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Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa



A.Y. Ozdemir, A. Ozdemir and C. Ozdemir
 
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ABSTRACT

The aim of this study is to describe and compare statistically the anatomical characters of mature leaves of Crocus fleischeri Gay, C. danfordiae Maw, C. chrysanthus (Herbert) Herbert, C. pulchellus Herbert, C. flavus Weston subsp. flavus, C. pallasii Goldb. subsp. pallasii, C. speciosus Bieb. subsp. speciosus Mathew, C. speciosus Bieb. subsp. ilgazensis Mathew, C. speciosus subsp. xantholaimos Mathew, C. olivieri subsp. olivieri. The anatomical variations in ten Crocus taxa have been investigated by means of nümerical methods (Analysis of variance and Pearson correlation). By the analysis of the investigated taxa from eight leaf anatomy related characters, it has been determined that palisade cell height and spongy cell width are the best character pairs which represent the variations in them. It has been also found that the results from numerical analysis of the leaf anatomy characters can provide additional evidences which correspond to the anatomy for the recognition of the taxa.

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A.Y. Ozdemir, A. Ozdemir and C. Ozdemir, 2010. Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa. Asian Journal of Mathematics & Statistics, 3: 16-24.

DOI: 10.3923/ajms.2010.16.24

URL: https://scialert.net/abstract/?doi=ajms.2010.16.24
 

INTRODUCTION

The Crocus genus belongs to Iridaceae which had a large and diverse family of about 92 genera and 1800 species and is mainly distributed in the Southern hemisphere (Ali and Mathew, 2000). It is represented by 37 species in Turkey (Guner et al., 2000). Many species of the family Iridaceae are grown in parks and gardens as ornamental plants due to their beautiful flowers (Baytop, 1984). Some Crocus species were used for making dye, perfume and medicaments since 1600 B.C. (Rudall and Mathew, 1990). The saffron Crocus (Crocus sativus L.) was the first to be cultivated and has been grown for economic purposes since ancient times. Abdullaev (2003) pointed out that the saffron could be useful in cancer chemoprevention in near future. Different studies on some Crocus species has been found in the literature (Mathew and Brighton, 1977; Dainauskaite et al., 2001; Halevy, 1990; Miroslavov et al., 2000; Loskutov et al., 2000). But there have been a few studies on the morphology and anatomy of Crocus species in Turkey (Ozdemir et al., 2004; Ozyurt, 1978; Akan and Eker, 2004; Akan et al., 2007; Ozdemir and Akyol, 2005). Recently, some researchers have reported that the extract of Crocus has antitumor, antimetagenic and cytotoxic activities and inhibits nucleic acid synthesis in human malignant cells (Nair et al., 1991; Abdullaev, 2003; Loskutov et al., 2000; Fatehi et al., 2003). The leaves of most Crocus species have a unique and distinctive shape in cross section.

Table 1: The investigated samples and their voucher specimens
Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
E. Endemic

We aimed to give detailed knowledge about the anatomical characteristics of leaves of ten Crocus taxa in Türkey.

MATERIALS AND METHODS

Plant samples were collected from natural populations between 1997-2006 (Table 1). Taxonomic description of the plant was made according to Davis (1984) and Mathew (1982). The experimental studies were conducted at Celal Bayar University, Faculty of Arts and Sciences Department of Biology. Anatomical works were carried out on the fresh samples preserved in 70% alcohol. Paraffin method was used for preparing cross sections of the tissues (Algan, 1981). Transverse sections, 15-20 μm were made using a sliding microtome and stained with Safranin-Fast Green. Microscopic examinations were made on an Olympus BX50 microscope. Micrometric ocular was used for the anatomical measurements. For the numerical analysis, 8 characters of the leaves were selected. This selection was based on the variations of the leaf anatomical data. Characters were coded as 1, 2, 3, 4, 5, 6, 7, 8 and the taxa were coded as A, B, C, D, E, F, G, H, I, J. Significance of the differences between the taxa and characters were evaluated by analysis of variance (regression analysis) and pearson correlation.

RESULT AND DISCUSSION

C. fleischeri
The leaves had a central rectangular keel and two lateral arms, with their margins recurved towards the keel with a pale stripe running axially along the center of the leaf formed by the parenchymatous chlorophyllous cells. Both adaxial and abaxial surface except the groove parts of leaf had the cuticle. Epidermis cells, except the groove parts of abaxial surface are with straight walls. The epidermal cells on the groove parts had walls with papillae. Stoma cells are present only on groove part of leaf.

Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
Fig. 1: General drawing of the Crocus leaves. (a) C. fleischeri, (b) C. danfordiae, (c) C. chrysanthus, (d) C. pulchellus, (e) C. flavus subsp. flavus, (f) C. pallasii subsp. pallasii, (g) C. speciosus subsp. speciosus, (h) C. speciosus subsp. ilgazensis, (i) C. speciosus subsp. xantholaimos and (j) C. olivieri subsp. olivieri (scale bar: 500 μm)

These cells are in sunken position between epidermis cells with papillae. Palisade parenchyma cells are 1-2 layered. Spongy cells are present on abaxial side. Vascular bundles are located in one row in arms of keel and extending round abaxial margin of keel, but not across adaxial side. Major bundles occur at angles of keel and towards arm margins (Fig. 1, 2a).

C. danfordiae
Leaves of C. danfordiae had outline with central keel and two inflexed arms. The large central area of keel had thin walled cells which usually break down to form air space. The abaxial side of arms had two major protrusions. The epidermal cells of this protrusion had white papillae. Epidermis is cuticled except the grooved parts, which is single layered with flat-ovoidal cells. In the grooved parts the epidermal cells are papillate. Stomata is usually absent except in the grooved parts. Palisade parenchyma is 1-2 layered with spongy cells on periphery of vascular bundles. Vascular bundles are in one row in margins of arms and keel. Major bundles occur at angles of keel and arms, small bundles are located between the major vascular bundles (Fig. 1, 2b).

C. chrysanthus
The leaves had central rectangular keel and two lateral arm, with their margins recurved towards the keel. The characteristic pale stripe runs axially along the centre of the leaf. Stoma cells are in sunken position between epidermis cells with papillae. Adaxial surface and abaxial outer edge of keel except the groove parts of these surfaces are without stomata. Epidermal cells are 4-sided and with straight walls except in the edges of groove parts of abaxial surface. Epidermal cells on groove part of abaxial surface of leaf keel had walls with papillae. Vascular bundles are located in one row in arms of keel and extending around abaxial margin of keel, but not across adaxial side. Major bundles occur at angles of keel and towards arm margins (Fig. 1, 2c).

Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
Fig. 2: Cross-sections of the Crocus leaves. (a) C. fleischeri, (b) C. danfordiae, (c) C. chrysanthus, (d) C. pulchellus, (e) C. flavus subsp. flavus, (f) C. pallasii subsp. pallasii, (g) C. speciosus subsp. speciosus, (h) C. speciosus subsp. ilgazensis, (i) C. speciosus subsp. xantholaimos and (j) C. olivieri subsp. olivieri, e: Epidermis, m: Mesophyll, c: Cuticle, p: Palisade parenchyma, s: Spongy parenchyma s: Stoma v: Vascular bundle, sc: Sclerenchyma

C. pulchellus
Leaveshad central rectangular keel and two lateral arms, with their margins recurved towards the kell. The characteristic pale stripe runs axially along the center of the leaf is formed by the parenchymatous cells in the keel, which lack chloroplasts and break down to create an air space. Adaxial surface and abaxial outer edge of keel except the groove parts of these surfaces are without stomata. The cuticle is present on abaxial and adaxial surface. Epidermal cells are 4-sided. These cells except the ones on the groove parts of abaxial surface are with straight walls. It is difficult to distinguish the cells of palisade parenchyma from the cells of spongy parenchyma on the mesophyll of leaf. Because the mesophyll cells are more or less uniform in shape. Vascular bundles are located in one row in arms of keel and extending around abaxial margin of keel, but not across adaxial side. Major bundles occur at angles of keel and towards arm margins. Stoma cells are present on the groove parts of keel. These cells are in sunken position between epidermis cells with papillae (Fig. 1, 2d).

C. flavus Weston subsp. flavus
The leaveshad central triangular keel and two long lateral arms with their margins recurved towards the keel. The characteristic pale stripe runs axially along the center of the leaf is formed by the parenchymatous cells in the keel, which lack chloroplasts and break down to create an air space. Both adaxial and abaxial surfaces, except the parts in the grooveshad a thick cuticle. The epidermal cells are slightly furnished with papillae on the groove parts of the arms; stomata are present on these grooved parts. Mesophyll cells are more or less uniform in shape. Vascular bundles are located in one row and below the abaxial epidermis. The bundle sheath consists of sclerenchymatic cells at the phloem pole of major bundles. Abaxial epidermis had stomata while the adaxial epidermis had no stomata (Fig. 1, 2e).

C. pallasii Goldb. subsp. pallasii
The leaf had central slightly square keel and two lateral arms with recurved towards the keel. There is a pale stripe lying axially along the center of the leaf. The leaveshad the cuticle except groove part. The epidermal cells had cuticle with papillae. Adaxial epidermis is thicker than abaxial epidermis. Stoma cells are present only on the groove parts of leaf and in sunken position between epidermis cells. Palisade parenchyma cells are single layered. Vascular bundles are located in one row in arms of keel and extending around abaxial margin of keel, but not across adaxial side. There are major bundles occur at angles of keel and towards arm margins (Fig. 1, 2f).

C. speciosus Bieb. subsp. speciosus
The leaves had central rectangular keel and two lateral arms, with their margins recurved towards the keel with a pale stripe running axially along the center of the leaf. Both adaxial and abaxial surface except the groove parts of leaf had the cuticle. Epidermal cells are 4-sided, except the ones on the groove parts of abaxial surface are with straight walls. Stoma cells are present only on the groove parts of leaf. These cells are in sunken position between epidermis cells with papillae. Palisade parenchyma cells are 1-2 layered. Spongy cells are present on abaxial side. Vascular bundles are located in one row in arms of keel and extending around abaxial margin of keel, but not across adaxial side. Major bundles occur at angles of keel and towards arm margins (Fig. 1, 2g).

C. speciosus subsp. ilgazensis
The leaves had central slightly square keel. Margins of lateral arms recurved towards the keel with a pale stripe running axially along the center of the leaf. Both adaxial and abaxial surface except the groove parts of leaf had the cuticle. The epidermal cells on groove part had walls with papillae. Stoma cells are in sunken position between epidermis cells with papillae. Palisade parenchyma cells are 1-2 layered. Spongy cells are present on abaxial side. Vascular bundles are located in one row in arms of keel and extending around abaxial margin of keel, but not across adaxial side. Major bundles occur at angles of keel and towards arm margins (Fig. 1, 2h).

C. speciosus subsp. xantholaimos
The leaves had central nearly triangular keel and two lateral arms. The margins of the lateral arms are recurved towards the keel. The keel of leaf is filled with large parenchymatous cells. Stoma cells are present on the groove parts of keel. These cells are in sunken position between epidermis cells with papillae Vascular bundles are located in one row in arms of keel and extending around abaxial margin of keel (Fig. 1, 2i).

C. olivieri Gay subsp. olivieri
Leaves had central and nearly rectangular keel and two lateral arms. The tips of the lateral arms are recurved towards the keel. The keel of leaf is filled with large parenchymatous cells that lacked chloroplasts and were broken down in places so as to form air space. Mesophyll is located in the lateral arms had chloroplasts and differentiated into palisade and spongy parenchyma. It was somehow difficult to distinguish the palisade cells from the others in the mesophyll. Vascular bundles are arranged in one row along the arms of keel and they are closer to the abaxial side. Major bundles occur at the angles of keel and at the tips of arms. The cuticle on adaxial epidermis is thicker than the cuticle on the abaxial. The outer surface of epidermis is straight everywhere except the groove part of leaf surface. Only the groove parts of the leaf surface had stomata. Stoma cells are in sunken position between epidermis cells with conspicuous micropapillaes (Fig. 1, 2j).

Statistical Analysis
The anatomical measuments of the investigated taxa were shown in Table 2. Significance of the differences between the Crocus taxa was evaluated by analysis of variance (regression analysis) and pearson correlation (correlation). And the statistical analysis of the results were given Table 3-6.

The differences among the investigated taxa are shown as A-C, A-E, A-F, B-C, B-F, B-J, C-E, C-F, D-H, G-I and J-A in Table 3 and 5, are significant at levels of 0.01 and 0.05. Furthermore, a significant difference has been found at levels of 0.05 between E and B according to Table 3 based of Pearson correlation method (correlation). According to Table 4 and 6, there are important correlations among the anatomical characters of the leaves of the investigated taxa, shown as (Table 2) 1-4, 1-6, 4-5, 4-6 and 7-8 at levels of 0.01 and 0.05.

Table 2: Anatomical measuments of the Crocus taxa
Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
Values are in Mean±SD. SD: Standard deviation; 1-8: Character codes

Table 3: Pearson correlation (correlation) based on anatomical characters of the leaves of the investigated taxa
Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
*Significant at the level of 0.05; **Significant at the level of 0.01

Table 4: Pearson correlation (correlation) based on 8 anatomical characters of the leaves of the investigated taxa
Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
*Significant at the level of 0.05; **Significant at the level of 0.01

Table 5: Correlation between 10 investigated Crocus taxa (regression analysis)
Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
MS: Mean square; NS: Not significant; *p<0.05; **p<0.01

Rudall (1994) has also pointed out that leaves of most Crocus species had a unique and distinctive shape in cross section, comprising a central square or rectangular keel and two lateral arms. The present investigations are in agreement with Rudall’s (1994) findings, except that the leaves of C. flavus subsp. flavus and C. speciosus subsp. xantholaimos.

Table 6: The results of analysis of variance among the10 investigated taxa (regression analysis)
Image for - Statistical Comparative Leaf Anatomy of Some Crocus L. Taxa
MS: Mean square; NS: Not significant, *p<0.05, **p<0.01

These taxa had central slightly triangular keel different from the types mentioned above. These differences may be caused by these taxa localized more arid region. Because triangular keel which exists in the leaves of C. flavus subsp. flavus and C. speciosus subsp. xantholaimos cause deeper groove parts in the leaf surface. Therefore stoma cells of these taxa are in sunken position between epidermis. It is observed that the leaves have a pale stripe running axially along the centre of the leaf. This is a common feature in the genus (Rudall and Mathew, 1990). The results of the study show that there is a single layered palisade parenchyma in mesophyll of C. pallasii subsp. pallasii and C. olivieri subsp. olivieri while there is a 2-3 layered palisade parenchyma in mesophyll of the other Crocus taxa. Akan and Eker (2004) pointed out that there is 4-5-layered palisade parenchyma in mesophyll of Crocus cancellatus subsp. damascenus and C. pallasii subsp. turcicus.

As shown in Table 3 and 5, there are important correlations between C. speciosus subsp. xantholaimos and C. speciosus subsp. speciosus which are subspecies of C. speciosus. On the other hand, there are no important differences between subsp. speciosus subsp. ýlgazensiz and subsp. xantholaimos which are subspecies of C. speciosus.

It is mentioned that there are close relationships between C. danfordia and C. chrysanthus (Davis , 1984) Also, according to the statistical results derived, there is a considerable relation between the two taxa, at the level of p<0.01. The results of the statistical analysis were presented in Table 4 and 6. It was found that there were statistically important differences between element 1-4, 1-6, 4-5, 4-6 and 7-8 at levels of 0.01 and 0.05. By the analysis of investigated taxa from eight leaf anatomy related characters, it has been determined that palisade cell height and spongy cell width are the best characters pairs which represents the variations in them. It has been also found that the results from nümerical analysis of the leaf anatomy characters can provide distinct evidences, which are corresponding to the anatomy for recognition of the taxa.

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