INTRODUCTION

Status of Sericulture in India

India is the second largest silk producing country in the world next to

China, and accounts for 16 % of the global production of silk. More than six

million people are engaged directly or indirectly in this highly remunerative agro-

based cottage industry (Dandin et al., 2003). Though India ranks second in the

world with regard to silk production, it is the highest producer of silk among the

tropical countries.

The agro climatic conditions in many parts of India are favourable for the

luxuriant growth of mulberry facilitating silkworm rearing throughout the year. The

major traditional mulberry growing states in India are Karnataka, Andhra

Pradesh, West Bengal, Tamil Nadu and Jammu and Kashmir apart from some

non-traditional states like Kerala, Uttar Pradesh, Maharashtra, etc. However, the

production of mulberry raw silk is mainly confined to the states of Karnataka,

Andhra Pradesh, Tamil Nadu, West Bengal and Jammu & Kashmir, which

together account for about 98 % of the country’s total mulberry silk production

(Dandin et al., 2003).

Silkworm (Bombyx mori L.)

Silkworms are monophagous insects, which complete their life cycle only

by feeding the mulberry leaves (Plate 1). It was estimated that about 60 % of the
cost of cocoon production goes towards mulberry leaf production (Yokoyama,

1962; Anonymous, 1997). The silkworm synthesizes the silk proteins, fibroin and

sericin, in the silk glands. These are directly derived from the proteins and amino

acids available in the mulberry leaf, on which the worm feeds during the larval

period lasting 25 - 30 days (Ullal and Narasimhanna, 1987). Similarly, 65 - 70 %

of the silk protein produced by the silkworm is directly derived from the protein of

the mulberry alone (Fukuda et al., 1959; Rangaswamy et al., 1976).

It is estimated that the silkworms emerging from 100 diseased free layings

(50,000 larvae), consume more than 1.5 tonnes of mulberry leaf to produce 65 -

70 kg cocoons (Kawakami, 2001).

Mulberry leaves of selected maturity and quality are repeatedly harvested

for feeding the silkworms in accordance with the requirements at different instars

of the silkworm larvae.

Mulberry leaf is a major economic component in sericulture since the

quality and quantity of leaf produced per unit area has a direct bearing on cocoon

harvest (Dandin et al., 2003). More than 60 % production cost of silk is attributed

to mulberry cultivation alone (Yokoyama, 1962; Anonymous, 1997). Hence,

cultivation of mulberry is the most crucial factor in sericulture for the production of

lustrous silk.

Mulberry (Morus spp.)

Mulberry (Morus spp.), the sole food plant of silkworm is a perennial,

deep-rooted, fast growing tree species and is widely adaptable to different agro
climatic conditions. It produces protein rich foliage and is believed to have had its

origin on the lower slopes of the Himalayas.

The genus Morus belongs to the family Moraceae, having 68 species

growing as tree in wild and cultivated forms in different countries of the world

(Sanjappa, 1989). But only five species of Morus viz., M. indica, M. alba, M.

bombycis, M. sinensis and M. multicaulis are potentially important as food for the

silkworm.

In India, mulberry is cultivated under a wide range of agro climatic and soil

conditions (Plate 1). Post 1990, the leaf yield level of mulberry has increased

from 30 - 35 tonnes/ha/yr and reached the present exorbitant level of 50 - 60

tonnes. This increase has been brought about by the development and release of

new high-yielding varieties, and has resulted in higher cocoon production and

crop stability (Dandin et al., 2003).

Further, mulberry is perhaps one of the very rare tree species, which can

serve all the important requirements of mankind namely food, medicine, fodder,

fibre, fuel etc., and is known a wonder plant (Chowdary et al., 2009a; Chowdary

and Mala, 2009b). The mulberry leaf plays a major role (38.2 %) among the

factors contributing for a successful silkworm cocoon crop to obtain good quality

of raw silk (Miyashita, 1986).

Various research institute of Central Silk Board in the country has evolved

high yielding mulberry varieties suitable for the different agro climatic conditions

of the country. Further, it is very well established the requirement of higher doe of

fertilizers for maximizing the production of quality mulberry leaves to the tune of
50 – 60 tonnes/ha/yr. This in turn has increased cocoon productivity and reduced

the leaf-cocoon ratio in southern states of India (Dandin et al., 2003).

For increased qualitative mulberry leaf production various organic

fertilizers (farm yard manure, green manures & oil cakes), chemical fertilizers

(nitrogenous, phosphatic and potassium), biofertilisers (nitrogen fixers &,

phosphate solubulizers) and plant growth promoters (salicylic acid, seriboost,

triacontanol, zinc sulphate, ferrous sulphate) have been recommended during

mulberry cultivation. Simultaneously, bioformulations of different antagonistic

microbes (Pseudomonas fluorescens, Trichoderma harzianum and T. viride)

have also been recommended for crop protection especially for soilborne

diseases (Dandin et al., 2003; Sharma et al., 2003).

The present project work has been carried out with the following objective:

 To evaluate the morpho physiolgoical variations in some of the elite

mulberry genotypes.
 MATERIALS, METHODS & RESULTS

I. General procedures of cleaning glassware

Glassware such as beakers, conical and round bottom flasks, test tubes,

watch glasses, pipettes, etc. were thoroughly washed in detergent solution (0.1

%) after rinsing with potassium dichromate (K2Cr2O7). Then this glassware were

washed with several rounds of clean water, was kept on a draining rack for a few

hours. Finally the drying of the glassware was done in an oven at 110 – 120°C

for 1h. Further, sterilization of glassware was done by keeping them in a hot air

oven at 160°C for 1h.

II. Morphological parameters

The plant growth parameters were recorded on 65th day after pruning of

experimental plot maintained under irrigated conditions as per the recommended

package of practices (Dandin et al., 2003). Data on plant growth parameters (5

plants randomly selected) includes average plant height (cm), number of

branches/plant, number of leaves/plant, average leaf yield/plant, moisture

content of leaf were estimated as follows.

(a). Average plant height (cm)

The length of each shoot of the plant was measured from the plant’s

crown (20 cm above the ground level 0) to the tip of each plant and the average

plant length was expressed in cm.

(b). Number of branches/plant

The total numbers of branches/plant were counter and tabulated.

(c). Number of leaves/plant

The total numbers of leaves/plant were counter and tabulated.

(d). Average leaf yield/plant

The leaf yield/plant assessed by harvesting all the leaves from plants and

fresh weight was taken. From the fresh weight the leaf yield/plant was calculated

by using the following formula:

Total fresh weight of leaves from five plants (g)

Leaf yield/plant (g) =
5

(e). Determination of leaf moisture content

Fresh mulberry leaves from the longest shoot of the plant of each

genotype were collected from ten randomly selected plants in pre-weighed

polythene bags and closed with rubber bands to avoid loss of moisture from the

leaves. Immediately after transportation to the laboratory, the samples were

weighed for fresh weight along with pre-weighed polythene bags. The fresh

weight of leaf samples was then determined by subtracting the weight of the

polythene bags. The leaf samples were then dried in a hot air oven at 65 – 70°C

temperature till a constant weight was arrived. Three replications were

maintained. The percentage of leaf moisture was then calculated by using the

following formula on fresh weight basis (Anonymous, 1990).

Fresh weight of leaf – Dry weight of leaf

Leaf moisture (%) = × 100
Fresh weight of leaf

Table1: Growth and yield parameters of elite mulberry

genotypes under irrigated conditions

SL NO VARIETIS HEIHGT NUMBER NUMBER LEAF MOISTURE

OF THE OF OF YIELD / %
PLANT BRANCHE LEAVES/ PLANT
(cm) S /PLANT PLANT
1 V1 126.7 16 247 0.650 75.5
2 S 1635 120.2 13 176 0.350 70.7
3 MYSORE 103.5 18 254 0.293 60.5
LOCAL
4 RC 1 102.5 15 182 0.385 70.2

5 RC 2 108.3 17 189 0.450 75.3

6 S 13 106.9 17 173 0.350 70.5
7 S 34 105.4 18 165 0.363 60.8

III. Physiological parameters

Measurement of leaf water potential in different
Mulberry varieties
Aim: To measure the leaf water potential energy of water\unit mass of water in a

biological system.

Principle In a thermodynamic sense it is an indicator of energy stands of water

in system. Water from soil absorbed by mulberry roots and conducted up warded

against gravitational pull to the apex of the plant. There by disturbing the water to

different organs only due to gradients in water potential. Water potential is a

drying force for SPAC< soil plant atmospheric condition>. Water potential is

indicated on “ψ. Water potential depends on y component potentials under the

same atmospheric pressure and temperature which are pressure potential;

osmotic potential or solute potential; gravitation potential and metric potential

shown as below ψw= ψgp+ψop+ψmp+ψpp

Gravitational potential: The position of water in the system.

Osmotic potential: Dissolved solutes in water.

Metric potential: Metric forces, which bind water in the system i.e., soil or leaf

Tissue.

Pressure potential: Hydrostatic pressure or trigger pressure in the system.

Overall water potential is a tool to axes water stress and plant metabolism. Water

potential controls the movement of water. Water potential is measured in terms of

pressure units like bars, MPa and Kilo Pascal’s

10 Bars =1 MPa

10 Mpa= 1 KPa

Water potential in the biological system is always a negative

(-), With reference to the pure water the water potential is “o” Hence,

measurement of leaf water potential in mulberry is important to understand

biomass production.

Materials:

 Index leaves of different mulberry varieties (V-1, S-36, S-34, Mysore

Local, S-13, RC-1 and RC-2).

 Blade

 Water potential cups (4 cm diameter)

Instruments:

Dew point potential meter (Model wp-4) Decagon devices. Inc.

This is the fastest instrument for measuring water potential giving reading

directly in MPa. It measures water potential of both leaf as well as the soil from

0to 60 MPa. With accuracy of MPa. WP-4 uses the chilled mirror dew point

technique to measure the water potential of the sample. Instrument contains led

indicator light on left side and functional keys on right side. Sample drawer and

LCD (liquid crystals display). It is a portable instrument.

Procedure:

Index mulberry leaves of different verities were harvested and kept on Plastic

covers and brought to the laboratory. After wiping the surface with tissue paper to
remove dust circular disc of 4cm diameter were cut with blade by the use of

sample cups from 3 sides of each leaf and 3 replication were maintained. Leaf

disc were kept in for temperature equilibrium on the instrument for 5-10 mins, by

keeping inside cups. Later on the lids are removed and cups were put into the

drawer and the water potential measurements were taken. The water potential of

leaf disc samples were measured by relating the sample water potential reading

to the vapour pressure of air in equilibrium. WP-4 measures water potential by

equilibrating the liquid phase of water with vapour stage of water in the closed

chamber with the help of sensor block, a dew point sensor, and temperature

sensor and infrared thermometer. From these measurement the vapour pressure

of the air is computed at dewpiont temperature in the water potential of samples

in equilibrium and water potential readings of samples displaying on LCD.

Result: The measurement of leaf water potential of different mulberry samples,

which are noted on mega, Pascal’s (Mpa) are below in the table.

Inference: It is inferred that Mysore local is having highest water potential

followed by S-13, S-36, V-1, RC-2, S-34 varieties.

Table2: Water potential in different elite mulberry genotypes

Varieties Tempe Avg PP Avg MPa Avg

rature
 V-1 31.0 31.1 40.83 4.84 -6.65 -6.64

31.1 40.85 -6.90

31.2 .40.83 -6.54

S-36 1.5 31.53 4.78 4.87 -5.78 -6.67

1.5 4.94 -8.45
1.6 4.90 -5.78

S-34 31.3 31.66 4.75 4.78 -5.48 -6.10

31.5 4.95 -8.66
31.3 4.63 -4.16

Mysore 31.7 31.7 4.93 4.93 -8.27 -8.15

local
31.7 4.94 -8.37
31.7 4.91 -7.83

S-13 31.8 31.9 4.86 4.91 -7.02 -7.83

31.9 4.93 -8.13
32.0 4.94 -8.34

RC-1 32.1 32.1 4.72 4.74 -5.05 -5.34

32.1 4.75 -5.06
32.1 4.76 -5.39
RC-2 32.1 32.03 4.80 4.82 -6.09 -6.44
32.1 4.82 -6.36
31.9 4.85 -6.86

b) Measurement of leaf relative water content and

water saturation deficit in different mulberry verities
Aim: To determine the leaf relative water content and water saturation deficit in
different mulberry varieties.

Principle:

Water is the important input for mulberry growth and crop productivity. The entire

plant water relation depends upon the contents of water on the plant system .All

physiobiochemicall activities depend on the plant water status. Water status in

the plant can be measured in terms of leaf moisture content and moisture

retention capacity and relative water content (RWC). Relative water content is

defined as water or moisture content of mulberry leafs in relation to absorption

and retention capacity of the plants for physiological efficiency. RWC is an

intergraded measure of plant water status in composition to turgidity. Relative

water content is a physiological tool to understand plant water status, which

directly reflects the physiological mulberry. Water saturation deficiency is a

indicator of percent water reduction to its total turgidity. Cell turgidity reflects

division and differentiates cells and growth of the plants.

Materials:
Index leaves of different mulberry varieties (V-1, S-36, S-34, Mysore local, RC-1,

RC-2)

 Sample Cup

 Petriplates

 Blade.

 Electronic weighing machine.

 Distilled water.

 Hot air oven.

Procedure:

Index leaves of different mulberry varieties are harvested and brought to the

laboratory by keeping in polythene cover. The leaves are wiped out with tissue

paper to remove dust. With the help samples cup 4 cm diameter. 6 leaf discs was

cut from each variety. Soon after this the fresh weigh of the leaf discs are

weighed in electronic beam balance and the leaf disc were immersed in distilled

water on petriplates under laboratory condition with ambient light for 3hours.

After, the leaf discs were taken out in the petriplates and water are wiped out by

using blotting paper. Then the turgid weights of leaf discs are recorded with the

help of electronic balance. Then the leaf disc are put in sample covers and kept

in hot air oven for 24 hours to get constant dry weight for each sample variety.

The oven dried (60 c) leaf disc was measured for the dry weight of the sample.

Result: The leaf relative water content and water saturation deficit of different

verities are as follows.

 Table 3: Relative water content water saturation deficit in
different elite mulberry genotypes

Sl. Varieties Fresh Turgid Dry weight

weight (gm) weight (gm) (gm)
No.

1 V1 1.395 1.686 0.496

2 S 13 1.270 1.485 0.416
3 S 34 1.365 1.536 0.404
4 S 36 1.209 1.363 0.384
5 Mysore 1.005 1.143 0.345
local

6 RC 1 1.223 1.339 0.393

7 RC 2 1.302 1.470 0.446

C) Determination of Transpiration Rate in Mulberry

Leaves
Aim: To determine the rate of transpiration in different mulberry varieties by

gravimetric method.

Principle: one of the most important process of plant function is uptake of water

from the soil .the mechanism for water uptake and transport with in the plant is

transpiration, the loss of water from aerial part of plant in the form of vapour.

Transpiration plays several vital roles in keeping the plant alive. For most the

evaporation of water from leaf surface allows plants to cool themselves. Without

this ability the plants would quickly over heat and die on bright sunny days. It also

facilitates of nutrients uptake into the plants. Transpiration of water from the plant

creates gradients that drive movement in the soil towards the root.

Materials required:

 Twigs of different mulberry varieties with uniform number of leaves (V1, S

36, Mysore local, RC 2, RC1, S 13, and S 34).

 Conical flask

 Beaker and water

 Non absorbent cotton

 Aluminium foil

 Electronic balance

 Blade

Procedure: A 500ml beaker was taken with water in the lab and went to the field.

In field the longest branch of different mulberry were selected. Those branches
were bent into the beaker and the twig was cut under water with the help of a

blade. Those twigs are taken and brought to the laboratory and kept in water in

the beaker. Then immediately the twigs were transfer with uniform number of

leaves into a conical flask with full of water. Then mouth of the conical flask was

plugged with non-absorbent cotton, and the sealed with alluminium foil by taking

care to avoid the evaporation. Next the entire setup was weighed in electronic

balance and recorded. It is the initial weight of this setup. Finally the entire setup

was kept in open field and bright sunlight and allowed it for 2hours. After exactly

2hours the setup was brought to the lab and the final weight was taken.

Calculation: The transpiration rate is calculated in the different mulberry variety

(V1, S 36, Mysore local, RC1, RC2, S 13, and S 34) by the following formula.

Transpiration rate = (initial weight – final weight)/initial weight

TR %=( IW– FW)/ IW * 100

Result: By the above formula transpiration rate measured individually for 7

mulberry varieties.

Table4: Transpiration rate in elite mulberry genotypes

 Sl. VARIETIES INITIAL WEIGHT FINAL WEIGHT
No. (gm) (gm)
1 V1 453.8 447.2
2 S 36 433.8 426.9
3 Mysore local 383 375.6
4 RC 2 383.4 375.8
5 RC 1 439.8 432.4
6 S 13 436.5 424.6
7 S 34 440.8 436.4

d) Study of phenomenon of osmosis in plant water

relation

Aim: To study the phenomenon, of osmosis in plant water relations.

Objective: To understand the phenomenon of exosmosis and endosmosis using

potato resin osmoscopes.

Principle: Plant water status decides that crop growth in biomass production.

Hence, study of plant water relation is most important to understand physiology

of crop productivity. Water moves from its high potential to the lower one in the

biological system by the phenomenon of osmosis. Mass flow and diffusion of

water between different plant cells takes place through a semipermiable cell

membrane from a higher concentration to lower concentration. Being a universal

solvent, water dissolves different solutes in the cell and hence decreasing in

water potential. Due to concentration gradient of different solutes water may be

absorbed by the cells, is called endosmosis, and if a cells loose water to the

outside medium is known as exosmosis. This phenomenon pertains in plant

water relations, most specially for entry of water into the mulberry roots in the soil

media and translocation between different cells.

Materials required:

 Petriplates

 Distilled water

 Potato tubers

 Resins

 Blade

 1% sugar solution

 Blue ink

Procedure:
Endosmosis: A petriplates was taken and poured distilled water and then resins

were kept in it at five numbers. Then the setup was leave for 6 hours.

Observations: After 6 hours flaccid resins were swollen, due to the phenomenon

of endosmosis, i.e., the entry of distilled water into the resins to the turgid

condition. Since the dry resins contain less water (adhered to the cell contents),

the water from the outside medium enters to the resins and then it become

turgid.

The water enters due to the phenomenon of osmosis by cell membrane.

Exosmosis: A potato tuber was taken; the skin was peeled off and cut to make a

block. Than it was scooped with a blade and made well. Than in the well-distilled

water was added with a drop of ink and the level of water was marked with a pin.

After that the entire setup was kept in 1% sugar solution on a petriplates. Next

the setup was leave for 6 hours.

Observation: after 6 hours it was observed that the water level of the potato well

has come down. Simultaneously, it was observed the volume of sugar solution

has raised and attain blue colour. Sugar solution contain low osmotic potential,

then the distilled water, has high osmotic potential.

e) Estimation of total nitrogen and crude protein

content in mulberry leaves

Aim; Estimation of leaf nitrogen was carried out by the Micro-Kjeldahal method

Principle:
 The sample is digested with concentrated H2SO4 in the presence of

catalyst to convert the nitrogen in protein or any other Organic material to

ammonium sulphate. By steam distillation of this salt in the presence of a strong

alkali, ammonia is liberated and collected in Boric acid solution as ammonium

borate, which is estimated against a standard acid by titration. On an average

most proteins have 16% nitrogen in their composition. In other words, 1mg

nitrogen equals 6.25mg of protein. Thus, by finding out the amount of ammonia

formed, from a known amount of sample, one can calculate the amount of protein

present (1ml of 0.1 N Acid =1.401mg N).

Reagent:

 Concentrated H2SO4.

 Mercuric Oxide.

 Potassium sulphate

 Sodium Hydroxide – SodiumThiosulphate solution; dissolve

600gms of NaoH and 50 gms of Na2SO3.5H2O in water and make
up to 1 lt.

 0.02N standard HCl or H2SO4.

 4% Boric Acid Solution; dissolve 4gms of H 3 BO3 in warm water

and dilute 100ml.

Mixed indicator solution: mix two parts of 0.2% methyl red in ethanol with

one part of 0.2 % methylene blue in ethanol (mix one part of 0.2% methyl red in

ethanol with five parts of o.2% bromocresol green in ethanol).

Procedure:
 Estimation of leaf nitrogen was carried out by the Micro-Kjeldahal method,

following the basic principles laid out by Anonymous (1990). One hundred mg of

leaf powder was placed in a long necked micro-Kjeldahal flask and 10 ml of

concentrated sulphuric acid was added to it. Samples were digested in the micro

digestion unit till the digested solution turned white in colour. Later, 2 ml of 30 %

hydrogen peroxide was added to it and digestion was continued till the sample

became clear. Two to three drops of methyl red and Bromocresol green mixed

indicators were added to 25 ml of Boric acid solution in a 250 ml conical flask and

steam distilled the digested samples in a Kjeldahal distillation unit. After

distillation, the boric acid solution was titrated against 0.1 N sulphuric acid

solution till the green colour of the boric acid changed to brick red. The total

nitrogen content of the leaf samples was estimated by using the following formula

and then the crude protein content of the mulberry leaf was determined

multiplying the total nitrogen with the conversion factor of 6.25.

Calculation:

TV * N of acid* 0.014* vol of digested sample *100

Total Nitrogen (%) =
Weight of the sample (g) × Aliquot taken

Crude protein (%) = Total Nitrogen × 6.25.

Reagents preparation:

Catalyst mixture: Mix 20 g CuSO4 and 100 g K2SO4 finely by using piton and

mortar
Mixed indicator: Mix 0.1 g bromo cresol green with 0.07 g methyl red and

dissolve in 100 ml ethanol.

N HCl: N1V1 = N2V2;

500 X 0.1 = 11.3

500 X 0.1 = 4.42 ml concentrated HCl in 500 ml distilled water

In Kjeldhal flask fill NaOH 40 % solution (40 g in 100 ml)

Milli equivalent weight of nitrogen is 0.014

Table: 5. Total nitrogen and crude protein content in some elite

mulberry genotypes

Variety Total Nitrogen (%) Crude protein (%)

K2 1.96 12.25
V1 2.63 16.4
S 36 2.12 13.25

f) Estimation of available phosphorous.

Aim: To estimate the phosphorus in mulberry leaves

Principle:
 Available phosphorus was estimated by the method of Olsen et al. (1954).

In this method the available phosphate reacts with ammonium molybdate, in an

acid medium to form molybdophosphoric acid. The molybdo-phosphoric acid

then reduces to a blue coloured complex through reaction with stannous chloride.

Absorbance readings were taken at 660nm wavelength using a

spectrophotometer. A standard curve was prepared from the absorbance

readings of different concentrations of potassium dihydrogen phosphate.

Procedure:

One gram of leaf was taken in a 250 ml conical flask. To this add 20 ml of

0.5 N sodium bicarbonate (pH 8.5) and a pinch of carbon black were added and

continuously shaken for 30 minutes and then filtered through Whatman No.1 filter

paper. Similarly, a blank was prepared without using leaf. 5 ml of clean filtrate

was placed in a 25 ml volumetric flask to which 5 ml of 1.5 % ammonium

molybdate solution was added. After this ,10 ml of distilled water and 1 ml of

stannous chloride solution (10 g dissolved in 25 ml conc. HCl and then 0.5 ml of

this diluted to 66 ml with distilled water) were added and a final volume was

made up of 25 ml, by adding distilled water and allowing up to 30 minutes for

colour development. This was read at 660 nm wavelengths in a

spectrophotometer. The phosphorus (P) content was calculated (kg/ha) by

comparing the spectrophotometer readings with the standard curve.

Calculation:

Available R × Total volume of extract × Volume made up × 106

P (kg/ha) = × 2.24
 106 × wt. of sample (g) × volume of aliquot taken

Where, R = µ g of P in the aliquot (obtained from the standard curve).

Table: 6. Available phosphorus content in some elite mulberry

genotypes

Variety Available
phosphorus (%)

K2 0.22
V1 0.28
S 36 0.26

(f). Estimation of available potassium

Aim: To estimate the potassium in mulberry leaves

Principle:

Potassium was extracted from the leaf with the help of saturated

ammonium acetate solution, by shaking, followed by filtration. The extract was

determined by using a flame photometer as described by Rao (1993). One gram

of dry leaf powder was placed in a 250 ml conical flask; 20 ml of neutral

ammonium acetate solution was added to it and shaken for 30 minutes.

Immediately after this, it was filtered through Whatman filter paper No.2.

Potassium (K) concentration in the extract was determined with the help of flame

photometer. The reading was compared with the standard curve to determine

the amount of available potassium.

Reagent: 1.Potassium standard 100ppm of potassium dissolve 0.191g of KCL in

some volume of distill water and than make up the volume to one liter.

Materials: Flame photometer, pipettes, volumetric flask, etc.

Procedure:

Preparation of standard curve:

Take 0, 1, 2, 3, 4 and 5ml of 100ppm potassium solution in a separate 50ml

volumetric flask. Make up the volume to 50ml distill water and mix well. After

adjusting the needle of flame photometer to zero by feeding blank, adjust the

needle to 100 by feeding maximum concentrated k solution. Then feed the

standard to record the flame photometer readings. Plot flame photometer

readings.

For plant sample:

1. Pipette out 5/10ml of digested sample into a 50ml volumetric flask

and add 5/10ml of vanadomolybdate reagent

2. Make up the volume to 50ml and mix.

3. Read the colour intensity at 430nm after 30min.

4. Compare the unknown sample absorbance with standard cure and calculate

percent of the sample

Calculation:

R × Volume of extract × 106

Available K (kg/ha) = × 2.24
106 × Wt. of the leaf sample taken (g)

Where, R = ppm or µ g of K (obtained from the standard curve).

Table: 7. Available potassium content in some elite mulberry

genotypes

Variety Available potassium

(%)
K2 1.01
V1 1.26
S 36 1.11

Elite Mulberry genotypes

 V1

Mysore Local
RC 1

RC 2
S 13
S 34
THEORY CUM DEMO ON

NUTRIENT DEFICIENCIES IN MULBERRY AND THEIR

MANAGEMENT

Mulberry (Morus spp.) is intensively cultivated for feeding of money

spinning insect, silkworm (Bombyx mori). Silkworm rearing under controlled

environment for production of lustrous silk is one of the major activities in India

providing ample employment opportunities. Hence maximization of quality leaf

production is the vital aspect. Seventeen elements (9 macro and 8 micro

nutrients) have been reported to be essential for the growth of mulberry (Table

1). Plants take carbon, hydrogen and oxygen mostly from air and water, while the

other nutrients are taken from the soil especially in the form of ions. If the soils

are deficient with these nutrients, the plants show the deficiency symptoms. The

deficiency symptoms can be categorized in following groups.

Diagnosis of nutrient deficiencies in mulberry

• Chlorosis

• Necrosis or death of plant tissue

• Lack of new growth of terminal growth, resulting in resetting

• Accumulation of anthocynin and an appearance of reddish colour

• Stunting or reduced growth with either normal of dark green colour or

yellowing.
 Nutrient deficiency symptoms can be easily diagnosed in mulberry by visual

observation (Table 2 & Fig. 1)

Role of different macro and micro nutrients for qualitative

mulberry leaf production

Nitrogen
Nitrogen is the building block to form amino acids, proteins, DNA, RNA,

amides, amines, chlorophyll and structural constituents of cell wall etc. Hence it

plays an important role in mulberry foliage production.

Phosphorous
Phosphorous constitutes of enzymes, phospho proteins, phospho lipids

and nucleic acids. It promotes early maturity of leaf. It is a part of NADP, ADP,

DNA and RNA, electron transport in oxidation-reduction process. Translocation

of sugar and starch etc.

Potassium
Potassium actively involves in maintenance of cell osmotic potential by

regulating through stomata. It also provides resistance to plant against lodging of

pests/ diseases. Besides these functions potassium involved in formation of ATP,

starch and protein synthesis.

Calcium
Calcium as calcium phosphate helps in cell elongation and cell division

and also involves in translocation of carbohydrates. It plays an important role in

enzyme activation.
Magnesium
Magnesium constitutes as an essential molecule in chlorophyll. It acts as a

co-factor of transphosphorylase, dehydrogenase and carboxylase enzymes.

Associates in formation of polypeptide chain from amino acids and also involves

in formation of sugars.

Sulphur
Sulphur constitutes in cysteine and methionine formation specifically and

vitamins and hormone formation generally. It also involves in oxidation-reduction

process.

Iron
Iron plays an important role in synthesis of chlorophyll, nitrogen fixation,

photosynthesis, electron transfer etc. Regulates respiratory enzymes, particularly

cytochrome enzyme.

Zinc
Zinc acts as an important component of number of enzymes, protein

synthesis and also regulates plant growth hormones i.e. auxins.

Manganese
Manganese helps for evolution of oxygen during photosynthesis and also

a component of number of enzymes. It is a structural component of certain

metallo proteins. Activates electron transport system.

Copper
Copper involved in several enzyme systems. It helps in formation of cell

wall, electron transport system and oxidation-reduction process.

Boron
Boron plays an important role in transport of sugars across the cell

membrane, controls formation of sugars and starch, transpiration, carbohydrate

metabolism, amino acid metabolism and protein synthesis.

Molybdenum
Molybdenum actively involved in Nitrogen cycle by formation of nitrate

reductase enzymes.

Chlorine
Chlorine involved in phosphorylation, photosynthesis and osmotic

pressure regulation by acting as an ion in counter balance to cat ions.

Silicon
Silicon helps in possessing rigidity of cell wall, acts as a protector and

assists in photosynthesis.

Sodium
Sodium helps in osmotic regulation and also fulfills the functions of

potassium in some cases.

Vanadium
Vanadium involved in oxidation-reduction process of cells.

Cobalt
Cobalt involved in nitrogen fixation process.
Conditions ideal for nutrient absorption

The ability of plants to absorb water and nutrients depends on permeability of

root surfaces, which in turn influenced by the metabolic activity of the roots. The

metabolic activity is increased by supplying oxygen to the roots and removing

carbon dioxide. The ionic absorption process of nutrients is most efficient under

the following conditions, when

 The soil has a high concentration of nutrient ions.

 The soil is well aerated allowing oxygen to diffuse readily into the soil and

carbon dioxide diffuse out.

 The soil is sufficiently moist to permit ions in solution to contact large

areas of root surfaces moisture are the medium for transport.

 The leaves are exposed to adequate light

 If the nutrients are not made available to the plants as per the requirement

some symptoms may visible on the leaves.

Management of nutrient deficiencies

• It is advisable to use the inorganic fertilizers after the soil testing along

with the organic manures and biofertilizers.

• An emphasis should be laid down for incorporation of green manure in the

soil so that balanced nutrient management can be followed with out

decreasing the leaf yield producing potentiality in mulberry crop.Foliar

 application of Seriboost – a micro nutrient formulation. Mix 500 ml of

Seriboost in 200 liters of water for 1 acre of garden. I spray – 23 to 25days

after pruning/ leaf picking. II spray – 30 to 32 days after pruning / leaf picking.

Table 1. Essential elements and its available form to Mulberry

Elements Available form

A. Macronutrients
1. Sulfur So4
2. Phosphorous HPo4
3. Magnesium Mg2+
4. Calcium Ca2+
5. Potassium K+
6. Nitrogen No3, NH4
7. Oxygen O2, H2O
8. Carbon Co2
9. Hydrogen H2O
B. Micronutrients
1. Molybdinum Mo O-2
2. Copper Cu+4, Cu+2
3. Zinc Zn+2
4. Manganese Mn+2
5. Iron Fe2+, Fe3+
6. Boron Bo3, B4O7
7. Chlorine Cl
8. Nickel Ni

Table 2. Visual symptoms of nutrient deficiencies on Mulberry leaves

Element Deficiency Symptoms

On terminal leaves
Calcium Plants remain dark green, young bud leaves chlorotic,
finally death of terminal bud.
Boron Young leaves of the terminal buds with loose colour at the
base, death of the terminal bud.
On young leaves
Sulfur Light green leaf, veins still paler, no dead spot.
Iron Chlorosis, no spots, main vein typically green.
Copper Interveinal chlorosis, resetting and permanent wilting, leaf
detaches easily.
Manganese Leaf chlorotic, main and small vein dark green.
 On old leaves
Nitrogen Dwarfing and abnormally light green plant leaf erect, light
green to yellow.
Phosphorous Dwarfing and abnormally dark green plant, leaf erect and
unusual narrow. In acute condition greenish brown to
black, bronzing on backside of leaf.
Potassium Chlorotic leaves, small dead spots or specks at the tips
and margins, rusty appearance, crimping and cupping of
margin and tips.
Magnesium Chlorosis starting from tips and margins, with no dead
spots, vein green, base necrosis in acute condition, leaf
detaches easily.
Zinc Leaf become narrow and small, lamina chlorotic, veins
green, dead spot develop all over the leaf including veins,
tips and margins.
Molybdenum Leaf lighter green, golden yellow to orange, dead spot all
over the area except veins, affected areas extrude.

Fig. 1. Nutrient deficiency symptoms in mulberry

Nitrogen deficiency Phosphorus deficiency

Potassium deficiency Calcium deficiency

Magnesium deficiency Zinc deficiency

Sulfur deficiency Iron deficiency

 Manganese deficiency Copper deficiency

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