Kanke, Ranchi, Jharkhand

( A State Government University )

Research Project, College of Bio-Technology, Ranchi

Tissue Culture and allied research:

In-charge: Dr.(Mrs.) Madhuparna Banerjee

Establishment of Protocol for micropropagation:

Protocols for micropropagation of different valued plants have been established. Biochemical assay of some medicinally important plants are also being carried out to assure the quality of the regenerants. A brief account of the work is as follows:

  • Turmeric is one of the most ancient and important spices. From the time immortal it has been used as traditional medicine in India. It has been an item of export as well as foreign exchange earning for the producing countries. Since conventional method of propagation has its own limitations in respect of poor multiplication rate; in vitro mass propagation has great potential for obtaining disease free true to type planting materials. This investigation deals with the standardization of the viable protocol for mass propagation of turmeric in vitro.
  • Floriculture is the boon to improve the economic condition of the country. The conventional method of flower production does not overcome the demand of market. Gladiolus is bulbous ornamental plant and top selling flower in the domestic market. The increasing demand of high quality plant material for export and marketing competition has necessitated its true to type, disease free propagation. Viable Protocol for large scale production of Gladiolus cormlets has been developed.
  • The uses of bamboo are well known. The traditional means of propagation are insufficient to meet up the increasing demand of this forest plant. Research has been carried out to establish the protocol for micropropagation of different species of bamboo.
  • Since traditional means of propagation provide insufficient number of plantlets, micropropagation is always a better option to be the source of plantlets for banana cultivation. It provides disease free plantlets with uniformity in growth and reproduction. The study dealt with the protocol establishment for micropropagation of variety “Dwarf Cavendish”.
  • Better strains of Pisum sativum L. can be utilized for bringing improvement in the crop. Protocol has been established for micropropagation of the two varieties (HuDP-15 and HFP-4) of this crop. The differences that we observed during the field trials of the two varieties of Pisum sativum L. and those were indicated by their differential response to the same treatment under culture conditions have been revealed through RAPD analysis. For Eurayle ferox Salisb, further studies have to be carried out to establish suitable protocol for micropropagation

Hardened plants of TC raised bamboo

  • Considering the importance of the forest species, present investigation designed to standardized protocol for micropropagation can provide disease free true to type progeny in large scale through out the year. Regeneration through callusing was also tried in case of Gmelina arborea Roxb.
  • Vanda is the most popular genus among the orchids in south-east Asiatic countries. It is also most popular in India and has great medicinal importance and demand in floriculture trade. Good bud breaking was induced by using the combinations of BAP and NAA. Multiplication was also obtained in the media containing same combination of Phytohormones.
  • Micropropagation protocols of Colchicum autumnale and Celastrus paniculatus, which are important and endangered plants, have been established with a view to conserve as well as propagate these valued plants.
  • The agro climatic condition of Jharkhand, especially Ranchi is much favorable for the growth of the tropical orchid (Dendrobium sp.) of commercial interest. Protocol for mass in vitro propagation of this plant has been established successfully.
  • The plant, commonly known as Ashwagandha, is also known as Indian ginseng for its medicinal importance. Requirement of this valued plants are increasing day by day. Protocol has been established for micropropagation to produce true-to-type plants in large scale.
  • Suitable protocol for in vitro regeneration of mentha plants was established. A through trial with different combinations as well as concentrations of phytohormones was carried out to get optimum rate of multiplication.
  • Phalaenopsis is a pot plant variety of most demanding tropical orchid. Protocol for rapid in vitro multiplication of this variety was established. Hardening of the tissue culture raised plantlets was done successfully.
  • Eleusine coracana is one of the important crops of this region. An effective protocol for in vitro regeneration was established with a view to improve the variety.
  • A viable protocol for regeneration of Asparagus racemosus, which is a valued medicinal plant, has been established.
  • Micropropagation of Aloe vera has been established by using different phytohormones and large scale production has also been taken up.
  • Protocol for in vitro multiplication of Stevia rebaudiana, a valued medicinal plant has been established.
  • Regeneration of plants through in vitro by shoot tip cultures of Phlogacanthus thyrsiflorus and Adhatoda vasica was achieved. Both the plants were hardened successfully. Antioxidant property of leaves of both in vivo and in vitro grown plants of P. thyrsiflorus and A. vasica was detected by free radical scavenging activity and DNA damage protection.
  • An efficient protocol for micropropagation of this most important medicinal plant (Rauvolfia sepentina) has been established. Phytochemical analyses of this valued plant were also done to evaluate whether there is any differences between in vivo and in vitro grown plants.
  • Micropropagation of this valued plant (Jatropha curcas) was found difficult due to exudation of latex. However, protocol has been established for in vitro propagation by using apical as well as axillary shoot bud of selected varieties. Callusing was induced on pollens but regeneration of plantlets from callus could not be achieved.
  • A comparative account of antioxidant potential of tissue cultured, non tissue cultured and in vitro grown roots of Withania somnifera has been studied. Out of these, root extract followed by tissue cultured plant extract was found most effective to protect DNA (pUC 18) damage as antioxidant. A detailed Karyotype study was also done to compare tissue cultured and non tissue cultured plants.
  • A protocol for rapid micropropagation as well as microtuber production of two varieties of potato: Kchip3 and MP-644/97 has been established. Artificial seeds of these two varieties have also been produced successfully.
  • Efficient protocol for micropropagation as well as microcorm formation of Crocus sativus has been established. The antioxidant potential of saffron leaves was reported for the first time. The leaf extract was found effective to protect DNA damage (pUC18) caused by H2O2 and UV irradiation. Karyotype study revealed 8 triplets of chromosomes.

Important to note:

The college through its different units at Headquarters, KVK Dhanbad, KVK Darisai and ZRS, Dumka is providing tissue cultured plantlets to the farmers. For bulk supply, advance confirmed order in the month of January/early February help us in meeting timely supply.

Molecular Biology

i) Proteomics
In- charge: Dr. Himanshu Dubey

A. Non-Curricular Re-search:

Title of the Project: All India Network Project on Soil Biodiversity-Biofertilizers.

Funding Agency: Indian Council of Agricultural Research (ICAR )

Principal-Investigator: Dr. Himanshu Dubey

Broad-Objective of Project: Proteomics of Rhizobium bacteria

B. Curricular Re-search:

Research interests and rationale behind the research activities:

  • In-depth analysis of the major causatives reasons for abiotic stress tolerance, and deciphering the biochemical pathways for such mechanisms.
  • The targeting of structural genes or trans-acting elements regulating the key steps in these mechanisms is being used to engineer plants towards such abiotic stresses
  • A combination of strategies incorporating different approaches of genetic and phenotypic analysis is currently being used to substantially enhance the process of gene tagging and candidate gene discovery of complex traits associated with abiotic stresses.

The research areas include:

Molecular biology of metal toxicity:

  • In most countries it is chemical fertilizers which are supplying most of the macronutrients needed by crops. Farmers seldom apply micronutrients, even though intensive modern agriculture has a depleting effect.
  • Deficiencies are shown in various kinds of physiological damage, all of which affect the quality and quantity of produce.
  • We are presently undertaking identification of proteins which are specifically altered (induced or suppressed) in response to micronutrients stress (deficiency and toxicity). One dimensional gel electrophoresis (followed by coomassie and silver staining) is used to identify proteins altered in rice tissue in response to micronutrients stress (deficiency and toxicity). Proteins are extracted from rice tissue at specific intervals and analysed for the stressed and the recovery samples in response to various metal concentrations by employing the protein gels (Sodium Dodecyl Sulphate Polyacrylamide Gels i.e. SDS-PAGE).
  • The analysis showed that several changes happen in the rice plants to adapt to the stress regimes with respect to the protein levels. Further characterization of the protein bands with respect to reveal their exact identities is currently being pursued.

Molecular Biology of “Drought Stress” and “Submergence Stress” in rice:

  • “Proteomics” approach is currently being pursued in my laboratory for characterization of “Drought Stress and Submergence Stress” response in rice.
  • Protein profiling and characterization by “Two Dimensional Gel Electrophoresis” and by “One-Dimensional SDS-PAGE”: Changes in the morphological patterns as-well as protein profile patterns due to onset of drought and submergence stress (S) / recovery (R) are currently being studied in the laboratory. Proteins are extracted from different stress and recovery samples. The total soluble proteins are separated by two dimensional gel electrophoresis (By employing IPG Immobiline Dry Strips) and / or one dimensional SDS-PAGE. To monitor lower molecular weight proteins Gradient SDS-PAGE are routinely prepared and analysed. The gels are analysed by either Silver Staining and / or Coomassie Staining and documented by using specialized gel scanners by employing gel documentation software. Several protein alterations at the level of two-dimensional electrophoresis, and at the level of one-dimensional SDS-PAGE have been documented in our laboratory.
  • Changes in protein profiles under stress conditions may relate to the effect of altered cellular condition on gene transcription, translation and post-translational modification of proteins.

Apart from proteomics approach other approaches are also pursued in the laboratory to characterize the drought / submergence stress phenomenon. These include:

  • Morphological observations:
  • Propidiun Iodide Staining
  • Chlorophyll Analysis
  • ADH Isozyme analysis by Native-PAGE

Future Directions

  • In the post-genomic era, “Proteomics” is positioned at the center of the “Functional Genomics” to study gene functions on a genome-wide scale. The unique feature of proteomics is its feasibility to analyze the changes occurring at the protein level that cannot be predicted from genomic sequence. Proteins undergo post-translational modifications, proteolysis, recycling, multi complex formation and sub cellular translocations that are key events to regulate protein functions in cellular processes. Proteomics can eventually reveal all proteins in a cell or tissue at any given time, including those with post-translational changes.
  • The process of identifying the identities of the studied proteins by employing Protein-Microsequencing analysis and / or Mass Spectrophotometry (MS) is in progress.

ii) Genomics
In-charge: Dr. Anita Pande

  • Inter varietal difference in DNA profile of finger millet was studied by utilizing RAPD technique.
  • A partial genomic library of finger millet was characterized by studying the different loci observed through gel electrophoresis.
  • Optimum activity of amylase was studied in finger millet by using enzyme kinetics.
  • Efforts have been initiated to generate specific fingerprints for particular genotype belonging to Eleusine for further identification of the respective genotype.

Inter and intravarietal difference among the genotypes has been studied by using RAPD marker. A phylogenetic tree has been generated on the basis of the findings through RAPD. Further development of SCAR marker can be generated on the basis of the present study.

C) Veterinary

  • The effect of Stresroak (A herbal formulation) on humoral and cell mediated immunity (CMI) was studied in comparison to Leva misole in broiler chicks. For humoral immune status monitoring, chick were vaccinated with Lasota strain of Ranikhet disease virus vaccine on 7th day and booster dose on 28th day for CMI the graft versus hot reaction was employed. Haemaglutination inhibition antibody titre was significantly increased after 1st vaccination in all the three vaccinated in all the three vaccinated group and peaked on 21st day. Then there was significant decrease of HI antibody titre from day 21 to day 28 which increased further in 35th and 42nd day after 2nd vaccination on 28th day. The non vaccinated control group showed zero titre from 14th to 42nd day.
  • RAPD study of blood samples of goats from different zones were carried out to observe the differences if any.
  • Immunomodulatory effect of herbal feed supplement in normal and immunocompromized Broiler chicks has been studied.

On going Projects:

  • Production and supply of Tissue cultured plantlets from KVK units at Dhanbad & Darisai Principle Investigator- Dr.(Mrs.) Madhuparna Banerjee Facility created with the financial assistance of the Ministry of Agriculture, Dept. of Agriculture & Cooperation, Govt. of India.
  • Research on evolving crop varieties tolerant to abiotic stresses using biotechnological techniques  Principle Investigator- Dr. Anita Saxena Funded by Ministry of Agriculture & Sugarcane Development, Govt. of Jharkhand
  • Establishment of Satellite Tissue Culture Unit of College of Biotechnology at ZRS,Dumka Project In charge – Dr.(Mrs.) Madhuparna Banerjee Funded by Department of Agriculture, Ministry of Agricultur e & Sugarcane Dev. Govt. of Jharkhand.
  • All India Network Project On Soil Biodiversity-Biofertilizers. Principal-Investigator: Dr. Himanshu Dubey Funding Agency: Indian Council of Agricultural Research (ICAR)