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Seed Germination and Seedling Growth of Two Vegetables in Responses to Aqueous Extract of Four Herbal Plant Leaves

B. Roy
Department of Agricultural Chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur

B. C. Sarker
Department of Agricultural Chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur

M. R. Ali
Department of Agricultural Chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur

S. R. Das
Department of Biochemisry and Molecular Biology,Hajee Mohammad Danesh Science and Technology University, Dinajpur

M. A. S. Sayed
Department of Biochemisry and Molecular Biology,Hajee Mohammad Danesh Science and Technology University, Dinajpur

Abstract/ Executive Summary:

An experiment was conducted on aqueous extracts of leaves of some herbal plants viz., Beleric myrobalan; bohera (Terminalia belirica), Chebulic myrobalan; horitoki (Terminalia chebula), Sweet basil; tulsi (Ocimum gratissmum) and arjunaa myrobalan; arjun (Terminalia arjuna) for investigating the presence of biologically active substance. The seeds of two vegetable crops such as, swamp cabbage (Impoea aquatica) and okra (Hibiscus esculentus) were tested for germination and seedling growth treating with aqueous extract of four herbal plant leaves. The chemical investigation on effective plant extract was also attempted. The aqueous extract of bohera leaves increased germination of seeds tested and enhanced the growth of shoot length and root length of swamp cabbage (Impoea aquatica) and okra (Hibiscus esculentus) whereas the aqueous extract of horitoki significantly reduced and delayed germination, growth of shoot length and root length of swamp cabbage seeds compared with control; water. The maximum germination was in okra and swamp cabbage seeds within 5 and 4.8 days respectively, treated with the leaves extract of bohera. Shoot lengths were 16.2 and 16.43 cm and root length were 8.74 and 8.91 cm for okra and swamp cabbage seedlings, respectively. The aqueous extract of horitoki showed the lowest rate and late germination at 6 and 5.8 days and minimum shoot lengths were 13.13 and 13.36 cm, and root lengths were 5.91 and 6.08 cm for okra and swamp cabbage seedlings, respectively. The thin layer chromatography (TLC) examination of chloroform extract of bohera leaves showed four distinct compounds at Hexane: Ethyl acetate (3:1, v/v) while horitoki leaves showed three distinct compounds at Hexane: Ethyl acetate (5:1, v/v).

Keywords: Aqueous extract, Germination, Seed, Vegetables

Location: Department of Agricultural Chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh

Start Date: 00-06-2009

End Date: 00-06-2010

Program Area: Seed Technology

Commodity: Okra, Cabbage

Objectives:

1. To investigate the effects of aqueous extract of Bohera (Terminalia bellirica), Horitoki (Terminalia chebula), Arjun (Terminalia arjuna) and Tulsi (Ocimum tenuiflorum) on germination and seedling growth of cabbage and lady’s finger seeds and isolate the different bioactive compounds from the effective aqueous extract.

Methodology:

The experiment was conducted at research laboratory, Department of Agricultural Chemistry, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh, during June 2009- June 2010 for the study of the effects of naturally produced growth substances in four herbal plant leaves viz. beleric myrobalan, chebulic myrobalan, sweet basil, arjuna myrobalan on country bean, lady’s finger, swamp cabbage and yard long bean along with the attempt for chemical investigation on the effective leaf extract. One hundred g (100 gm) of fresh and clean leaves was taken and cut into smaller pieces, it was then blended by using blender and was taken in a 500 ml reagent bottle and 400 ml of water was added to it. It was then kept for 72 hours at room temperature of 29±20C and relative humidity of 85±5% with regular interval of stirring. After 72 hours the aqueous slurry was filtered through Whatmann filter paper No.1 and was taken in another 500 ml bottle. The filtrates of individual plant extract were stored and used for treating the seeds of vegetable crops along with water as a control and other comprehensive study. Laboratory experiments in petridishes were done for swamp cabbage and lady’s finger seeds for the observation of germination percentage; shoot growth and root growth, plant height etc. Clean petridishes with two sheets of filter papers were used. For the investigation of germination percentage, growth and development of vegetable seeds, 15 ml of each aqueous extract was put in each petridish. In control, only distilled water was used and amount of distilled water was also same. Then twenty five (25) seeds of each vegetable crop were kept in each petridish and each treatment was replicated three times. The petridishes were kept in natural diffused light under laboratory conditions at 29±2°c temperature and relative humidity of 85±5% after placing. 5 ml of water was used per day per petridish to keep constant moisture. In control, only water was added if necessary per day per petridish. In this experiment, all subsequent observations were recorded and it was started from 16th June, 2009. After setting the experiment, the germination percentages, shoot length, root length and completion of germination were recorded. Effects of different treatments on morphology of seedlings were also recorded. The collected data were analyzed statistically and the co-efficient of variance and means were compared by using Duncan’s New Multiple Range Test (DMRT). For isolation of crude compounds of the individual herbal plant, 100 g of the leaves of effective herbal plant’s powder was taken in a 2.5 L reagent bottle and 250 ml chloroform was added to the powder and it was then kept 72 h with regular interval of shaking. Then it was filtered by using Whatman filter paper No.1. The extract was collected in 500 ml reagent bottle and 200 ml of chloroform was added to the residue again, the reagent bottle was again kept for next 72 h with also regular interval of shaking. After 72 h it was filtered. The extracting processes were repeated for at least three times. The chloroform extracts of individual plant were combined together. The solvent was evaporated from the extract by using rotary film evaporator under reduced pressure. The different types of plant extracts made by chloroform were Horitoki (18.67 g) and Bohera (18.38 g). After evaporation the semisolid crude was stored in refrigerator at 0?C for further investigation. To the residue after chloroform extract, 200 ml of ethyl-alcohol was added to it. It was kept for 72 h with several interval of shaking. After 72 h it was filtered by using Whatman filter paper No.1. The extract was collected in 500 ml reagent bottle and 200 ml of ethyl-alcohol was added to the residue again, the reagent bottle was again kept for next 72 h with also regular interval of shaking. After 72 h it was then filtered. The process was also repeated for three times. The ethyl-alcohol extracts of individual plant were combined together. The solvent was evaporated by using rotary film evaporator under reduced pressure. The different types of plant extracts by using ethyl-alcohol were Horitoki (18.67 g) and Bohera (18.38 g). Both the extracts were also stored in refrigerator at 0?C for further investigation. Thin Layer Chromatography (TLC) was applied to detect or identify the number of compounds or number of components present in a crude extracts. TLC was prepared for the bohera leaves extract as the most potential and compounds were detected by using the solvent hexane and ethylacetate in the ratio of 5:1 (v/v).

Source of Information: J. Environ. Sci. & Natural Resources, 5(1): 141-150, 2012; ISSN 1999-7361

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Funding Source:

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Achievement/Findings:

The seeds of two vegetable crops such as, swamp cabbage (Impoea aquatica) and okra (Hibiscus esculentus) were tested for germination and seedling growth treating with aqueous extract of four herbal plant leaves. The chemical investigation on effective plant extract was also attempted. The aqueous extract of bohera leaves increased germination of seeds tested and enhanced the growth of shoot length and root length of swamp cabbage (Impoea aquatica) and okra (Hibiscus esculentus) whereas the aqueous extract of horitoki significantly reduced and delayed germination, growth of shoot length and root length of swamp cabbage seeds compared with control; water. The maximum germination was in okra and swamp cabbage seeds within 5 and 4.8 days respectively, treated with the leaves extract of bohera. Shoot lengths were 16.2 and 16.43 cm and root length were 8.74 and 8.91 cm for okra and swamp cabbage seedlings, respectively. The aqueous extract of horitoki showed the lowest rate and late germination at 6 and 5.8 days and minimum shoot lengths were 13.13 and 13.36 cm, and root lengths were 5.91 and 6.08 cm for okra and swamp cabbage seedlings, respectively. The thin layer chromatography (TLC) examination of chloroform extract of bohera leaves showed four distinct compounds at Hexane: Ethyl acetate (3:1, v/v) while horitoki leaves showed three distinct compounds at Hexane: Ethyl acetate (5:1, v/v).

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