IJBPAS, May, 2015, 4(5): 2690-2702 ISSN: 2277–4998

THE RELATIONSHIP BETWEEN PALYNOLOGY AND CLASSIFICATION OF SOME SPECIES (CHENOPODIACEAE)

*1SOMAYEH SAFIALLAH AND 1SARA JALILI Department of Biology, Islamic Azad University of Garmsar Branch, Garmsar, Iran * Corresponding Author: E Mail: [email protected] ABSTRACT The genus Bassia of Chenopodiaceae, has geographically vast distribution. This genus in Iran is distributed in desert zones and is known as Halophyte. In new classification Bassia genus includes genera Londesia, Bassia, Panderia and Kochia. This study goal is discussing importance of taxonomic morphologic relationships of pollen grains in recent natural classification of this genus. In this study pollen grains of genera Kochia and Panderia is examined using Electronic Microscope (SEM). Results from this study showed that pollen grains in examined species were pantoporate, isopolar, spheroidal and exine surface is with spinules ornamentation. Also micromorphology planes are very effective in examining and recognizing kinship relations and can be useful in distinguishing a natural from artificial classification. Keywords: Pollen morphology, Chenopodiaceae, Bassia, Kochia, Ornamentation, Panderia

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INTRODUCTION system [15]. In Bassia photosynthetic type Chenopodiaceae is one of the biggest is Kochioid [16]. From point of view of families of order [1]. This anatomic and morphologic evolution C4 family has geographically worldwide dynasties has limited materials [17], [18]. distribution [2]. Most versatile group is Kochia and Bassia with a genus from

Halophyte that has large expanding in salty relative family named Amaranthus have C4 and desert soils [3]. Flora Iranica set forth photosynthetic system [19], [20]. useful information about various genera of The conducted study can reveal kinship this family [4]. In flora Iran genera relationships. For the same reason anatomic Panderia, Bassia, Kochia are placed and micro-morphologic investigation of separately. [5] These genera stand Amaranthus indicate that leaf has C4 Kranz separately in some floras of [6] & [7] Anatomy and spheral pollen has pores [21]. according to [8]. From floristic point of Taxonomic investigation indicates close view they are located in Australian area and relationship of and in Eurasia Bassia hyssopifolia and Kochia Chenopodiaceae [22], [23]. In new scoparia has most similarity [9]. investigated genus stand in These genera often combine with each other Bassia. Exine surface of some genus has and are distributed in Europe, Australia, pores, small spines and hole [24]. Sequence North America and Asia. Its most diversity analysis using atpB-rbcl markers in tribes is in Irano-Touranian [10]. The genera Champhorosmeae has indicated Kochia, Bassia, Panderia, Kochia stands separately Panderia and Bassia genus with other in accordance with grain position stand and investigated genera are monophyletic [26]. shape of perianth [11], [12]. Kochia and (With referring to [24] and [20]). Similarity Bassia were located in separate subfamily between these genus morphology and from Panderia. From morphologic point of anatomic research and sequence view most species of tribes Camphorosmeae determinations is a reason of a loose has covered by thricome. Morphological relationship between these genera. Bassia investigation on this tribes has most has most bootstrap. [20] Palynologic homoplasy [13]. Anatomic and micro- morphology in Chenopodiaceae has taken morphological investigation on pollen grain place very rarely [27], [28]. For the First has considered these genera as completely time, Moquin-Tandon in 1849 described separate [14]. Leaf anatomy in Bassia and pollen in this family Chenopodiaceae

Kochia affirms C4 and C3 photosynthetic

2691 IJBPAS, May, 2015, 4(5) Somayeh Safiallah et al Research Article palynology by [29] [30] [31] [32] [33] [34] size and flower. Regarding low efficacy of [35] [36] (with referring to [25] and [27]). micromorphological features in genus From morphologic viewpoint pollen grain distinction, palynology features is used to Chonopods is polyporat and from strengthening taxonomy and boundary taxonomic viewpoint pollen grain size is determining. This study goal is to very important [37], [38]. Camphorosmeae understand better importance of palynology indicates that Bassia genus with morphologic obvious features of pollen species Bassia dasyphylla and Bassia grain in natural classification and its eriphora has Atriplex type and with very connection with taxonomy. Obtained result different sizes and spheroidal and is compared with recent classification in this pantoporate has pores exine and has Scabrat genus. [39]. Chenopodiaceae usually has spherical MATERIAL AND METHODS and pantoporate pollens [40]. Exine surface Distribution and material is coated with spines [41]. Electronic Distribution information of Bassia (Kochia microscope can set forth useful information & Panderia) is on the basis of Iran Flora. in palynology field. This information After gathering series distribution on a includes phylogeny and evolution in geographical map (Figure 1) is depicted dicotyledons [42]. Wodehouse palynology gathered samples is preserved in Islamic studies using electronic microscope has Azad University of Garmsar Herbarium indicated a correlation between pollen grain (IAUG).

Figure 1: Distribution map of Kochia and Panderia (Flora Iran) Garmsar: Garmasar to Varamin road 7 km, VardAbad, Tehran-Karaj freeway, 1320 m, 1000 m, Safiallah 2013. Semnan: Near to Safiallah 2013; Sefid Bridge 13 km, Lasjerd- 32 km of southwest of Semnan, Firouzkouk road, 1000 m, Safiallah 2013; 1400 m, Safiallah 2013. Tehran:

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North east of Firouzkouh toward Sefid ornamentation in these species didn't show (white) bridge, 2000m, Safiallah 2013. any separable feature (Figure 2). Pollen morphology Pollen shape and diameter Under examination samples after Investigated species like other genus of recognition and gathering from various Chenopodiaceae are observed as spherical, areas have been taken out of samples in polar symmetry and numerous pores. order to investigate features and grain Taxonomic important feature that make surface structure, shape and other features. possible to recognize type was pollen grain Samples were deterging by alcohol 70% to size. All P. turkestanica, K. Prostrata. K. clean from dust. Ready Pollen grains were stellaris, K. scoparia species were different located on aluminum studs. After coating by size. On this basis of two type of pollen with gold by electronic microscope [43] the were recognized in these species. Type 1) model was photographed by Philips- X130. includes 10-25 µm small pollens and type 2) In specialized terminology [40] & [44] was includes 25-30 µm middle pollen. K. used. stellaris by size is smaller than others. On RESULTS this basis with 19 µm size stands in first Obtained result from palynology research type. indicated that among the investigated P. turkestanica, P. pilosa, K. prostrata, K. species, pollen grain size, pores number and scoparia with diameter of between 25-30 size and ornamentation of exine surface µm stands in second type. Therefore biggest were different. Most important result is pollen is K. scoparia with diameter of 30.4 shown in Table 1. Exine surface µm. ornamentation, pores size and chord Pore diameter and its proportion to pollen All investigated species has numerous diameter is important investigated features. spherical pores on their surface. A number In general, pollen grains are in shape of of these pores are different in species. K. isopolar, pantoporate, spheroidal, radial scoparia has most pores and K. stellaris has symmetric. Surface Exine ornamentation is less pores. Important point about pollen microechinate( spinules) and polyporate. pores is its shape. All pores are spherical Similarity between species is very high. Of and have prominent ornamentations. the fixed ornamentation is pollen exine Distance between pores was concerned too surface that can be useful in distinguishing and no significant difference observed. in parents or genera level. Exine surface Investigations on pores diameters of species

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indicate small difference that is not a new species standing in Bassia genus comparable feature. Evident difference in indicate that there is no Polymorphism in convexity of pore shape in P. turkestanica is pollen and can be very useful in taxonomy. observed. Most important point in this research are Exine ornamentation features such as pollen shape, pollen Exine surface in species Panderia and ornamentation surface and pollen size, but Kochia is plane with pores. In addition in recognizing in genus level this features exine surface has spine prominent are not appropriate. But in order to ornamentation. These spinules are less than recognize kinship relations of species at 1 µm by height. By size microspines of P. genus level they are very useful. On the turkestanica and P. pilosa has smaller size same basis close relation of old genera and other species K. scoparia, K. prostrata, Kochia and Panderia can be observed. K. stellaris with little difference with Species with kinship relations has the same Panderia were bigger. type of pollen. Species that in the past stood Chord diameter separately on the basis of palynology and Importance of number of pores and C/D kinship relationship are not separable in proportion with pollen diameter indicate genus level but could have difference in that number of pores has direct relation with species level. Therefore investigation of C/D proportion and less number of pores these two genera from palynology point of more C/D proportion takes place. In K. view confirms their standing in same genus stellaris exactly this issue is indicated. with name of Bassia and creating a natural Difference among species that can bring classification with same features. about distinction in species level is size and number of pores that is different in each species. Results obtained from research on

Table 1: Comparison of morphology measurement of pollen grains Kochia and Panderia; PD: Pollen Diameter, Pored: pore Diameter, CD: Chord/Diameter, Pore No: Pore None Taxa PD Chord C/D µm Pore Pore No Spinules Pollen Surface Exine µm µm µm Shape ornamentation Kochia scoparia 28 3/35 0/119 1/79 44 0/17 Spheroidal Pore- spinules

Kochia prostrata 30/4 3/35 0/110 1/85 30 0/20 Spheroidal Pore- spinules Kochia stellaris 19 2/45 0/128 1/63 21 0/17 Spheroidal Pore- spinules

Panderia pilosa 27/2 3/08 0/113 2/52 26 0/12 Spheroidal Pore- spinules Panderia turkestanica 25/2 2/88 0/114 2/14 24 0/13 Spheroidal Pore- spinules

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A

B

C

D

E

Figure 2: SEM micro-morphological of pollen grain in Kochia and Panderia species: A) Panderia pilosa B) Panderia turkestanica C) Kochia prostrata D) Kochia scoparia E) Kochia stellaris DISCUSSION similarities and differences. Use of Applying micromorphology method palynology features in addition to especially pollen grain electromicroscopic recognition at genus level can be a great studies help effectively in confirming help in palynological in Chenopodiaceae

2695 IJBPAS, May, 2015, 4(5) Somayeh Safiallah et al Research Article subfamily. Number of pores on pollen as morphology load and morphologic periporate is used as a distinctive feature in similarity was consistent and it’s a reason analysis and classification of pollen for confirming new classification under the Caryophyllaceae and Chenopodiaceae. [45] name of Bassia. According to [28] Palynology research indicates that number researches and [13] Bassia pollen grain are and diameter of pores in various species are spherical, pantoporate and periporate and different and pores shape is similar and exine surface has spinulate. Difference in circular. There is little difference in pollen diameter and number of palynology features in Chenopodiaceae ornamentation and pores caused [13] to family. From taxonomical viewpoint pollen consider pollen grain Bassia and Kochia as grain size and pores shape is important [37], separated from each other and locate this [38]. species in two different genera. Most On this basis our studies on Kochia and features at genus level are polarity and Panderia indicate that two types of pollens pollen shape and exine ornamentation type are recognizable. First type was small pollen that in these genus didn't create separation and second was middle pollen. Diameter boundary. C/D proportion is another feature of pollen grain according to [37] and distinctive feature in Chenopodiaceae [38] theory is an appropriate feature and can family that is used in classifications [28]. be useful in introducing pollen type at genus C/D feature in Panderia, Kochia has small level indicated that the correlation between versatility and could not be an effective pollen grain size and flower. Regarding distinctive feature. On the basis of palynology researches in this genus and its conducted morphologic researches some species it was indicated that there is no species of Kochia are Halophytic. For the correlation. Some species had small size reason of ecologic conditions and pollen and small flower, but this feature is adaptability with environment lamina of not stable and does not have classification leaf is disintegrated and photosynthetic value. Conducted palynology researches on system is C4 , small flowers, leaf, stem and Kochia and Panderia are very limited and it perianth is covered with trichomes and roots can be said no study in taxonomic level is are long and deep [21]. taken place. Palynological studies are in Pollen grain morphological studies on these order to understand species morphologic species and other studied species know features [46]. On the same basis pollens features such as pollen diameter, number of indicated special similarities that species

2696 IJBPAS, May, 2015, 4(5) Somayeh Safiallah et al Research Article pores, and ornamentation exine type conditions and are not appropriate feature at according with morphological features. genus distinction level, in other word they Morphological structure and pollen are due to ruptures resulting from geological morphology array pattern are due to events and intense weather change. These mechanical adaptability with ecologic ruptures can produce highest level of conditions. Conducted phylogenetic homoplasy. Investigated species on the basis researches on Chenopodiaceae, of pores and pollen shape indicated no Amaranthaceae confirm that they are polymorphism and can be a description of monophyletic [47] [48] [49] [50] (with path of evolution and kinship continuity in referring to [20]). Palynology research on species. Our palynology researches revealed genus Amaranthus introduces pollen grains it properly. as spherical and poriporate and having ACKNOWLEDGEMENTS spinules ornamentation on exine surface We are grateful to Dr. E. Hosseyni for his [21]. Palynology researches on some species support on our study. of genus Bassia (Kochia & Panderia) from REFERENCES Chenopodiaceae family indicate kinship [1] Cuenoud, P., Savolianeu, V., relation between these species and genus Chatrou, L.W., Powell, M., Grayer Amaranthus. Great palynological similarity R. J. & Chase, M.W. (2002) can be known an appropriate reason for molecular phylogenetics of the emphasizing kinship relation. New Caryopyllales based on I & s rdna , classification of Bassia include genera and plastid rbcl , atpb, and matk Kochia and Panderia [24]. sequences. Am,7 Bot., 89, 132-144 Palynology similarity of these genera from [2] Kuhn, U (1993). Chenopodiaceae.In taxonomical viewpoint can confirm results the families and Genera of vascular obtained from recent research in position of 2 a natural classification, and distinction on (kubitzki,k.,Rohwer,J.G.,and the basis of morphological researches Bittrich,v.eds), Berlin, Heidelbery : feature such as verticality or horizontality of springer.pp.200-280 grain, perianth ornamentation, perianth [3] Akhani.H , Ghasemkhani. M trichomes and perianth outgrowths are not ,Chuong sdx, ed wards known as natural classification [52] (with GE.(2008).Occurrence and forms of referring to [7], [11]). These features can be kranz Anatomy in photosynthetis in affected by ecological and regional Ble-pharis ciliaris(l) b.l.Burtt

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