M.Sc CBCS IIIrd Semester
Avanced Plant Pathology I (DSE1)
Unit V
Topics: Anatomy of galls.
What you will learn?
The basic structure of various galls.
The tissue organization in galls.
Specialized tissues in galls.
Just like normal plant organs and structures, galls induced by insects present anatomic and
histologic characteristics of their own, which vary greatly in their diversity and degree of
complexity. The galls may vary from simple and isolated cytological transformations to a new
arrangement of plant tissues. The degree to which the insect manipulates the plant’s growth to
form the gall varies considerably and involves changes ranging from the induction of cell
proliferation to the formation of a complex structure that the plant does not produce under
normal conditions.
“Crown galls” induced by the genus Agrobacterium are an example of structures formed due to
the proliferation of cells with a low level of differentiation; hence, they are considered the
simplest and least derived plant gall. On the other hand, galls induced by insects are very well
organized structures showing different degrees of differentiation, the reason why they are
considered as the most complex and derived structures.
Both systems require a previous state of “conditioning” towards the development of gall. In the
case of insect galls, the “conditioner” is the insect itself, which modulates the tissue that will
form the structure through mechanical action and the secretion of chemical substances. In crown
galls the conditioning factor is given by a series of metabolic events prior to the genetic
transformation of plant cells by the bacterium.
After the initial stimulus, cell proliferation in both systems develops in a different way; in the
case of bacteria-induced crown galls, cell proliferation occurs in an uncontrolled way and does
not require the continuous presence of bacteria once the process is initiated. In contrast, for
adequate and complete development of galls induced by insects, in general, the continuous,
active presence of the insect is required
Just like normal plant organs and structures, galls induced by insects present anatomic and
histologic characteristics of their own, which vary greatly in their diversity and degree of
complexity . Galls induced by insects are distinct from those induced by fungi and bacteria in
their form, organization, and complexity.
The gall has a chamber called gall cavity which is lined by small closely packed mass of cells
lacking in chlorophyll, rich in cell contents. This zone constitutes the nutritive zone and is
surrounded by thick sclerenchyma followed by parenchyma with pigments and vascular supply,
epidermis and cuticle. There is usually abundance of polyploidy cells with nuclear gigantism.
Gall cells are also emlarged. The inside of insect induced galls is usually composed of highly
nutritious cells. This allows the insect to feed without having to chew through plant tissue or
hard to digest plant chemicals like oils and tannins.
Things that effect gall formation and structure are:
1. Type of gall inducer
2. Mode of feeding: biting , sucking, chewing, piercing etc.
3. Salivary secretions: contain growth promoters, abnormal amino acids etc.
4. Life stage: larva, adult
5. Type and age of plant part
6. Plant species
7. Position of cecidozoa
8. Mode of development
More complex and diverse galls are induced by insects such as those of the Cynipidae and
Cecidiomyiidae families, which show extreme examples of radial symmetry, belonging to the
orders Hymenoptera and Diptera, respectively
The major anatomical changes that take place are:
Hypertrophy
Hyperplasia
Infiltration
Anomalies during cell division
Tissue degeneration and necrosis
Hypoplasy i.e. inhihibition of normal development.
Hypertrophy is cell enlargement without cell division. Practically all cells of gall show
hypertrophy including epidermis, vascular bundles etc. It is an important event in gall
development. In some cases it is the predominant process. It also represents the last phase of gall
M.Sc CBCS IIIrd Semester
Avanced Plant Pathology I (DSE1)
Unit V
Topics: Physiology of gall formation.
What you will learn?
What metabolic changes take place in gall tissues
Which metabolites are responsible for abnormal growth
The variation in morphological complexity is also followed by a variety of physiological traits.
Galling insects do not only control the developmental patterns of the host plant, so as to define
the gall phenotype, but also its physiology. Galls are sinks of photoassimilates. Galls may
produce photoassimilates, but in such low values that it seems improbable that they could
guarantee gall maintenance without draining resources from their host organ. The physiological
gradients inside and outside the gall tissues revealed specific enzymatic activities and are also
accompanied by cytological peculiarities .The galls function as sinks of nutrients mobilized from
the other host plant parts . A large set of evidences support that the galling insect is able to
manipulate the host plant, inducing the formation of nutritionally superior cells in comparison to
the other healthy plant tissues, the nutritive tissue. The cells of this tissue have a high
concentration of lipids, glucose, amino acids, and a high enzymatic activity, including
phosphatases, proteases, and aminopeptidases rich in RNA and ribosomal RNA of the nucleolus
(Bronner 1992). On the other hand, the parenchyma cells of the outer cortex form a reserve tissue
characterized by a high concentration of starch, low concentration of lipids and glucose, and low
enzyme activity. As the larva feeds on the cells of the nutritive tissue, there is a replacement of
substances by the cells of the reserve tissue. The translocation of substances between the two
tissue zones has been proven to need an intense enzymatic activity. Also, the substances
accumulated may be diverse, such as proteins, carbohydrates, and lipids
The most general hypothesis suggests that gall formation is triggered by the action of chemical
substances secreted by the gall inducer, including plant growth regulators such as auxins,
cytokinins, indole-3-acetic acid (IAA), and other types of compounds. However, the mode of
action of these chemical substances and the general mechanism by which the insect could control
and manipulate plant development and physiology is still not known.
Gall formation is a defense reaction of the host plant and is related to either the feeding activity.
The prime aim of host is to restrict the proliferation and the main aim of insect is to suck
nutrients from the host. In many cases the stimulus appears to come from the saliva of the
developing larva, while in others it seems to be chemically injected during egg laying. Like
effective parasite the gall insects do not kill their hosts but they draw the resources of the host
cells for themselves. Galls restrict the injury by gall maker. Insect gets food , shelter and place to
reproduce.
Various theories exist regarding gall development and induction mechanism:
1. Insect synthesizes cecidigen (gall forming metabolite) which causes gall formation.
2. Insect stimulates plant hormones. Deranged metabolism leads to gall formation.
3. Enzymes or enzyme like secretions in insect saliva are responsible for gall induction.
For successful survival the cecidozoa utilizes two basic adaptive strategies:
development of abnormally large cell (hyperptrophy) or excessive multiplication of cells
(hyperplasia)
Transformation of differentiated tissues into meristematic tissues.
The cellular realignment due to hypertrophy in early stages of feeding often result in the
establishment of galls. The gall inducer exploits the resources of the host maximally by
synchronizing the time of active growth in the plant with the appearance of gall insect.
Gall morphogenesis is a complex phenomenon, which involves reorientation of the plant’s
development by the inducing insect. The degree to which the insect manipulates the plant’s
growth to form the gall varies considerably and involves changes ranging from the induction of
cell proliferation to the formation of a complex structure that the plant does not produce under
normal conditions.
Several theories have been proposed to explain the physiology of gall formation.
1. Insect saliva alters the subcellular environment of cells and thus places it in a state of
chemical shock which induces osmotic changes. This establishes the first recognizable
stage in gall induction. Plant produced metaplasied cells to repair the wound and
neutralize the osmotic-change induced stress. Localized metabolic changes spread, from
these cells, not throughout the involved plant organ, but in a limited manner around the
immediate site of insect occurrence. The osmotic-change related stress prevails till the
insect feeds. Osmotic stress affects electrical properties of the plasma membrane and
impacts on IAA activity, which in turn, alters H + -transport systems. During the physical
action of insect feeding, the host-cell wall breaks down, and the degenerated wall
materials act as elicitors.
2. The most general hypothesis suggests that gall formation is triggered by the action of
chemical substances secreted by the gall inducer, including plant growth regulators such
as auxins, cytokinins, indole-3-acetic acid (IAA), and other types of compounds.
However, the mode of action of these chemical substances and the general mechanism
by which the insect could control and manipulate plant development and physiology is
still not known.
3. The inducing insect can modify the expression of genes within restricted areas of the host
plant, thereby producing new developmental events in the tissues under its influence.
4. Insect synthesizes and secretes cecidogen; the gall forming metabolite which induces gall
formation.
5. The feeding /ovipositon activity stimulates the growth hormones and or cause altered
synthesysis of carbohydrates, proteins, hormones, nucleic acids of the host plant which
causes gall formation.
6. Enzymes or enzyme like secretions in the saliva are responsible for gall induction.
7. Salivary secretions of the insect serve as precursors to growth promoters or salivary
proteolytic enzymes convert plant protein into amino acids by hydrolysis.These abnormal
amino acids like nopaline, octopine etc. lead to abnormal growth.
8. Insect produces specific cecidotoxin which is responsible for gall formation.
9. Presence of Tumor inducing plasmid or tumour inducing principle in the salivary
secretions causes abnormal growth.
10. The tumour has the capacity to synthesize auxin or salivary secretion contain substances
that prevent degradation of auxin. High concentration of auxins in the gall tissue leads to
hypertrophy and hyperplasia and gall formation.
11. Amino acids present in the salivary secretions of gall-inducing insects, essentially lysine,
histidine, and tryptophan, could function as “preconditioners” for gall induction. It seems
that these amino acids could cause major plasticity and would increase the sensitivity of
the plant tissue to the action of the corresponding inducing insect.
12. It has been speculated that polyphenol oxidase (PPO), also present in the saliva
secretions of insects and the phenolic compounds derived from its enzymatic action,
could increase plant tissue sensitivity to the stimulus of the inductor insect. It has also
been suggested that the complex interaction between the host plant tissue and polyphenol
oxidase might be of fundamental value in gall formation. Interactions and the balance
between insect polyphenol oxidase and the host plant could determine whether the
“attack” of an insect causes injury (necrosis) or gall development.
13. The modulation of redox potential has been related to gall initiation and
establishment, especially concerning the accumulation of reactive oxygen species .
14. Different studies have reported that indoleacetic acid (IAA) could be a powerful
gall- inducing agent, and it has also been speculated that this compound could interact
with other plant growth regulators, like cytokinins and gibberellins, or in a synergistic
way with other chemical substances, to promote the induction and maturation of these
structures.
15. Symbiotic relationships between gall-inducing insects and microorganisms have
been hypothesized to be involved in plant gall development. Some researchers have
reported that simultaneous infection with different species of endosymbionts in the same
host organism is a common phenomenon in several insect groups.
16. Secretions of phytohormones, such as cytokinins, by endosymbiotic
microorganisms have also been associated with the plant–galling insect interaction. The
inducing insects obtained their ability to induce galls via endosymbiotic microbes, which
have acquired the biosynthetic pathways to produce IAA and trans-zeatin family
cytokinins from plants.
17. It has been proposed that galling insects acquired genes from symbiotic
microorganisms through horizontal gene transfer . Horizontal gene transfer (HGT) is the
movement and transference of genetic information between different organisms, and it is
a common phenomenon between pathogens of animals and plants, and between
symbionts and pathogens. The mechanism through which Agrobacterium
tumefaciens transfers genes from the bacterium to plant cells occurs through the action of
the T-DNA segment present in the Ti plasmid.
General characters of Cecidogenesis (Gall Development)
Gall is a result of specialized reaction of plant cell to the changes in its internal environment
caused by gall inducing organism. The major anatomical changes that take place are:
Hypertrophy
Hyperplasia
Infiltration
Anomalies during cell division
Tissue degeneration and necrosis
Hypoplasy i.e. inhihibition of normal development.
Hypertrophy is cell enlargement without cell division. Practically all cells of gall show
hypertrophy including epidermis, vascular bundles etc. It is an important event in gall
development. In some cases it is the predominant process. It also represents the last phase of gall
development. Hyperplasia or increased cell division is perhaps the most important histogenic
event during cecidogenesis. Intensive cell proliferation is usually found in parenchyma,
cambium, medullary rays. Epidermis shows weak reaction. In some cases cell fusion is also seen
eg. Synchytrium, Heterodera galls. In crown gall three important changes occur during
tumorigenesis;
The transformation period: resulting in primary tumour
The duplication period: the primary tumour undergoes repeated cell divisions leading to
an undifferentiated mass of tumour.
The organisation and differentiation period; during this time the structural differentiation
of tumour occurs.
Work done on viral tumours also indicates that pericycle is stimulated to divide due to viral
activity which results in tumour formation. In wound tumour of clover roots, tangential division
of pericycle opposite to primary phloem results in tumour development=The reason for
hypertrophy and hyperplasia may be presence of proteolytic enzymes in the saliva of gall
forming insect that convert plant proteins into abnormal amino acids. The insect saliva may also
contain growth hormones which may cause cell proliferation.
Various studies indicate that the gall formation involves:
1. Altered synthesis of carbohydrates, proteins, hormones and nucleic acids.
2. Salivary secretion of insect e.g. aminoacids that serve as precursors to growth promoters
or salivary proteolytic enzymes convert plant proteins into amino acids by hydrolysis.
3. Insects produce specific cecidotoxin.
4. Presence of Ti Plasmid.
5. Deranged auxin –cytokinin metabolism:
tumour has the capacity to synthesize auxin in vitro
tumour synthesizes more auxin
t here is less destruction of auxin protectors or absence of auxin inhibitors.
J Indian bot Soc Vol 74 (1995) 129-133
HISTOPATHOLOGY OF STEM GALL OF PROSOPIS CINERARIA (LINN.)
DRUCE INDUCED BY AN UNKNOWN CHALCID
KANKIKA SHARMA*, PAYAL LODIHA AND U. KANT
Department of Botany, University of Rajasthan, Jaipur-302 004, India.
(Accepted December 1994)
The present investigation deals with the histopathology of stem gall of Prosopis cineraria induced by an unknown chalcid. These
acrocecidial galls are globose, solid, hard and woody. Structure of gall tissue differs from nomal stem in several features. In young galls
proliferated cortex forms the bulk of galls, however in mature gall the hypertrophicd vascular tissue adds to its width. Large number of gall
cavities are situated towards the periphery in the xylem tissue. These cavities are surrounded by thin walled cells containing dense cytoplasm
and promincnt nuclei which constitutes the nutritive zonc. In very old galls the bark ruptures to expose these gall cavities.
Key Words: Prosopis cineraria, chalcid, nutritive zone, hypertrophy, hyperplasia, gall cavity.
Plants galls are abnormal, uncontrolled, largely normal stem the gall tissue presents an abnormal
independent growlh of the host tissues which arise
under the influence of an external stumulus. Develop- ment and anatomy of several inscct induccd galls on
certain plant species has been worked out by many
workers (Kostoff and Kendall, 1929;Lewis and Walton
1947;Mcyer, 1965; Kant and Arya, 1971; Raman etal,
1978; Kant and Sharma, 1981 Ananthakrishnan and
Raman, 1988; Kant and Karnawat, 1989; Kant et al.
1994; Ramani et al. 1994). Mani (1973) has reporled
brief description of siem gall suructure of Prosopis
cineraria indced by an insect, but there is no work on
development and anatomy of this gall. This investiga- tion is therefore concerned with the Histopathology of
the stem gall produced on Prosopis cineraria by an
unknown chalcid.
appcarance. Normal young stem exhibits a single layer
of epidermis made up of rectangular cells and coated
with thick cuticle. Cortex is 5-9 layered and is made up
of thin walled, oval to polygonal cells with very few
intercellular spaces. A sclerenchymatous pericycle
surrounds the stele. Vascular bundles are conjoint,
collatcral and open. In the cenure lies a parenchymatous
pith. (Fig. 1 A)
In a mature stem cpidermis is replaced by cork
tissue. Secondary growth is typical dicotyledonous
type.
Gall morphology Stem galls of Prosopis are
acrocecidia, These are globose, solid, hard and woody.
Young galls are smooth and greenish yellow, but matre
galls are brown in colour. In this hard and woody solid
gall as many as 200-500 oval or spherical larval cavities
are prescnt. Rupture of bark in mature galls exposcs
these cavities which contain bright, metallic grecn
MAERIALS AND METHODS
Gall and normal stem material was collected from
infcsted plants growing in Jaipur and adjoining arcas
and fixed in F.A.A. (formalin acetic acid alcohol) and
70% ethyl alcohol. Dehydration, clearing and embed
ding were donc following the tertiary butyl alcohol
method as recommended by Johansen (1940). Mic- rotome scctions were cut at a thickness of 7-14 um and
stained withh Johansen's safranin and fast green com- binations. External morphology of the gall was studied
under the stereoscopic microscope.
coloured chalcid.
Gall anatomy: In a young gall the cpidermal cells
are rectangular, parenchymatous, and ccovered by a
thick cuticle. Cortex is characterizcd by hypertrophy
and hyperplasy. It is gencrally 12-20 laycred. Cells are
polygonal. Thin walled and closcly packed without any
intercellular spaccs. This highly prolifcraled cortex
contributcs to the bulk of the gall. Poricycle is not very
conspicuous though it is composcd of sclerenchyma- Lous cclls. (Fig. 1, 1B) In a mature gall epidermis is
rcplaccd by cork. Cork cclls arc irregular in shape and
OBSERVATIONS
Structure of normal stem : Normal stem shows
typical dicotyledonous structure. As comparcd to thc
Reccived June 1994
Department of Botany, Mohanlal Sukhadia University, Udaipur -
313 001, India.
Sharma et al.
ck- C
P
- ph- nz
- md
-xy
-
Higure I. a-b. Sem gall of Prosopis cineraria (Lim.) Druce. Fig. 1.a 'Transverse section (T. S.) of nonal stem x 369, lig, 1.6 1.>.o
gall. x 369.
Abbreviation: CK-cork, C-conex, P-pericycle, ph-phlocm, nz-nurinive zone, md-medullary ray, xy-xylem, v-vessel, lc-larval cas
are compacly arranged. In a very old gall even the
cortex is replaced by phelloderm cells. Highly proliferated vascular cylinder is the m Contributor to the width of the gall. Cells ot
Histopathology
of stem gall ol iPPOSD, nesia
h nu-
(196
als
at
sto
59 .
:
Se
OT
-lc
,1l
Figure 2. A-G Stem galls of Prosopis cineraria (L.inn.) Druce. Fig. 2. A-D T. S. of galls showing comparative stages in development. x 12; Fig.
BLongitudinal section of gall showing larval Cavities. x 12. Fig. F Cross scction of gall. x 12; Fig. GT. S. of nomal stem. x 12. Abbreviation: 1 c-larval cavity, i-insect.
secondary vascular tissues are predominantly thin walled show cnormous cells prolifcration in galls. (Fig. 1 B)-
and closely packed. As compared to the normal stem, Largenumbcr of larval cavitics are situated towards the
xylem vessels are smaller in size and medullary rays periphery in the xylem tissue and each cavity contain