Cladistic analysis of Taxaceae s. l.

Taxaceae s. l. is a wider concept of classification treating five genera of Taxaceae s. str. and Cephalotaxus together. Cephalotaxus is morphologically very similar to the five genera of Taxaceae s. str. Various models of classification for six genera have already been published. However, the phylogenetic position and genuine relationships of these genera and species are still confusing. A cladistic analysis of Taxaceae s. l. has been carried out to resolve the problem existing in their phylogeny and to provide a new approach regarding the relationships of these six genera. Parsimony analyses were based on 28 characters and eight genera including two outgroups Agathis and Sciadopitys. The most parsimonious tree retained with branch length 38 and 194 rearrangement trials. Consistency index was 0.68 and retention index was 0.66. Principally, two main clades were found: one represented by Austrotaxus forming the base of the tree and another by the remaining five genera. Taxus + Pseudotaxus clade split after Austrotaxus, and Cephalotaxus was sister to Torreya + Amentotaxus clade. Taxus + Pseudotaxus clade was supported by the highest bootstrap value. Finally, cladistic analysis does not support existence of Cephalotaxaceae. Therefore, it would be better to classify Cephalotaxus within Taxaceae s. l. with the other five genera.     

Plastome phylogenomics of Cephalotaxus (Cephalotaxaceae) and allied genera

Background and Aims Cephalotaxus is a paleo-endemic genus in East Asia that consists of about 7–9 conifer species. Despite its great economic and ecological importance, the relationships between Cephalotaxus and related genera, as well as the interspecific relationships within Cephalotaxus, have long been controversial, resulting in contrasting taxonomic proposals in delimitation of Cephalotaxaceae and Taxaceae. Based on plastome data, this study aims to reconstruct a robust phylogeny to infer the systematic placement and the evolutionary history of Cephalotaxus.MethodsA total of 11 plastomes, representing all species currently recognized in Cephalotaxus and two Torreya species, were sequenced and assembled. Combining these with previously published plastomes, we reconstructed a phylogeny of Cephalotaxaceae and Taxaceae with nearly full taxonomic sampling. Under a phylogenetic framework and molecular dating, the diversification history of Cephalotaxus and allied genera was explored.Key ResultsPhylogenetic analyses of 81 plastid protein-coding genes recovered robust relationships between Cephalotaxus and related genera, as well as providing a well-supported resolution of interspecific relationships within Cephalotaxus, Taxus, Torreya and Amentotaxus. Divergence time estimation indicated that most extant species of these genera are relatively young, although fossil and other molecular evidence consistently show that these genera are ancient plant lineages.ConclusionsOur results justify the taxonomic proposal that recognizes Cephalotaxaceae as a monotypic family, and contribute to a clear-cut delineation between Cephalotaxaceae and Taxaceae. Given that extant species of Cephalotaxus are derived from recent divergence events associated with the establishment of monsoonal climates in East Asia and Pleistocene climatic fluctuations, they are not evolutionary relics.     

Diversity and evolution of leaf anatomical characters in Taxaceae <Emphasis Type="Italic">s.l.—</Emphasis>fluorescence microscopy reveals new delimitating characters

Taxaceae s.l. comprise six genera (including Cephalotaxus) and about 35 species; The present study aims to give new insights into the evolution of this family, especially into the phylogenetic position of Cephalotaxus. Moreover, only little is known about comparative leaf anatomy of this family and this study aims to expose and interpret the diversity and evolution of leaf anatomical characters and to assess their applicability to identify taxa at the generic and species level. A detailed phylogeny was reconstructed, using both maximum likelihood and Bayesian inference, with a combined dataset of four molecular markers from the plastid and nuclear genomes. Leaf sections from 132 specimens, representing 32 species and four varieties (fresh and herbarium material) were inspected, using fluorescence microscopy. Ancestral characters were reconstructed using Mesquite. The phylogenetic analyses provided full support for Cephalotaxus as sister group to Taxaceae s.str. Within the latter, two monophyletic tribes Taxeae (comprising Austrotaxus, Pseudotaxus, and Taxus) and Torreyeae (comprising Amentotaxus and Torreya) were fully supported. Fluorescence microscopy was shown to be very useful for identifying leaf tissues and their constitution. We were able to show that particularly sclerified tissues have highest potential for the discrimination of both freshly collected samples and rehydrated herbarium specimens at the generic and species level. A correlation between the presence of different sclereid types could be shown and sclereids were hypothesized to pose a primitive trait in the evolution of Taxaceae s.l. New identification keys were generated on the basis of leaf anatomical characters. The microscopic method presented here is applicable for further studies within gymnosperms and probably in angiosperms, as well.     

Phylogeny of Taxaceae and Cephalotaxaceae Genera Inferred from Chloroplast matK Gene and Nuclear rDNA ITS Region

Phylogeny of the Taxaceae genera and the monotypic family Cephalotaxaceae has been extraordinarily controversial. In this paper chloroplast matK genes and nuclear ITS sequences were determined for all six genera of the two families and representatives of other conifer families. Analysis using either the nonsynonymous sites or the deduced amino acid sequences of matK genes strongly indicates that taxad genera and Cephalotaxaceae are monophyletic, with the Taxodiaceae/Cupressaceae clade as their sister group. Cephalotaxus is basal to the taxad genera, among which two clades, Torreya/Amentotaxus and Taxus/Pseudotaxus/Austrotaxus, are resolved. They correspond to Janchen's two tribes, Torreyeae and Taxeae. In Taxeae, Austrotaxus is the first to branch off. Analyses of the nuclear ITS sequence data corroborated the topology of the matK gene tree. These results refute the views that Cephalotaxaceae has no alliance with Taxaceae and that Austrotaxus and Amentotaxus should be excluded from the Taxaceae. We estimated the divergence time between the Taxodiaceae/Cupressaceae and the Cephalotaxaceae/Taxaceae clades to be 192–230 Myr ago and the divergence time between taxads and Cephalotaxus to be 149–179 Myr ago. Soon after the latter divergence event, within 6–8 Myr, the two taxad tribes originated. In conclusion, our data do not support Florin's claim that taxads could be traced to Devonian psilophytes (359–395 Myr ago).     

Morphology and anatomy of male cones of Pseudotaxus chienii (W.C. Cheng) W.C. Cheng (Taxaceae)

Results of the present study indicate that male cones of Pseudotaxus chienii are representing inflorescences with strongly reduced flowers. The results fit quite well with investigations showing that sporangiophores of Taxus and also of Pseudotaxus comply with reduced flowers. The only difference between male cones in Taxus and Pseudotaxus is the absence of pherophylls in Taxus. Furthermore our results complete a transition series beginning with Cephalotaxus going on to Pseudotaxus and ending with Taxus and Torreya. In this progression Pseudotaxus can be regarded as an intermediate link between the inflorescences of Cephalotaxus and the simple, unbranched cones of Taxus. The entire transition series shows that sporophyll-like sporangiophores can be derived by reduction of lateral cones. There is however no sign that a similar process has occurred in other conifer groups.     

Interspecific relationships and origins of Taxaceae and Cephalotaxaceae revealed by partitioned Bayesian analyses of chloroplast and nuclear DNA sequences

The precise delimitation of Taxaceae and Cephalotaxaceae is not totally resolved. Some contradicting taxonomic proposals have been published, which demonstrates the difficulties in establishing a natural classification of the families and especially in proposing a relevant treatment within the genera Taxus and Cephalotaxus. The aims of this study are to contribute to the phylogeny and specific delineation of the two conifer families on the basis of molecular data. A cladistic analysis of the sequences of five chloroplast (matK, rbcL, trnL, trnL-trnF spacer, and psbA-trnH spacer) and one nuclear (ITS) molecular markers was carried out, both individually and in combination, by distance, parsimony, likelihood, and Bayesian methods. The results confirm that the two families are monophyletic. In the genus Taxus, T. floridana is the first-branching taxon; T. brevifolia and T. globosa cluster together and are sister to T. baccata; the endemic T. yunnanensis clusters with T. wallichiana in subclade B and is only distantly related with the other four Taxus species in China (subclade A); T. fuana is closer to T. baccata than to other Taxus species. Torreya jackii and A. formosana are the first-branching species within Torreya and Amentotaxus, respectively. C. koreana and C. wilsoniana could be treated as two varieties of C. harringtonia. The ancestral distribution area of Taxaceae and Cephalotaxaceae is restricted either to Southwest China or Southeast China by DIVA analysis. The relaxed molecular clock indicates that the deepest divergences in Taxus go back to the late-Cretaceous. psbA-trnH, rbcL third codon position, and matK first codon position contributed most to the separation of taxa in Discriminant function analysis. Our results confirm, on a basis of multiple molecular markers and a complete sampling of basic species, the suggested monophyly of Taxaceae and Cephalotaxaceae and propose interspecific relationships within each group, with profound nomenclatural and taxonomic implications. Combination of partitioned Bayesian analysis and likelihood-based methods produced a more robust phylogenetic hypothesis for the two studied families.     

The phylogenetic positions of the conifer genera Amentotaxus,Phyllocladus, and Nageia inferred from 18s rRNA sequences

To determine the evolutionary positions of the conifer genera Amentotaxus, Phyllocladus, and Nageia, we obtained 18S rRNA sequences from 11 new taxa representing the major living orders and families of gymnosperms. With the published Chlamydomonas as an outgroup, phylogenetic analyses of our new data and available sequences indicate that (1) the Gnetales form a monophyletic group, which is an outgroup to the conifers, (2) the conifers are monophyletic, (3) Taxaceae, Cephalotaxaceae, Cupressaceae, and Taxodiaceae form a monophyletic group, (4) Amentotaxus is closer to Torreya than to Cephalotaxus, suggesting that Amentotaxus is better to be classified as a member of Taxaceae, (5) Phyllocladus, Dacrycarpus, Podocarpus, and Nageia form a monophyletic group, and (6) Pinaceae is an outgroup to the other families of conifers. Our finding that Phyllocladus is a sister group of the Podocarpaceae disagrees with the suggestion that the phylloclade of the genus is an ancient structure and that the genus is a terminal taxon within the Podocarpaceae. The genus Nageia is more closely related to Podocarpus than to Dacrycarpus and was derived from within the Podocarpaceae. In conclusion, our data indicate that in conifers, the uniovulate cone occurred independently in Taxacaeae and Cephalotaxaceae, and in Podocarpaceae after the three families separated from Pinaceae, and support the hypothesis that the uniovulate cone is derived from reduction of a multiovulate cone.Correspondence to: S.-M. Chaw     

Comparative palynological and wood anatomical studies of Indian Taxus wallichiana Zucc., Cephalotaxus mannii Hook. and Cephalotaxus griffithii Hook.

A comparative study of Taxus wallichiana Zucc., Cephalotaxus mannii Hook. and Cephalotaxus griffithii Hook. of two families Taxaceae and Cephalotaxaceae has been carried out in detail to support the taxonomic existence of two families. The comparative studies have been carried out on the basis of wood anatomy and palynology. The anatomical properties of wood including the tracheids, ray parenchyma, axial parenchyma and number of cross-field pits have been described in detail. The palynological studies include the shape, size and ultrastructure of pollen grains. These studies give a taxonomic support for the recognition of two different genera of families Taxaceae and Cephalotaxaceae which are closely related.     

Leaf anatomy as an additional taxonomy tool for 18 taxa of Matricaria L. and Tripleurospermum Sch. Bip. (Anthemideae-Asteraceae) in Turkey

This study concerns the evaluation of leaf anatomical characteristics of the closely related genera Matricaria (4 taxa) and Tripleurospermum (14 endemic taxa, 15 accessions) in Turkey using cluster analysis (CA) and principal component analysis (PCA). All taxa have the same basic leaf structure composed of lower and upper epidermis including anomocytic or anomocytic-anisocytic stomata, parenchymatic mesophyll, and vascular bundle. Statistically significant differences based on ploidy level have been found without generic delimitation. The differences mainly concern stomatal length and the relative size of the vascular bundle. These anatomical characters can be used to distinguish diploid from polyploid taxa of the genera.     

The course of resin canals in the shoots of conifers

The course of resin canals in stem cortex and the continuity between resin canals in leaves and those in stem cortex were investigated. The present paper is the first of three parts of the investigation. In this paper, fundamental features of resin canals and actual resin canal patterns in the Taxaceae, Cephalotaxaceae and Podocarpaceae are reported. From the observation of serial transections of shoots, composite diagrams and three-dimensional models of resin canal patterns are drawn. Central canals, if present, run vertically in stem cortex and sometimes divide, end blindly or unite each other. The distance between two adjacent central canals fluctuates rhythmically in connection with the vascular supply from the stem to leaves. The resin canal patterns of the families are classified into four types. Those ofTaxus, Nothotaxus and three species ofDacrydium belong to the Taxus type, those ofTorreya andCephalotaxus to the Torreya type, those ofDacrydium elatum, Podocarpus alpinus, P. elatus, P. elongatus andP. neriifolius to the Dacrydium type, those ofP. macrophyllus, P. nagi andP. koordersii to the Podocarpus type.     

Middle Miocene Cephalotaxus (Taxaceae) from Yunnan,Southwest China,and its implications to taxonomy and evolution of the genus

The infrageneric taxonomy of the genus Cephalotaxus Siebold et Zuccarini ex Endlicher (Taxaceae) is controversial as morphological characters have been doubted to change coherently among species. Although the epidermal features are considered as important characters in the taxonomy of conifers, they have not been systematically studied in this genus. Neogene foliage shoots with well-preserved epidermal structure not only can provide systematic links to the extant species, but also have implications for the taxonomical definition of the extant species by their epidermal features. In this paper, the morphologies of leafy shoots combining epidermal structure of a Neogene Cephalotaxus, which was collected from the middle Miocene of Southwest China, and ten extant species (variants) were studied. Morphology as well as epidermal structures was compared among species (variants) in Cephalotaxus. Based on the comparisons, the taxonomical definition of extant species in genus Cephalotaxus are discussed. Our results indicate that leaf morphological characters in combination with the features of epidermal structure can distinguish species in genus Cephalotaxus. The fossil foliage shoots from the middle Miocene of Southwest China were assigned to a new species, C. maguanensis n. sp., which shows close affinities to the extant C. hainanensis. On the basis of leaf and epidermal information, we suggest there are seven extant species in Cephalotaxus. With global cooling and the uplift of the Qinghai-Tibet Plateau since the middle Miocene, the cooling climate induced the differentiation and southward expansions of C. maguanensis, and its extant offspring, C. hainanensis, is distributed to Hainan Island and the Indo-Chinese peninsula.     

从幼苗形态学特征探讨红豆杉科各属间的系统演化

本文根据幼苗的形态学特征,尤其是子叶的结构、说明在红豆杉科中最原始的属是榧树属,其幼苗为留土萌发,植株粗壮;子叶肥厚,有吸收功能,叶肉不分化,其细胞中充满淀粉粒：维管束在子叶的上部或顶部为单束2条,下部合并成双束1条。其次是穗花杉属,幼苗为出土萌发,植株粗壮,下胚轴的下部肥厚;子叶稍肉质,叶肉不分化,质体淡绿,有一定的光合功能,但主要是积累淀粉,维管束双束1条,在子叶的顶端趋于分裂。最进化的是红豆杉属和白豆杉属,可能还有澳洲红豆杉属,其幼苗为出土萌发,植株纤细;子叶薄,叶状,叶肉分化成栅栏和海绵组织,维管束单束1条。本科中族的划分和属的系统排列,从原始到进化,应该是：族1．榧树族(1．榧树属);族2．穗花杉族(2．穗花杉属);族3．红豆杉族(3．澳洲红豆杉属,4．红豆杉属,5．白豆杉属)。     

八角目(Illiciales)植物花被片表皮比较形态

在光学显微镜和扫描电镜下,观察了八角科(Illiciaceae)八角属(Illicium Linn.)2组11种20个材料、五味子科(Schisandraceae)南五味子属(Kadsura Ksempf. ex Juss.)2亚属2组8种15个材料和五味子属(Schisandra Michx.)2亚属4组6种17个材料的花被片表皮形态特征。首次报道了八角目2个科（八角科和五味子科）3个属（八角属、南五味子属和五味子属）植物花被片表皮细胞的形状、分泌细胞的形状及分布、气孔器的形状及分布、花被片表面的纹饰。通过与八角目2个科3个属植物的叶表皮形态比较,发现花被片表皮气孔器外拱盖均为单层,与叶表皮气孔器外拱盖层数（常绿种类为双层和落叶种类为单层）之间没有相关性,还在五味子科中发现2个新的性状（气孔对和环列型气孔）。通过对两性花、雌花和雄花花被片表皮观察,发现花被片表皮形态与花的性别之间没有相关性。通过对八角属、南五味子属和五味子属花被片表皮比较,发现五味子属与南五味子属相比,其花被片表皮表现出更多的衍生性状;南五味子属与八角属相比,前者花被片表皮具有更多的衍生性状;而南五味子属花被片表皮形态与五味子属的相似性程度较大,支持五味子科包含南五味子属和五味子属、八角科包含八角属的观点。     

Taxonomic and ecological implications of leaf cuticular morphology in Castanopsis, Castanea, and Chrysolepis

Leaves of 41 species of Castanopsis, six species of Castanea, and Chrysolepis chrysophyllum Hjelmq. were examined. In Castanopsis, all species possessed cyclocytic stomata with thickened subsidiary cells; thin-walled peltate trichomes are the most frequent type on the abaxial surface of the leaves of this genus. In Castanea, stomata are transitional between cyclocytic and anomocytic; thin-walled peltate trichomes were recorded for the first time on the abaxial surface of Castanea leaves. In Chrysolepis, cyclocytic stomata with non-thickened subsidiary cells and thick-walled peltate trichomes were observed. The thickened subsidiary cells support the placement of the “fissa-group” in Castanopsis. The results of this study support the idea that Castanopsis and Castanea are sister groups. Thick-walled peltate trichomes were only recorded in Chrysolepis, thus supporting its taxonomic separation from Castanopsis. The phylogenetic distribution of trichome types among genera of Fagaceae is summarized. The evolutionary trends of trichome types in Castanopsis are discussed, as are the implications of stomatal and trichome features on fossil identification and ecology.     

Phylogeny of taxaceae and cephalotaxaceae genera inferred from chloroplast matK gene and nuclear rDNA ITS region

Phylogeny of the Taxaceae genera and the monotypic family Cephalotaxaceae has been extraordinarily controversial. In this paper chloroplast matK genes and nuclear ITS sequences were determined for all six genera of the two families and representatives of other conifer families. Analysis using either the nonsynonymous sites or the deduced amino acid sequences of matK genes strongly indicates that taxad genera and Cephalotaxaceae are monophyletic, with the Taxodiaceae/Cupressaceae clade as their sister group. Cephalotaxus is basal to the taxad genera, among which two clades, Torreya/Amentotaxus and Taxus/Pseudotaxus/Austrotaxus, are resolved. They correspond to Janchen's two tribes, Torreyeae and Taxeae. In Taxeae, Austrotaxus is the first to branch off. Analyses of the nuclear ITS sequence data corroborated the topology of the matK gene tree. These results refute the views that Cephalotaxaceae has no alliance with Taxaceae and that Austrotaxus and Amentotaxus should be excluded from the Taxaceae. We estimated the divergence time between the Taxodiaceae/Cupressaceae and the Cephalotaxaceae/Taxaceae clades to be 192-230 Myr ago and the divergence time between taxads and Cephalotaxus to be 149-179 Myr ago. Soon after the latter divergence event, within 6-8 Myr, the two taxad tribes originated. In conclusion, our data do not support Florin's claim that taxads could be traced to Devonian psilophytes (359-395 Myr ago).     

MODIFIED REMNANTS OF CENTRIPETAL XYLEM IN THE LEAF TRACE OF CEPHALOTAXUS

A study of the leaf traces at the nodes in various species and varietal forms of Taxus, Torreya, Amentotaxus and Cephalotaxus reveals, only in Cephalotaxus, an unusual type of parenchymatous tissue associated with the xylem of the leaf trace. The cells of this tissue occur in one to three layers, have abundant cytoplasm and conspicuous nuclei. The thin walls of these cells are devoid of pits and show spiral or spiral-reticulate thickenings. These thickenings, although readily taking the counterstain, reveal the presence of lignin as determined by the phloroglucin test and by fluorescence microscopy. This tissue is always internal—that is, adaxial—to the protoxylem of the trace. From the node it accompanies the trace for a short distance into the leaf base, where it gives way centrally to the typical leaf parenchyma and laterally to the transfusion tissue on the flanks of the bundle. In the basipetal direction it is in contact with the inner face of the annular protoxylem of the stele, eventually disappearing in the typical pith parenchyma. This tissue occasionally reveals instances of well-lignified tracheid-like centripetal elements. On the basis of the characteristics of this tissue, it is suggested that its origin lies in former centripetal xylem. The significance of this tissue to the evolution of the stele and the systematic position of Cephalotaxus is discussed.     

遮荫对金莲花光合特性和叶片解剖特征的影响

以金莲花为材料,系统研究了不同遮荫处理(全光照、40%、60%和80%遮荫)对金莲花叶片光合特性和叶解剖结构的影响,为金莲花引种驯化和抚育管理提供理论依据。结果表明:全光照下金莲花叶净光合速率有明显光合"午休"现象,遮荫处理下"午休"现象不明显。其中,40%遮荫处理显著促进了金莲花叶片净光合速率,80%遮荫处理则起抑制作用;40%和60%遮荫处理促进金莲花叶蒸腾速率和气孔导度的提高,降低了蒸汽压亏缺,有利于光合作用的进行;80%遮荫处理下胞间CO2浓度提高,气孔限制值和蒸汽压亏缺均显著高于其它处理,抑制了金莲花叶蒸腾作用和光合作用;遮荫处理降低了金莲花叶光补偿点和暗呼吸速率,40%和60%遮荫处理下表观量子效率和最大净光合速率提高,说明金莲花有一定耐荫性;金莲花叶光合特性与解剖结构关系密切。遮荫降低了金莲花叶气孔密度和气孔指数,但气孔纵轴长和横轴长显著高于对照。40%和60%遮荫处理下,单叶气孔数量没有明显变化;遮荫下金莲花叶片增厚。栅栏组织厚度和细胞层数减少,栅栏细胞宽度加大,且第二层栅栏细胞有向海绵组织过度的趋势。海绵组织厚度增加,栅栏组织/海绵组织比值减小。随遮荫度增高,栅栏组织和海绵组织细胞间隙增大。该结构有利于提高对光能的利用效率;遮荫处理促进维管束的发育,维管束数量增加,木质部、韧皮部分化明显,其中中脉导管直径和导管数目增加,有利于蒸腾作用和光合作用的进行。     

New insights into seminal and stomatal morphology and their contribution to the taxonomy of the Old World succulent perennial Salicornioideae

Seminal and stomatal morphology of three succulent halophyte genera (Sarcocornia, Halocnemum and Arthrocnemum) around the Mediterranean basin and surroundings areas were investigated. Scanning electron microscopy (SEM) investigations showed well-different seminal coat ornamentations among the studied taxa. Both Arthrocnemum macrostachyum and Halocnemum strobilaceum do not show trichomes. Conversely, the species Sarcocornia perennis and S. fruticosa can be easily identified by the presence of long curved hairs against papillae or short conic hairs, respectively. Because most of the herbarium vouchers of Sarcocornia lack seeds, and thus the permanency of seeds on plants is quite short, an alternative vegetative character as stomata was also studied using SEM and light microscope. The detailed stomatal morphological features were analysed for the first time to infer taxonomic relationships among these perennial halophytes. Stomata are constantly paracytic for the four studied species, but two morphotypes were identified related to the level arrangement of guard cells, named as sunken and prominent stomata, respectively. Sunken stomata are found in A. macrostachyum, H. strobilaceum and S. fruticosa, whereas prominent stomata are only characteristic for S. perennis. Besides, we have found somehow relationships among the ecology of these plants based on the frequency of floodings and the observed morphology and density of stomata. Nonetheless, each observed morphotype is a stable character, without any influence of climate or environmental conditions. As a result, seminal coat ornamentation and stomata morphology are the two most reliable characteristics for discriminating Mediterranean halophytes taxa, especially related to Sarcocornia taxa.     

Action of growth regulators on the cotyledonary stomata ofCucumis sativus L.: Structure and ontogeny

Anomocytic stomata and stomata with single subsidiary cells are commonly observed Sometimes a stoma appears anisocytic. Double cytoplasmic connections between nearby stomata and division of guard cells with persistent or degenerating nuclei are seen in GA. One or more divisions of guard cells, displaced guard cells and single guard cells with or without pore are noticed in SUC. Formation of single guard cells is a common feature in TIBA. Paracytic stomata, one and a half stomata and persistent stomatal initials are seen in SUL. COUM seems to be not inhibitory inCucumis sativus. In COL stomata with unequal guard cells, unequal stomatal cells with thickening in between but without intervening pore, stoma with double pores, persistent stomatal initials which may be solitary or in groups with varying shapes and with one or two nuclei of different shapes are noticed. The growth regulators affect the frequency of stomata, epidermal cells; stomatal index; size of guard and epidermal cells.