University of Sulaimani School of Science Department of biology

A Comparative study of Leaf Surface Appendages and stomatal distribution of Alcea kurdica
-------------------------------------------------------------------------------------------------------------------------------A project submitted to the Department of Biology, School of Science, University of Sulaimani in partial fulfillment of the requirements for the degree of B. Sc. in Biology Abu Bakr Sidiq Sergeti & Yousuf Abdul Qadir Supervised by Heyder Usman Fetah, M.Sc in Botany -------------------------------------------------------------------------------------------------------------------------------Abstract: stomata numbers on upper and lower leaf surface were studied in Alcea kurdica of three different localities (Sulaimani, Dukan and khurmall).also epidermal appendages (trichomes) were studied in the same sample on leaf, petiole and stem.it was observed that both stomata and trichomes were more abundant on the lower leaf surface than the upper ones. Dissecting and compound light microscopes and also used ordinary camera.

Introduction
Order Malvales (mallows) includes only a large family (Malvaceae) of herbs or shrubs with stellate hairs, more than (90) genera and (1600) species found mostly in the tropics and temperate places of all continents. Some of them are native to Iraq. Malva (the common mallow, used as food in rural dishes), Gossypium (cotton), Abolmoschus (okra), and some ornamentals are members.( Erkan 2012) The genus named Alcea (mallow, hollyhock) by the ancient Greek botanist Dioscorides, has more than (50) species, mainly in South West & Central Asia, only (five) of them are found in Iraq and Kurdistan as wild plants and also may be useful for gardening. Alcea kurdica is a perennial herb about 30-80 cm tall, with few or several simple or branched stems. Branches are rather long ascending in the lower half, and densely tomentose (hairy), or more rarely glabrous (naked, no hairs). Trichomes (hairs) are stellate (star-shaped), sometimes with long rigid branches (Townsend 1980). Leaves are very variable in form; cordate (heartshaped), orbicular, (circular) with a wide basal sinus. They have 5-7 broad, shallow, obtuse (without tip) lobes, or are digitately (finger-like) divided 3/4 of the way to the petiole with 5-7 oblong or obovate-oblong lobes up to about 3-3.5 times as long as broad. Margins are crenate-serrate. Upper leaves are frequently more deeply devided. Leaf indumentum (hairs) densly softly stellatetomontose, more rarely rather sparsely stellate hairy. Petioles of lower leaves are 7-18 cm long, but upper leaves have gradually shorter stalks. Stipules are caducous (quickly falling), 5-7 mm long, deeply bifid (branched) with one or both lobes occasionally toothed. Flowers are deep violet-pink to rose-pink, white or white with a yellow center. Inflorescence is an elongating raceme, on short (0.5-3cm), densly tomentose peduncles (stalks). Bracteoles maybe 68, from 1/3 to 2/3 as long as the calyx. Calyx is 1.62 cm. long, the sepals are lanceolate-oblong, usually very distinctly striate (ribbed), stellate-tomentose. Petals are 4.25-4.75 cm long, broadly cuneate (triangular) obcordate (inversed heart-shaped), with apexes emarginate (lowered tips). Fruits are about 2 cm in diameter; carpels yellowish-brown, 5-9 mm in diameter, deeply sulcate (grooved) dorsaly. Each dorsal margin is broadly winged. Seed is reniform (kidney-like), 3-5 mm long, with the persistent bases of mostly deciduous hairs. (Essau 1974). This plant mostly grows in mountain slopes, in denuded oak forests, on limestone, igneous and metamorphic rocks, sometimes in fields and vineyards in the mountain and on the upper plains. It prefers altitudes of (150 1500) m, and may be found at (35 -1900) m. Its flowering and fruiting are in May July or Aug. The plant is common in the lower forest zone, and

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occasional in the upper forest zone of Iraq, (mountains of Kurdistan). Some common places to be found in are:- Ahmad awa (Atarat 2009), Sinjar, Zakho, Gare Dagh (Duhok), Amadiya, Sersink, Zawita, Atrush, Shaqlawa, Bradost, Halgurd, Rayat, Haji Umran, Qandil, Dokan, Qarahanjir, Derbendi Bazian, Sulaimani, Kani Spi, Kirkuk, Qara Tepe, Kifri, Jabal Hamrin, Baghdad, Mosul, etc. It is also present in some other countries like Crete (Greece), Cyprus, Syria, Lebanon, Palastine, Turkey, Iran (Muhammad Karami 2001), and Afghanistan. (Townsend 1980). The stomata (sing. Stoma, in Greek means mouth) are apertures in the epidermis, each bounded by two guard cells which by changing their shapes control the size of the stomatal opening, regulate the flow of gases between the leaf and its environment and control the amount of water passing through a leaf. Plants typically close their stomata at daytime to avoid too much water loss. The aperture leads to a space (the substomatal chamber), which is continuous with other intercellular spaces in the (6) mesophyll. The stomata are most common on green aerial parts of plants, particularly the leaves, not found in chlorophyll-less parasitic plants, nor in roots of any plants. They occur on some submerged aquatic plants, and on variously colored petals of the folwers, sometimes nonfunctional, and also on (5) stamens and gynoecia. In green leaves stomata may be on both surfaces (amphistomatic leaves) or on one only, but commonly the lower surface has more. This plant, Alcea kurdica shows this condition. Guard cells may occur at same level as the adjacent epidermal cells, or they may protrude above or be sunken below the surface of the epidermis. In some plants stomata are restricted to the epidermis that lines depressions in the leaf, the stomatal crypts. The guard cells are generally crescent-shaped with blunt ends (kidney-shaped) and often have ledges (shelves) of wall material on the upper and lower sides. In sectional views such ledges appear like horns. sometimes a ledge occurs only on the upper side, or non is present. (Essau 1974). Microbial entry into host tissue is a critical first step in causing infection in animals and plants. It has been assumed formerly that microscopic surface

openings, such as stomata, serve as passive ports of bacterial entry during infection. Surprisingly, it is found recently that stomatal closure is a part of plant innate immune response to restrict bacterial invasion.(Melotto 2006) Trichomes (Greek for hairs) are epidermal appendages of diverse forms, structures, and functions. They may occur on all parts of the plant, either they persist throughout the life of an organ, or they are ephemeral (shortly falling). Some persisting hairs remain alive; others become devoid of protoplasts and are retained in dry state. The epidermal trichomes usually develop early in relation to the growth of the organ. Trichomes are morphologically different, one type is the common hair. In structure they are unicellular or multicellular, branched or unbranched, uniseriate or multiseriate, and glandular or nonglandular. Some trichomes are branched in dendroid (tree-like) manner; others have branches oriented largely in one plane (stellate hair). Commonly a multicellular hair can be divided into a foot, which is embedded in the epidermis, and a body projecting above the surface. Cells surrounding the foot are sometimes morphologically distinct from other epidermal cells. Another common type of trichome is the scale, also called peltate hair. It consists of a discoid plate of cells, often born on a stalk or attached directly to the foot. The trichome cell walls are commonly of cellulose and are covered with cuticle, they may be lignified, or impregnated with silica or calcium carbonate. The trichomes of Alcea kurdica leaves belong to the branched hairs (stellate) (Essau 1974). Trichomes play major roles such helping increase the rate of water absorption, reducing the rate of water loss due to evaporation in order to keep the leaf surface cool, and providing a defence against insects, since the "hairiness" of the leaf impedes insect infestation. This is mostly what happens but maybe some insects prefer it? Glandular or secretory hairs provide a chemical defense against potential herbivores.

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Materials & Method:

Ten plant samples of A. kurdica from each three localities; Sulaimani University campus (the old campus), Dukan district, and Khurmal district, were taken randomly. Leaves of different sizes, ages and positions on the plants, from different plants (in size, age, and location in the habitat) were studied for stomata and trichomes on both sides. Specimens were brought to laboratory fresh or kept cool in the fridge. Preparations were done by hand striping-off the epidermal tissue with simple scalpels. Only water was used in mounting on glass slides, and both dissecting and compound microscopes were utilised to observation. Photographs were taken by ordinary cameras of suitable resolution.

Results and Discussion

Table (1): Average number of stomata in lower and upper surfaces of Alcea kurdica leaves in three different locations. Locality Sulaimani Dukan Khurmal Lower leaf surface stomatal average (of 10 samples) 20 16 19 Upper leaf surface stomatal average (of 10 samples) 9 11 10

Figure (1): Average number of stomata in the lower and upper leaf-surfaces of Alcea kurdica in three different localities.

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Figure(2) :Lower leaf-surface trichome.

Figure (3): Upper leaf-surface Trichome.

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Figure (4): Trichomes, lateral views. 10 X.

Figure (5): Trichomes on petiole 10X.

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Figure (6): Trichomes, light Microscope 10 X

Figure (7): Stomata on upper leaf surface of sample in Zanko campus. 40x

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Figure (8): Stomata on lower leaf surface of sample in Zanko campus. 40x

Figure (9): Stomata on lower leaf surface of sample in Zanko campus. 40x

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Figure (10): Stomata on lower leaf surface of sample in Zanko camp, 100x

Figure (11) :Stomata on upper leaf surface of sample in Khurmal 40X .

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Figure (12) :Stomata on lower leaf surface of sample in Khurmal 40X .

Figure (13) :Stomata on upper leaf surface of sample in Dukan 40X .

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Figure (14): Inflorescence of sample plants.

Figure (15): Flower of sample plant.

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Figure (16): Leaf of sample plant.

Figure (17): Sample plant habit.

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Table (1) gives average number of ten samples for each three localities, we can observe that in all the samples lower leaf surface have more stomata than the upper ones (Shaheen 2010). The difference is near to be 2:1 there are no significant difference between samples of the three localities. The same difference has been observed between trichomes, numbers of both leaf surfaces but according to figure (4) the trichomes are stellate branched and multicellular with articulate septa, same trichomes are crowded in the petiole surfaces and also on the stem in figure (5).Stomata and trichomes are shown in the figures by Dissecting microscope and both high and low power of compound microscope. And according to this observation we have not seen any subsidiary cells, because all the cells surrounding the guard cells are alike figure (10)

Conclusion:
After much observation, comparison and according to references this plant is one of the components of the dry habitat flora having characteristic xerophytic features such as abundant trichomes, robust habit, stomatal concentration mostly in the lower leaf epidermis.

References:
1. Essau, K. (1974). Plant anatomy. Wiley Eastern, NewDelhi, 2 ed. pp. 158- 170. 2. Townsend, C.C. & E. Guest. (1980). Flora of Iraq. V.4. Iraqi ministry of agriculture &Kew
nd

Royal Botanical Gardens. pp. 252-254. 3. Melotto M., William Underwood, Jessica Koczan, Kinya Nomura,and Sheng Yang He. (2006). Plant Stomata Function in Innate Immunity against Bacterial Invasion. Cell 126. p.269. 4. Ararat, K., N. Abdul Hassan and S. Abdul Rahman. (2009), Key Biodiversity Survey of Kurdistan, Northern Iraq, Edited by A. Bachmann, p. 55. 5. Erkan M. Uzunhisarcikli and Mecit Vural.(2012). The taxonomic revision of Alcea and Althaea (Malvaceae) in Turkey.p.614. 6. Shaheen, N., M.A. Khan, G. Yasmin, M.Q. Hayat, S. Munsif and K. Ahmad, 2010. Foliar epidermal anatomy and pollen morphology of the genera Alcea and Althaea (malvaceae) from Pakistan. Int. J. Agric. Biol., vol.12.p.329334. 7. M. Karami, M. E. Kasmani and A. A. Alamesh. Plants of Hashilan Wetland, Kermanshan, Iran.(2001). J. Sci. I. R. Iran. Vol. 12, No.3. p.206. 8. http://www.gwannon.com/species/Alcea-kurdica 9. http://www.eoearth.org/article/Stomata?topic=49510. 10. http://www.marietta.edu/~spilatrs/biol103/photolab/stomata.html 11. http://evolution.berkeley.edu/evolibrary/article/mcelwain_03

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