MULBERRY PROPAGATION

Dr. Rohith L. Shankar

Professor
Department of Sericulture
Yuvaraja’s College (Constituent Autonomous College with Potential for Excellence)
University of Mysore Mysore-570005
rohithlsycm@gmail.com / rohithlsycm.weebly.com
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The two methods of propagation practiced for mulberry cultivation are: 1) Seedling
propagation [sexual method] and 2) Vegetative propagation [asexual method]. Recent advances in
biotechnology have now brought in a third method of propagation, namely, micropropagation
[tissue culture].

I. SEEDLING PROPAGATION

Seedling propagation is rarely practiced by commercial rearers because the quality of the
variety cannot be maintained as mulberry is – normally a wind – pollinated plant. Moreover, this
method involves a long gestation period before leaves can be harvested for silkworm rearing.
However, this is the normal method of propagation in research centers where breeding studies are
carried out. Here the flowers are protected from cross – pollination and only controlled pollination
is allowed. Normal cross – pollinated plants are used to produce a varied population for selection
and hybridization. Some exotic varieties of mulberry can be propagated only by seedling
propagation.

The nursery for seedling is selected in a shady area to protect the seedling from direct
sunlight and excessive temperature. The soil is prepared to fine tilth by ploughing and breaking
the clumps. Equal quantities of red soil and farmyard manure are applied and mixed by ploughing.
Convenient – sized seed – beds, normally 0.9m square, are made.

The seeds are extracted from ripe fruits available in March – April in the tropical regions
and May – June in the temperate regions. As there is no dormancy period for seeds, freshly
collected seeds have a high germination percentage. The germinating ability gradually gets lost
and it is not desirable to store the seeds for more than three months. If the seeds are to be preserved
for a longer period, they should be stored in a sealed, airtight contains and kept in a cool place.

Seeds are soaked in water for 24hours to soften the testa and at the same time to eliminate
unviable seeds, insect – infested seeds and unfertilized seeds etc. which float on the water. This is
called the flotation test of testing seed viability. The other method of testing the viability of the
seeds are: 1) use of vital dyes, 2) electrical conductance test and 3) embryonic behavior method.
The viable seeds are either broadcast in the field or sown individually in holes in a line marked
with rope, at a depth of less than 25mm. if sown in deeper layers, the seeds fail to germinate due
to lack of oxygen.

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 Germination is also affected by environmental temperature and light. The minimum,
optimum and maximum temperature ranges for germination are: 18 – 21oc, 33 - 36oc and 36 - 39oc
respectively. Violet blue and bright light are harmful for germination. In the dark, germination is
controlled by temperature. The time taken for germination is ten days at 27oc, eight days at 30oc
and six days at 33 - 36oc in dark condition.

Water is applied through a can so that the seeds are not drifted away towards one side or
drawn to deeper layers. Mats of bamboo strips or pleated coconut leaves are placed on the seed-
beds to protect the germinating seeds from excessive sunlight. The coverings are placed one foot
above the seed – beds. Seeds germinate in 10 days. After three months, the first transplanting is
done to give a wider space of 22.5 cm between them. After two years, they are once again
transplanted to their final locations or used to prepare cuttings or as stocks for grafting methods.

II. VEGETATIVE PROPAGATION

This is most popular method used commercial plantation. Its chief advantages are the
following.
1. The desired hereditary characters can be maintained throughout.
2. Large number of plants can be raised quickly and commercially.
3. Pest and disease resistant plants can be raised.
4. Plants adapted to specific localities can be grown.

The vegetative propagation methods fall under three main categories:- Cutting, Grafting and
Layering.

i. CUTTING: This method is adopted for growing varieties fully acclimatized to local conditions.
This is the most popular method of cultivation in South India.

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 Shoots of proper maturity and thickness with active, well developed buds are chosen for
preparing cutting. The tips of very tender branches and the bases of over – mature branches are
rejected. Pencil – thick branches (10 – 12 mm dia) from eight to ten – month – old plants of the
desired variety are used for preparing cuttings. The branches cut into 18 – 20 cms (7” to 8”) long
cuttings with a minimum of three internodes with well – developed buds are used for irrigated
plantation and with 5 to 6 internodes are used for rain – fed plantation. Care should be taken that
ends are cut cleanly with sharpen knives with no splits or peelings in the bark. The cuttings are
placed in a previously prepared nursery with about 2.50 cm of the cutting with one node projecting
above the soil. The cuttings are watered and in ten days, roots develop from buds in the internode
below the soil and leaves from the buds in the internode above the soil. It is reported that parts
containing high carbohydrate root more readily than those rich in nitrogen.

Certain varieties, particularly those evolved from temperate regions, do not produce roots
from cutting easily. Rooting is induced in these by the use of root hormones and growth regulators
like indole acetic acid (IAA), indole butyric acid (IBA), L-Naphthalene acetic acid (NAA), 2, 4 –
dichlorophenoxy acetic acid (2,4-D) or commercial products like rootone, Seradix etc. these

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growth regulators accelerate callus formation and root initiation by increasing the rate of cell
division. These root promoters may be applied directly to the cuttings before planting either as a
lanolin paste or as a powder. Alternatively, the cuttings may be given a quick dip in a concentrated
solution or a prolonged dip in a weak solution of these substances.

The cuttings may be planted directly in the fields or may be grown in a nursery and ten
transplanted. In the later case, after two or three months, they are transplanted to the fields.

The cuttings are supplied at nominal cost by the State Sericultural and Central Sericultural
Departments to the farmers. During the transport, care must be taken that the cuttings do not dry
up and to prevent this, they are packed in wet gunny cloth or polythene gags. If the field is not
ready to plant the cuttings, they are temporarily stored in deep pits one foot deep in a bunch and
water is sprinkled. As soon as the field is ready, the cuttings are planted in the field.

Cuttings prepared in India are from young plants and are called soft-wood cuttings. In
Japan, cuttings are prepared from hard – wood branches.

Though it is the easiest of the vegetative propagation methods, it is not very popular in
Japan because rooting and establishment from cuttings are slow due to the temperate climate.
However, with the introduction of rooting promoters, cutting propagation has come to be widely
practiced in that country. In addition to the simple cutting method followed in India, the following
special types are followed in that country.

Kiban method of cutting propagation uses cuttings which are short and placed vertically
in shallow furrows. In subuse method scion are used as cuttings. The beds are specially prepared
with furrow with a width and depth of 20m alternating with ridges. The scion of about 40cm length
are arranged horizontally in the furrows and the two edges of the scion are inserted into the sidewall
of furrows. The central portion of the scion is kept exposed and covered with straw mat or husks.
In this method two or three plants are obtained from each scion. Shirodoshi method uses the left
– over shoots of layering with well – developed roots or accessory buds as cutting materials. In
mulching accelerated cutting method, thick shoots are collected from the base which develop
into roots before the buds in them sprout. Cuttings are prepared from these shoots and held upside
down and covered with sandy soil of about 5cm height. After twenty to thirty days mulching tissues
form and they are taken out and then planted in the nursery. In order to prevent drying of the soil
in the cutting beds water is sprayed. Warm bed cutting method is one in which root development
is accelerated by using a warm bed before sprouting in spring.

ii. GRAFTING: Grafting is a technique of joining the parts of two plants in such a way that they
unite and grow as one plant. This method is practiced when cutting propagation fails due to poor
rooting ability. The part of the graft combination which is to become the upper portion of the shoot
system of the new plant with the desired characters is called scion. The part which becomes the
basal or root portion of the future plant is called stock. The scion is, therefore, selected from the
desired variety and the stock is selected from hardy, local variety which takes root easily. The
stock should be in a more advanced stage of growth than the scion. It should be in a more advanced
stage of growth than the scion. It should also be selected carefully as it alone influences the growth
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and strength of the plant growing from the scion. Basic principles to be observed while practicing
grafting are:-

 Local adaptability of the stock and the superiority of the scion.

 Compatibility of the stock and the scion.
 Maximum cambial contact between stock and scion to facilitate the formation of graft
union.
 Production and interlocking of parenchyma cells (callus tissue) of both the stock and the
scion.
 Differentiation of new cambium across the callus bridge.
 Formation of xylem and phloem from the new vascular cambium in the callus bridge.

Grafting is called root grafting, shoot grafting or bud grafting depending upon the scion material
used. In all grafting methods, the stock is of the indigenous variety and the scions is of the desired
variety.

A. Root grafting: Root of 0.6 to 2.5cm thickness are selected from one to two – year old
seedlings of the indigenous plant, and cut into pieces of 5 to 7.5cm length. The top end of the
root is cut obliquely. Scion nits of 80 to 10cm length with two to three buds are prepared from
the desired variety. The top end of the stock is given an oblique cut and a corresponding oblique
cut is given to basal end of the stock in between the wood and bark the bark of the scion is
removed. Before insertion, care is taken to see that the cortical layers of the stock and the scion
are in close proximity to each other. The two are tied and covered with grafting wax. Organic
fusion take place between the stock and the scion and the plant which develops from the buds of
the scion has the desired characteristics. Root grafting is most successful of the grafting methods.
It can be done very quickly as it does not require any bandaging. From a single one – year plant,
5 to 6 stocks can be prepared and 5 to 6 plants raised.

Root grafting when done on plants without removing them from the soil and effecting the
grafting at the transitional zone below the ground is called “in situ” grafting. If it is done by
preparing the short cuttings of roots and planted in a separate place like nursery bed or pots etc. it
is called “ex situ” grafting.

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B. Shoot grafting: When the scion is inserted into the stem portion, it is called shoot or stem
grafting. When more than one scion is inserted into the stock, to get a bushy growth, it is called
crown grafting. It is called whip grafting when the stock of 1.2 to 2.5cm thickness is given a
sloping cut to a length of 3.5 to 5cm and the scion of same thickness cut in a similar way is fitted
exactly and tied with fiber and sealed with grafting wax. The grafting used to renovate old plant is
called wedge grafting. In this method, the old plant is pruned at a convenient height and a V –
shaped incision is made at the cut – surface. The basal portion of the scion is cut obliquely to fit
the notch in the stock. After inserting the scion, grafting wax is applied.

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C. Bud grafting: This method is resorted to when the scion material is in short supply. This
method involves the removal of only one bud and a small section of the bark without wood and
grafting it to another stock. This method avoids the transmission of disease through scion as it uses
only a small portion of it. But the main disadvantage of this method is that it takes a very long time
for plants to grow. Three types of bud grafting are commonly practiced for mulberry.

a. Patch budding: in which a portion of the bark with a bud is removed from the stock and
a scion bud is inserted in its place and tied with fibres and sealed with grafting wax.
b. T- budding: in which a T – shaped incision is made in the stock at the nodal region into
which the scion bud is inserted and bandaged and sealed, and which, from its appearance
like a shield, is also known as shield budding.
c. Flute budding: in which the bark of the stock is removed to a length of 2.5 to 3.5cm round
the plant. The bud of scion with same length of bark is placed in this region and bandaged.

Other methods practiced in Japan are bark grafting and venner grafting.

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iii. Layering: this method of propagation involves the development of roots from a stem while it
is still attached to the mother plant. The rooted stem is then detached, to be grown as a new plant.
Such a rooted stem is known as a layer. Root formation on the stem during layering is stimulated
by various treatments which interrupt the downward translocation of organic materials from the
leaves. These materials (carbohydrates, auxins and growth regulators)
Accumulate near the point of treatment and cause the formation of the callus where the
rooting occurs. The major advantage of this method is simplicity. There is no fear of the roots
getting dried up as in cuttings or grafting. This is safer than other methods of vegetative
propagation. The disadvantages include the time involved, the expenses and its unsuitability for
large – scale multiplication. Poor rooting varieties cannot be layered. This method is used to obtain
a large – sized plant in a short time. It is also used to fill the gaps in the field where cuttings have
failed to grow. There are many methods of layering.

a. Simple layering: From a lower branch of bush mulberry, the bark is removed from a 2.5
to 3cm portion in the middle of the branch. The branch is bent in such a way that this portion
comes in contact with the soil and it is covered with soil while the tip of the branch emerges out
of the soil. Roots develop in the covered region and the branch is cut from the parent plant to
lead an independent existence.

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Slight deviations from simple layering are practiced in Japan.

Magedori method is used for low – cut trained mother plants. This is practiced in early spring.
The new branches are bent in a radial manner and their middle portions are covered with soil.
Roots develop from the portion kept under the soil. The branches are cut from the mother plant
and new saplings are transplanted. Chiba type is only a modification of magedori. Makadedori,
Shumkudori, Maritosi, Yokobuse and Mari are some of the other modified methods.

b. Trench layering: This method is popular in Japan. A branch of the plant is bent
horizontally. A considerable length of the middle of the branch is covered with mud and manure.
New shoots develop from the buds and root develop below the soil. Thus, a number of plants can
be grown the same branch.

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c. Air layering: This is also known as gooting. In this method, 1 to 2 cm circular bark is removed
from the middle of an erect branch. Peat mass or well – decomposed organic manure with a little
root hormone is placed in this region and tied securely with as polythene cover. Water is sprinkled
in this region. Roots develop in 1 to 2 months and the branch is severed from the parent and planted
to develop into a new plant.

III. MICROPROPAGATION METHODS

Conventional vegetative propagation methods like cutting, grafting or layering require a

locally adapted variety for rooting to take place. They also take a long time for establishment. They
do not allow any room for genetic manipulation or improvement of the varieties. In order to
develop new varieties as well as to propagate them in as short a period as possible and also in as
large numbers as possible, new micro propagation methods involving tissue cultures have been
evolved for other crops. These have been tried for mulberry as well. Some of these new in vitro
techniques are explained below.

Tissue Culture:
This is followed for some elite cultivars which do not root easily meristems from apical or axillary
buds are excised from the desired variety and grown in a defined nutrient medium. The meristems
are then allowed to form callus and from the callus new plants are allowed to develop. By this
method, some eight to ten plants can be regenerated from a single bud in about three months.

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Production Of Haploids and Isozygous Lines:
Haploids cells of the desired variety are obtained by culturing anther cells in tissue culture medium.
The chromosomes of these anther cells are doubled by cholchicine treatment. The resulting cells
isozygous diploids as the chromosomes of the haploids are doubled. The fertile isozygous diploids
can be exploited for cultivation and breeding trials.

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Endosperm Culture:
The method is used for producing triploid plants which are known to be high yielders and rich in
nutritive value for silkworms rearing. The endosperm tissue of the desired variety is cultured in
tissue culture medium and the plants are propagated from this callus tissue. This method is easier
than the conventional method of crossing a tetraploid and diploid plant to obtain a triploid plant.

Synthetic Seeds:
This popular biotechnology technique for micropropagation has also been tried for mulberry. In
this process, the somatic embryos or meristems are encapsulated in a gel which mimics the seed
coat and these seeds are generated into complete plantlets in a defined nutrient medium. In a
vegetatively propagated plant like mulberry in which the percentage of success with cuttings is
about 40 – 50% and the time involved in pruning, transportation and transplantation is very long,
the method of artificial or synthetic seed is very desirable. The axillary buds are encapsulated in a
gel, packaged and transported safe. The seeds can be germinated in vitro or in the soil directly.

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Induction Of Somoclonal Variation:
Explants are treated with mutagens to induce mutations and then cultured to produce callus. Due
to altered karotype, the callus may show somoclonal variation. From this callus culture with
somoclonal variation, the desired plant can be propagated. SVI is one such variety showing
somoclonal variation.

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Protoplast Culture And Somatic Hybridization:
Protoplast of efficient and undesirable plants can be fused and such somatic hybridization may
eventually result in a hybrid plant with the desired qualities of the efficient plant and the hardy
nature of the other plant. Somatic hybridization has the additional advantage that it produces, in
addition to nuclear hybrid, a hybrid of other cytoplasmic organelles including mitochondria. Such
a hybridization and recombination of non – nuclear factors can be brought about only by this
method.

Genetic Engineering:
Genetic gene responsible for improved quality of the leaves can be isolated and cloned. They can
be introduced into the plants indirectly by using plasmids or other vectors, or directly by using
techniques like PEG mediated DNA uptake, electroporation, microinjection, and microprojectile
bombardment.

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These new techniques offer great scope for the genetic improvement of mulberry in a short time.

The various applications of micropropagation are:

• Plant tissue in small amounts is sufficient for the production of millions of clones in a year
using micropropagation. It would take a great deal of time to produce an equal number of plants
using conventional methods.
• The technique of micropropagation provides a good alternative for those plant species that
show resistance to practices of conventional bulk propagation.
• An alternative method of vegetative propagation for mass propagation is offered through
micropropagation. Plants in large numbers can be produced in a short period. Any particular
variety may be produced in large quantities and the time to develop new varieties is reduced
by 50%.
• Large amounts of plants can be maintained in small spaces. This helps to save endangered
species and the storage of germplasm.
• The micropropagation method produces plants free of diseases. Hence, disease-free varieties
are obtained through this technique by using meristem tip culture.
• Proliferation of in vitro stocks can be done at any time of the year. Also, a nursery can produce
fruit, ornamental, and tree species throughout the year.
• Increased yield of plants and increased vigor in floriculture species are achieved.
• Fast international exchange of plant material without the risk of disease introduction is
provided. The time required for quarantine is lessened by this method.
• The micropropagation technique is also useful for seed production in certain crops as the
requirement of genetic conservation to a high degree is important for seed production.
• Through somatic embryogenesis production of synthetic artificial seeds is becoming popular
nowadays.
With micropropagation having various advantages over conventional methods of
propagation, this method holds better scope and future for production of important plantbased
phytopharmaceuticals. Independent of availability of plants, micropropagation offers a lucrative
alternative approach to conventional methods in producing controlled amounts of biochemicals.
Therefore, intense and continuous efforts in this field will direct controlled and successful
production of valuable, specific, and yet undiscovered plant chemicals.

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References:

• https://www.youtube.com/watch?v=tWQGJ7ELA7Y

• https://mulberrytrees.co.uk/propagation/

• https://agritech.tnau.ac.in/sericulture/seri_mulberry%20cultivation.html

• http://www.fao.org/3/x9895E/x9895e04.htm

• http://egyankosh.ac.in/bitstream/123456789/9099/1/Unit-1.pdf

• A LABORATORY MANUAL FOR SERICULTURE- II semester practical. Mulberry

Cultivation and Silkworm Rearing, edited by Dr. Rohith. L Shankar, Assistant Professor,
Yuvaraja’s College, Mysuru-570005

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