ISSN 1021-4437, Russian Journal of Plant Physiology, 2016, Vol. 63, No. 4, pp. 542–548. © Pleiades Publishing, Ltd., 2016.
Gene Expression at Suaeda salsa Seed Germination under Salinity
J. Jia, Y. Zhou, L. H. Dai, G. Z. Wang, J. Feng, Y. T. Shao, X. Y. Bian, and G. P. Wei
School of Mathematics, Physics and Biological Engineering, Inner Mongolia University of Science and Technology,
Baotou, Inner Mongolia, China
Received July 9, 2015
Abstract—Suaeda salsa is a succulent halophyte belonging to Suaedoideae; sodium, chloride and sulfate ions
or combination of the ions were useful to its growth. The seed germination is important event in plants life
cycle, and it is sensitive to salt stress. To further understand the molecular mechanisms of S. salsa seed ger-
mination under salt stress, cDNA-AFLP and qRT-PCR were used to identify genes differentially expressed
during seed germination under 100 mM NaCl stress (0, 12, 24, 36, 48, and 60 h), and 113 transcript derived
fragments (TDFs) over 80 bp in length were identified using cDNA-AFLP. Gene ontology analysis revealed
that these genes were involved in stress response, signal transduction and protein synthesis, etc. Further, four
genes closely related to salt resistance were selected and their expression during S. salsa seed germination were
investigated by qRT-PCR. The result showed differential gene expression at various germination stages under
salt stress. Further work will be focused on cloning these genes and identifying their functions in S. salsa ger-
mination process under salt stress.
Keywords: Suaeda salsa, cDNA, AFLP – qRT, PCR, сandidate gene, NaCl stress, halophytes
Salt stress is a one of the major abiotic stresses
affecting on plants. Soil salinity has been considered as
a result of irrigation and excessive use of fertilizers.
About 20% of the world’s irrigated lands have been
affected by salinity. Currently, salinity limits produc-
tion of crop from 21% of 10 million hectares irrigated
land in China . The majority of crops belong to gly-
cophytes, whose growth is dramatically limited in soils
with high salt . However, it is well known that
growth of some halophytes, especially dicotyledon
halophytes, is stimulated by moderate NaCl , such
as Suaeda salsa.
Although halophytes are naturally adapted to salin-
ity, their salt tolerance is greatly influenced by genetics
and developmental stage . The seeds are sensitive to
salinity and the ability to germinate under saline con-
ditions is obviously essential for plant establishment in
saline environments . Seed germination is influ-
enced by osmotic effect or ion toxicity . To improve
seed germination in salinized soil, the identification of
key genes during seed germination is a first step.
The S. salsa plant is an annual halophyte, and occurs
in both saline inland and intertidal zones . It can tol-
erate seawater and salinity fluctuation resulting from
water evaporation and tidal inundation . There is lit-
tle information available on seed germination of S. salsa
for physiological and biochemical levels . However,
there are no reports regarding molecular level.
cDNA amplified fragment length polymorphism
(cDNA-AFLP) is one of molecular techniques, and is
widely available at various plant species . This
technique can provide rapid and multiple comparisons
of the plant response to different stress , and was
applied to identify the transcripts potentially involved
in barley seed dormancy, in Nicotiana tabacum dor-
mancy breaking and in Arabidopsis thaliana seed ger-
The optimum salt concentration for S. salsa seed
germination was 100 mM NaCl , and it was used in
experiment. In this study, to identify the candidate
genes involved in seed germination under NaCl treat-
ment genes were screened by cDNA-AFLP and then
further analyzed by qRT-PCR.
MATERIALS AND METHODS
Plant material preparation. The seeds of Suaeda
salsa used in this study were harvested from Bayannur
city, Inner Mongolia, China. The seeds were sterilized
for 30 s with 70% ethanol, then 5 min with 0.1% (w/v)
, then washed for three times in sterile dis-
tilled water, then were germinated in 100 mM NaCl
 in the dark at 30°C, and were collected at 0, 12,
24, 36, 48, and 60 h.
The article is published in the original.
Abbreviations: cDNA-AFLP—cDNA amplified fragment length
polymorphism; SDS—sodium dodecyl sulfate; TDFs—tran-
scripts of different fragments.