Research Article

Morpho-Anatomical Alterations in Asthama Plant (Euphorbia hirta L.) in Various Regions of Faisalabad, Pakistan

Shahid Ali Khan1*, Farooq Ahmad1, Muhammad Zubair Akram2, Yan Tongyu3, Fatima Urooj1, Kamran Ahmad1, Saeed Ullah4, Sharmeen Zulfiqar5, Tahira Batool6, Aqsa Sarwar1, Yaqoob Sultan7 and Samreen Nazeer8*

1Department of Botany, University of Agriculture, Faisalabad, 38000, Punjab, Pakistan; 2School of Agricultural, Forest, Food and Environmental Sciences, University of Basilicata, Potenza 85100, Italy; 3Department of Life Sciences, Northwest Agriculture and Forestry University, China; 4Shandong Provincial Key Laboratory of Plant Stress, Shandong Normal University, Jinan, China; 5Department of Chemistry, Superior University Lahore, 54000, Punjab Pakistan; 6Department of Botany, Bahauddin Zakariya University, Multan, 60000, Punjab, Pakistan; 7Department of Grass Breeding, Lithunian Reseach Center for Agriculture and Forestry, 58344, Kedainiur, Lithunia; 8Department of Food and Drug, University of Parma, 43124 Parma, Italy.

Received | July 11, 2024; Accepted | August 30, 2024; Published | October 04, 2024

*Correspondence | Shahid Ali Khan and Samreen Nazeer, Department of Botany, University of Agriculture, Faisalabad, 38000, Punjab, Pakistan; Department of Food and Drug, University of Parma, 43124 Parma, Italy; Email: [email protected], [email protected]

Citation | Khan, S.A., F. Ahmad, M.Z. Akram, Y. Tongyu, F. Urooj, K. Ahmad, S. Ullah, S. Zulfiqar, T. Batool, A. Sarwar, Y. Sultan and S. Nazeer. 2024. Morpho-anatomical alterations in asthama plant (Euphorbia hirta L.) in various regions of Faisalabad, Pakistan. Sarhad Journal of Agriculture, 40(4): 1144-1152.

DOI | https://dx.doi.org/10.17582/journal.sja/2024/40.4.1144.1152

Keywords | Euphorbia hitra, Morphological and Anatomical characteristics, Faisalabad

Copyright: 2024 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Euphorbia hitra is belong to Euphorbiaceae, family and most commonly referred to as the spurge family and ranking as the sixth biggest family of flowering plants. It consists of 334 genera that are organized into 52 tribes and 5 subfamilies (Bueno et al., 2023). In terms of Angiosperms, Rubber plant (Hevea), castor oil (Ricinus communis), cassava and tapioca (Manihot), and tung oil (Aleurites fordi) are commercially valuable members of this family, which ranks among the top 25 economically significant plant families (Alyas et al., 2020). Furthermore, it is also at the forefront of the ethno-medicinal sector. Ancient Chinese medicine utilized approximately 33 species, belonging to 17 genera, of the Euphorbiaceae family for herbal medicinal purposes (Laldingliani et al., 2022). Pakistan is well-known around the world for its Ayurvedic treatments (Mussarat et al., 2021). Euphorbia hirta has traditionally been used to treat feminine diseases, respiratory ailments (cough, coryza, bronchitis, and asthma), worm infestations in children, dysentery, jaundice, acne, gonorrhea, digestive issues, and tumors (Amtaghri et al., 2022). The castor oil plant (Ricinus communis), the Barbados nut (Jatropha curcas), the cassava (Manihot esculenta), and the Pará rubber tree (Hevea brasiliensis) are all examples of species belonging to the Euphorbiaceae family that have significant economic importance (Chan et al., 2010).

The Euphorbia hirta plants are widespread in the hotter regions of Pakistan and Australia, and it is frequently discovered in waste spots along the sides of roadways (Shaheen et al., 2018). Euphorbia exhibits a considerable number of species in regions outside the tropics, including the Mediterranean Basin, the Middle East, South Africa, and the southern United States (Pahlevani et al., 2020; Riina et al., 2013) and low-growing weeds with a prostrate habit (McDonnell et al., 2018; Yang et al., 2018). Primarily found in tropical regions, the majority of these species are concentrated in the Indo-Malayan area and tropical America (Ali et al., 2021).

In terms of economic significance, members of the Euphorbiaceae family are of tremendous significance to our nation (Ernst et al., 2015). Throughout the many regions of Faisalabad, which is a popular city in Pakistan, they have been discovered to be valuable in a variety of ways, including the production of lumber, the provision of food and fodder, the development of hedges, the landscape and beautification, and the formation of hedges (Alam et al., 2017). It is possible to find them in the regions of Jaranwal, Manawala, Gatwala, Samundri, and Sahianwala, as well as throughout the entirety of the southern region of Pakistan (Nawaz et al., 2021). The range of this species can be found in many different regions of Pakistan and West Africa. Because of the economic significance of these Euphorbiaceae family in Pakistan (Ullah et al., 2014). It is necessary to conduct an investigation using anatomical lines of evidence. The primary objective of this investigation is to characterize, identify, and validate the classification that is already in place for these taxa.

In order to attain favorable taxonomic decision making, plant anatomical investigations, which are considered a systematic line of evidence in Plant Taxonomy, are utilized in conjunction with other systematic lines (Rouhan and Gaudeul, 2021). The rationale for this is that anatomical characteristics are both stable and maintained, and as a result, they are utilized as taxonomic characteristics in the context of plant systematic and taxonomic research (Ding et al., 2024). Anatomical characteristics, such as root anatomy, trichome anatomy, stem anatomy, stomata and epidermal anatomy, wood anatomy, nodal anatomy, sclereids and fibers, cambium anatomy, and leaf anatomy, must be utilized in biosystematics and taxonomic research in order to identify plants, determine genetic relationships, and resolve taxonomic disputes. Our experiment will help the researchers to identify the various physiological, biochemical and anatomical attributes of Asthama plant for better understanding the mechanism involved in altering these features of plants.

Materials and Methods

Collection and identification of plant samples

Euphorbia, a native species, has been collected from various biologically distinct regions. A selection of plant species was made from eight distinct geographical areas. Collection was conducted based on the habitat and ecology of the selected species. Euphorbia hirta was obtain from several ecological regions in Faisalabad, such as Jaranwal, Manawala, Gatwala, Samundri, Sahianwala, Old Botanical Garden, New Botanical Garden, and Khurianwal.

Section cutting

The roots, stems, and leaves were finely sectioned into little pieces using an aluminum blade. To prevent desiccation, the sections were kept in petri dishes containing water. The slides were scrutinized using a light microscope to ascertain their suitability for staining. To achieve desirable outcomes, the most suitable components that may be stained were transformed into ceramic plates.

Staining

The highest quality sections were preserved in a solution containing 30% alcohol for a duration of 12 to 15 minutes. Subsequently, the piece was immersed in a solution consisting of 50% and 70% alcohol for 15 minutes. The dried alcohol sections were subjected to a brief exposure of approximately 15 minutes to a small amount of safranin by following (Ruzin, 1999). The samples were pigmented or dyed. The additional strain was subsequently eliminated from the sections by introducing them to 95% alcohol. These portions had a duration of approximately five minutes. A single drop of quick green was applied to the sections, and because of its reactivity, it was promptly eliminated. Fast green was fully eradicated using 100% ethyl alcohol after being administered three or four times. Administer a small amount of xylene solution at the conclusion.

Preparation of slides

Class images were taken to prepare slides. The slides were coated with Canada balsam using a needle to make them permanent. The parts were placed on slides coated with Canada balsam, then they were shifted and covered with a cover slip. The slides were anatomically analyzed using a light microscope. The photos were designed to possess intricate details.

Morphological and anatomical characteristics

By meter ruler, shoot and root lengths were estimated. Leaf count was calculated via counting. Leaf area was determined using fast and accurate method. An image captured with a digital camera was saved in JPEG format. The RGB image was then converted into the CIELAB color space. The color-transformed image was segmented using a threshold technique. The algorithm’s accuracy exceeds 99%, as demonstrated by comparing its results to those obtained using the grid count method.

Electrical balance measured shoot, fresh weight, and root fresh weight. Electrical balance measured shoot, root, and dry weight. Root dries and Such as root anatomy root diameter epidermis thickness Sclererenchyma thickness epidermal cell area cortex thickness cortex cell area endoderm thickness. The vascular bundle including stem anatomy stem radius epidermis thickness epidermal cell area vascular bundle numbers vascular bundles phloem area metaxylem leaves anatomy midrib thickness lamina thickness mesophyll thickness vascular bundles many vascular bundles phloem area metaxylem.

Statistical analysis

Data underwent one-way ANOVA (Analysis of Variance) to examine the morpho-anatomical characteristics of the Euphorbia helioscopia species by using Statistix 8.1..

Results and Discussion

The morphological characteristics of Euphorbia species collected from various ecological regions in Faisalabad, including Gatwala, New Botanical Garden, Old Botanical Garden, Sahianwala, Jaranwala, Khurianwala, Samundri, and Manawal ecotypes, were analyzed using Analysis of Variance (ANOVA).

Morphological attributes

The results of ANOVA showed significant difference in Euphorbia hirta root length. E. hirta from New Botanical Garden had the longest roots and Khurianwala the shortest. Euphorbia hirta shoot length varied greatly, with Samundari showing the longest shoot length. Euphorbia hirta shoots were the shortest in Sahianwala (Figure 1C). The graph for E. hirta root fresh weight changed significantly with Sahianwala indicated the highest. Manawala has the lowest Euphorbia hirta root fresh weight. Euphorbia hirta shoot fresh weight varied greatly, with Samundari showing the maximum. Euphorbia hirta shoot fresh weight was lowest in Sahianwala and Gatwala (Figure 2A). Analysis of variance (ANOVA) showed that Samundari has the most Euphorbia hirta leaves. New Garden has the fewest Euphorbia hirta leaves. The results for E. hirta root dry weight varied significantly indicating that Samundari has the highest root dry weight. Sahianwala has the lowest Euphorbia hirta root dry weight (Figure 2C). Likewise Euphorbia hirta shoot dry weight varied greatly, with Samundari showing the maximum. Sahianwala had the lowest Euphorbia hirta shoot dry weight (Figure 2D). The highest leaf area of Euphorbia hirta was found in the New Garden using ANOVA. A minimal Euphorbia hirta leaf area was found in Manawala.

 

 

Anatomical parameters

According to graphical data the largest root radius of Euphorbia hirta root samples showed non-significant variations. Sahianwala papulations had the smallest root radius (Figure 3A). ANOVA results found that Euphorbia hirta root samples from Manawala had the highest epidermal thickness. Epidermal thickness was lowest in Khurianwala E. hirta. The greatest epidermal cell area of Euphorbia hirta root samples were found in Khurianwala following Analysis of Variance (ANOVA) and Sahianwala papulations have the fewest epidermal cells (Figure 5B). The Euphorbia hirta root samples from Manawala had the highest sclerenchymatous thickness; ANOVA showed significant difference. Shianwala and Gatwala papulations have the thinnest sclerenchyma (Figure 3C). Euphorbia hirta root samples from Old Botanical Garden had the highest endodermis thickness; ANOVA indicated no significant variation. Endodermal thickness was lowest in New Garden ecotypes. ANOVA showed that Euphorbia hirta root samples from Khurianwala had the highest cortical cell area. New and Old Botanical Garden papulations have the smallest cortical cell area. ANOVA showed that Euphorbia hirta root samples from New Garden and Samundri had the highest vascular bundle area. Jaranwala papulations had the smallest vascular bundle. ANOVA from New Garden indicated the greatest variation in Euphorbia hirta root cortical area thickness. Manawala papulations had the thinnest cortical area.

 

Results for Euphorbia hirta stem radius samples had significant variantions in different regions. ANOVA showed no significant differences. Sahianwla had the smallest stem radius. Euphorbia hirta root samples from Manawala had the highest epidermal thickness (Figure 3B). Minimal epidermal thickness was found in Sahianwla and Gatwala. Vascular bundle counts of Euphorbia hirta root samples from Khurianwala were highest. Gatwala had the fewest vascular bundles (Figure 4). ANOVA revealed that Euphorbia hirta root samples from Manawala had the most vascular bundle area. Jaranwala papulations contained the smallest vascular bundle (Figure 5). ANOVA revealed that Euphorbia hirta root samples from Sahianwala had

 

 

the highest phloem area. Jaranwala papulations contained the least phloem. ANOVA revealed that Euphorbia hirta root samples from Sahianwala had the largest metaxylem area, while new garden papulations had the smallest metaxylem (Figure 6). Graphical data showed that Euphorbia hirta leaf samples from new garden had the highest lamina thickness. Samundri and Manawala populations have thin lamina. Euphorbia hirta leaf samples from Gatwala had the highest midrib thickness by analysis of variance (ANOVA) (Figure 7). Midrib thickness was lowest in old botanical garden papulations. ANOVA indicated the maximal mesophyll thickness of Euphorbia hirta leaf samples from Gatwala without significant variation. Mesophyll thickness was lowest in old botanical garden ecotypes. Khurianwala Euphorbia hirta leaf samples had the highest vascular bundle counts. The Gatwala populations had the fewest vascular bundles. Jaranwala Euphorbia hirta leaf samples had the highest vascular bundle area according to Analysis of Variance (ANOVA). The old botanical garden ecotypes had the smallest vascular bundles. Euphorbia hirta leaf samples from Gatwala had the highest phloem area by Analysis of Variance (ANOVA). The phloem area was lowest in Manawala papulations. ANOVA showed that Euphorbia hirta leaf samples from Samundri and New Garden had the highest metaxylem area. Metaxylem area was lowest in old botanical garden and Sahianwala papulations.

 

Pakistan’s landscape is characterized by environmental variety, which governs species diversity through habitat, biotic interactions, productivity, and water resources (Khan et al., 2023). Plants’ genetic diversity makes them adaptable to many environments. Plants adapt to different environments through unique anatomical modifications like epidermal thickness, cell area, endodermis thickness in roots, sclerenchymatous thickness, midrib thickness, lamina thickness, vascular bundle numbers, area, metaxylem and phloem areas of leaves, deep or vast vascular bundles, and cell area (Iqbal et al., 2023). Several environmental conditions caused the entire Euphorbia hirta population to change their anatomy and morphology (Qasem and Foy, 2001). Thus, adjustment can help survive the changing environment. Many morphological characteristics of angiosperms’ vegetative portions are studied to answer taxonomy issues, such as, these features include epidermal thickness, cortical cell area, phloem area, xylem area, number of vascular bundles, pith area, and root and stem radius (Carlquist, 2012). These traits help plants live in varied environments. Leaf midrib and lamina thickness are typical (Liu et al., 2020). Trichrome, stomatal densities, phloem and xylem areas, and the number of adaxial and abaxial stomata assist Euphorbia hirta live in halophytic conditions (El-Hamid and El-Bous, 2019).

 

Euphorbia hirta species has a crucial environmental gradient reaction that may help them avoid, tolerate, or survive harsh environmental conditions (Salama et al., 2019). In the dry habitat, E. hirta showed a thick epidermis, sclerification of tissue, osmolyte accumulation, expanded phloem and metaxylem vessels, oil gland formation, photosynthetic activity, and improved antioxidant formation (Iqbal et al., 2022). However, saline environmental conditions significantly improved biomass productivity and growth parameters (Egamberdieva et al., 2019). The increased epidermal thickness, inner and outer phloem, sclerenchyma fibers, lamina thickness, orientation, density, and size of stomata on the surface of leaves were all characteristics of the habitats (Filartiga et al., 2022). The population of salt marshes had larger root cross-sectional area, metaxylem vessels, and number of vascular bundles, cortical tissues, and endodermal thickening in terms of anatomical changes all these anatomical modifications of Euphorbia hirta species help to survive in different environmental conditions such as saline, drought, and in waterlogging conditions (Yang et al., 2013).

According to (Nowak et al., 2021) the structure of the leaf epidermis can uncover numerous distinctive characteristics that are useful for classifying organisms. The previous research shown the need of using scanning electron microscopy (SEM) to analyze leaf epidermal anatomical characteristics in classifying certain Euphorbiaceae species. Acalypha wilkesiana Müll. Arg., Bischofia javanica Blume, Codiaeum variegatum (L.) A. Juss., Euphorbia cotinifolia L., Euphorbia helioscopia L., Euphorbia hirta L., Euphorbia milii Des Moul., Euphorbia neriifolia L., Euphorbia prostate Aiton, and Euphorbia pulcherrima the following plant species are included: Willd. ex Klotzsch, Euphorbiaroyleana Boiss., Euphorbia trigona Mill., Excoecaria cochinchinensis Lour., Jatropha integerrima. The following plant species are depicted in Plate 1: Jacq., Phyllanthus emblica L., Putranjiva roxburghii Wall., R. communis L., Sapium sebiferum (L.) Roxb., Trewia nudiflora L., Vernicia fordii (Hemsl.) Airy Shaw). The epidermal cell size varied among the examined Euphorbiaceae taxa, in addition to other leaf morphological characteristics. Abaxially, epidermis cells ranged in length from 68.25 ± 1.25 to 142.4 ± 0.45 μm. T. nudiflora had the smallest size, whilst A. wilkesiana had the greatest (Alyas et al., 2020). Trichomes have been seen to assist in withstanding heat stress and cold temperatures illustrated by (Naidoo et al., 2023). He also contributes to seed dissemination, enhance water absorption, and provide protection against the detrimental effects of UV-B radiation. Trichomes were found in a limited number of species in the current investigation. Trichomes were observed on the abaxial surface of E. hirta, E. prostate, E. pulcherrima, and P. roxburghii. Trichomes were observed on the upper surface of E. pulcherrima and P. roxburghii. There were no detected changes in the types of trichomes present in the leaves of Euphorbiaceae.

Conclusions and Recommendations

The current study on Euphorbia hitra collected from the Faisalabad region confirms that this species exhibits adaptations in its morphology and anatomy to survive in diverse environmental conditions. More specifically, these adaptations include thicker epidermal and endodermal layers in roots, increased stem area, and a larger cortical region. These modifications contribute to the plant’s tolerance of water deficit and its overall success in various habitats.

Acknowledgements

All the authors are thankful to the University of Agriculture, Faisalabad, Pakistan for providing lab space to conduct the analytical activities.

Novelty Statement

These adaptations allow Euphorbia hitra to thrive in diverse environments.

Author’s Contribution

Shahid Ali Khan: Conduct experiment and data collection.

Farooq Ahmad: Supervise the whole experiment as project head.

Muhammad Zubair Akram: Initial drafting and finalizing the MS.

Yan Tongyu: Statistical analysis.

Fatima Urooj: Statistical analysis.

Kamran Ahmad: Statistical analysis.

Saeed Ullah: Data collection.

Sharmeen Zulfiqar: Data collection.

Tahira Batool: Helped with relevant literature.

Aqsa Sarwar: Data collection.

Yaqoob Sultan: Data collection.

Samreen Nazeer: Reviewed final draft of MS.

Conflict of interest

The authors have declared no conflict of interest.

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