Chapter –II

MORPHOLOGY AND ANATOMY

MORPHOLOGY

Introduction

A critical survey of concerned literature has been made on Abutilon

ranadei of the world in general and India in particular, in the beginning

of the proposed work during 2008 and then updated in consecutive years.

Detailed information on Abutilon ranadei in Maharashtra, its distribution,

status, diagnostic features etc. was collected through referring systematic

account of the species in various districts, state and National Floras and

research publications. A critical note on the species was prepared by

consulting literature and herbarium at Botanical Survey of India, Western

Circle, Pune and BAMU Herbarium, Department of Botany, Dr.

Babasaheb Ambedkar Marathwada University, Aurangabad (M.S.).

Geographical Distribution

Abutilon ranadei is a Critically Endangered shrub with great

potential as an ornamental; it is restricted to Maharashtra State in Western

India. Where it occurs at 600-1,200 m above sea level, between 16.4 –

19˚ N and 72 – 74˚ E on the crest line of the Western Ghats. It is

restricted to highly fragmented populations in ten localities, in moist

deciduous forest on hill slopes, especially in thickets or stands of

Strobilanthes callosa (known locally as ‘karvi’), but some localities of

this species grown in tree shade (Shelimb Pune district) or open place,

road side (Kas plateau). Detailed information of these localities is

27

represented in (Table-1 and Graph-1). From these localities, seed, stem

cuttings, floral buds, nodal segments of Abutilon ranadei and also soil

samples were collected for the investigation. Table-1: Localities in Western Ghats from which Abutilon ranadei collected.

Sr.

No.

Date Locality Field No. No. of

Individ

ual.

Longitude Latitude Altitude

1 30/11/2008 7702 Kas Plateau

Satara

01 E17047’89” N17043’34” 2222 ft.

2 02/12/2008 7704 Amba Ghat

Ratnagiri

15 E16058’80” N73046’78” 2029 ft.

3 11/01/2009 7712 Rajgad 20 E73044’37.5 N18018’55.3” --

4 15/01/2009 7707 Torana fort

Pune

23 E73037’39.3” N18016’77.3” 3897 ft.

5 26/01/2009 77011 Torana fort

Pune

175 E73037’39.3” N18016’77.3” 3897 ft.

6 15/09/09 7722 Ghotne

Plateau

75 E 073044.732” N 17004.521” 2409 ft.

7 14/08/2010 7751 Amba Ghat

Ratnagiri

50 E17046’638” N17059’769” 2003 ft.

8 13/02/2011 7759

7761

Velhe Pune 60

30

E 73037’440”

E 73037’427”

N 18016’763”

N 18016’768”

3885 ft.

3930 ft.

9 26/02/2011 7762 Dongerwadi,

Pune

30 E73026’ 399” N18037’ 419” 1844 ft.

10 26/02/2011 7765 Shelimb, Pune 45 E73026’611” N18037’533” 1881 ft.

Species Information

Scientific Name : Abutilon ranadei Woodr. & Stapf.

Common name(s) : Ghanti Mudra, Son-Ghanta (in Marathi)

Conservation Status : Critically Endangered (CR) according to

IUCN Red List criteria.

Habitat : Edges of moist deciduous forest.

Key Uses : Potential ornamental.

28

Known hazards : None known.

Taxonomy (Bentham & Hooker 1862; Wafaa, 2009)

Division : Phanerogamus

Sub-Division : Angiosperms

Class : Dicotyledons

Subclass : Polypetalae

Series : Thalamiflorae :

Order : Malvales

Family : Malvaceae

Genus : Abutilon

Species : ranadei Woodr. & Stapf.

Abutilon ranadei was first collected at Amba Ghat in the Kolhapur

District of Maharashtra State by Namdeorao B. Ranade, sometime

Keeper of the Herbarium at the College of Science, Pune Punekar (2001).

Kew botanist Otto Stapf and G. M. Woodrow described it as a new

species in 1894 and named it in honour of Mr. Ranade. Because of its

narrow range and extreme rarity Woodrow (1894). In the past, it has been

rated as Endangered or even presumed extinct, and recently been

assigned to the Critically Endangered category. In addition to its type

locality, A. ranadei has now been collected in 10 new locations: in Pune

district (Shelimb, Dogarwadi, Rajgad, and Torna Fort), Satara district

(Kas plateau and Vasota Fort), Ratnagiri district (Amba ghat and

Gothne), Sindhudurg (Amboli) Districts of Maharashtra State and

Kolhapur district (Radhanagari) and morphological studied of different

localities (Table- 2).

29

Abutilon ranadei Woodr. & Stapf. In Kew Bull.1894:99.1894. Cooke, Fl.

Pres. Bombay 1: 101. 1958 (Repr.); Almeida & Almeida J. Bombay Nat.

Hist. Soc. 86: 478-479. 1989; Paul in Sharma et. al. Fl. India 3: 273.

1993; Almeida Fl. Maharashtra 1: 101-102. 1996; Bachulkar & Yadav J.

Bombay Nat. Hist. Soc. 94: 591-592. 1997; Venkanna & Das Das in

Singh et. al. Fl. Maharashtra St. Dicot. 1: 299. 2000; Punekar et. al. J.

Econ. Tax. Bot. 25: 261-263. 2001; Sardesai & Yadav Fl. Kolhapur 68.

2002. Tetali et. al. J. Bombay Nat. Hist. Soc. 101: 3. 344-352. 2004;

(Photo plate No. -2. a-k)

It is a small sized tree or shrub, height measuring 2.5-3.5 m high.

The vegetative plant parts bear star-shaped hairs. Leaves are simple, long

petiolate, stellately hairy on both sides, ovate to rounded-ovate, with tips

that taper to a point, heart-shaped bases and scalloped to toothed margins.

Flowers are solitary, axillary, large, campanulate in shape and about 2.5

cm in diameter. The calyx is bell-shaped, with lobes free up to the

middle. The size of the calyx is 3.5 x 1 cm, and it is covered with glands

and star-shaped hairs. The corolla is bell-shaped with pale purple petals

with orange-yellow tips and size of the corolla is 4.7 x 1.5 cm. The petals

are twice as long as the calyx. Stamens are many and united to form

staminal column of 2-3.5 cm. It is hairless with purple lines. The

filaments are white with a reddish base, 3-5 mm long, and have dumbell-

shaped glandular hairs in the upper part. The anther lobes are kidney-

shaped and are initially green, turning dark rose at maturity and

brownish-violet at dehiscence. The five carpels have a sharp point and are

densely hairy throughout (style). There are five styles, which are up to 7

mm long and sparsely hairy. The fruits are schizocarpic (split into a

number of seed-containing parts). Capsules are few seeded.

30

Fls. and Frts: – September to March

Exsiccata: –SAS; 7702, 7704, 7707, 7711, 7722, 7751, 7759, 7761, 7762

and 7765

Localities: Torna, Rajgad, Shelimb, Dongarwadi, Kas plateau, Vasota,

Amba ghat and Ghotne.

Distribution: – Kolhapur; Pune; Satara; Ratnagiri and Shindudurg

Flowering and pollination

Flowering begins in early November and continues until the end of

March. During the pollination stage the glandular hairs on the calyx tube

emit a strong odour and secrete nectar from the nectaries, which are

located at the base of the petals. These attract insects such as honey bees

(Apis mellifera) and certain fast-moving Dipteran flies, which are most

likely to be the pollinators. The time of flower anthesis seems to be

temperature and light dependent. The flowers are protandrous and anther

dehiscence takes place just prior to flower opening but this protandrous

condition is not so pronounced. This protandrous condition is observed in

most of the members of the family Malvaceae (Faegri & Pijl, 1980;

Dawar et. al., 1994). Though the upper anthers mature first and till the

maturation of all anthers stigmas also become mature and. Mainly Bees

(Hymenoptera) and Butterflies (Lepidoptera) are found to visit the

flowers of Abutilon ranadei. Api ssp. (Honey bee) seems to be

responsible for pollination either by bringing the stigmas near to the

anthers or by transferring pollen grains from their body parts which are

adhered while visiting the flowers. Similar type of pollination was

observed by Gottsberger (1967) on some Brazilian genera of Malvaceae

and Dawar et. al., (1994) on Sidaovata complex. Breeding experiments

revealed that A. ranadei is self compitable and facultative autogamous

31

taxon and no significant difference was found in fruit set of open

pollination and autogomy Rubina et. al. (2010), Rubina, (2010). While,

fruit setting was significantly reduced in geitonogamy and xenogamy.

Besides this, pollen/ovule ratio also supports the facultative autogamous

nature of the species. Thus in A. ranadei both selfing and insect mediated

crossing occur.

Abutilon ranadei faces both man-made and natural threats.

Anthropogenic threats include the periodic harvesting of firewood from

the edges of the forests where it occurs, forest fires, and the collection of

Strobilanthes callosa stems for house-building and agricultural practices,

which disturb the habitat of this dwindling species. A. ranadei also faces

natural pests such as tropical red spider mites, striped mealy bugs,

cabbage semi-loopers, aphids, purple scale insects, leaf miners and snails.

Amongst the known natural pests, Mealy bugs present the most

common threat (Photo plate No. 3.).

Previous literature are studied on reported by hamming bird feeder

of Abutilon (Catlin, 1976), and some Pest and diseases Abutilon ranadei

by Tetali et al. (2004),

1. Tetranychus cinnabarinus (Tropical Red Spider Mite)

Family: Tetranichidae

Description: An oval shaped mite. Tiny red or greenish with four

pairs of legs. Polyphagus, common serious pest of greenhouse plants and

other cultivated crops.

Feeding habit: External feeder. All stages of insects feed on the

lower side of the leaf surface.

Damage: Scarification, leaf silvering and appearance of yellow

patches.

32

Control: Biological:

Phytoseilus riegeli (Predaceous mite). Chemical: 1) Foliar spray of

Dimethoate (0.5 ml/liter), 2) Carbaryl (2 ml/liter) + Neemmarin (3 ml/

liter), 3) Sulphur (3 gm/liter), 4) Kelthane (2ml/liter).

2. Ferrisia virgata (Ckll.) (Striped Mealy bug)

Family: Pseudococcidae

Description: An ellip0tic shaped mealy bug with a pairof

conspicuous longitudinal submedian dark stripes, promounced long tail

and long glassy wax threads.

Feeding habit: The most serious polyphagous pest. Sucking type,

feeds on tender parts and leaves.

Damage: Wilting, infestation by sooty mouds growth retardation.

Control: Biological:

Crytoleumus monterouzuni (6 per 100 sq. m) Chemical: 1)

Malathiuon (1 ml/liter) + fish oil resin soap Azinphos-methyl 92 ml/liter).

3. Trichoplusia ni (Hb.) (Cabbage Semi-looper) Family: Noctuidae

Description: Green with a thin, white lateral line, and two white

lines along the middle of the back. There are two pairs of prolegs.

Feeding habit: Larva feeds on young leaves. Active at low

temperature, makes irregular holes in the leaf.

Damage: Irregular holes in the leaf lamina.

Control: Biological: 1) Bacillus thuringiensis, 2) Trichogramma

(parasitoid eggs 2000-3000 per 100 sq. m) Mechanical; Ultraviolet light

traps. Chemical: Carbaryl (2 ml/liter).

4. Unidentified (Aphids) Order: Homoptera Family: Aphididae

Description: A small soft-bodied, sluggish insect, with piercing

and sucking mouth parts.

33

Feding habit: Usually attacks tender parts. Feeds on the lower

surface of the leaves.

Damage: Leaf curling, Infection with sootymoud, presence of ants.

Control: Biological: 1) Ladibird (Coccinellidae), 2) Crysoptera

carnae (adults 400 per 100 sq. m), 3) Hymenopterous parasites.

Chemical: 1) Soap water, 2) A number of systemic insectisides

(Permethrin, Pirimicarb).

5. Chrysomphalus aonidum (L.) (Florida Red scale or purple scale)

Family: Diaspididae

Description: Adult female is purplish and circular, with a reddish-

brown boss or nipple in the center.

Feeding habit: Feeds on leaves, young shoots and twigs.

Damage: Saliva is toxic, causing necrosis.

Control: Biological control: Chilochorus nigritus Chemical: 1)

Carbary (3%), 2). Parathion (0.5%), 3) Malathiuon with white oil.

6. Unidentified Leafminer (Microlepidoptera)

Description: Minor pest. Tunnel leaf mine with no central line of

faecal pellets.

Feeding habit: Attacks during rainy season.

Damage: Leaf tunnels. Destroys the photosynthetic structure.

Control: Chemical: 1)Triozophos (Hostathion 3 ml/liter), 2)

Phosphamidon (1 ml/liter) + Fish resin oil soap (1 ml/liter)

7. Unidentified Snail. Phylum Mollusca

Description: Nil

Feeding habit: Nocturnal feeders. Feed on young leaves, flower

buds.

Damage: Minor damage. Young leaves are affected.

34

Control: Mechanical: Hand pick, Cabbage and Papaya-yellow

leaves for trapping Chemical: 1) 2-4% salt water, 2) Lime treatment

Although critically endangered in the wild, Abutilon ranadei can

be propagated by seed and vegetative propagation under nursery

conditions. However, the percentage of seeds germinating is very low.

Vegetative propagation methods such as air layering are more successful,

and this method is used at the Nauroji Godrej Centre for Plant Research

(NGCPR, Shindewadi), Satara District, Maharashtra State of India. A.

ranadei plants produced by air layering are then planted out into their

natural habitat in Maharashtra.

Uses: Abutilon ranadei is an attractive plant with showy, nectar-

producing flowers and could be cultivated as an ornamental

35

Table –2. Distributional variations on different localities of Abutilon ranadei Woodr. & Stafp. In Western Ghats

Sr. No.

Name of Character

Name of localities

Torna fort-1

Kas Rajgad Amba ghat

Torna fort -2

Shelimb-1

Sherval shrusti

Ghotane Shelimb-2

1 Distribution Rare Rare Rare Rare Rare Rare Rare Rare Rare

No. of individual

80 1 10 28 30 29 15 21 33

Habit Shrub Shrub Under shrub Shrub Shrub Small tree

Shrub Small tree Small tree

2 Hight 1-5 m 2-3 m 2-3 m 2-3.5 m 2-3.5 m

1-5 m 3-5 m 2-5 m 3-5 m

Stem White White Green Green Green Stem infected

Green Green white

Stem infected

Association Carvia callosus

No specific

Carvia callosu

Carvia callosus

Shade of tree

Cultivated Carvia & other

Shade of tree

Status of plant Well fruiting compare other

Single plant

Poor fruit setting

Poor fruit setting

Equal to Torna

Very poor fruit setting

Cultivated Poor fruit setting

Very poor fruit setting

No. of sites 3 1 1 3 2 1 1 2 1

36

ANATOMY

INTRODUCTION

Any report on the anatomical study on Abutilon ranadei not

appeared but a brief description about the genus is given in Anatomy of

Dicotyledons (Metcalf & Chalk, 1965), Esau (1977) and other species

Abutilon theophrasti anatomically studied root, stem and leaf (Aysegul,

2003; Yun and Taylor 2006), Abutilon indicum morphol anatomical

studies of leaves (Ramadoss et. al., 2012) and also some other genera of

family Malvaceae Anatomical description of Hibiscus (Adedeji and Dloh,

2004).

Several anatomical features are specific to specific taxa. Hence

these may be used for delimitation of the species. These anatomical

features having taxonomic values are used as criteria for separating the

species, genera and even families. The anatomy of plant gives the criteria

of epidermis, cortex, secondary phloem, medullary rays, crystals, fibres

and tanniniferous cells, which forms the important parameters in

standardizations.

Maceration is the separation of the cells which gives the idea of

complete cell regarding their size, shape etc. Maceration also gives the

idea regarding the cell inclusions like starches, crystals (raphids and

spherophides) and association of the cells. In anatomy, only the outline of

cell is known in transverse section. While maceration, complete and

isolated cells ready for the observation.

For the first time, the anatomical characteristics of the root, stem,

petiole and leaf of Abutilon ranadei were studied during the present

investigation.

37

MATERIAL AND METHODS

Anatomy and Maceration: The plants were collected from different

localities of Western Ghats, Maharashtra and authentically identified. The

exact location of the plant whose samples were collected is given in the

form of longitude, latitude and altitude. The date of their collection and

field numbers are also provided (Table No. 2). The samples were

collected by large knife, chisel and saw without damaging the plants. The

plant specimen was preserved in 70% alcohol for their maceration and

anatomical work. The sample were collected in polythene bags or zip

lock bags and brought to the laboratory within 2-3 days.

The morphological characters of the plant were studied in detail

and their herbarium sheets were prepared which are preserved in the

Herbarium of Department of Botany, Dr. Babasaheb Ambedkar

Marathwada University, Aurangabad. Fresh and dried plant samples were

studied morphologically in the field as well as in the laboratory.

The anatomical characters of the plant were studied with the help

of free hand transverse sections, taken with blades. From each part some

sections were unstained while others were double stained. Both unstained

and stained sections are permanently preserved. Permanent preparations

were observed under microscope. Photographs were taken with the help

of digital camera (Sony cyber) by micro photographic techniques. The

stem was also studied by maceration techniques. The pieces of stem were

boiled in Jeffery’s fluid (Chromic acid 10% and Nitric acid 10% in equal

proportion) as well as by Schultz’s method. The macerated cells were

studied in detail. Their photos were taken with the help of digital camera

(Sony cyber). The dimensions of the cells in sections and those obtained

during maceration were measured by ocular.

38

Dermatology

Dried leaves of representative specimens of the Abutilon ranadei in

Botany garden, Department of Botany, Dr. Babasaheb Ambedkar

Mrathwada University, Aurangabad were used for dermatological

studies. Leaf samples were prepared according to the Clark’s (1960)

technique as modified by Cotton (1974). Dried leaves were placed in

boiling water for few minutes until unfolded. The leaves were placed in a

tube filled with 88% lactic acid kept hot in boiling water bath for about

30 to 40 min. Lactic acid softens the leaf due to which it was possible to

scrap the leaf surface with sharp scalpel. Slides of both abaxial and

adaxial surface of leaf were prepared and mounted in clean 88% lactic

acid. Both qualitative and quantitative micro morphological

characteristics of foliar epidermis were observed using LM. Micro

histological photographs of both surfaces were taken by Nikon (FX-35)

camera equipped on light microscope. Basic terminology used in stomata,

trichome classification and description is that suggested by Harris and

Harris (2001), Inamdar (1983) and Distribution of stomata and its relation

to plant habit Raja et. al., (1981), Inamdar (1969a). However, simple self

explanatory terms are added to identify the specific types of trichomes

and stomata.

i) Trichomes: -

Tichomes are outgrowths of epidermal cells, (Roy, 2006). For

studies of trichomes following procedure was adopted-

1. Scrap the trichomes from leaf surfaces with the help of razor.

2. Stain trichomes in safranin and mount in glycerin on a slide.

3. Observe slide under microscope and mention the type of trichome.

4. Take the dimensions with the help of ocular micrometer.

39

5. Take Photograph.

6. Determine the observations for upper and lower epidermis separately.

ii) Stomata: -

Stomata are microscopic pores on the epidermal surface of aerial

parts of higher plants formed by a pairs of specialized epidermal cell

termed guard cells, which control opening and closing of the pore by

changing their turgidity and thus regulates the gaseous exchange between

plants and environment (Roy, 2006).

For studying stomata following procedure was adopted-

1) Peel out upper and lower epidermis separately by means of forceps.

Keep it on slide and mount in glycerin water.

2. Take Photograph.

3. Mention the type of stoma and occurrence of stoma (amphistomatic /

epistomatic / hypostomatic).

4. Measure the length of stoma, dimension of guard cell and dimension of

subsidiary cell with the help of ocular micrometer.

5. Determine the values for upper and lower epidermis separately.

6) Record the result for ten fields and calculate the average number of

stomata per square mm.

iii) Epidermal cells: -

1. Peel out upper and lower epidermis separately by means of forceps.

Keep it on slide and mount in diluted glycerin.

2. Take Photograph.

3. Record the nature and outline of epidermal cells.

4. Determine the values for upper and lower epidermis separately.

5. Record the result for each of the ten fields and calculate the average

number of stomata per square mm.

40

iv) Leaf constants / quantitative microscopy: -

The leaf constants or dermatological characters of leaves like

stomatal number, stomatal index, palisade ratio, vein-islet number, vein

termination number, and trichomes were studied.

a) Determination of stomatal number:-

Definition: It is average number of stomata per square mm of the leaf.

Procedure:

1. Peel out upper and lower epidermis separately by means of forceps.

Keep it on slide and mount in glycerin water.

2. Place the slide with epidermal peel on the stage.

3. Count the stomata present in the area of 1 mm square. Include the cell

if at least half of its area comes within the square.

4. Record the result for each of the ten fields and calculate the average

number of stomata per square mm.

b) Determination of stomatal index:-

Definition: Stomatal index is the percentage, which the number of

stomata forms to the total number of cells, each stoma being calculated as

one cell. Stomatal index can be calculated by using following equation.

S

I = ----------- x 100

E + S

I= Stomatal index, S= No. of stomata per unit area, E= No. of epidermal

cells in the same unit area

Procedure:

1. Peel out upper and lower epidermis saperately by means of forceps.

Keep it on slide, mount in glycerin water and observe under microscope.

41

2. Draw the diagram with help of camera lucida by drawing square of 1

mm by stage micrometer.

3. Count the number of stomata, also the number of epidermal cells in

each field.

4. Calculate the stomatal index using above formula.

5. Determine the values for upper and lower epidermis separately.

6. Record the result for each of the ten fields and calculate the average

stomatal index for both epidermises separately.

c) Determination of palisade ratio: -

Definition: The palisade ratio is the average number of palisade cells

beneath one epidermal cell of a leaf. It is determined by counting the

palisade cells beneath four continuous epidermal cells.

Procedure:

1. Clear a piece of leaf with 10 % KOH solution.

2. Trace off the outlines of four cells of the epidermis with the help of

camera lucida.

3. Then, focus down to palisade layer and trace off sufficient cells to

cover the tracings of the epidermal cells. Complete the outlines of those

palisade cells, which are intersected by the epidermal walls.

4. Count the palisade cells under the four-epidermal cells. (Include the

palisade cell in the count when more than half is within the area of

epidermal cells and exclude it when less than half is within the area of

epidermal cells.)

5. Calculate the average number of cells beneath a single epidermal cell;

this figure is the palisade ratio.

6. Repeat the determination for ten groups of four epidermal cells from

different parts of leaf. This average is the palisade ratio of the leaf.

42

d) Determination of vein-islet number:-

Definition: A vein-islet is the small area of green tissue surrounded by

the veinlets. The vein-islet number is the average number of vein-islets

per square millimeter of a leaf surface. It is determined by counting the

number of vein-islets in an area of one square millimeter of the central

part of leaf between the midrib and margin.

Procedure:

1. Clear a piece of leaf with 10 % KOH solution.

2. Draw a square of 1 mm by stage micrometer.

3. Place the slide with cleared piece of leaf on the stage.

4. Trace the veins, which are included within the square with the help of

camera lucida.

5. Count the number of vein-islets in the square millimeter. Where the

islets are intersected by the square, include those on two adjacent sides

and exclude those islets on other sides.

6. Record the result for each of the ten fields and calculate the average

number of vein-islets in an area of one square millimeter.

e) Determination of vein termination number: -

Definition: Veinlet termination number is defined as the number of

Veinlet terminations per square millimeter of leaf surface midway

between the mid rib and margin.

Procedure:

1. Clear a piece of leaf with 10 % KOH solution.

2. Draw a square of 1 mm by stage micrometer.

3. Place the slide with cleared piece of leaf on the stage.

4. Trace the veins, which are included within the square with the help of

camera lucida.

43

5. Count the number of vein terminations in the square millimeter.

6. Record the result for each of the ten fields and calculate the average

number of vein terminations in an area of one square millimeter.

Result

Dermatology of leaves

1)Trichomes :-

Leaflet shows-presence of unicellular trichomes (133.28 to

399.81µ long) on both surfaces but more common on lower epidermis

(Photo plate No. 4g; Table-3).

Table – 3a. Diversity of foliar trichomes of Abutilon ranadei Woodr. &

Stapf.

2) Stomata :-

The stomata are anomocytic, hypostomatic, with stoma length

23.32 µ (average) and 16.65 to 29.97 (range). The average size of guard

cell is 16.65 X 4.995µ and range is between 13.32 X 1.665 to 19.98 X

Type of Trichomes Description

Stellate More than two ray cells held together in the same

cell cavity, quite variable in number of ray cells

and their relative length and thickness.

Flask-Shaped Three diverse morphological are identified Type-I.

Unicellular, elongated, basal portion slightly

dilated gradually narrowing upwards with an

apical opening. Type-II. Basal swollen portion

multicellular, with unicellular neck like portion

having an apical opening.

44

8.325µ. Subsidiary cells are wavy in outline with average cell size 32.96

X 21.30 µ and range between 29.97 X 18.31 to 36.63 X 24.97 (Table-5).

3) Subsidiary cells: -

The epidermal cells near guard cells are termed as subsidiary cells. It

determines type of stoma (Metcalfe and Chalk 1950; Roy 2006). Shape,

size and number of subsidiary cells can be used for standardization.

4) Epidermal cells: -

In surface view the upper epidermal cells (average cell size 45.78

X 28.13µ, range 43.29 X 29.97 to 59.94 X 36.63µ) are slightly bigger in

size as compared to lower epidermal cells (average size 45.78 X 28.13µ,

range 43.29 X 24.97 to 49.95 X 31.63µ). Epidermal cells are wavy in

outline with irregular shape (Photo plate No. 4e-f; Tables-5).

The details on micro morphological features of the foliar epidermis

of Abutilon ranadei may serve as a useful taxonomic tool to delineate the

species studied (Photo plate No. 4e-f; Table-5).

The vein islet number: Abutilon ranadei, mean value was 21.8, range

was 20 -23. There was a large difference in the value of vein islet number

of the species of Abutilon. Therefore we can use as a criterion to

differentiate the species on the basis of vein islet number. (Photo plate

No. 4e-f; Table-4).

Veinlet termination number: The veinlet termination number of

Abutilon ranadei mean value was 5.4 and range was 5-6. There is a large

dfference in the leaflet termination number of species of Abutilon ranadei

therefore we can use this criterion to delimit the taxa on the basis of

veinlet termination number. (Photo plate No. 4e-f; Table -4).

45

Table – 4. Vein islet number of Abutilon ranadei Woodr. & Stafp.

Sr. No. Vein islet number Vein termination no.

1 21 6 2 22 5 3 23 6 4 20 5 5 21 6 6 23 5 7 23 5 8 22 5 9 22 6 10 21 5

Mean 21.8 5.4

Range 20-23 5-6 SE 0.33 CD 0.74

46

Table – 5. Dermatology of Abutilon ranadei Woodr. & Stapf.

Sr.No. Epidermal Cell Stoma

Guard Cell

Upper µm Lower µm Upper µm

Lower µm

Upper µm Lower µm

Length µm Width µm Length µm Width µm Length µm

Width µm

Length µm

Width µm

1 37 27 39.5 30.5 15.5 15.5 20.5 9.5 5 24.5

2 39.5 25.5 37 27 20.5 19 23 7 23.5 13

3 37 25.5 30 25.5 20.5 20.5 23 9.5 23 15.5

4 39.5 25.5 37 25.5 23 15.5 25.5 7 20.5 13

5 37 25.5 37 23 20.5 15.5 23 9.5 23 9.5

6 30.5 23 27 20 23 19 25.5 9.5 23 9.5

7 30.5 23 255 190 23 15.5 25.5 13 25.5 13

8 37 23 30 23.5 15.5 20.5 25.5 9.5 23 13

9 30.5 25.5 37 20.5 15.5 20.5 20.5 13 25.5 15.5

10 30.5 23 39.5 27 23 15.5 20.5 9.5 23 13

Range 30.5-39.5 23-27 27-39.5 20-190 15.5-23 15.5-20.5 20.5-25.5 7-13 5-25.5 9.5-

24.5

SE 1.24 0.47 22.06 16.56 1.04 0.75 0.69 0.64 1.89 1.33

CD 2.79 1.07 49.85 37.42 2.35 1.70 1.56 1.44 4.27 3.02

47

Table -6. Quantitative foliar epidermal features of investigated taxa of Abutilon ranadei (Stomata)

Table -7. Quantitative foliar epidermal features of investigated taxa of Abutilon ranadei (Trichomes)

Sr. No. Epidermal features Measurment Surface

1 Ordinary Epidermal Cells 29.5 (32.3 + 2.83) 40 x 18 (24.3 + 2.13) 29.5

Adaxial

2 L. x W. µm Min. 27 (30 + 2.24) 27 x 15 (22.6 + 2.87) 33

Abaxial

3 Stomata L. x W. µm 23 (20 + 0) x 12.5 (13.8 + 0.56) 17

Adaxial

4 Min. (Mean ± S.E) Ma. 23 (23 + 0) x 16 (16.6 + 0.24) 17

Abaxial

5 L. Sto. opening µm 17 (18 + 0.45) 19 Adaxial 6 Min. (Mean ± S.E) Ma. 17 (18 + 0.45) 19 Abaxial 7 Stomatal Complex L. x W. µm 33 (43 + 2.74) 48 x

22 (27.2 + 2.24) 33 Adaxial

8 Min. (Mean±S.E) Ma. 33 (40.6 + 2.74) 51 x 25 (28.4 + 1.57) 33

Abaxial

Flask-Shaped H. x W. µm Stellate L. x W. µm Min. (Mean ± S.E.)Ma. Min. (Mean± S.E.) Ma. Type-II S.c.r. N.r.c. 123(138.8 130 (250.72 ±35.94)375 x more than 5 ±3.3140)x22 25 (25.5 ±1.12) 27 (23.8±1.0)27

48

A. Lamina

1. Leaf Anatomy:

The leaf is bifacial. The transverse section of the leaf exhibited the

following Characters:

i) Epidermis: The upper epidermis consists of a single layer of

rectangular cells with a fairly thin, smooth cuticle. Numerous hairs and

stellate glandular hairs of different types cover it. The covering hairs are

generally tufted with straight walls and acute apices. Glandular hairs can

be differentiated in two types the long ones, with a unicellular stalk a

multicellular glandular head; the short ones with multicellular stalk and

glandular head. The former have a globe-like composed of 10-12 layers

of cells. The latter with two celled stalk and 2-3 celled head are similar to

the ones of the leaf of other Althea, other species of Abutilon and Malva

species Warszawa et al. (2006) and Nighat et al. (2010).

Stomatal number is approximately same on both epidermises.

Small epidermal cells and 2-4 subsidiary cells around the stomata were

observed (Photo plate No. 4 and Table-5).

II) Mesophyll: The mesophyll is clearly differentiated into palisade and

spongy parenchyma. Under upper epidermis, the mesophyll contains 2

layers of palisade which is composed of compactly arranged long

cylindrical cells. The spongy mesophyll, being thinner than the palisade

layer, is formed of thin walled, isodiametric paranchymatous cells with

few intercellular spaces. Mucilaginous cells are observed generally in the

palisade and occasionally in the spongy parenchyma structure. Numerous

cluster crystals are scattered in the mesophyll (Palisade has mostly the

bigger crystals) and they are the most characteristics elements of the

mesophyll (Photo plate No. 4).

49

B. Midrib

The upper and lower epidermis of the midrib is similar to that of

lamina except that the cells are similar. Trichomes and glandular hairs are

also densely observed on the midrib. Under the upper epidermis a

projecting prominent part, consisting of 7-8 layers of collenchymatic

cells, is observed as the most striking characteristics of the leaf. Under

this part, palisade parenchyma is suddenly interrupted. Between this

prominence and the vascular bundle, parenchymatous cells and a big

mucilaginous cell can be observed.

A crescent-shaped vascular bundle is present in the center of midrib. The

vascular bundle contains 2-4 layers of llignfied radiating xylem with an

arch of phloem consisting of thin walled, compactly arranged, small cells.

The rest of the midrib, composed of parenchymatous cells, contains

starch starch grains, cluster crystals eith rare mucilage.under the midrib,

close to lower epidermis groups of corner – collenchyma cells are also

observed (Photo plate No. 4).

2. Petiole

The upper and lower epidermis of the petiole is similar to that of

lamina except that the cells are similar. Trichomes and glandular hairs are

also densely observed on the midrib. Under the upper epidermis a

projecting prominent part, consisting of 7-8 layers of collenchymatic

cells, is observed as the most striking characteristics of the leaf. Under

this part, palisade parenchyma is suddenly interrupted. Between this

prominence and the vascular bundle, parenchymatous cells and a big

mucilaginous cell can be observed.

A crescent-shaped vascular bundle is present in the center of petiole. The

vascular bundle contains 2-4 layers of llignfied radiating xylem with an

50

arch of phloem consisting of thin walled, compactly arranged, small cells.

The rest of the midrib, composed of parenchymatous cells, contains

starch starch grains, cluster crystals eith rare mucilage.under the midrib,

close to lower epidermis groups of corner – collenchyma cells are also

observed (Photo plate No. 4c).

A crescent-shaped vascular bundle is present in the center of

midrib. The vascular bundle contains 2-4 layers of lignified radiating

xylem with a rich of phloem consisting of thin walled, compactly

arranged, small cells. The rest of the petiole, composed of

parenchymatous cells, contains starch grains, cluster crystal with rare

mucilage. Under the midrib, close to lower equipment groups of corner-

collenchyma cells are also observed (Photo plate No. 4c).

3. Stem

A transverse section of the stem is somewhat rounded and exhibits

the following chracters.

i) Epidermis : Epidermis is composed of a single layer of isodiametric

cells with convex outer and inner walls. Cuticle is thin and smooth

.Glandular hairs are similar to those of the . leaf with respect to form and

abundance. Cluster crystals, generally scattered all over are occasionally

found under the epiderimis as uniseriate lines .Epidermis o stem also

contains many of glandular and covering hairs, E glandular hairs are

generally observed as unicellular and simple, or bicellular and clustered

Number of multicellular tufted hairs is less than moocellular and

bicellualr ones (Photo plate No. 4b).

ii) Cortex : Adjacent to the epidermis, the cortex contains a thick layer of

collenchyma cells. The remainder of cortex is composed of 5-6 layers of

parenchyma cells of different sizes. Cortical parenchyma, a thinner layer

51

than collenchyma, contains cluster crystals in the cells or in the cortex

(Photo plate No. 4b).

iii) Vaseular cylinder : Underneath the cortical parenchyma,6-7 layers of

phloem sclerenchyama, consisting of more or less a complete ring of

fibres, which are sometimes interrupted by the rays are present. The

phloem is composed of crushed , irregular cells. Between the phoem and

xylem layers, cambium with 5-6 layers of thin walled, crushed,

rectangular cells are clearly observed.

Xylem contains a contunuos ring of 11 -12 layers of cells.

Tracheae with large central spaces are few, but tracheids with small

central spaces are in compact groups of cells Medullary rays are either

uniseriate or biseriate.

The pith is composed of thin walled and large, rounded

parenchyma cells. Some of these cells are transformed into

mucilaginous cells .Some big cluster crystals are scattered also in pith

.Mucilaginous cells are less and small in cortical parenchyma but

numerous and big in pith parenchyma (Photo plate No. 4b).

4. Root

The root is composed of unicellular, rectangular, suberized ,

sometimes deformed epidermal cells. Some simple and 2-4 celld tufted

hairs are observed .Under this 2-3 layers of hypodermis is seen. A thick

layer of cortical parenchyma, consisting of thin walled rectangular cells

of different sizes is observed. The cells contain starch grains and cluter

crystals. In the cortex, phloem sclerenchyam makes a ring of triangle

towers which is sometimes interrupted by phloem parenchyma.

Under these characteristic sclerenchymatous structures, phloem is

observed with little and crushed cell groups. Adjacent to phloem,

52

cambium is well marked by 7-8 layers oc cells. Medullary rays are

forwarded into the cortical parenchyma forming triangles. The cells

contain either a few starch grains or none.

Xylem consists of radilly arranged vessels which nearly cover all of the

inner part of the root Tracheae with big central spaces are few. The

smaller tracheids area abundant The rays are composed of 2-3 cell –width

parts pith is not very broad and consists of rounded parenchymatous cells

containing starch grains and cluster crystals. Mucilaginous cells are also

present among the pith cells. But the number of the mucilaginous cells is

fewer with respect to those found in leaf and stem because of the

restricted area of the pith (Photo plate No. 4a).

In this study the anatomical structure of the root stem and leaf of

Abutilon ranadei the only species of Abutilon growing in Turkey were

investigated for the first time.

Maceration

Vessels:

Very small (25-50 µ mean tangential diameter) in Abutilon

ranadei, in irregular clusters and in radial multiples of 2 or 3; multiples

seldom more than 20 per sq. mm. spiral thickening in small vessels of the

species observed. Perforations simple. Inter vascular pitting alternate;

small to minute; pits to ray and wood parenchyma typically similar to

intravascular pitting but some times simple and with some horizontally or

obliquely elongated pits in the species. Mean member length 0.2-0.7 mm,

mostely 0.33-0.45 mm (Photo plate No. 5).

53

Parenchyma:

Rather scanty to abundant; vasicentric to slightly aliform in the

species. Terminal parenchyma present. Strands most commonaly of 2-4

cells.

Very variable on type, ranging from a) high multiseriate rays

composed mainly of narrow upright cells, to there with numerous

uniseriate rays, to b) large homogeneous rays with few uniseriate, or c)

short, heterogeneous, storied rays. High multiseriate rays relatively few

uniseriates, both upright cells tend to be more nearly square; up to 4-9

cells wide, commonly with tendency to be of 2 distinct sizes in woods

with larger rays. 12-15 per mm; markedly heterogeneous, with square or

upright cells intermingled with procumbent cells tending to in groups

(Photo plate No. 5).

Fibers:

Typically with small simple pits. Commonly storied in woods in

which the parenchyma is distinctly storied. Mean length 0.36-2.33 mm;

usually of medium length 0.9-1.6 mm (Photo plate No. 5).

Tracheids

Tracheids were found which was pitted, wall ruptured and

measured 42 µ. Pitted vessels rounded at both end and measured as 57 µ.

Tracheids were pitted and measured 35 µ. Tracheids were reticulate and

measured 35 µ. Fiber tracheid, septate lumen present and measured 54

µ.On fiber tracheid, pits elongated, wall ruptures and measured 30

µ(Photo plate No. 5).

54

DISCUSION

Root anatomy has the characteristic layers of the dicotiledoneous

plants the most characteristic feature being phloem sclerenchyma in

cortex. Parenchymatous cells of cortex and the pith are rich in starch

grians and clustered crystals like the elements of the root of Altheae

species. Moreover, 1-2 mucilaginous cells are present in the pith.

Leaf anatomy is very similar to that of Malva sylvestris; another

member of the Malvaceae family (Yazgan et. al., 1986). But it can be

differentiated from Malva sylvestris by its 2-yalered palisade

parenchyma with big clustered crystals, mucilaginous cells and

multicellular, long glandular hair. Nether the leaf of Althea nor Malva

have these kinds of glandular cells (Baytop, 1981). However, the

characteristic elements of Malvaceae, such as short, multicellular

glandular hairs, simple unicellular and multicellular tufted eglandular

hairs, tufted crystals and mucilaginous cells are observed in this species.

Clustered crystal and simple and clustered eglandular hairs are

observed in all of the studied organs of the plant. Multicellular and long

eglandular hairs of leaf and the stem are interesting. The towers of

phloem sclerenchyma in the root are the characteristic elements to the

plant.

55