eChapters

Summary

Tropical forests are declining sharply and continuously growing demand of wood leads to removal of trees from their natural habitats. Clonal propagation of superior trees is must for further exploration and conservation of these genetic resource. Tissue culture technique offers rapid and large scale propagation of desired traits. However, micropropagation of adult trees is difficult; many problems are associated like- maturity, contamination, phenolic leaching, season, variants, long life cycle, etc. Shoot bud culture is the easiest and successful tissue culture technique for the toughest material to clone- mature trees. The chapter deals with the bud culture method of heartwoods and associated problems at different stages – explant establishment, shoot multiplication, rooting and acclimatization. Also the possible solutions of the problems are discussed on the basis of previous reports.

Abstract

Quercus species of family Fagaceae are very important from both economic and ecological perspectives. These exhibit irregular fruiting, and produce seeds recalcitrant to storage, thereby difficult to propagate through conventional procedures. Again because of their very difficult to root nature, extremely limited success has been observed in vegetative propagation. Application of new technological approaches like in vitro propagation may be of interest for accelerating tree improvement programs to these hardwood species. This review provides a summary of the advancement in the application of in vitro propagation to oak species of Kumaun West Himalaya India.

A. In vitro Rumen Fermentation Technique for Forage Quality Evaluation

The term in vitro means “in a glass” in contrast to in vivo which is defined as “within the living body”. In vitro implies a close relationship to an in vivo measurement an in vitro reaction is used as a substitute for a reaction which normally occurs and is observed within a living body. the terms  “artificial rumen” and “in vitro rumen fermentation” are often used interchangeably.

The main advantage of the in vitro techniques rests in the ability to use them to study activity of micro-organisms away form the control and influence imposed by the host animal.

ABSTRACT

The in vitro culture studies in crop plants show that there is a differential response of various explants to different concentrations of auxins and cytokinins. This indicate that hormonal requirement is specific to the genotype and developmental stage of the explant. Therefore standardisation of media and explants were done for in vitro callus culture and plant regeneration in L. esculentum and L. pimpinellifolium at the Department of Olericulture, College of Horticulture during 2000-2002.

The source materials selected for the study were L. esculentum var. Sakthi and L. pimpinellifolium. The explants were leaf segments (0.5 cm²) from fully opened leaves, nodal segments (0.5 cm) and internodal segments (1.0 cm). The media consisted of the basal MS and ½ MS media along with different auxins and cytokinins either singly or in combination. The auxins used were NAA (0, 1, 2 and 3 mg l^-1 and 2,4-D (0.5, 1, 1.5 and 2 mg l^-1) and the cytokinins used were BA and kinetin (1, 2, 3, 4 and 5 mg l^-1).

ABSTRACT

Advances in biotechnology, particularly methods for culturing plant cells and tissues, can provide new means for the commercial processing of even rare plants and the chemicals they produce. These new technologies will extend and enhance the usefulness of plants as renewable resources of valuable chemicals. Aristolochic acid is mainly found in the genus Aristolochia. Aristolochic acid has been shown to have anti-tumour activity and has been recommended for its activity against various effects of snakebite and its capacity to inhibit phospholipase A₂ or other enzymes. Considering the importance of medicinal use of A. bracteolata and A. indica, in the present study, they were selected for in vitro development and rapid multiplication of plant through tissue culture. The study was undertaken to enhance the in vitro production of Aristolochic acid in different tissue cultures. Cell suspension culture was established from friable calli of both the species in MS liquid medium. Bergmann plating technique was carried out for cell line selection for more production of Aristolochic acid. Primary transformation was carried out by Agrobacterium tumefaciens strain LBA4404 / pTOK233 with Gus and hpt genes to confirm the transgenic potential of A. bracteolata and A. indica. In order to enhance the production of aristolochic acid in in vitro culture condition, the physical, chemical and biological conditions were altered. Aristolochic acid was isolated, identified and purified from naturally growing plants as well as from tissue culture plants and all other culture staes

1. Introduction

Atalantia monophylla (L.) DC. is a small tree belonging to the family Rutaceae; resembles Citrus in general aspects and is considered a close wild relative of Citrus. The species is commonly called as wild lime, in Malayalam it is called as Kattunaragam and in Tamil Katuuelumichai. A. monophylla is used in Ayurveda, Folk and Siddha systems of medicine. It is used for the treatment of rheumatoid pain and glandular swelling (Sankaranarayan et al., 2010). The root is believed to be antispasmodic (Kiritikar and Basu, 1999), the leaves are often applied for itching and other skin complaints (Panda, 2004) and acridone alkaloids isolated from roots were reported to have anti-allergic activity (Chukaew et al., 2008). The chloroform extract of leaves has shown antiviral activity (Chansakaow et al., 1996). Besides its medicinal value A. monophylla leaves are used as flavoring agent, berries are used for making pickles, juice of berries are used for dyeing purpose and wood as timber. It is also useful as a rootstock for breeding new cultivars of Citrus Linn. (Guha Bakshi et al., 1999). This species has high medicinal value and the natural population is under threat due to habitat destruction, use of wood as firewood, collection of fruits for making pickles and harvesting of leaves, roots for medicinal purposes by the local communities. Hence there is a need for developing an appropriate conservation strategy for this species.

Before applying fungicides in field at large areas, it is required to be thoroughly investigated under laboratory conditions for its efficacy on pathogen, subjected. Basic methods of testing these chemicals against subjected pathogen, under laboratory conditions are described here.

1. Poisoned Food Technique

Principle

The principle involved in this technique is to poison the medium with fungicides and then allow a teat fungus to grow on such medium. The test may be conducted both on solid and liquid medium.

Principle

Orchid seeds are produced in large numbers but are exceedingly small, hence are also called “dust seeds” may be as small as 0.01 by 0.05 mm in dimension and weigh as little as 10-30 ug. Seedling development is generally a slow process in orchids, with the seedlings passing through an initial stage, termed the protocorm stage, prior to the production of distinct vegetative organs (leaves, roots, etc). The protocorms of terrestrial orchids are subterranean and entirely heterotrophic, with all of the nutritional needs of the orchid being supplied by the mycorrhizae (the symbiotic fungus). The length of the protocorm stage is variable, however, in many cases, months, or even years, elapse before leaves and roots are produced.

The combination of a small seed size and fungal symbiont requirement largely precludes the propagation via conventional seed propagation protocols. Accordingly, most of the early research on the development of orchid seed germination protocols focused on in vitro germination with the seeds and fungal symbiont being sown on the same petri plate. Successful protocols were developed for several species; however, the process was complicated by the diversity of fungal symbionts required (across orchid taxa) and the difficulty in isolating and culturing these fungi.

In the 1920’s the process of orchid seed propagation was revolutionized by Lewis Knudson with the demonstration that nutrient media that could be used as a substitute for fungal infection. The isolation and culture of multiple fungal strains was no longer required, and improvements in germination frequency, germination speed, and seedling vigor were generally observed. To date, successful asymbiotic germination techniques, termed ‘flasking’, have been developed for many epiphytic orchids, with several species being saved from extinction via the application of these techniques.

Aplant collector and conservationist faces many problems. These include adverse weather, limited availability of personnel, insufficient material because of poor growing season, general scarcity of the plant in the target region, grazing of the plant by animals, damage by pests or diseases or immaturity of the material. These problems are common to both seeds and vegetative propagules. The vegetatively propagated plant material face another unique problem related to excessive weight and bulk. The cost and inconvenience of transporting large amounts of material can be a limitation on the scope of a plant exploration mission. There is additional need to maintain selected clones in a disease-free condition for distribution to national programmes.

The knowledge and principles from interconnected disciplines of bioengineering, material science and life sciences are combined in tissue engineering with the aim of developing a construct that can wholly or partly restore/maintain/augment the function of a damaged tissue or organ. Tissue engineering construct generally comprises cells on a suitable scaffold. The ultimate functionality of a tissue engineering construct is mainly dependent on the cell behavior on scaffolds. The proliferation and cell viability on scaffolds varies from one cell type to another and from one species to another. So it is important to investigate individual cell-scaffold combinations.

The similarity in structure and composition to bone mineral, osteoconductive properties, ability to integrate with the bony tissue and absence of immune response makes hydroxyapatite [Ca10 (PO4)6(OH)2] the choice of calcium phosphate biomaterial with only disadvantage of slow rate of resorption. On contrary, silica–calcium phosphate composite in comparison to calcium phosphate is rich biomaterials having a faster resorption rate. Studies have shown that coating of hydroxyapatite with a calcium silicate containing layer encourages cell proliferation and osteogenic differentiation of human bone marrow-derived stromal cells.

In this chapter a composite scaffold (calcium silicate, hydroxyapatite and tricalcium phosphate), namely HASi with elements in the following percentages: 66.36 % - calcium, 25.35 % - phosphorus and silicon - 8.29 % and porosity of 50 – 500 µm is undertaken to evaluate the rBMSC proliferation and differentiation properties in vitro with the aim of assessing cytocompatibility of HASi and rBMSC in terms of cell attachment, cell morphology, cell proliferation and osteogenic differentiation.

The estimation of digestibility by feeding trials with sheep or other ruminants is laborious and requires large quantities of feed. Therefore, the chemical composition of feedstuffs are used to predict digestibility by making use of established mathematical relationship between digestibility and composition. However, since the relationship between digestibility and chemical composition varies with the type of feedstuffs, for each group of feedstuffs, mathematical equations will have to be developed by relating chemical composition with in vivo digestibility. In spite of this, the errors associated with this approach of predicting digestibility is large. This has led to the development of techniques that would mimic digestive function of the ruminant in the laboratory.

In vitro methods of feed evaluation using rumen fluid and/or digestive enzymes are the most widely used techniques in ruminant feed evaluation.Although,the technique adopted in different laboratories differ, the principle of the technique is essentially a partially simulated rumen environment in vitro by incubating rumen fluid anaerobically in buffered medium at 39°C for variable period of time. Currently the most commonly used rumen in vitro techniques are,

The Indian centre of origin and diversity of cultivated plants represent an outstandingly rich reservoir of nature’s wealth of plant species which include a large number of economically important species including Vegetable Crops. Common vegetables like okra and brinjal are native to this region. Out of the recorded 110 genera and 640 species of cucurbits, 100 species representing 36 genera occur in India. Twenty of these species are endemic. Adoption of high yielding crop varieties and hybrids along with changing land use patterns have led to rapid replacement of native cultivars in most cases. Conserving and sustainable utilization of locally adapted types and primitive landraces are, hence, now major concerns at the national and global levels.

The National Bureau of Plant Genetic Resources (NBPGR) at New Delhi is the nodal organization in India with the national mandate to plan, conduct, promote and coordinate all activities concerning plant exploration and collection, germ plasm import and exchange, plant quarantine, germplasm multiplication and distribution, evaluation and characterization, documentation and conservation of both indigenous and introduced genetic variability in cultivated plants and their wild relatives.

The challenging job of collecting, genetic evaluation and utilization of plant genetic resources cannot be satisfactorily accomplished by one organization. The enormous task of safeguarding and promoting greater utilization of these valuable resources can be best achieved through a networking approach. Professional development is the key to success of this endeavour. NBPGR organizes regularly short-term training courses on various aspects of PGR to impart latest knowledge and techniques to scientists and technicians engaged in crop improvement programmes and handling germpalsm collections.

Introduction

Vegetable crops include a large number of species, mainly used as an essential complement to the daily diet, providing vitamins, minerals, fiber, specific amino-acids and other active metabolites. Largely emphasizing the need to accumulate and conserve in gene banks the genetic diversity that is most useful to breeders. One of the primary reasons to sustain conservation of plant genetic resources in gene banks is to prevent the loss of genetic diversity. The great diversity of types in cultivation is also considered a genetic resource itself, to be maintained or increased, since it adds to the diversity of vegetables being grown and consumed and could serve to replace more established but similar crops in case of need (Crisp and Astley, 1985). The risk of genetic erosion due to the introduction of single new cultivars was considered especially high by Crisp and Astley (1985) for those vegetables that are built on a narrow genetic base, such as garlic, broccoli, tomato, cucumber, etc. Genetic erosion is difficult to document with solid data available in vegetable crops. On a global level, efforts of the International Board for Plant Genetic Resources (IBPGR, now IPGRI) to conserve vegetable crop germplasm began in 1980, with the definition of a number of priority crops for conservation, according to their importance for rural development and to their economic value for farmers in the tropics (Abelmoschus esculentus and related species, Allium spp., Amaranthus spp., Brassica spp., Capsicum spp., Cucurbita spp., Lycopersicon esculentum, Momordica charantia and related species, and Solanum melongena). During last six–seven decades, germplasm of promising and profitable varieties have been built up and developed by different agencies, initially assembling local types and selecting from them and by introducing promising types from other parts of the world. The introduction and acclimatization of vegetable germplasm from within the country and all over the world is important for exploitation and incorporation of useful characters such as early maturity, high yielding capacity, resistance to diseases and pests and tolerance to different climatic hazards, viz., drought, heat, cold and flood situations.

ABSTRACT

Dendrophthoe falcata (L.f.) Ettingsh, a unique hemiparasitic taxa belongs to the family Loranthaceae, spreads over the entire forest in the South West Bengal. It is generally stem hemiparasitic taxa, as it is found mostly on aerial part of the host plant due to lack of root system. It depend on the host plant for mainly water and to some extend carbon. It synthesizes its own food. This plant has haustoria instead of root, they penetrate their haustoria into the vascular bundle mainly in the xylem tissue of the host plants. This species has numerous medicinal values with a long history of its use in Chinese traditional medicines. The present work deals with the anti-diabetic, phytochemicals and silver nanoparticles properties of D. falcata. The responsible enzyme i.e. Alpha-amylase breaks the large starch molecules which produces free glucose and simultaneously increases the blood sugar level and as a result hyperglycemia occurs. This study also reveals that the plant’s leaves extracts have inhibitory activities on this key alpha amylase enzyme and apart from that aqueous extract of this plant contains some secondary metabolites which is very crucial data for drug preparation and any other research purposes. The maximum amount of silver nanoparticles is present in this plant which is valuable as a silver line in near future implications for different drugs preparations

26.1 Introduction

We have already discussed the basics of tissue culture techniques in Chapter 2. Plant tissue culture techniques in combination with recombinant-DNA technology are essential requirements for development of transgenic plants (will be discussed in next chapter). However, tissue culture techniques like anther, pollen, ovule and meristem culture can themselves be used for improvement of different traits or as an aid to conventional breeding methods. In recent years, isolated microspore culture has been preferred as a breeding tool and an experiment system for various genetic manipulations. Besides, using protoplast fusion and hybrid embryo rescue, distant hybridization has been successfully achieved in many crops. Protoplast fusion technique can be used for the transfer of cytoplasmic male sterility from one species to another in very short duration. Somaclonal variation generated during tissue culture can also be utilized for recovery of variants showing disease resistance or other desirable characters.

India over several millenniums has been the treasure land of biological wealth, intellectual knowledge and spiritual wisdom. During both pre and post Vedic periods, arts, science and technology had flourished well in various fields. Indian agriculture to some extent is much traditional and dates back to about 10,000 years. India with its traditional crop varieties and crop production technologies was able to feed its population and also exporting some selected produces to other continents. All the Indian villages had been the self sufficient, self sustained and self reliant units. The development of modern agricultural production was achieved by creating large scale specialized farm production units, increased mechanization and use of chemical inputs.

Family: Convolvulaceae

Flowering: Most part of the year

Fruiting: Most part of the year

Botanical description: The plant is a semi-aquatic tropical herbaceous plant. The stem of the plant is thick, fistular or spongy, rooting at the nodes trailing on moist soil or mud, or floating on water. Leaves are variable in shape and size,such as ovate-oblong, ovate- triangular, lanceolate, elliptic-oblong.The flowers are trumpet-shaped, 3-5 cm diameter, usually white in colour with a purple center.Capsule ovoid, glabrous and 2-4 seeded.

Common name    :    English- Scarlet morning glory         
        Hindi- Lalpungli
        Odia- Panikod

Family    :    Convolvulaceae
 
Habit & Distribution
The plant is (Fig. 53 a) a herbaceous climber, common in open forests and open places over hedges, bushes, fairly naturalized. It is naturalized throughout India.

General Characters
It is a twining, smooth to hairy annual plant. Leaves are ovate or broadly ovate, heart shaped having bases with pointed tips. The flowers are red in colour with a longer stalk than the leaves. Each flower stalk may bear a single flower or may have several flowers. The fruit is a round capsule up to 8 mm in diameter containing few seeds. Flowering and fruiting occurs during September-January.

Common name    :    English- Yellow-Throated Morning Glory     
        Hindi- Neelkalme
        Odia- Kaladana

Family    :    Convolvulaceae

Habit & Distribution
The plant is an ornamental climber (Fig. 54a), common in waste places and road sides. It is widely distributed throughout the tropical countries.

General Characters
It is a climber. Stems trail for several metres and climb over low bushes, reddish, muriculate, hairless or velvety. The Leaves are ovate, 5-15 × 5-15 cm, blunt or long-pointed at the tip and heart-shaped. Flowers are funnel-shaped, blue with purplish tinge, 3.5-4.5 cm long, with a yellow center and borne in several-flowered cymes. The cyme is carried on a stalk 5-25 cm long. Capsule is ovoid to globose, hairless. Seeds are trigonous, brownish, hairless or very shortly velvet-hairy.

Common name    :    English- Royal jasmin         
        Hindi- Chameli
        Odia- Niali/Banamali

Family    :    Oleaceae

Habit & Distribution 
It is a shrub (Fig. 55 a), commonly found in forests and distributed in Central and Southern India.

General Characters
The plant is nearly erect, not twining or climbing. Branchlets are hairy. Leaves are simple, opposite, variable in size up to 18 cm long and 10 cm broad with long pointed end. Flowers are white, fragrant, three forked clusters with flowers stalk of 5 mm long. Berry is mostly simple, 1-1.5 cm long, ellipsoid and black when ripe. The plant flowers during March-May and bears fruits during June-July.