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 VOLUME Vol.05 Issue03 2025 

PAGE NO. 28-31 

DOI 10.37547/ajahi/Volume05Issue03-08 

 
 
 
 

Amplification of the dreb2a transcription factor gene 

from salsola richteri (moq.) Kar. Ex litv. Growing in the 

southern Aralkum 
 

Doston Rizaev 

Junior Researcher Institute of Bioorganic Chemistry of the Academy of Sciences of the Republic of Uzbekistan, Uzbekistan 

 

Jamoliddin Ziyavitdinov 

Doc. chem. sciences, prof. Institute of Bioorganic Chemistry of the Academy of Sciences of the Republic of Uzbekistan, Uzbekistan 

 

Sanjar Sherimbetov 

Doc. biol. sciences, prof. Institute of Bioorganic Chemistry of the Academy of Sciences of the Republic of Uzbekistan, Uzbekistan 

 

Bahtiyor Adilov 

Doc. biol. Sciences, Institute of Genetics and Experimental Plant Biology of the Academy of Sciences of the Republic of Uzbekistan, 

Uzbekistan 

 

 

 

Received: 29 January 2025; Accepted: 28 February 2025; Published: 31 March 2025 

 

Abstract: This study investigates halophytic plants of the genus Salsola L., which are widely spread in the southern 
part of the Aralkum Desert. During the research, a PCR product of approximately 1200 bp associated with the 
DREB2A gene was amplified in one of the species, Salsola richteri. This gene plays a key role in plant adaptation 
to drought and salt stress. The obtained data will be used for sequencing the DREB2A gene and analyzing its 
expression.   

 

Keywords: Salsola richteri, DREB2A, transcription factor, salt tolerant plants, primer. 

 

Introduction: Salsola richteri is a shrub or a small tree 
ranging from 1 to 3 meters in height. In its young stage, 
it is covered with finely tuberculate (or papillose) 
leaves, which later become leafless. The stem is weakly 
branched, with smooth gray bark, up to 5 cm thick at 
the base, and its woody branches soon acquire a milky-
white color [1]. This species is promising for 
phytoremediation practices due to its good growth on 
sandy soils, high seed productivity, ability to propagate 
by seeds and cuttings, and tolerance to significant 
salinity. Its powerful root system makes it effective for 
sand stabilization [4, 5]. The high protein content and 
the formation of substantial organic mass with 
economical water use allow S. richteri to be utilized as 
a valuable forage plant for autumn-winter pastures [2, 

3, 5]. 

Karakalpak scientists studied the growth and 
development of S. richteri under the ecological 
conditions of the Karakalpak part of the Kyzylkum 
Desert from 2014 to 2018. Fruiting of S. richteri began 
as early as the first year of life, in the third decade of 
June. Fruits were mainly formed on fourth-order 
shoots, with the number of fruits per shoot ranging 
from 16 to 38 and the number of leaves from 14 to 40. 
Fruiting depended on the age of the shrubs; in the 
second year of cultivation, a single shrub could produce 
between 3,000 and 6,000 seeds [5]. 

S. richteri belongs to the groups of euhalophytes and 
hemixerophytes, which are adapted to soils with a 
moderate level of salinity (1.8–2.3%; Cl 0.1–0.23%—

 

https://doi.org/10.37547/ajahi/Volume05Issue03-08
https://doi.org/10.37547/ajahi/Volume05Issue03-08
https://doi.org/10.37547/ajahi/Volume05Issue03-08
https://doi.org/10.37547/ajahi/Volume05Issue03-08


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American Journal Of Agriculture And Horticulture Innovations (ISSN: 2771-2559) 
 

 

chloride-sulfate and sulfate salinization or 1.3–1.8%; Cl 
0.1–0.23%—sulfate-chloride and chloride salinization). 
This species is capable of successfully growing in such 
soils, maintaining its dominant position in the 
phytocenosis even as salinity levels decrease. It forms 
plant associations and formations over vast areas that 
have been exposed after water receded. In his 
research, S. G. Sherimbetov analyzed the chemical 
composition of plants found in the desiccated regions 
of the Aral Sea. Among the studied halophytes 
(Climacoptera aralensis, Kalidium capsicum, 
Halostachys belangeriana, Salsola richteri, Haloxylon 
aphyllum, Tamarix hispida), a high content of minerals 
such as Ca, Cl, K, Mg, and Na was observed.  

The plant Salsola drummondii Ulbr., belonging to the 
genus Salsola L., is capable of successfully growing and 
completing its life cycle under high salinity conditions 
(500–800 mM NaCl). This plant adapts to salt stress by 
reducing the level of photosynthetic pigments, 
decreasing carotenoid content, and increasing the 
activity of antioxidant enzymes [6]. Based on the genes 
of Salsola iberica, which are responsible for resistance 
to abiotic stress factors, a bank of expressed sequence 
tags (EST—Expressed Sequence Tag) was created to 
analyze the molecular mechanisms of adaptation. This 
bank included 377 ESTs, which were grouped into 227 
unique fragments. Similarities were found between S. 
iberica ESTs and stress-resistance genes, including salt-
induced proteins, betaine-aldehyde dehydrogenases, 
and calcium-binding proteins [7]. Salsola ferganica is a 
desert herbaceous plant that grows in arid regions of 
western and northwestern China. 

To normalize gene expression in S. ferganica under 
abiotic stress, nine reference genes (TUA-1726, TUA-
1760, TUB, GAPDH, ACT, 50S, HSC70, APT, and U-box) 
were tested under six stress conditions. The analysis 
revealed that ACT and U-box exhibited the highest 
stability among all tested variants [8].  

A review of scientific literature has shown that despite 
S. richteri is tolerance to salinity and drought, there is a 
lack of data on transcription factors responsible for 
these adaptation mechanisms. Abiotic stress plays a 
crucial role in plant growth and development, as plants 
are exposed to various adverse factors such as drought, 
low and high temperatures. Under stress conditions, 
several stress-resistant genes are activated, among 
which DREB (Dehydration Responsive Element Binding) 
genes play a particularly important role. These genes 
encode proteins of the Apetala2/ethylene (AP2/ERF) 

family, which bind to the dehydration-responsive 
element (DRE)/C-repeat in the promoter regions of 
stress-resistance genes. The DRE cis-element, located 
near the promoter regions of stress-associated genes, 
serves as the binding site for DREB transcription 
factors, which regulate osmotic stress in plants. 
Drought and high salinity levels induce the expression 
of the DREB2 gene, which plays a key role in regulating 
abiotic stress-responsive genes. 

The expression of OsDREB2A in Oryza sativa is 
enhanced under salt stress and dehydration, but the 
gene exhibits low sensitivity to low temperatures and 
abscisic acid (ABA) [9]. Similarly, in Zea mays, the 
transcript level of ZmDREB2A increases under high-
temperature stress. Arabidopsis thaliana demonstrates 
DREB2A activation primarily in response to drought and 
salt stress [10]. 

The aim of the study is to design a new primer for 
obtaining the full DREB2A gene sequence from S. 
richteri, a plant from the Chenopodiaceae family. 

METHODS 

As research objects, plant biomaterials of S. richteri 
collected from the Southern Aralkum in 2021 were 
used. The plant materials were identified by staff of the 
Institute of Botany, Academy of Sciences of the 
Republic of Uzbekistan. 

Total DNA was extracted from the leaves of the plant 
using the PureLink Plant Total DNA Purification Kit 
(Invitrogen by Thermo Fisher Scientific). 

Primer Design 

To amplify the DREB2A gene from plants of the 
Chenopodiaceae family, a search for the nucleotide 
sequence of the DREB2A gene was conducted in the 
NCBI database (NCBI - www.ncbi.nlm.nih.gov) using the 
BLASTN algorithm in the BLAST web application within 
the GenBank database. Based on the nucleotide 
sequences of the DREB2A gene from Salicornia 
brachiata (ID: GU809211.1), Haloxylon ammodendron 
(ID: KP765243.1), Beta vulgaris (ID: XM_010694618.3), 
and Chenopodium quinoa (ID: XM_021876866.1) for 
the reverse primer, as well as Chenopodium album (ID: 
OX_419225.1) and Salicornia ramosissima (ID: 
OX_596239.1) for the forward primer from the NCBI 
database, a design of specific degenerate primers was 
created for the amplification of approximately 900 and 
1500 nucleotide sequences using the CLUSTAL O(1.2.4) 
Multiple Sequence Alignment program (Table 1). 

Table 1. 

Oligonucleotide sequence of primers for amplification of the DREB2A gene of some 

species of the Chenopodiaceae family 



American Journal Of Agriculture And Horticulture Innovations 30 https://theusajournals.com/index.php/ajahi 

American Journal Of Agriculture And Horticulture Innovations (ISSN: 2771-2559) 
 

 

 

 

 

 

 

Polymerase chain reaction 

The amplification of DREB2A gene fragments from the 
studied halophytic plant Salsola richteri was performed 
using a MiniAmp™ Plus Thermal Cycler (Applied 
BioSystems, USA) with specific primers. The 
polymerase chain reaction (PCR) of the DREB2A gene 
was carried out according to the protocol of the 
Phusion™ High-Fidelity DNA Polymerase kit (Thermo 

Fisher 
Scientific, 
USA) [7]. 

For the 

quantitative determination of nucleotides in the PCR 
products, electrophoresis was performed in a 2% 
agarose gel. The length of the PCR products was 
determined using a 100 bp DNA Ladder marker 
(Invitrogen, USA) (Fig. 1). 

 

 

 

 

Figure 1. Electropherogram of the PCR product of the DREB2A gene 

M - DNA marker; 1. S. richteri; 2. S. richteri 

 

RESULTS AND DISCUSSION 

As a result of the study, a PCR product with an 
approximate length of 900 and 1200 base pairs was 
obtained from Salsola richteri. It was established that 
the PCR product length corresponds to the predicted 
size during primer design, confirming their specific 
binding to the complementary sequence. According to 
NCBI, the length of the DREB2A gene in the registered 
species Salicornia brachiata (ID: GU809211.1), 
Haloxylon ammodendron (ID: KP765243.1), Beta 
vulgaris (ID: XM_010694618.3), and Chenopodium 
quinoa (ID: XM_021876866.1) ranges from 1100 to 
1200 base pairs. The length of the PCR products 
obtained from S. richteri also falls within this range, 
confirming their affiliation with the DREB2A gene. The 
obtained results enable further investigation of the 
nucleotide sequence of the DREB2A gene in S. richteri  

 

and the study of its expression levels under drought 
and salinity conditions. 

CONCLUSION 

DNA fragments were isolated from plants of the 
Chenopodiaceae family distributed in the Southern 
Aralkum. Using bioinformatics online resources,  
specific DREB2A primers were designed. The DREB2A 
gene was successfully amplified from the DNA of 
Salsola richteri using PCR. It was established that the 
PCR products obtained from S. richteri were suitable 
for sequencing nucleotide pair sequences. The results 
of this study provide a foundation for further analytical 
research aimed at understanding the functionality of 
the DREB2A gene in S. richteri. 

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American Journal Of Agriculture And Horticulture Innovations 31 https://theusajournals.com/index.php/ajahi 

American Journal Of Agriculture And Horticulture Innovations (ISSN: 2771-2559) 
 

 

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