and attracts lot of insects, ants and contaminates from pollen and dust particles. In order to collect the sap fresh and hygienic, a simple device was developed.

Crop protection research has brought out tangible and effective methods to suppress various pests in major agro-ecosystems. Indigenous knowledge in this regard has been tested and validated for its suitability.

Among the 137 Phytophthora isolates collected from bud rot/fruit rot affected coconut gardens in Tamil Nadu, Kerala and Karnataka, four isolates from bud rot affected gardens were identified as P. nicotianae and one as P. capsici. Since, P. nicotianae is found associated with bud rot disease of coconut in three districts representing three states, it is expected that this may emerge as a major pathogen of coconut. Further, placing Trichoderma coir pith cake in the innermost leaf axils of coconut palm in disease endemic areas just before the onset of southwest monsoon (May-end) and thereafter at two months interval was found to be very effective in the management of bud rot disease of coconut.

Cost-effective farming systems for plantation crops cultivation for increasing production and productivity in different agro ecologies have been developed.

In coconut-based cropping system, elephant-foot yam cv. Gajendra yielded high amount of corm. Guinea grass (var. GGCo 3) produced higher green fodder (89.2 tonnes/ ha/year) under husk application. There was lower specific volume of soil and soil porosity (higher soil compaction), and soil dehydrogenase activity (an index of biological activity) as important abiotic pre-disposing factors in incidence of root wilt disease. Growing Gomphrena globosa as intercrop in root wilt- affected coconut gardens has been shown to be cost-effective and sustainable. In areca-based mixed farming system, total cash inflows and outflows from arecanut + dairy (3–4 milch cows) amounted to ‘4.41 and ‘5.96 lakh, with a net profit of’1.55 lakh. The system productivity of arecanut + cocoa system (3,127 kg/ha) was 30% higher per unit area than arecanut alone (2,405 kg/ha). Integrated nutrient management on four improved arecanut varieties recorded highest chali yield (4.19 kg).

High-yielding Plantation crops varieties and hybrids with tolerance to pests / diseases and to various farming constraints such as soil and water salinity, soil acidity, drought, flood and such other factors have been developed.

Three coconut varieties (IND 045S, IND 048S and IND 058S), two arecanut varieties (VTL 62-Shriwardhan selection and VTL7-Nalbari) and two selections of cocoa (VTLC 1 and VTLC 57) were identified for commercial cultivation. Coconut variety, Kalpa Samrudhi, was recommended for commercial cultivation in Kerala and Asom, and Kalpa Sankara and Kalpasree for coconut root(wilt)- affected tracts in Kerala. Molecular-markers associated with hybridity in coconut, dwarfness and resistance to root (wilt) and yellow leaf diseases were utilized. Homology-basedmodelling of somatic embryogenesis receptor-like kinase (SERK) protein in coconut, development of a computational tool for detection of microsatellites in whole genome sequences, development of algorithm for gene annotation and genome-wide analysis of microsatellites in strains of plant growth promoting (P. fluorescens) are achievements in bioinformatics.

The term plantation crops refer to those crops which are cultivated on an extensive scale in contiguous area, owned and managed by an individual or a company.

There are two groups of plantation crops namely coconut, arecanut, oil palm, cocoa, cashewnut dealt by Ministry of Agriculture and tea, coffee, rubber dealt by the Ministry of Commerce.

These are high value commercial crops of greater economic importance and play a vital role in improving Indian economy, especially in view of their export potential, employment generation and poverty alleviation particularly in rural sector.

Common name, symptoms of damage and management of diseases of major spices and condiments

Recent Initiatives by National Research Centre on Seed Spices, Ajmer

Standardization of the drying techniques

In fennel five methods of grain drying were employed viz., shade dryer (30°C), sun dryer (35°C to 37°C), solar dryer (40°Cto 42°C), vegetable dryer (50°C) and multipurpose grain dryer (60°C). Fennel grains which were dried under solar dryer, gave maximum test weight (5.17 g), recovery (39.82 %) with lowest moisture content (61.30%) and highest taste marks (9.3) and colour marks (9.2).

A protocol for SYBR green based real-time RT-PCRfor detection of Piper yellow mottle virus and Cucumber mosaic virus infecting black pepper and Cardamom mosaic virus (CdMV) and Banana bract mosaic virus (BBrMV) infecting cardamom was developed. A native isolate of Phytophthora capsici (Is. No. 98-93) infecting black pepper was completely sequenced using next generation sequencing platform, Illumina-Solexa GA II. The sequence data was assembled by taking Joint Genome Institute’s P. capsici as reference genome. The treatment of black pepper cuttings with Carbendazim + Mancozeb (0.1%) was effective for the management of anthracnose disease in nursery.

The black pepper accession, 1114 (Kumbhachola), was tolerant to P. capsici. The accession was earlier found resistant to pollu beetle and drought. A new bacterial wilt disease on small cardamom was recorded in Wayanad, Kerala. The phenotypic and genetic characterization revealed that causative organism is Ralstonia solanacearum (Biovar 3 phylotype 1) and was 100% similar to ginger strain of R. solanacearum. Bacillus amyloliquefaciens (GRB 35) and Serratia marcescens (GRB 68) were promising for enhancing sprouting of rhizomes, growth promotion and reducing the incidence of soft rot and bacterial wilt diseases in ginger in the field.

Cost-effective farming systems for spices cultivation for increasing production and productivity in different agro ecologies have been developed.

An organic package for production of black pepper, ginger and turmeric by applying farmyard manure, vermicompost, ash, rock phosphate, Azospirillum sp. and phosphobacteria and Trichoderma sp. and Pseudomonas sp. as biocontrol agents for disease control was developed. Further, a foliar spray of 1% solution of complex fertilizer 19 : 19 : 19 (N : P : K) during spike initiation period (April second week, May first week and May fourth week) during lean cropping season under irrigated condition enhanced black pepper (cv. Panniyur I) yield by 20–25%. Soil application of zinc up to 10 kg/ ha or foliar spraying of ZnSO4 (0.25%) and Borax (0.2%) twice (60 and 90 days after planting) was recommended for higher yield and quality for turmeric in zinc and boron deficient soil.

Among all the treatments, sprinkler and drip irrigation methods caused early sprouting; early flowering with increased plant height and more number of leaves and flowers/plant as compared to the control (rainfed). Also, stiga fresh weight, dry weight, length and saffron yield were improved in sprinkler and drip irrigation methods over the control. Raised beds resulted in early sprouting, early flowering with increased plant height and more number of leaves and flowers/plant.

High-yielding crop varieties and hybrids with tolerance to pests /diseases and to various farming constraints such as soil and water salinity, soil acidity, drought, flood and such other factors have been developed.

Eight cardamom hybrids, NKE 12 × MB 5 (1,499 kg/ha), MB 5 × NKE 19 (1,461 kg/ha), GG × NKE 12 (1,350 kg/ha), RR 1 × CCS 1 (1,245 kg/ha), CCS 1 × RR 1 (1,022 kg/ha), ASH (1,930 kg/ha), NKE 12 × GG (1,746 kg/ha), GG × NKE 19 (1,635 kg/ha), were identified for high yield and resistance to mosaic and rhizome rot diseases. Two unique mutants, AFg-3 and AFg-4, for powdery mildew resistance were isolated in fenugreek. In ajwain, AA 93 has been developed which flowers 40 days earlier compared to other varieties. Thirty- two elite saffron clones were selected and evaluated for fresh and dry weight of pistil, stiga and style length, and weight. CITH 125 (4.5 kg/ha), CITH 123 (4.3 kg/ha), CITH 124 (4.3 kg/ha), CITH 122 (4.0 kg/ha), CITH 12 (4.5 kg/ha), CITH 121 (3.9 kg/ha), CITH 107 (3.8 kg/ha), CITH 120 (3.8 kg/ha) and CITH 104 (3.7 kg/ha), having higher saffron yield were identified.

• The term Spice and Condiments applies to natural plant or vegetable products or mixtures in whole or ground form, which are used for imparting flavour, aroma and piquancy to the food items.

• Under the act of Parliament, a total of 52 spices are brought under the purview of Spices Board.

• However 109 spices are notified in the ISO list.

• India is the largest producer, consumer and exporter of spices and spice products in the world and produces more than 50 spices.

• The spices production in India is of the order of 5.74 million MT from an area of about 3.07 million ha.

• Chilli is the major spice crop occupying about 25.8 percent of area under cultivation and contributing about 22.7 percent of total spices production in the country.

• Garlic accounts for 21.9% of production and 8.1% of area Turmeric accounts for 16.9% of production and 6.3% of area.

• Rajasthan occupies major area under spices owing to seed spices cultivation in the state followed by Gujarat, Andhra Pradesh, Madhya Pradesh and Karnataka.

• Andhra Pradesh, which is the foremost state producing chilli and turmeric in the country, ranks first in terms of production.

The Distributed Information Sub Center at Kerala Agricultural University, Thrissur was set up in May 1995 as the 23^rd center of the BTISNet of DBT, Government of India. The area of specilisation of the Centre is Agricultural Biotech- nology. The Centre passed through several phases involving Internet connectivity through the ordinary C band connectivity, then the Ku band connectivity etc. Now the commu- nication facilities of the Centre have been strengthened by procuring an ISDN Internet connection and a connection through ASIANET. The Center caters to the needs of the students (UG, M Sc & Ph D) and faculty of all departments of the Colleges in the main campus as well as of the other nearby academic and research centers of the University. Over the years, DISC has come to occupy a position in the KAU in the area of Infor- mation Technology , information sources and services based on Internet, CD-ROMs etc. The Centre has procured databases in the field of Biotechnology and developed several databases in the field of Agriculture and related fields. The center regularly conducts training programmes in Bioinformatics. The research activities in the field of Bioinformatics have been enhanced. The Centre has now been upgraded to DIC to facilitate intensive research on Bioinformatics in an effective manner. During the year 2004, the Centre has also started offering a course on Bioinformatics for the M.Sc Biotechnology students of the College of Horticulture.

Biotechnology research in KAU spread over various research stations and cam- puses includes Plant Biotechnology, Animal Biotechnology and Fisheries Biotechnol- ogy. Thrust areas for Plant Biotechnology include developing in vitro protocols for mass multiplication of elite types of fruit plants, ornamentals, medicinal plants, spices, plantation crops and forest species. The other thrust areas include checking clonal stability and genetic fidelity of in vitro derived plants using molecular markers, DNA finger printing and characterisation of germplasm with molecular markers, imparting biotic and abiotic stress tolerance in otherwise accepted crop varieties, genetic trans- formation for imparting/ improving expression of desired traits, in vitro germplasm conservation, enhancement of secondary metabolites production etc.

The crops identified for biotechnology research at KAU include spice crops like black pepper, ginger, turmeric, vanilla, cardamom; plantation crops like cashew, coco- nut and medicinal & aromatic plants such as Kaempferia, Trichopus, Gymnema, Holostemma and Coscinium. For improvement of such crops through molecular breed- ing, it is highly essential to identify/design specific probes/primers, micro-satellites, etc. By combining the research results generated at the Biotechnology Centre and informa- tion retrieved from the Bioinformatics Centre, suitable probes, primers are being de- signed for characterization and improvement of these unique crops through molecular breeding.

ABSTRACT

The Cry2A proteins are promising candidates for managing the development of Cry1Ac resistance in insects. The Cry2A class of proteins is different from that of Cry1A, and experiments have shown that insects resistant to Cry1Ac are not cross-resistant to Cry2A proteins. However, the Cry2A proteins are less toxic than Cry1Ac to the polyphagous pest Helicoverpa armigera. Structure-based protein engineering of Cry toxins may pave the way for directed evolution of variants with broad spectrum of insect species specificity, enhanced potency, and stability. In the present study homology modeling of Cry2Ac was done using DeepView software. The template for Cry2Ac protein sequence (accession number X57252) was selected using ExPdb template search. The raw sequence of the Cry2Ac was superimposed over the Cry2Aa PDB template (1I5P). The superimposed model was submitted to the SWISS Model server. After receiving the Cry2Ac model, energy minimization was done using Gromos96. The threading energy and force field energy were calculated. The homology model of Cry2Ac made by us along with the Cry2Ac model from the SWISS-MODEL Repository was used for comparative analysis of receptor binding regions of Cry2Aa. Results showed variation between the two highly homologous Cry2A proteins in their receptor binding regions which may account for differences in spectrum of insecticidal activity.

ABSTRACT

Astudy was undertaken at Department of Soil Science & Agrl. Chemistry, College of Horticulture, Vellanikkara to standardize different amelioration techniques to prepare enriched coirpith compost. Twelve treatments were tried using different combinations of coirpith, Pleurotus spp., Schizophyllus spp., urea, rock phosphate, cowdung slurry, mushroom waste, goat manure, glyricidia and water hyacinth. The results of the various enrichment techniques revealed that the temperature in the compost in general, increased to a peak value of 47oC on the 22nd day of composting and later decreased to a stabilized value. The observed temperature of 40-50 °C found during the thermophilic stage is essential for the rapid degradation of lignocellulose, as the thermophilic microfungi and actinomycetes involved in this process thrive at that temperature. Though the pH show a hike at the initial stage of composting, it decreased and stabilized at the later stage of composting which is conducive for growth and proliferation of microbes. The treatment coirpith with Pleurotus spp., glyricidia +20% cowdung slurry and goat manure was found to be ideal for compost preparation.