Studies on antimicrobial, biochemical and image analysis in Mirabilis jalapa

Master's Thesis, 2013

48 Pages












Mirabilis jalapa Linn. (Family: Nyctaginaceae) is a widely used in conventional medicine in many parts of the world for the treatment of various diseases viz. virus inhibitory activity, anti-tumor activity, etc. Very few reports are available on Architecture of pollen grains, image analysis, Antimicrobial activity, pharmacognostic and phytochemical nature of Mirabilis jalapa Linn. The genus Mirabilis contains 350 species in 34 classifications. The common garden-variety four-o’clock (Mirabilis jalapa) is also known as Marvels of Peru. Four o’clock received its name because of its habit of opening in the late afternoon. It is not actually the time of day that causes the flowers to open, but the drop in temperature. The flowers close the next morning, except on dull, cloudy days.

An interested aspect of this plant is that flowers of different colors can be found simultaneously on the same plant. Additionally, an individual flower can be splashed with different colors. Another interesting point is a color-changing phenomenon. For example, in the yellow variety, as the plant matures, it can display flowers that gradually change to a dark pink color. Similarly white flowers can change to light violet.

Mirabilis jalapa (Plate 1) is said to have been exported from the Peruvian Andes in 1540. Around 1900, Carl Correns used the four o'clock as a model organism for his studies on cytoplasmic inheritance. The flowers are used in food colouring. An edible crimson dye is obtained from the flowers to colour cakes and jellies. In herbal medicine, parts of the plant may be used as a diuretic, purgative, and for vulnerary (wound healing) purposes. The root is believed an aphrodisiac as well as diuretic and purgative. It is used in the treatment of dropsy. Powdered, the seed of some varieties is used as a cosmetic and a dye. The seeds are considered as poisonous can cause vomiting and diarrhea and, in large quantities, death.

Each plant may have hundreds of two-inch, trumpet-shaped blossoms that will last throughout the summer until the first frost. Even on a single plant, these blossoms may be pink, yellow, white, salmon, red, or striped or blotted with any of these colors. Because of their variety of shades, though, they are hard to co-ordinate with other garden flowers and are best used as bedding, border, background, or in containers.

In mining, geology, and quarry production, it is well known that the properties of aggregates, such as size and shape, are very important information for particle characterization and the optimization of production. For the last fifteen years, image analysis techniques have been used for aggregate particle measurement, which increases speed and accuracy of analysis. There are a number of methods for measuring aggregate particle size and shape in image analysis, the stability of measurement methods is very important.

Palynology is the study of plant pollen, spores and certain microscopic plankton organisms (collectively termed palynomorphs) in both living and fossil form. Botanists use living pollen and spores (actuopalynology) in the study of plant relationships and evolution, while geologists (palynologists) may use fossil pollen and spores (paleopalynology) to study past environments, stratigraphy (the analysis of strata or layered rock) historical geology and paleontology.

Melissopalynology is the study of pollen in honey, with the purpose of identifying the source plants used by bees in the production of honey. This is important to honey producers because honey produced by pollen and nectar from certain plants as mesquite, buckwheat, tupelo or citrus trees demand a higher price on the market than that produced by other plant sources. Some plants may produce nectar and pollen that is harmful to human health. A careful monitoring of the pollen types found in honey may identify these toxic sources and the honey produced may be kept out of the commercial market.

Pollen grains are showing two nuclei. The smaller (generative) nucleus forms two sperm cells. Endoplasmic reticulum threads present throughout the pollen grain and several mitochondria and plastids containing starch.


Fig: 1. Mother Plants

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Fig: 2. Mother Plants with Scale Reading

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Mirabilis jalapa Linn. belongs to the family Nyctaginaceae and is a large herbaceous plant grown in gardens throughout India. The present project work entitled “Studies on antimicrobial, biochemical and image analysis in Mirabilis jalapa varities” contains following objectives:

1. Staining of pollen grain and observation of meiotic stages.
2. Phytochemical studies of 3 different colored plant varieties of M.jalapa by TLC
3. Comparative Evaluation of Antimicrobial Activities of plant leaf Extract (3 different colored varieties) of Mirabilis jalapa.
4. Image analysis of data got by experimentation using software.


Nature has been a source of medicinal agents for thousands of years and an impressive number of modern drugs have been isolated from natural sources, many based on their use in traditional medicine. Various medicinal plants have been used for years in daily life to treat disease all over the world. They have been used as a source of medicine. The widespread use of herbal remedies and healthcare preparations, such as those described in ancient texts like the Vedas and the Bible, has been traced to the occurrence of natural products with medicinal properties. In fact, plants produce a diverse range of bioactive molecules, making them a rich source of different types of medicines. Higher plants, as sources of medicinal compounds, have continued to play a dominant role in the maintenance of human health since ancient

times [1]. Over 50% of all modern clinical drugs are of natural product origin [2] and natural

products play an important role in drug development programs in the pharmaceutical industry [3].

There has been a revival of interest in herbal medicines. This is due to increased awareness of the limited ability of synthetic pharmaceutical products to control major diseases and the need to discover new molecular structures as lead compounds from the plant kingdom. Plants are the basic source of knowledge of modern medicine. The basic molecular and active structures for synthetic fields are provided by rich natural sources. This burgeoning worldwide interest in medicinal plants reflects recognition of the validity of many traditional claims regarding the value of natural products in health care.

The relatively lower incidence of adverse reactions to plant preparations compared to modern conventional pharmaceuticals, coupled with their reduced cost, is encouraging both the consuming public and national health care institutions to consider plant medicines as alternatives to synthetic drugs. Plants with possible antimicrobial activity should be tested against an appropriate microbial model to confirm the activity and to ascertain the parameters associated with it. The effects of plant extracts on bacteria have been studied by a very large

number of researchers in different parts of the world [4] [5] [6]. Much work has been done on

ethnomedicinal plants in India [7] [8] [9]. Interest in a large number of traditional natural products has increased [10]. It has been suggested that aqueous and ethanolic extracts from plants used in allopathic medicine are potential sources of antiviral, antitumoral and antimicrobial agents [11], [12]. The selection of crude plant extracts for screening programs has the potential of being more successful in initial steps than the screening of pure compounds isolated from natural products [13].

This plant is 50-100 cm high. It has antifungal, antimicrobial, antiviral, antispasmodic, antibacterial, diuretic, carminative, cathartic, hydragogues, purgative, stomachic, tonic and vermifuge properties. [14] This plant contains alanine, alphaamyrins, arabinose, beta-amyrins,

campesterol, daucosterol and dopamine [15], and is used to treat conjunctivitis, edema, fungal

infections, inflammation, pains and swellings.

Mirabilis jalapa L. (Nyctaginaceae) is a tropical American herb that is commonly cultivated in North America where it is perennial in the south and warm west and annual in the north. In Mirabilis jalapa pollen performance was influenced by the number of competing pollen

grains or pollen tubes, but was not influenced by potential genetic differences with load diversity [16].

B P Cammue et al., has isolated from seeds of Mirabilis jalapa L. two antimicrobial peptides, designated Mj-AMP1 and Mj-AMP2, respectively. These peptides were also active on two tested Gram-positive bacteria but were apparently nontoxic for Gram-negative bacteria and cultured human cells [17].

J Kataoka et al., 1991, cloned a cDNA for Mirabilis antiviral protein (MAP), a ribosome- inactivating protein (RIP), which inhibits the mechanical transmission of plant virus and the in vitro protein synthesis of both prokaryotes and eukaryotes [18]. A potent antiviral activity was found in extracts from a yellow flower cultivar of Mirabilis jalapa L. (Nyctaginaceae) in root, leaf, and stem tissues and in in vitro cultured cells [19],[20].

Extracts of Mirabilis jalapa (Nyctaginaceae), containing a ribosome inactivating protein (RIP) called Mirabilis antiviral protein (MAP), were tested against infection by potato virus X, potato virus Y, potato leaf roll virus, and potato spindle tuber viroid. Several plants, such as Pelargonium hortorum, Chenopodium album, C. amaranticolor, Capsicum frutescens, Azadirachta indica, Vitis vinifera, and Rosa banktia, possess antiviral factors. Plant-derived antiviral compounds are active against plant, animal, and human viruses. Plant antiviral compounds are grouped as furocoumarins, alkaloids, terpenoids, lignins, and specific proteins. Among plant-derived antiviral proteins, a group called ribosome-inactivating

proteins (RIPs), which are widely distributed in higher plants, hold promise for agricultural and pharmaceutical applications [21].


Study species

Mirabilis jalapa has tubular flowers are fragrant and vary in color among plants. The self- compatible, perfect flowers each have 5–6 stamens and a single-ovulate ovary. An individual flower opens for one night in the early evening, the exact time depending on temperature and relative humidity, and closes early the next morning. An individual plant produces between 25 and 75 flowers in one flowering season.

Growth form: Annual or perennial herb.

Size: 0.5 - 2 m tall.

Root: a swollen and somewhat tuberous taproot.

Stems: usually several, erect to slightly decumbent, branching, light or bright green, sometimes with a yellow or pink hue, mostly hairless, but sometimes hairy near the base or even glandular-hairy further up; if hairy, often in two lines.

Leaves: about midstem and above, opposite, on 1 - 7 cm long stalks, somewhat elongate triangular to egg-shaped or lance-shaped, 4 - 14 cm long, 2 - 9 cm wide, with blunt or indented bases, and non-toothed edges.

Inflorescence: of several, terminal or axillary, compact clusters with one to fifteen flowers on short, 0.5 - 5 mm long stalks, and each cluster subtended by a pair of 2 - 17 mm long leaf- like bracts. Each flower sits atop a green, 0.5 - 1.5 cm tall, bell-shaped cup (involucre) formed by five fused bracts with triangular tips.

Flowers: pink or yellow or orang, sometimes white or striped, usually hairless, 3 - 5 cm long, radially symmetric, funnel-shaped with a long, narrow tube, and five, abruptly flared lobes.

Sepals: showy, brightly colored, not green, and mimicking petals. The five sepals are fused for most of their length, constricted above the ovary into a long, narrow tube, then separated into five, abruptly flared lobes.

Petals: none.

Stamens: five, long, and extending beyond the sepal tube.

Pistil: with one, single-chambered, superior, somewhat globular ovary; one, long, threadlike style, which extends beyond the stamens; and a rounded, head-like stigma.

Fruit: a hard, dark brown or nearly black, 0.7 - 1.1 cm long, broadly ellipsoid to inversely egg-shaped, one-seeded, nut-like achene, which is tightly enclosed by the remnant bract cup (involucre).


[1]. Farombi EO. African indigenous plants with chemotherapeutic potentials and biotechnological approach to the production of bioactive prophylactic agents. African J Biotech 2: 662-671, 2003.

[2]. Stuffness M, Douros J. Current status of the NCI plant and animal product program. J Nat Prod 45: 1-14, 1982.

[3]. Baker JT, Borris RP, Carte B, Cordell GA, Soejarto DD, Cragg GM, Gupta MP, Iwu MM, Madulid DR, Tyler VE. Natural product drug discovery and development: New perspective on international collaboration. J. Nat. Prod. 58: 1325-1357, 1995.

[4]. Reddy PS, Jamil K, Madhusudhan P. Antibacterial activity of isolates from Piper longum and Taxus baccata. Pharmaceutical Biol 39: 236-238, 2001.

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[7]. Maheshwari JK, Singh KK, Saha S. Ethnobotany of tribals of Mirzapur District, Uttar pradesh, Economic Botany Information Service, NBRI, Lucknow. 1986.

[8]. Rai MK. Ethnomedicinal studies of Chhindwara district (M.P.). I. Plants used in stomach disorders. Indian Medicine. 1: 1-5, 1989.

[9]. Negi KS, Tiwari JK, Gaur RD, Pant, K.C. Notes on ethnobotany of five districts of Garhwal Himalaya, Uttar pradesh, India. Ethnobotany 5: 73-81, 1993.

[10]. Taylor RSL, Manandhar NP, Hudson JB, Towers, G. H. N. Antiviral activities of Nepalese medicinal plants. J Ethnopharmacol 52:157-163, 1996.

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[12]. Vlietinck AJ, Van Hoof L, Totté J. Screening of hundred Rwandese medicinal plants for antimicrobial and antiviral properties. J. Ethnopaharmacol 46: 31-47, 1995

[13]. Kusumoto IT, Nakabayashi T, Kida H. Screening of various plant extracts used in ayurvedic medicine for inhibitory effects on human immunodeficiency virus type 1 (HIV-1) protease. Phytotherapy Res 9: 180-184, 1995.

[14]. Dimayuga RE. Antimicrobial activity of medicinal plants from Baja California Sur/Mexico. Pharmaceutical Biol 36: 33-43, 1998.

[15]. Yang SW. Three new phenolic compounds from a manipulated plant cell culture, Mirabilis jalapa. J Nat Prod 64: 313-17, 2001.

[16]. Richard A. Niesenbaum, The effects of pollen load size and donor diversity on pollen performance, selective abortion, and progeny vigor in Mirabilis jalapa (Nyctaginaceae), American Journal of Botany. 86:261-268, 1999.

[17]. B P Cammue, M F De Bolle, F R Terras, P Proost, J Van Damme, S B Rees, J Vanderleyden and W F Broekaert. Isolation and characterization of a novel class of plant antimicrobial peptides form Mirabilis jalapa L. seeds. The Journal of Biological Chemistry 267: 2228-2233, 1992.

[18]. J Kataoka, N Habuka, M Furuno, M Miyano, Y Takanami, A Koiwai. DNA sequence of Mirabilis antiviral protein (MAP), a ribosome-inactivating protein with an antiviral property, from mirabilis jalapa L. and its expression in Escherichia coli. The Journal of Biological Chemistry 266: 8426-8430, 1991.

[19]. Kubo S., Ikeda T., Imaizumi S., Takanami Y., Mikami Y. A potent plant virus inhibitor found inMirabilis jalapa L. Ann. Phytopathol. Soc. Jpn. 56:481–487, 1990.

[20]. Ikeda T., Takanami Y., Imaizumi S., Matsumoto T., Mikami Y., Kubo S. Formation of anti-plant viral protein by Mirabilis jalapa L. cells in suspension culture. Plant Cell Rep. 6:216–218, 1987.

[21]. Jorge M. Vivanco, Maddalena Querci, Luis F. Salazar, Antiviral and Antiviroid Activity of MAP-Containing Extracts from Mirabilis jalapa Roots, Plant Disease, 83(12): Pages 1116-1121, 1999.

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Studies on antimicrobial, biochemical and image analysis in Mirabilis jalapa
Plant studies
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Dr. Kaladhar DSVGK (Author), 2013, Studies on antimicrobial, biochemical and image analysis in Mirabilis jalapa, Munich, GRIN Verlag,


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