❖ Leaf primordial meristem already contains procambial

cells
❖ Note early development of procambium at the leaf base
(leaf trace)

Median longitudinal section through the growing point of Carya Buckleyi var, arkansana, showing an early
developmental stage of a foliage leaf. Note that derivatives of both the tunica (f) and corpus (cs) are present in
the primordium. Legend: arb, primordium of staminate catkin bud; c, cortex of internode; fl, foliage leaf
primordium; p, pith of stem; pc, procambial tissue; sc, portion of bud scale. (x900).

❖ Leaves emerge from groups of founder cells (immediate

precursors of the leaf primordia) that originate from the
peripheral zone of the shoots meristem
❖ The newly-formed leaf primordium consists of about 100
cells, in full grown leaf estimated to be 130,000. Every one
of the primordium cells divides about 10 times throughout
leaf development.
❖ The total number of cells involved has been estimated:
about 100 in cotton & Impatiens, to 250 in maize, 30-100 in
Arabidopsis.

Corn SAM
Apical meristem is elevated above organogenic region so apex
does not undergo plastochronic changes

Development of Leaf Axis

Longitudinal section of a shoot apex of maize (Zea mays), a panicoid grass, with
a one-layered tunica. Parts of each leaf occur on both sides of the axis because
the leaves encircle the stem in their growth
Phyllopodium Plate Meristem
- undifferentiated leaf primordium, leaf axis without blade and - A meristematic tissue (plate meristem) consisting of parallel
petiole layers of cells dividing anticlinally.
Apical meristem = primordium elongation - Brings about increase in surface area of the leaf.

❖ Cell division and cell growth in young primordium forms

primordial leaf axis or phyllopodium (has dorsiventral
symmetry, will become petiole and midrib)
❖ Phyllopodium increases in thickness and forms marginal
meristem on either side

❖ Marginal meristems (around 2 or more layers thick) outer

layer (like protoderm), short-lived
- Marginal meristem
= lateral extension that produces the 2 panels of blade
- B & C growth of blade initiated by marginal meristems,
initiation of major veins and elongation of petiole
- D. Cross section of blade showing position of adaxial
and marginal meristem

Origin of leaf primordium in flax

A-D longitudinal sections, E-H cross sections

A,B emergence of primordium thru enlargement and periclinal
Generalized diagram for leaf growth in a dicot. Apical growth (increases length), divisions of subsurface cells
marginal growth (increases width). C,D upward growth
E. primordium before initiation of blade
❖ Leaf lamina forms as outgrowth resulting from
F-H growth of blade: products of marginal meristematic
cytokinesis in marginal meristems
activity forming plate meristem with intercalary anticlinal
❖ Division concentrates on certain regions (depends upon
division predominating
the species), leads to the typical shape of the dicot leaf
blade
Expansion and Secondary Morphogenesis
Young leaf continues growth and differentiation then achieves
Lobed Leaves
mature size and form. Leads to increase in surface area and
- Development of lobed leaf is a result of differential activity
volume.
of leaf margin
- cells that comprise the mature leaf are formed
- In regions of future lobes, meristematic activity is
- mesophyll, provascular tissues, system of veins
considerable, while in the regions of future sinus there is little
develop
or no activity
- characteristic margin develops
- characteristic lobing develops

Development of provascular strands

After cessation of marginal growth, further growth of the
lamina is brought about by cell division in the various layers of
the lamina.
Development of lamina in Malus pumila Development of Procambial Strands
❖ Precedes development of lamina
❖ Acropetal direction

A & B 1st and 2nd leaf show marginal meristems developing

along ventral flanks of midrib
C & D 3rd and 4th leaf, midrib and veins are differentiating

Origin of internal layers of leaf

Longitudinal section of primordium showing provascular strand of midrib

Vein Development
- Polar transport of auxins possibly from leaf margins
influences differentiation of provascular tissue and ultimately
mature veins

A. Subepidermal initial from which B has arisen

❖ Anticlinal division of cells in the epidermal and

subepidermal layers.
❖ New cells that are added to the different layers of the
lamina originate from rows of marginal and submarginal
initials.

Tissue differentiation in the leaf

Development of veins containing vascular tissues

Last order veins

enclose small regions
of parenchyma called
areoles into which
extend one or more
vein endings

a. In dicots, venation is initiated by acropetal differentiation of

a central provascular strand in the phyllopodium.
Development of mesophyll and provascular strands b. As lamina expands, various branch orders of venation
system differentiate in a hierarchical manner with 2nd order
veins developing first, preceded by second order provascular
strands.
c,d. 3rd, 4th and higher order veins develop beginning in apical
regions of leaf and proceed basipetally.
 ❖ Leaf arises as a dorsiventral protuberance that grows in
height and width. Grow more actively on abaxial side than
on adaxial side
❖ Curves toward the apical meristem
❖ Procambium present before leaf primordium has
emerged as protuberance.

Stages in the ontogeny of a monocot leaf

- Piece of tobacco leaf to show minor vein

- The ultimate branching of veins in reticulate venation
patterns form meshes delimiting small areas called AREOLES A1, A2. Emergence of protuberance on the side of the apical
meristem
Development of veins in a monocot leaf B,C,D extension of protuberance laterally around meristem to
form a collar or sheath.
E. Enclosure of apical meristem and elongation of "blade" and
sheath by intercalary meristems

e. Initial "midvein" or initial provascular strand differentiates

acropetally and basipetally in the base of the leaf primordium.
f. provascular strands of large veins first differentiates
acropetally, then basipetally.
g. intermediate veins are initiated in the apical region of the
young leaf and differentiates basipetally.
h. small longitudinal and transverse veins are formed in the
Stages of leaf development in Arabidopsis
apical region of the leaf with transverse vein
formation proceeding basipetally.

Summary of early leaf development

Diagrams illustrating stages of leaf development in Arabidopsis. (A) Leaf

primordium. (B) Blade morphogenesis associated with marginal meristem
activity (blue). (C) Blade expansion associated with plate meristem activity
(yellow). (D) Cell pattern formation and cell differentiation within tissue layers,
including stomatal and vein precursors (red)
Genes involved in acquisition of adaxial-abaxial asymmetry
❖ Adaxial] PHANTASTICA (PHAN) in Antirrhinum
- PHANTASTICA (PHAN)
- ARGONAUTE1(ARG1)
- PINHEAD (PNH)
- PHABULOSA (PHAB)
❖ Abaxial
- YABBY gene family
- FILAMENTOUS FLOWER (FIL)
- YABBY2 (YAB2),
- YABBY3 (YAB3)

PNH and FIL expression during leaf initiation. The distribution of PNH and FIL
mRNA, respectively, is indicated in shades of red. (a) PNH is expressed
uniformly across the Il site and becomes restricted to adaxial leaf tissue by the
P2 stage. PNH is also expressed in developing vascular tissue. (b) FIL is
expressed uniformly across the I2 and Il sites, and is restricted to abaxial leaf
tissue by the Pl stage. Note that in a real Arabidopsis apex, the leaf primordia
form a spiral so that 12, 11, Pl and P2 cannot be captured in a single cross
section.
 ❖ SPCH - speechless: drives first asymmetrical division
that initiates stomatal development
❖ MUTE - acquisition of GMC identity
❖ FAMA ("Goddess of Rumor)" - promote final guard cell
differentiation
Abscission
- active separation of a leaf, flower, fruit from a stem without
injury
- the site where this happens is called the abscission layer
- High levels of auxin keep leaf tissue in an ethylene
insensitive state, but as auxin levels drop, the abscission-
promoting and auxin-repressing effects of ethylene become
stronger

Hypotheses for the Generation of the Stomatal One-Celled

Spacing Pattern. Hypotheses tested by analysis of dental resin
impression series
 Trichome Development

Schematic representation of key stages of stomatal development in Arabidopsis

thaliana. The stomatal lineage initiates with the entry division - postprotodermal
cells (grey) convert to meristemoid mother cells (blue) that undergo an
asymmetric division to generate meristemoids (red) and stomatal lineage
ground cells (SLGC) (white). The meristemoids differentiate into guard mother
cells (pink) and undergo symmetric cell division to generate mature guard cells
(green), or continue asymmetric division, in order to undergo up to three
amplifying divisions. In addition, the stomatal lineage ground cells undergo
asymmetric spacing division, during which new meristemoids are produced, in
order to prevent the direct contact of adjacent stomatal lineages.

1ST PHASE - initiated when trichome precursor stops dividing,

begin endoreplication

❖ Small protuberance
❖ Grows out after 2-3 endoreplications and undergo 2°
successive branching
❖ Last round or 4th endoreplication occurs after 1st
branching. Now 16 fold increase from 2C to 32C.
❖ Trichome increases size and undergoes tip growth and
diffuse growth

2ND PHASE - rapid expansion; cell wall thickens & becomes

covered with papillae; the base has ring of cells called socket
or accessory cells

Diagrammatic representation of types of stomatal development in

Commelinaceae, not to scale. Sequence of divisions is variable, those
represented are the most commonly observed. Abbreviations used: t.m.c.,
terminal subsidiary cell mother cell; g.m.c., guard cell mother cell; l.m.c., lateral
subsidiary cell mother cell; t.s.c., terminal subsidiary cell; 1.s.c. (I), innermost
lateral subsidiary cell; l.s.c., (2), outermost subsidiary cell.
Trichome Development and Mutant Phenotypes Flower Development
❖ Glabra 1 (GL1) and Transparent testa glabra (TTG) - both
required for trichome development. Control of Flowering
- GL1 product is a transcription factor that accumulates in ❖ Flower formation involves a phase change from
trichome cells-to-be vegetative growth to reproductive growth
- Trichome cell enlarges, DNA increases (8X more than - it is triggered by a combination of environmental cues
surrounding cells) and internal signals
- Enlarging cell grows outwardly, nucleus is pushed into ❖ Transition from vegetative growth to flowering is
subapical location associated with the switching on of floral meristem
- Then volume has increased 10-fold, it branches. identity genes.
❖ TRIPTYCHON (TRY) - these are transcription factors that regulate genes
try - lead to formation of "nests" of 2 or 3 neighbouring required for the conversion of the SAM into the floral
trichome meristem
❖ gl2 - are flattened and integrated into the epidermis
Wildtype trichome - GLABRA 2 promotes outward growth Transcription factors regulate conversion:
of trichome cell from epidermis Indeterminate vegetative SAM → Determinate floral meristem
❖ STICHEL (STI) – product is needed for branching, sti ❖ Production of flowers by the SAM stops primary growth
mutant trichomes are unbranched of the shoot. The apical meristem is consumed in the
❖ gl3 - smaller trichomes, later endoreplication promoted production of flower parts.
by GLABRA 3 gene
❖ kak (kaktus) – enlarged trichomes with several branches
❖ an (angustifolia) – normal stalk, long branches
❖ zwi (zwichel) - has 2 branches with unequal length
❖ cdo (chardonnay) – glass-like
❖ cha (chablis) – glass-like

Genes in Trichome Development

Specification of trichome cell
- GLABROUS 1 (GL1)
- TRANSPARENT TESTA GLABRA (TTG)

Progressive morphogenesis, control enlargement

Process of Flower Formation:
- GLABROUS 3 (GL3)
1. Induction: the decision to switch from vegetative to
- KAKTUS (KAK)
generative phase
Controls local outgrowth of committed cells
2. Evocation: transition from vegetative shoot meristem
- GLABROUS 2 (GL2)
to the floral meristem with the associated generation of
the primordia for flowers, and other lateral organs
Control length
- DISTORTED (DIS)
3. Actual Flower Formation
- GNARLED (GRL)
- KLUNKER (KLK)
4. Functional Phase of the flowering process begins i.e.
when the flower organs have gained their characteristic
Control branching
forms and functions. The maturation of the reproductive
- ANGUSTIFOLIA (AN)
organs and pollination and fertilization take place.
- STICHEL (STI)
- ZWICHEL (ZWI)
Formation of Floral Primordia Sepals arise where only A genes are active
Petals arise where A & B are active
Stamens where B & C are active
Carpel where only C genes are active

Viewed from above, the meristem can be divided into four

concentric circles, or whorls, each of which develops into a
circle of identical organs.

A simple model explains how the three classes of genes can

direct the formation of four organ types.

Each class of genes affects two adjacent whorls

4 different floral organs are initiated sequentially

❖ 5 sepal primordia
❖ 5 petal primordia
❖ 10 stamen primordia
❖ Carpel primordium undifferentiated

When a SAM is induced to flower, the relative position of each

primordium determines its development into specific type of
floral organ- a sepal, petal, stamen or carpel.
Floral organs develop in 4 concentric whorls.

ABC genes regulate expression of other genes

By combining the ABC hypothesis with the rule that if A-gene

or C-gene activity is missing, the other spreads through all
A genes are switched on in the 2 outer whorls (sepals & four whorls, explain the phenotypes of mutants lacking a
petals) functional A, B, or C gene.
B genes on 2 middle whorls (petals and stamens)
C genes on the inner (stamens and carpels)
- Where gene A activity is present, it inhibits C and vice versa.
If either A or C is missing, the other takes its place. A and C
genes are cadastral genes

Six key genes initially were identified in Arabidopsis that specify floral organ
identity: APETALA1 (AP1), APETALA2 (AP2), APETALA3 (AP3), PISTILLATA (PI),
AGAMOUS (AG) and SEPALLATA 1-4 (SEP)
 Root Development

Facets of Root Growth and Development

❖ Root apical meristem
❖ Differentiation of primary tissues
❖ Vascular tissue differentiation
❖ Root hair formation
❖ Lateral root formation
(Nodule development)

Fate Map for Root Development

To track down which tissue or cells are expressing certain

genes: track down the location of specific mRNAs using
labeled probes in place or in situ in the intact organism

❖ Evidence that A,B, and C genes are active in particular

whorls come from in-situ hybridization:
Nucleic acid from cloned genes labeled with dyes
was used to tag the location of complementary
mRNA in developing flowers.
❖ Nucleic acid probes will hybridize with mRNA transcribed
from the gene.