VCSG Uttarakhand University of Horticulture and Forestry

College of Horticulture, Bharsar

MAP-509: Master’s Seminar TOPIC: Role of elicitors in production of plant secondary

metabolites under in-vitro condition Presented By: Yashwant Singh Tariyal 13162 Department of plantation crops, Spices, Medicinal and aromatic plants Yashutariyal50@gmail.com

Secondary metabolites o They are biosynthetically derived from

primary metabolites. o Produces product that aid in the growth

and development of plant but not required for the plant to survive.

o They facilitate the primary metabolism in plant.

o They are the major component of plant defense mechanism against herbivores, pest and pathogen.

Classification of Plant secondary

metabolites

oNitrogenous secondary metabolites. Alkaloids Non-protein amino acids (NPAAs) Amines Cyanogenic glycoside Glucosinolates Alkamides Lectins, peptides, polypeptide

Kumar et al. 2014

o Non-Nitrogenous secondary metabolites Terpenes including iridoids Steroids, saponins Flavonoids, tannins Phenylpropanoids, lignin, coumarins,

lignans Polyacetylenes, fatty acid, waxes Anthraquinones and others

polyketides Carbohydrates, organic acids

Kumar et al. 2014

Economical importance of plant

secondary metabolites

o Used in preparation of various dyes. o Used in medicine industry. o Used for making agrochemicals

insecticides and raw material for industries.

o Used as perfume and cosmetics. o Used as food colour and flavoring

agent.

Advantages of in-vitro production of

secondary metabolites

oHelpful in conservation of RET plant species. oCompounds can be produced under controlled

conditions as per market demand. oCulture system are independent of environmental

factors, seasonal variations, pest and microbial diseases and geographical constraints.

oCell growth can be controlled to facilitate improved product formation and consistent quality of the product.

oIt is very useful in the plants which are difficult and expensive to grow in field.

oThe production time is less and labour cost is minimal. oIn vitro production of natural material is cheaper is

compared to synthetic production.

Disadvantages of in-vitro production of

secondary metabolites

oIn general, in-vitro production of secondary metabolites is lower when compared to the intact plant.

oMany time secondary metabolites are produced in differentiated tissue/organ . In such cases the culture cell which are non-differentiated produce less quantity.

oCultured cells are genetically unstable and may undergo mutation this may cause in reduced production, as culture ages. oVigorous stirring is necessary to prevent aggregation

of cell this may often damage cells. oStrict aseptic conditions have to be maintained

during culture technique.

Elicitors o Elicitors are substances which induce

physiological changes in the plant cells. o Plants cells respond to these stressors by

activating an array of mechanisms, similar to the defense responses to pathogen infections or environmental factors.

o They affects the metabolism. o Enhances the synthesis of phytochemical/

plant secondary metabolites.

Moreno et al. (2014)

Elicitors

Plant Tissue Culture

Investigation of Plant Defense Mechanisms

Investigation of Enzymology of

Secondary Metabolism

Investigation of Regulation of

Secondary Metabolism

Increase Yield of Target Substance

Utilization of elicitation of plant tissue cultures in various areas of research.

Veersham, 2004

Elicitors

Physical Chemical

Light, Temperature Abiotic Biotic

Complex Defined Composition

Metal Ions, Oxalates

Yeast Cells Wall, Mycelia Cell Wall, Fungal Spores Carbohydrates, Proteins,

lipids, Glycoprotein, Volatiles

Radman et al. (2003)

Classification of elicitors

Case Study

Table:1 :-Effect of β–phenylalanine on growth of tissue and production of quercetin on liquid culture of Citrullus colocynthis.

Meena et al. 2014

Table:2:- Effect of elicitors on the secondary metabolites content in leaf derived callus cultures of Centella asiatica.

Usha et al. 2015

Elicitor (g L-1)

Growth Index

Hyoscyamine (mg/g dw)

Scopolamine(mg/g dw)

Aspergillus niger

0.0 5.79 1.39±0.004 0.069±0.001

0.1 6.32 1.46±0.003 0.077±0.0004

0.5 5.61 1.75±0.005 0.082±0.0005

1.0 5.07 1.77±0.005 0.087±0.001

1.5 4.34 1.63±0.004 0.081±0.001

Table: 3 (a)-:Effect of biotic elicitor on the alkaloid content in the callus and root culture of Datura metel L.

Barmukh et al. 2009

The values represent the mean ± SE of three independent experiment each performed on 21 replicates.

Elicitor (g L-1)

Growth Index

Hyoscyamine (mg/g dw)

Scopolamine(mg/g dw)

Alternaria sps.

0.0 5.79 1.39±0.004 0.069±0.001

0.1 5.96 1.43±0.006 0.074±0.001

0.5 5.43 1.59±0.004 0.079±0.0008

1.0 4.89 1.64±0.003 0.084±0.001

1.5 4.24 1.52±0.004 0.078±0.001

Table: 3 (b)-:Effect of biotic elicitor on the alkaloid content in the callus and root culture of Datura metel L.

The values represent the mean ± SE of three independent experiment each performed on 21 replicates.

Barmukh et al. 2009

Elicitor (g L-1)

Growth Index

Hyoscyamine (mg/g dw)

Scopolamine(mg/g dw)

Fusarium monoliforme

0.0 5.79 1.39±0.004 0.069±0.001

0.1 6.14 1.44±0.003 0.072±0.014

0.5 5.61 1.63±0.005 0.075±0.0007

1.0 5.25 1.66±0.004 0.081±0.0006

1.5 4.92 1.55±0.006 0.072±0.0006

Table: 3 (c)-:Effect of biotic elicitor on the alkaloid content in the callus and root culture of Datura metel L.

The values represent the mean ± SE of three independent experiment each performed on 21 replicates.

Barmukh et al. 2009

O. basilicum L.: Elicitation 1 - MeJA 25μM, 12 h, Elicitation 2 - Chitosan 200 mg/L, 12 h, Elicitation 3 - MeJA 25μM + chitosan 200 mg/L, 8 h

O. sanctum L.: Elicitation 1 - MeJA 25μM, 48 h, Elicitation 2 - Chitosan 50 mg/L, 24 h, Elicitation 3 - MeJA 100 μM + chitosan 200 mg/L, 4 h

O. gratissimum L.: Elicitation 1 - MeJA 50 μM, 8 h, Elicitation 2 - Chitosan 50 mg/L, 24 h, Elicitation 3 - MeJA 25 μM + chitosan 100 mg/L, 24 h

Figure: 1:-Quantification of total alkaloid content in Ocimum species

Mathew and Sankar 2014

Figure: 2:-Quantification of total terpenoid content in Ocimum species.

O. basilicum L.: Elicitation 1 - MeJA 25μM, 12 h, Elicitation 2 - Chitosan 200 mg/L, 12 h, Elicitation 3 - MeJA 25μM + chitosan 200 mg/L, 8 h .

O. sanctum L.: Elicitation 1 - MeJA 25μM, 48 h, Elicitation 2 - Chitosan 50 mg/L, 24 h, Elicitation 3 - MeJA 100 μM + chitosan 200 mg/L, 4 h .

O. gratissimum L.: Elicitation 1 - MeJA 50 μM, 8 h, Elicitation 2 - Chitosan 50 mg/L, 24 h, Elicitation 3 - MeJA 25 μM + chitosan 100 mg/L, 24 h .

Mathew and Sankar 2014

Conclusion

o According to WHO report about 70-80% 0f world population depends on the herbal based drugs.

o The demand for the plant and their purified product is continuously increasing day by day. This puts a lots pressure on the agricultural land for the production of medicinal plants among which some of the plants are difficult or expensive for cultivation due to various biotic abiotic and economical factors.

o The plant tissue culture techniques are the best alternatives for the production of economically important secondary metabolites and elicitors plays a key role in increasing the in-vitro production of these compounds with consistent quality as per market demand.

Thank you!!!!