Puccinia

Systematic Position

Kingdom : Fungi
Sub Division : Basidiomycotina
Class : Pucciniomycetes
Order: Pucciniales
Family: Pucciniaceae
Genus : Puccinia
Habit and Habitat :
Puccinia graminis is an obligate parasite, polymorphic, macro cyclic and
heteroecious rust (Figs. 11, 12). It effects wide range of hosts including wheat, barley,
oats and rye. Grass hosts include Agrostis, Dactylis and Agrophyron. P.
graminis tritici involves in its life cycle two distinct alternate host plants i.e.,
wheat (Triticum vulgare fam. Poaceae) and Barberry (Berberis vulgaris fam.
Berberidaceae).

The wheat plant is called the primary host and the barberry plant is secondary or
alternate host. In Northern India, the black rust appears after March, but in South
India, the rust may appear as early as in the fourth week of November thus, causing
great loss to wheat crops.

2. Symptoms of Puccinia Graminis:

On Wheat:
The symptoms of the disease appear as large, elongated and brown pustules
(uredosori) on the stem, leaf sheath and leaf (Fig. 1 A). Later on these brown
pustules change into black coloured large pustules (teleutosori, Fig. 1 B). Grains of
the infected plants are shriveled, much lighter in weight and thus reducing the yield.
The pathogen shows a balanced host parasitic relationship. Even in the severe
infection, the parasite does not cause much serious damage except that the growth of
the plants may be somewhat retarded and the grains may be of reduced size and of
poor quality.

On Barberry:
Infection first starts on the dorsal surface of the leaf in the form of minute, dark
coloured and flask shaped pycnia which appear as yellow spots (Fig. 2 A). Beneath
Pycnia, on the ventral surface, appear cup like projections of aecia (Fig. 2 B) or
aecidia.
3. Vegetative Structure of Puccinia Graminis:
The mycelium is dikaryotic (each cell of the mycelium bears two nuclei) on primary
host (wheat) and monokaryotic (each cell of the mycelium bears only one nucleus)
on the secondary or alternate host (barbery). The monocaryotic mycelium is also
called hapiomycelium or primary hyphae and the dikaryotic mycelium is called
secondary hyphae.

The mycelium is well developed, branched, septate and does not spread throughout
the host, but is localised to isolated patches. It is either intercellular or intracellular,
with the former producing bulbous, branched or knotted haustoria into the cells for
obtaining nourishment.

The transverse septa are present or long intervals between the cells. Each septum
contains a simple central pore. In contrast with many Basidiomycetes the dolipore
parenthesome complex is completely absent. Cell wall is made up of fungal cellulose.
The cytoplasmic membrane surrounds the granular cytoplasm and reserve food
material remains in the form of glycogen bodies and oil globules.

4. Life Cycle of Puccinia Graminis:

Puccinia graminis is long cycled rust (macro cyclic). At the time of reproduction
it produces five distinct stages in a regular sequence.

These are as follows:

Stage 0: Spermogonia bearing spermatia and receptive hyphae.
Stage I: Aecia bearing aeciospores.
Stage II: Uredia bearing uredospores.
Stage III: Telia bearing teleutospores.
Stage IV: Promycelia bearing basidiospores.
Out of these five stages, Uredo stage, Teleuto stage are produced on the primary host
(wheat) and remaining two stages, (spermogonial and aecial stages) are produced on
the secondary host i.e., barberry.

Stages on Primary Host (Wheat):

Uredospore’s or Uredo stage:
This stage is formed by the infection of the aeciospores brought from the infected
barberry plants or by the uredospore’s themselves coming from the neighbouring
wheat plants infected earlier. Both the spores are bi-nucleate and on germination,
produce a germ tube on wheat leaf.

The germ tube grows over the surface of the epidermis of the host and on reaching a
stoma its tip swells up and forms a vesicle like structure called appressorium (Fig. 3).
The protoplasm of the germ tube migrates into the appressorium. Now it is cut off
from the germ tube by a septum. The appressorium produces a narrow hypha. It
enters inside the sub-stomatal chamber through stoma.

Its tip again swells up and forms a sub-stomatal vesicle. The contents of the
appressorium migrate to vesicle through narrow hypha. A hypha of the dikaryon
(two nuclei) cells develops from this vesicle (Fig. 3). It branches and produces
hyphae which spread in between the cell (intercellular) but occasionally produce
hausotria.
Development of Uredospore’s:

Within 5-6 days, the mycelium absorbs sufficient food from the host. It begins to
aggregate near the surface of the infected organs and forms a compact mass. These
are called uredia. From these uredia arise vertically a layer of bi-nucleate parallel
cells known as basal cells. The basal cells elongate vertically and divide transversely
into a lower cell (foot cell) and an upper cell (uredospore mother cell).
The upper cell divides again and its upper daughter cell swells to form a single,
bi-nucleate, oval, uredospore or uredinospore, while the lower daughter cell matures
into a stalk (Fig. 4 A). Thus, the uredospore’s are formed in a group and each such
group is called as uredospore’s or uredinium (Fig. 4 A). The developing uredospore’s
exert pressure on the over-lying epidermis. By this pressure the epidermis bulges out
and later breaks up and the uredospore’s get exposed.

The uredospore’s are golden brown and oblong, ovate or ellipsoidal in shape. They
are double walled, echinulate, binucleate (the two nuclei belonging to opposite
strains) and possess four equatorially arranged germ pores (Fig. 4 B).

A uredospore can infect only a wheat plant. After falling on a suitable host it
germinates within a few hours and produces a dikaryotic mycelium. The mycelium is
capable of producing uredospore’s again within 10-12 days after germination. Thus,
these spores cause several successive infections during the season, and spread the
fungus and the disease from field to field provided the environmental conditions are
favourable (sufficient moisture). On the basis of this behaviour these uredospore’s
are also known as repeating spores.

Teleutospores or Telial Stage:

Towards the end of the growing season of wheat crop, the environmental conditions
become unfavorable (hot and dry) for the growth of the uredospore’s. Now uredosori
produce another kind of spores called teleutospores. First, they develop among the
uredospore’s within the same sorus, but later they develop in separate sori known as
teleutosori or teleutopustule (Fig. 5 A).

As the crop matures, the number of uredospore’s is reduced and the sori contain
only teleutospores. This stage is known as the black stage and hence the name black
rust is given to the disease (Fig. 5 A, B). The teleutospores are dark brown or black in
colour. They are bi-celled and spindle shaped structures (Fig. 5 C) with a pointed
apex and thick smooth wall.

Each cell of a teleutospore has a single germ pore and two nuclei (one of plus strain
and the other of minus strain, Fig. 5 C). As the teleutospores reach towards maturity,
karyogamy takes place and the two nuclei fuse to form a diploid nucleus (Fig. 5 C).
The development of teleutospores is entirely similar to the uredospore’s.
At this stage the teleutospores undergo a period of rest. During resting period they
lie on the ground or still attached to the host. These are the dormant cells and are
capable of tiding over unfavorable period.

Basidial Stage:
After the resting period, the teleutospores germinate during the early part of spring.
They germinate in situ and either one or both of its cells give rise to a germ tube.
known as promycelium. The promycelium together with the teleutospore cell is
called basidium. However, many authors prefer to call the teleutospore cell as the
hypo-basidium and the promycelium as the epibasidium (Fig 6 A-C).

The diploid nucleus of the teleutospore migrates into the promycelium and divides
meiotically into four haploid nuclei (Fig. 6 C). The septa appear between the nuclei
and divide the promycelium into four haploid cells. Each haploid cell of the
promycelium produces a slender, short, lateral, tube-like structure known as
sterigma (Fig. 6 D). The sterigma swells up at the end to form a spore like cell.
The haploid nucleus from each promycelium cell migrates into this developing spore
cell through its respective sterigma. Thus, at the tip of each sterigma, a minute spore
is formed.

This spore is called basidiospore (Fig. 6 D). Each cell of promycelium produces a
single basidiospore. Thus, from a single cell of teleutospore four haploid, unicellular,
uninucleate basidiospores are formed. Two, out of the four telutospore basidiospores
are of ‘+’ strain and the other two of ‘-‘strain.

Soon after the basidiospore formation they are forcibly ejected by the ‘water droplet
method’. (In this method a liquid begins to collect in the form of a droplet at the base
of the basidiospore. This droplet gradually attains a bigger size and suddenly pushes
off the basidiospore forcibly into the air to a short distance.) The basidiospores are
carried away by wind. They are capable of germinating only on Barberry plants
(Berberis vulgaris) available on hills. They perish soon if the alternate host is not
available.

Stages on Barberry Plant:

The basidiospores, which fall over the upper surface of barberry leaf start
germinating soon. They germinate by giving out a germ tube which penetrates
through the epidermis.

The germ tube elongates and divides inside and develops into hyphae. These hyphae
grow between all the cells lying in between the lower and upper epidermis. The
hyphae are composed of uninucleate haploid cells (primary mycelium) and are of ‘+’
or strain.

Several basidiospores of different strains may infect the same Berberis leaf. Thus,
haplomycelium of two different strains (‘+’ or ‘-‘) is formed. It remains haploid for
sometime and the fusion between these two strains (‘+’ or ‘-‘) may occur at a later
stage.

Spermogonial or Pycnidial Stage:

This stage is also known as Pycnial or spermatial stage. After about four days of the
infection, the haplomycelium collects and forms dense mats both beneath the upper
and lower epidermis. The mycelial mats beneath the upper epidermis are known as
primordium of spermogonium while the mats beneath the lower epidermis are
known as primordium of aecidium or protoaecidium.

In 7 to 10 days after infection, each primordium of spermogonia matures into a small

flask shaped structure called spermogonium or pvcnidium. The pycnidia appear as
minute yellowish specks on the upper surface of the leaf (Fig. 2 A). A vertical section
through these specks reveals that each spermogonium opens on the upper surface of
the host leaf through a pore like structure known as ostiole (Fig. 7).

Its wall consists of three kinds of hyphae:

(i) Periphysis:
The ostiole is surrounded at the fringe by the long, delicate, sterile hyphae known as
periphysis. They develop near the ostiole from the spermogonial wall and project
from the ostiole.

(ii) Flexuous or Receptive Hyphae:

They also arise from the lateral wall of the spermogonium. They are slender, delicate,
cylindrical, septate, simple, branched or un-branched with blunt ends. They are
present amidst periphysis and sometimes it is difficult to distinguish between the
two.

(iii) Spermatiophores or Pycnidiophores:

These are slender, short, vertical, uninucleate hyphae which arise form the base of
the spermogonium (Fig. 7). Each spermatiophore (or sporophore) produces several
small uninucleate spermatia or pycnidiospores at its tip by abstraction method (Fig.
8 A).

The spermatia are unicellular, small, oval to spherical, hyaline and smooth walled
structures. The spermatia fill the spermogonial cavity and are exuded from the
ostiole in a droplet of nectar, which is a thick, sticky, sweet liquid.
According to Craigie (1927) and Butler (1927) the spermatia function as male cell
while receptive or flexuous hyphe represent the female sex organs (Fig. 8 B). The
spermatia may be ‘+’ or in their sexual nature depend r.g upon the mycelium,
produced by the basidiospores ‘+’ or ‘-‘. The insects are attracted by this nectar.

The spermatia are dispersed from one spermogonium to another spermogonium on

the same leaf or to another leaf. As a result, the ‘-‘ spermatia are transferred to ‘+’
receptive hyphae and are transferred to receptive hyphae. Now the spermatization
takes place.

The spermatia of strain come in contact with the tip of the receptive hyphae of
opposite strain. The intervening wall at the one stage of contact between these two
dissolves and the spermatium nucleus passes downwards through septal pores and
form a bi-nucleate cell (Fig. 9 A-C). This pair of nuclei of opposite strains is called a
dikaryon and this process is called dikaryotization.
Aecial or Aecidial Stage:
The haplomycelium forms the primordium of aecidium or protoaecidium beneath
the lower epidermis. The further development of protoaecidium into aecidium takes
place only after the dikaryotization.
The spermatial nucleus (male nucleus) by mitotic division forms a second male
nucleus, which moves to the next cell, through septal perforation. In this way the
male nuclei produced by successive mitotic divisions pass down and all the cells of
primary mycelium are dikaryotized.

The dikaryotic basal cells of the protoaecidium arrange themselves vertically beneath
the lower epidermis and are called as sporophores. Each bi-nucleate basal cell then
cuts off a chain of bi-nucleate cells in basipetal succession on the side towards the
lower epidermis of the host.

These cells are the aecidiospore mother cells (Fig. 7). These cells further divide
transversely to form a large cell and a small cell. The large cell develops into
aecidiospore while the small cell remains sterile and is known as disjunctor or
intercalary cell. The latter dissolves and sets free the aeciospores.
With the development of the aeciospores some of the basal cells lying at the
periphery of protoaecidium mature into a one-celled thick protective layer called
peridium.

This entire structure is cup shaped and is known as aecium. The developing
aeciospores rupture the peridium by exerting a pressure on it. Thus, the aeciospores
are liberated. They are unicellular, polyhedral, thin walled, bi-nucleate and orange
yellow coloured.

Germination of Aeciospores:
The aeciospores are disseminated by wind. They are capable of immediate
germination but cannot infect barberry plants. Falling on suitable host i.e., wheat
leaf they germinate by producing a germ tube or primary hyphae.

The further development of the germ tube is similar as described in the uredinal
stage and ultimately the dikaryotic mycelium is produced. This is the mycelium
which produces the uredospore’s and later the teleutospores on wheat. In this way,
the life cycle of Puccinia graminis is completed.