







































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. 

 

https://theusajournals.com
https://doi.org/10.37547/ajahi/Volume04Issue09-03
https://doi.org/10.37547/ajahi/Volume04Issue09-03


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 vitro 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 дасянтҳус палл), занесеного до 

червоної книги України. Матеріали ХХИІІ 

Міжнародної науково-практичної конференсії 

«Екологія. Людина. Суспілство» (м. Київ, Україна, 

7 грудня 2023 р.) 40-42.  

 


