Effect of Amaranthus Viridis Extract on Growth as Well As Induction of Defense in Chickpea

Effect of Amaranthus Viridis Extract on Growth as Well As Induction of Defense in Chickpea

Sheeba Menon1

1Biology Department,

K K Shah Jarodwala Maninagar Science College, Maninagar, Ahmedabad 380008,

Gujarat, India,

Janki Thakker2

2Department of Biotechnology,

P D Patel institute of Applied Science, Charotar University of Science and Technology,

Changa, Anand-388421, Gujarat, India,

Abstract:- Amaranthus viridis L is sometimes eaten as cooked vegetable, fodder for cattle and green manure; the leaves are diuretic and purgative and are used for treating inflammations, boils and abscesses, gonorrhoea, orchitis and haemorrhoids. Cicer arientinum L (Chick pea) seeds were treated with extract, leachate and dried leaves of the plants. After 2 hours, coated Chick pea seeds were transferred to individual pots and kept for germination. The vegetative parameters were calculated after fifteen days. The study of these vegetative parameters indicated that there is positive plant to plant interaction. Treated Chick pea plants showed significant increase in root- shoot length, number of leaflets, fresh and dry weight as compared to plant grown with distilled water. Change in defense enzymes like Peroxidase, Phenylalanine ammonia lyase and phenolics were noted in extract treated plants. Extract can be used for growth promotion as well as defense induction.

Key Words – Amaranthus viridis, vegetative parameters, Peroxidases, Phenyl ammonia lyase, Poly phenol oxidase

INTRODUCTION

A family of about 65 genera and 900 species, Amaranthaceae are mostly distributed in tropical but also in temperate regions. About 18 genera and over 50 species have been reported from India (1). Amaranthus viridis.L grows annually as an erect, monoecious herb up to 100- 300cm tall, much branched. Leaves are alternate, simple, petiolate, base tapering, margins entire, apex acute or obtuse or retuse or emarginated, Blade glabrous (2). Amaranthus viridis is also been used as a medicinal herb (3).

Chick pea is an important source of cheap protein with high energy and nutritive value (4). Chick pea is the 3rd most important pulse crop (5). Chick pea seeds contain Malic and Oxalic acids which lower blood cholesterol levels (6).

Chemicals with Allelopathic potential are present in a variety of plant tissues (7). Allelopathy effects specific plant processes such as cell division and elongation, action of inherent growth regulators, mineral uptake, photosynthesis, respiration, stomatal opening, protein synthesis and membrane permeability (8).

The aim of the present study was to analyse the effect of the extract of Amaranthus viridis L leaves on germination and growth as well as on the defense enzymes of Chick pea. The Bioassays was carried out to study the effect on Chick pea leaves and compared with control.

MATERIALS AND METHODS

Preparation of Plant extracts

Disease free and fresh Amaranthus viridis L. plant was collected from Changa campus. The leaves were separated and washed under tap water. The fresh leaves were used to prepare 4% fresh leaf extract and leachate. 4% fresh leaf extract was prepared by crushing 4 g fresh leaves in distilled water. For Leachate preparation 250 g of Fresh Amaranthus leaves were soaked for 24 h in 500 ml distilled water. Similarly, 10 g of leaves were oven dried at 55-60º C for 24 h and used to make dried leaf paste with required quantity of water.

POT CULTURE

Chick pea seed were purchased from D Mart store, Ahmedabad. Ten non-sterilized Chick pea seeds each were soaked in 4% Amaranthus fresh leaf extract, Amaranthus fresh leaf leachate and coated with Amaranthus dried leaf paste for 2 h (9). After 2 h the treated Chick pea seeds were shifted to pots and kept for germination under natural conditions. A total of three replications of treated Chick pea seeds were kept undisturbed and watered at regular intervals. Chick pea seeds with distilled water were kept as control. The readings for vegetative parameters of Chick pea plant like germination percentage, root length, shoot length, number of lateral roots and leaflets, fresh and dry weight was taken after 15 days regularly.

ENZYMATIC (DEFENSIVE) ASSAYS

The Enzymatic (Defensive) Peroxidase activity in Chick pea leaves were determined by using method developed by (10) and absorbance was measured at 420nm by UV visible Spectrophotometer (Shimadzu). Phenylalanine ammonia lyase were determined by using method developed by (11) and absorbance was measured at 270nm by UV visible Spectrophotometer (Shimadzu). Poly phenol oxidase were determined by using method developed by (12) and absorbance was measured at 280nm by UV visible Spectrophotometer (Shimadzu).

RESULTS

The germination per cent increase in Chick pea seeds after treatment was 90% with Amaranthus dried leaf paste, 80% with fresh leaf extract and 70% with fresh leaf leachate. Meanwhile the germination per cent was 30% with distilled

water (control) (Table-1). There was a gradual increase of treated Chick pea plant in all the vegetative parameters after 15 days as compared to control (Table-2). Increase in POX,

PAL and PPO was found in treated Chick pea leaves with 4% fresh leaf Amaranthus extract, fresh leaf leachate and dried leaf paste as compared to control (Table-3,4)

germination (%)

germination (%)

100

80

60

40

20

0

I N C H I C K P E A S E E D S A F T E R 1 5 D A Y S

CONTROL AMARANTHUS 4%FRESH LEAF EXTRACT

AMARANTHUS FRESH LEAF LEACHATE(1:2 w/v)

AMARANTHUS DRIED LEAF PASTE(1:10 w/v)

parameters

Graph-1 Germination (%) in Chick pea seeds after 15 days

vegetative parameters of Chick pea seeds after 15 days

parameter readings

parameter readings

60

40

20

0

Root length(cm) Shoot length(cm) Nos of lateral roots Nos of leaflets Fresh wt(gm) Dry wt(gm) vegetative parameters

CONTROL AMARANTHUS 4%FRESH LEAF EXTRACT AMARANTHUS FRESH LEAF LEACHATE (1:2 w/v) AMARANTHUS DRIED LEAF PASTE(1:10 w/v)

50

50

parameter readings

parameter readings

Graph 2 Vegetative parameters of Chick pea seeds after 15 days

Defense Enzymes on Chick pea in Amaranthus Extracts (After 15 Days)

Defense Enzymes on Chick pea in Amaranthus Extracts (After 15 Days)

Peroxidase (POX) (U/ml)

Poly phenol oxidase (PPO)enzyme (U/mg)

Peroxidase (POX) (U/ml)

Poly phenol oxidase (PPO)enzyme (U/mg)

0

0

CONTROL

CONTROL

AMARANTHUS 4%FRESH LEAF AMARANTHUS FRESH LEAF AMARANTHUS DRIED LEAF EXTRACT LEACHATE(1:2 w/v) PASTE(1:10 w/v)

parameters

AMARANTHUS 4%FRESH LEAF AMARANTHUS FRESH LEAF AMARANTHUS DRIED LEAF EXTRACT LEACHATE(1:2 w/v) PASTE(1:10 w/v)

parameters

Graph- 3 Defense enzymes (POX and PPO) on Chick pea in Amaranthus extracts (after 15 days)

Phenyl ammonia lyase (PAL) enzyme (U/ml)

0.15

parameter readings

parameter readings

0.1

0.05

0

CONTROL AMARANTHUS 4%FRESH LEAF EXTRACT AMARANTHUS FRESH LEAF

LEACHATE(1:2 w/v)

AMARANTHUS DRIED LEAF PASTE(1:10

w/v)

parameters

Graph- 4 Defense enzymes (PAL) on Chick pea in Amaranthus extracts (after 15 days)

DISCUSSION

According to study conducted by Jabeen et al., (13) the low amount (10gm) of weed powder increased the amount of organic matter in soil, resulting in the better growth than the control. Under the present study the vegetative parameters of treated Cicer seeds showed better growth than the control.Allelochemicals can stimulate or inhibit plant growth depending on their concentration (14). The presence of defence enzymes like peroxidase, poly phenol oxidase and phenylalanine ammonia lyase was analysed and was more in treated Cicer leaves. The defence enzymes extend mechanical support to the plant. Peroxidases play important role in lignifying cell walls and thus increasing plant resistance (10). Phenyl ammonia lyase is involved in phenolics synthesis including phytoalexins and suberin (11). Poly phenol oxidase helps in the production of brown pigments (12).

CONCLUSION

Based on the analysis carried out Amaranthus viridis fresh and dried leaves have the potential to promote germination. It shows that Amaranthus fresh and dried leaves have both the potential, one is germination and exceptional defence parameters.

ACKNOWLEDGEMENT

First of all, the authors are thankful to the Principal and staff members of K K Shah Jarodwala Maninagar Science college, Ahmedabad for allowing to finish my research work. The authors are also thankful to University Grants Commission (UGC) for awarding Teacher fellowship. The authors are obliged and thankful to Charotar University of Science and Technology for providing necessary facilities

REFERENCES

1. Rahman A H M M and Gulshana M I A, 2014. Taxonomy and medicinal uses on Amaranthaceae family of Rajshahi, Bangladesh. Applied Ecology and Environmental Science, 2(2): 54-59

2. Shah G L, Flora of Gujarat, 1978, Published by S P University,

   Anand

3. Malik K; Nawaz F and Nisar N, 2016. Antibacterial activity of Amaranthus viridis. Bulletin of Environment, Pharma and Life Science, 5(4): 76-80

4. Cokkizgin A, 2012. Botanical characteristics of chick pea genotype (Cicer arientinum L.) under different plant densities in organic farming. Scientific research and essays, 7(4): 498-503

5. Roy I; Biswas P K; Ali M H; Haque M N and Parvin K, 2016. Growth and reproductive behaviour of chick pea (Cicer arientinum L.) as influenced by supplemental application of Nitrogen, irrigation and hormone. Plant Science today,3(1): 30-40

6. Snafi A E A, 2016. The medical importance of Cicer arientinum- A review. International Organization of Scientific Research Journal of pharmacy, 6(3): 29-40

7. Dhole J.A; Lone K.D, Dhole G.A and Bodke S.S, 2013. Allelopathic effect of aqueous and ethanolic extracts of some common weeds on seed health of Triticum aestivum.L (Wheat). International J of current Microbiology and applied sci, 2(6):254- 260

8. Deka S.J and Rao.S, 2015. Allelopathic effects on herbaceous weed plant seeds germination. International Journal of Phytopharmacology, 6(4): 225-228

9. Lalitha P; Shivani K and Rama Rao R, 2012. Parthenium hysterophorus-An economical tool to increase the agriculture productivity. International Journal of Life Science Biotechnology and Pharma research, 1(1): 113-127

10. Jagdale G.B; Kamoun S and Grewal P. S, 2009. Entomopathogenic nematodes induce components of systemic resistance in plants:

    Biochemical and molecular evidence. Biological control, 1-8

11. Sija S L; Potty V P and Santoshlal P S, 2016. Detection of Phenyl ammonia lyase activity in different plant parts of Anacardium occidentale L. International journal of pharma and Bio Sciences. 7(4), 100-104

12. Palmer J K, 2017.Banana polyphenol oxidase: Preparation and properties. American society of plant Biologists.508-513

13. Jabeen N; Ahme M; Shaukat S. S and Iram-Us- Slaam, 2013. Allelopathic effects of Weeds on Wheat (Triticum aestivum L.) germination and growth. Pakistan Journal of Botany, 45(3): 807- 811

14. Babar H B; Tanveer A; Muhammad T; Ahsan A; Ahmad A U H; Muhammad, A N and Muhammad M J, 2009. Allelopathic potential of wild onion (Asphodelus tenuifolius) on the germination and seedling growth of chick pea (Cicer arientinum). Weed Biology and Management,9: 146-151