Efficiency and quality indicators of in vitro propagation of high-yielding mulberry varieties and hybrids

Scientific research results in pandemic conditions (COVID-19)

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5.

Столяров Владислав Олимпийское воспитание. Теория и

практика. [Книга]. - Москва : Национальное образование, 2014. - стр.
402.

6.

Столяров Владислав Современный олимпизм и олимпийская

педагогика: достижения, проблемы, перспективы [Книга]. - Москва :
ПЛАНЕТА, 2018. - стр. 528.

Yorkinoy Mirzaeva - Senior Lecturer, Department of Sericulture and

Mulberry growing, Tashkent State Agrarian University Uzbekistan

Nortoji Khujamshukurov - Professor of Tashkent University of Chemical

Technology, Doctor of Biological Sciences Uzbekistan

Nigora Sokhibova - Tashkent State Agrarian University, Department of

Sericulture and Mulberry growing 2nd year PhD student Uzbekistan

EFFICIENCY AND QUALITY INDICATORS OF IN VITRO PROPAGATION OF

HIGH-YIELDING MULBERRY VARIETIES AND HYBRIDS

Yorkinoy Mirzaeva, Nortoji Khujamshukurov, Nigora Sokhibova

Abstract: This article describes the hybrid of Oshima I2N2 (Morus

bombycis Koidz.) × SANIISH-25 (Morus multicaulis Perr.), Obtained by
crossbreeding of mulberry tree, which is widely used in production in
Uzbekistan. The specificity of the variety and the development of each clone
were studied in the process of different effects on the growth and
development of mulberry in standard nutrient media in vitro from the
selected Jarariq 9 varieties.

Our study found that when we planted 3 clones in a new environment,

leaving one joint, the third apical part grew better than the 2nd medium
clone, but the 3rd clone growth and root rate, which was cut close to the 3rd
nutrient medium, was higher than the 1st and 2nd clones.

Key words: Mulberry (Morus sp.), micropropagation, in vitro screening,

M.Bombycis Koidz, M.Multicaulis Perr., M.Nigra L., M.Alba L.

It is no secret that today there are all the conditions for scientific and

practical research using the domestic potential for further development of
the economy of Uzbekistan. Our research is in vitro micro propagation of
mulberry tree varieties and hybrids suitable for our climatic conditions. Our
results are a scientific and practical demonstration of the advantages and
effectiveness of traditional multiplication methods and high quality
indicators.

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51

Due to the fact that silkworm breeding in the country can be exported

in a very short time - 30-35 days, now it is planned to re-feed silkworms 3-
4 times. In order to fulfill the set plans, there is a need to further increase the
number and variety of mulberry trees. It is not necessary to use fertile land
to expand mulberry fields, as the mulberry tree is drought tolerant and can
grow even in areas with high salinity or salinity. Farmers always plant
mulberry trees on both sides of the roadside ditches and on the other side
of the border to prevent cotton and grain fields from being washed away by
the ditches in the downpour. In the provinces, you have seen mulberries
planted in two rows on both sides of the bush, because the mulberry tree is
planted to consume the fruit and, most importantly, to use the leaves to feed
the silkworm. Due to the efforts of representatives of the silk industry, great
opportunities are opening up for the development of silkworm breeding.

However, there are a number of problems in the process. The scarcity of

mulberry plantations, which are the main food for silkworms and the fact
that their existing ones have become unusable, is one of them. It is no secret
that in some districts of the country the productivity of mulberry trees has
significantly decreased due to negligence in the implementation of agro-
technical measures. In addition, mulberry disease and pest control
measures are not carried out in a timely and quality manner.

The head of state instructed the Ministry of Agriculture,

"Uzbekipaksanoat" Association, regional governors and heads of relevant
departments to allocate land for the establishment of mulberry groves,
renovate the old ones, and use water-saving technologies. The goal is to
increase cocoon production to 21,000 tons next year and 30,000 tons by
2025.

Based on this, by the end of 2022, it is planned to plant 20,000 hectares

of new mulberries and renew 10,000 hectares of obsolete mulberries.

Mulberry propagation is mainly of two types: from traditional

vegetative cuttings or by grafting and from seed. In these methods, it is not
possible to grow any amount of high-yielding plants without the diseases we
expect. The traditional method of propagating mulberries does not give an
increase in economic efficiency. A group of scientists from the University of
Taif in Saudi Arabia (2014) (Attia O Attia, Eldessoky Sdessoky, Ehab I El-
Hallous & Hanan F Shaaban) also pointed out that reproduction through
breeding work is also time consuming.

In India, Bangladesh, Pakistan and a number of other Asian countries,

the mulberry tree is the only food for silkworms, longevity, soil erosion
protection, plant protection from downpour, the use of fruit as a natural
nutrient for small animals and birds during ripening considering the
economic potential of mulberry , a group of Indian scientists have pointed
out that most research is being done on mulberry trees to create high-

Scientific research results in pandemic conditions (COVID-19)

52

yielding, disease-resistant, nutritious mulberry varieties adapted to the
natural climate. Kunjupillai Vijayan. A. Tikader and others. 2011. Article ·
June 2011 DOI: 10.1007/978-3-642-20447-0_5

Researchers in China, India, Japan and Korea, where silkworm breeding

is practiced, are also working on genetic improvement through mulberry
selection, genomic and breeding resources, as well as conservation,
collection, use and transmission of valuable wildlife to future generations.

Propagation of mulberry varieties is one of the effective methods of

micro clonal propagation by modern advanced in vitro method. Micro clonal
reproduction allows reproduction at any time of the year.

Over the past decade, scientists have advocated the high efficiency of

propagation of several tropical and subtropical fruit trees from cell tissue (
Agarwal et al., 2004; Tsukamoto et al 2007 ; Perez Tornero et al..,2010;
Skiada et al ., 2010).

In our research, we began with the development of in vitro method,

which is effective and of great importance in the development of mulberry
breeding, selection of high-yielding disease-resistant mulberry varieties and
hybrids in the regions of the country.

Jarariq-9 variety and Uzbek hybrid were selected for the experiment.
Several experiments were carried out to introduce the selected

mulberry varieties and hybrids into the culture.

Cut green shoots of mulberry are left in running water overnight and

then rotated in a magnetic stirrer in a soapy solution for 30 minutes.

To sterilize the cut green shoots for propagation, cleaning was carried out

in several different solutions and an acceptable one was selected.

After cleaning the cut green shoots from bacteria and fungi, experiments

on the introduction of culture began in the shortest time period possible.

The work process was carried out in a special laminar-box. Pre-distilled

water and various prepared nutrient media were sterilized and prepared in
an autoclave for the pre-work process.

The next task we need to determine is to select the optimal one for in

vitro propagation of mulberry in a medium prepared at different
concentrations.

When preparing the nutrient medium in a volume of 1 liter, 20 mg / l of

each mother solution is taken and mixed with half the amount of distilled
water. The sucrose is dissolved in the rest of the water. Macro- and micro-
nutrients and sucrose solutions are poured together, physiologically active
additives are added, the pH is increased to 5.6-5.8, and agar is added.

Samples from all mulberries were selected according to the age of the

mulberry. Samples were taken from young seedlings planted from fresh
seeds, 2–3-year-old and 10–15-year-old mulberries. As a result of the
experiments, it was determined that the most suitable specimen for

Scientific research results in pandemic conditions (COVID-19)

53

propagation was taken from 2-3-year-old mulberries, and the process was
planned to continue in these samples. This is because the results of in vitro
propagation of 2-3-year-old mulberry tree green shoots with a high rate in
our experimental results were studied.

Modifications to various components in our prepared nutrient medium,

such as phyto-hormones, vitamins, sucrose, and other supplements, have
different effects on the growth and development of mulberry.

It has been pointed out that in plant tissue proliferation; a number of

experiments are required to select the optimal norm for the growth of each
plant cell tissue (Skoog & Miller 1957).

Muhammad Akram and Fahim Aftab, both scientists at the Department

of Botany at the University of Punjab in Pakistan, also cited Morus alba L.'s
2012 study on in vitro reproduction of white mulberry.

At the time of introduction of mulberry into the culture, it was found

that the varieties depend on the varietal characteristics. Although the
growing parts of the mulberry were taken at the same time and introduced
in the same environment, the thickening of the stem of the Balkhi mulberry
and Shotut variety caused tissue damage and this affected the stem.

Mulberry soaked in special solutions is transplanted into a sterilized

nutrient medium in a laminar-box. Before planting, mulberry is sterilized,
soaked in water and the crushed areas are cut off. The nutrient medium was
also in different concentrations and the effect of mulberry on the sterile
environment was studied.

The cut genotype of the Uzbek hybrid was transplanted into the nutrient

medium at 7 days. In this case, at 12–15 days, bluntness was observed in the
root part, and the root began to emerge. Only the transition from a hormone-
free environment to a sterile state was used.

The plant, which was planted on April 6, was cloned on May 16. Leaving

one joint, 3 clones were removed from each seedling in the jar and planted
in the prepared nutrient medium. The original material was put back into
growing.

On May 16, 3 cuttings were taken from each shoot at 3 weeks of age from

the mother material in the arrow state, where the in vitro micro-
reproduction of the mulberry was transferred to the new medium and we
were able to get 12 cuttings from 4 seedlings.

In the process of micro propagation of mulberries, the specificity of the

variety and the development of each clone have been studied and the study
continues. Because after the apical part of the plant was cut, the 2nd joint
showed slow growth, the 3rd joint developed from the lateral bud, and the
growth from the lateral bud in the 4th joint also gave a very good indication.
Micro-cloning was performed at 3 weeks and 4 weeks after the introduction
of mulberry into the culture. Different sections of mulberry were used as

Scientific research results in pandemic conditions (COVID-19)

54

planting material. The apical part of the mulberry, the first joint and the
second joints were planted separately.

Our research showed that when we planted 3 clones in a new

environment, leaving one joint, the third apical part grew better than the
2nd medium clone, but the 3rd clone growth and root rate, which was cut
close to the 3rd nutrient medium, was higher than the 1st and 2nd clones.
These indicators are illustrated in the table. The results of our research in
Table 1 show the growth rates of the apical part of the 1st end of the hybrid
of Uzbekistan.

Table 1

Analysis of growth rates when cloning Uzbekistan hybrids.

Table 2

The growth rates of the 2nd middle part of the hybrid of Uzbekistan are

covered.

№

Chosen
for the
experime
nt

Repit
ition

The
time
of
planti
ng

Initial
state

Results obtained
after 14 days
20.04

Results obtained after 24
days
30 .04

Heig
ht,
sm

Root
lengt
h, sm

Numb
er of
roots,
piece
s

Height
, sm

Root
lenght,
sm

Number
of rots,
pieces

1

st

part

apical

1

06 0.4 1.5

6.6

1.1

4

10.2

2.3

6

2

06.04

1.5

7

0.9

4

10.9

2.1

7

3

06.04

1.5

6.5

0.8

5

10.1

2.0

7

Average

06.04

1.5

6.7

0.93

4.3

10.9

2.13

6.66

Scientific research results in pandemic conditions (COVID-19)

55

Table 3

The growth rates of the hybrid part of Uzbekistan close to the 3rd lower

environment are given.

Each jar was filled with 100 ml of nutrient medium and 25 ml of nutrient

medium was calculated for each clone rod. In this case, phytohormones were
used. In the second experimental variant, the growth rates of seedlings
planted on May 3 were found to be higher than on April 6. Growth rates were
found to be higher as the transplants adapted to the nutrient medium. Table
4 shows a comparative table of averages of clones planted in 3s. The highest
rate was the 3rd clone growth rate, which was close to the nutrient medium.

Table 4

This table shows the average values of Tables 1-2-3 from the hybrid of

Uzbekistan

№ Chosen

for the
experime
nt

Repitio
n

The
time of
plantin
g

Initial
state

Results obtained after
14 days 20.04

Results obtained after 24
days 30 .04

Heigh
t, sm

Root
lenght,
sm

Numb
er of
roots,
piece
s

Heigh
t, sm

Root
length,
sm

Num
ber
of
roots,
piece
s

3

rd

part

close to
the lower
environm
ent.

1

06 0.4

1.5

7.1

1.5

4

11.2

3.3

8

2

06.04

1.5

7

1.3

5

10.9

2.9

7

3

06.04

1.5

6.9

1.4

5

11.1

3.1

8

Average

06.04

1.5

7

1.4

4.6

11.06 3

7.6

№ Chosen for the

experiment

The time of
planting

Initial
state

Results obtained after 14
days 20.04

Results obtained after 24
days
30 .04

Height,
sm

Root
length,
sm

Number
of roots,
pieces

Heigh
t, sm

Root
length,
sm

Numbe
r of
roots,
pieces

1

The average of
the 1

st

part

apical

06.04

1.5

6.7

0.93

4.3

10.9

2.13

6.66

2

The average of
the 2

nd

medium

part

06.04

1.5

5.7

0.67

2.66

9.9

2.1

5

3

The average of
the 3

rd

part

close to the
lower
environment

06.04

1.5

7

1.4

4.6

11.06 3

7.6

Scientific research results in pandemic conditions (COVID-19)

56

The process of rooting of mulberry can last for 20-25 days if the nutrient

medium is chosen correctly, if the nutrient medium for the plant is chosen
correctly, but there are not enough conditions and the specificity of the
variety is not taken into account this figure might get delayed to 30-35 days.
It has also been discovered that it is also important to take into account the
room temperature.

Acknowledgements
We express our gratitude to the staff of the Scientific Research Institute

named after O. Sodikov of the Academy of Sciences of the Republic of
Uzbekistan (E.G Khalmuratov, G.B Nasyrova) and the Tashkent Institute of
Chemical Technology, Scientific Laboratory "Biotechnology" (Prof. N.A.
Khojamshukurov).

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