Volume 04 Issue 09-2024
14
American Journal Of Agriculture And Horticulture Innovations
(ISSN
–
2771-2559)
VOLUME
04
ISSUE
09
Pages:
14-18
OCLC
–
1290679216
Publisher:
Oscar Publishing Services
Servi
ABSTRACT
The global decline of plant species exerts a significant detrimental impact on the stability of natural ecosystems.
Endemic species with restricted geographical distributions are at risk of extinction in their natural habitats.
Consequently, the investigation of rare plant species, the analysis of their current status, and the development of
conservation measures are of substantial scientific and practical significance. Astragalus knorringianus Boriss.
represents one such critical plant species.
KEYWORDS
In vitro technology, rare species, nutrient medium, sterilization, seed germination.
INTRODUCTION
Astragalus knorringianus Boriss. is a perennial herb
belonging to the Fabaceae family. It flowers in March-
April, and its seeds mature in May-June. The peduncle
measures 2-5 cm in length, while the corolla is 2-2.5 mm
long. The plant produces 2-3 flowers, situated at the
terminus of a single stem, exhibiting pale yellow or
dark red coloration, and measuring 32-37 mm in length.
The calyx is 18-21 mm, with teeth 1-3 mm long,
predominantly 0.2-0.3 mm long, dark pubescent, and
tubular in form. The vexillum component measures 33-
36 mm in length, with an arcuate lamina 13 mm in
width. The upper portion is dentate, while the lower
portion tapers. The wings are approximately 30 mm
long, with an elongated lamina of about 11 mm. The
keel measures 28 mm in length, featuring a sharp
lamina of 9 mm in the upper section. Pollination occurs
Research Article
STERILIZATION AND STUDY OF GERMINATION OF ASTRAGALUS
KNORRINGIANUS SEEDS IN VITRO CONDITIONS
Submission Date:
Sep 20, 2024,
Accepted Date:
Sep 25, 2024,
Published Date:
Sep 30, 2024
Crossref doi:
https://doi.org/10.37547/ajahi/Volume04Issue09-03
Daniyorova Shakhnoza Olimjon’s daughter
Jizzakh state pedagogical university, Uzbekistan
Journal
Website:
https://theusajournals.
com/index.php/ajahi
Copyright:
Original
content from this work
may be used under the
terms of the creative
commons
attributes
4.0 licence.
Volume 04 Issue 09-2024
15
American Journal Of Agriculture And Horticulture Innovations
(ISSN
–
2771-2559)
VOLUME
04
ISSUE
09
Pages:
14-18
OCLC
–
1290679216
Publisher:
Oscar Publishing Services
Servi
via insects. The pod attains a length of up to 7 cm and
a width of 4 mm, possessing a short beak and covered
with white and black trichomes. The pod morphology
is arcuate or straight, narrowing towards both
extremities. Upon maturation, the seed coat assumes
a brown coloration. Reproduction occurs through
seeds. The seeds are of moderate size, with a
thousand-seed weight of 5.95 g.
Astragalus knorringianus is a rare endemic species that
occurs in northwestern Pamir-Aloy. Its distribution
encompasses the mountains of Nurota, Molguzar, and
Turkestan in the Jizzakh, Samarkand, and Navoi
regions. In addition to its presence in Uzbekistan, the
species is also found in Tajikistan and Kyrgyzstan.
The Red Book of Uzbekistan editions (2016, 2019)
enumerate 54 species from the sedge family, of which
34 belong to the Astragalus genus. The flora of certain
mountainous regions in Central Asia is characterized by
an abundance of Astragalus L. species. Significant
research has been conducted on rare and endemic
species of the Astragalus genus. Notably, Kamelin
(1990) identified 14 specific endemic taxonomic
features of Astragalus in Syrdaryo Karatov.
Populations of some rare Astragalus species, including
Astragalus abolinii, have been observed to be in
satisfactory condition in the region [5].
Astragalus centralis, as reported by Sh.U. Saribaeva [6]
in South-West Kyzylkum. Sheld., has been the subject
of senopopulation studies. The current state and
viability of this plant species were evaluated, and the
primary factors contributing to the reduction of its
range were identified, along with the development of
protective measures. In recent years, new species
(Astragalus belolipovii Kamelin ex F. O. Khass. et N.
Sulajm., A. russanovii F. O. Khass., Sarybaeva et
Esankulov, A. zaaminensis F. O. Khass. & Esankulov)
have been described in the Kokhistan region, enriching
the flora inventory with new discoveries. Information
regarding the assessment of vitality status,
morphogenesis, ontogenesis periods, and types of
senopopulations of Astragalus holargyreus Bunge is
reflected in the works of K.F. Shomurodov (2018) and
Sh.U. Saribaeva (2009). Astragalus belolipovii was
initially discovered by I.V. Belolipov near the Kulsay
forest cottage and subsequently cultivated in the
Botanical Garden in 1975.
Plant biotechnology facilitates the preservation of rare
and endangered plant species.
Regarding in vitro technology, it is crucial to emphasize
its efficacy:
1. Rapid propagation: The process of plant propagation
utilizing the in vitro method is significantly more
expeditious and efficient than conventional methods.
This technique enables the production of millions of
seedlings within a single year.
2. Genetic stability: This technology maintains the
genetic stability of plants, ensuring that the resulting
seedlings possess identical quality and characteristics.
3. High yield: In vitro propagated plants exhibit high
yield and enhanced resistance to diseases.
Numerous studies have been conducted on the
cultivation of various Astragalus L. species under in
vitro conditions [7,8]. However, research on the in vitro
cultivation of Astragalus knorringianus from tissues
and organs has not been undertaken.
Research results. For this purpose, we conducted
studies on in vitro sterilization and fertility of
Astragalus knorringianus seeds. A review of the
literature indicated that sterilization and fertility
determination of this species' seeds in vitro have not
been previously investigated.
Volume 04 Issue 09-2024
16
American Journal Of Agriculture And Horticulture Innovations
(ISSN
–
2771-2559)
VOLUME
04
ISSUE
09
Pages:
14-18
OCLC
–
1290679216
Publisher:
Oscar Publishing Services
Servi
Astragalus knorringianus seeds were collected from a
community of plants from diverse herbaceous astragali
almond orchards in the vicinity of Karasoy village (N
082738 E 739259 h=485 m) (near Temurlang gate) of
Jizzakh region.
Seed germination biology is a critical stage in a plant's
developmental cycle, wherein a seed emerges from
dormancy and initiates growth to form a new plant.
This
process
comprises
several
interrelated
physiological and biochemical stages. The biology of
seed germination is of significant importance in plant
reproduction, ecological adaptation, and dispersal,
facilitating the transfer of genetic material to
subsequent generations.
For the cultivation of Astragalus knorringianus seeds,
fully matured seeds were selected. The sorted seeds
were sterilized in the in vitro scientific laboratory of
Jizzakh State Pedagogical University to examine seed
viability and viability.
In isolated tissue culture, strict adherence to sterility is
essential. The nutrient-rich composition of the medium
is conducive to microbial growth, which can readily
damage plant parts (explants). Consequently, both
explants and nutrient medium must be thoroughly
sterilized. All tissue-related processes (culture transfer,
transfer to new nutrient medium) are conducted in a
sterile environment, utilizing laminar flow hoods and
sterile instruments. Maintaining sterility during the
growth period is crucial, as microorganisms can enter
and contaminate the test tube through the moist
stopper of the container due to temperature
fluctuations or humidity [1]. Typically, seeds are
sterilized for 10-20 minutes, while vegetative parts are
sterilized for 5-10 minutes [2]. For the sterilization of
plant organs, R.G. Butenko's method was employed.
Various methods were utilized for the in vitro
sterilization of Astragalus knorringianus seeds.
The seeds stored for three weeks were prepared for
collection in the Invitro scientific laboratory. For this
purpose, the nutrient medium was first prepared.
276.25 mg of Murasiga - skuga feed was measured into
250 ml of distilled water. Subsequently, 1.875 g of
sucrose was added to the mixture and homogenized
with a magnetic stirrer until dissolved. The pH indicator
of the prepared solution was adjusted to 5.8. At the
conclusion of the process, 2 g of agar-agar
polysaccharide obtained from seaweed was added, the
container was covered with tissue paper and filter
paper, and sent to the autoclave for sterilization. This
nutrient medium was sterilized in an autoclave at a
temperature of 120° C and a pressure of 0.75 atm for 15
minutes. In the subsequent step, the explant is
sterilized.
Volume 04 Issue 09-2024
17
American Journal Of Agriculture And Horticulture Innovations
(ISSN
–
2771-2559)
VOLUME
04
ISSUE
09
Pages:
14-18
OCLC
–
1290679216
Publisher:
Oscar Publishing Services
Servi
Figure 1. Germination of Astragalus knorringianus seeds in vitro conditions
For this study, mature seeds of Astragalus
knorringianus were selected. Initially, the seeds were
washed in distilled water, followed by a 10-minute
wash in soapy water, and subsequently rinsed in
distilled water 3-4 additional times. Further procedures
were conducted in a laminar flow hood. The seeds
were immersed in a 4 percent sodium hypochlorite
solution for 10 minutes. They were then rinsed 5-6
times in distilled water and submerged in a 70% solution
of ethyl alcohol for 30 seconds, followed by 2-3 rinses
in distilled water.
Upon complete sterilization, the seeds were
inoculated onto Murashige and Skoog nutrient
medium in glass vessels within a laminar flow hood,
adhering to aseptic techniques. The inoculated seeds
were placed in a dark environment at a temperature of
4 C for 14 days. Subsequently, the seeds were
transferred to an incubator maintained at a
temperature of 22 C under light conditions.
Figure 2. Washing seeds in soapy water; Placing samples in the thermostat.
Volume 04 Issue 09-2024
18
American Journal Of Agriculture And Horticulture Innovations
(ISSN
–
2771-2559)
VOLUME
04
ISSUE
09
Pages:
14-18
OCLC
–
1290679216
Publisher:
Oscar Publishing Services
Servi
Germination of seeds commenced 16 days after
sowing. During the second week of seed germination,
55% of the sown seeds germinated.
CONCLUSION
The rate of seed germination exhibits variability. It was
observed that the germination rate of the cultivated
seeds attained 80 percent. The obtained results
pertaining to the seed germination of this species will
serve as foundational material for the investigation of
ontogenetic morphogenesis of the species within a
brief temporal period in vitro.
REFERENCES
1.
Misirova, S.A., Ernazarova N.N. Fighting measures
the disease causes a very dangerous fungal species
widespread in Tashkent region. International
journal of botany and research (ijbr) 6 (2016): 5-12.
2.
2. Davronov K, Biotechnology, scientific, practical
and methodological foundations. Tashkent, 2008.
p. 77-78.
3.
F.O. Khassanov (Eds). Red data book of
Uzbekistan. Vol. I. Tashkent. Chinor ENK 336 p.
(2019)
4.
Podlech, D. and S. Zarre. 2013 A taxonomic revision
of the genus Astragalus L. (Leguminosae) in the
Old World. vols. 1-3. Naturhistorisches Museum,
Wien, 2439 pp.
5.
Тожибаев Комилжон Шаробитдинович флора
Юго
-
Западного
Тян
-
Шаня
(в
пределах
республики узбекистан). 03.00.05 –
ботаника
автореферат диссертатсии на соискание ученой
степени доктора биологических наук Ташкент –
2010.
6.
6. Saribaeva Sh.U. Astragalus centralis YE. Sheld.
description of bioecological characteristics and
senopopulations in South-Western Kyzylkum:
Autoref. dis. ...b.f.n Tashkent, 2009. - 22 p.
7.
J. B. Šenkyřík. Exploring in vit
ro oryzalin-induced
polyploidy
in
Astragalus
membranaceus:
implications for gene expression. Plant cell, Tissue
and Organ Culture (2024). 1-14 p.
8.
О. Манжура, О. Кваско. Култивування In vitro
рослин
астрагалу
шерстистоквіткового
(Астрагалус дасянт
ҳ
ус
палл), занесеного до
червоної книги України. Матеріали ХХИІІ
Міжнародної науково
-
практичної конференсії
«Екологія. Людина. Суспілство» (м. Київ, Україна,
7 грудня 2023 р.) 40
-42.
