Progressive Horticulture, Vol. 53, No.

1, June 2021
© Copyright ISHRD, Printed in India
DOI : 10.5958/2249-5258.2021.00010.5

[Research Article]

Constitutive over-expression of Osmotin improves seed germination under salt and

osmotic stress in transgenic tomato (Solanum lycopersicum L. cv. Pusa Ruby)

Vikas Yadav Patade, Deepti Khatri, Sanjay Mohan Gupta* and Madhu Bala
Defence Institute of Bio-Energy Research, Haldwani-263 139, Uttarakhand, India.
*E-mail: smg555@rediffmail.com

Abstract
Transgenic tomato (Solanum lycopersicum L. cv. Pusa Ruby) was developed through constitutive over expres-
sion of Nicotiana tabacum Osmotin gene. Our earlier studies have validated improved cold tolerance in the trans-
genic tomato. The present study reports salt and osmotic stress responses in the transgenic tomato at germination
stage. Transgenic and wild type tomato seeds were placed for germination in presence of salt (NaCl; 0, 100, 150,
200 mM) or polyethylene glycol induced osmotic stress (PEG 6000; 15, 20 %). The seed performance in terms of
germination and seedling establishment under control (non-stressed) condition was significantly (P ≤ 0.05) im-
proved in the transgenic (99.0 + 1.6 %) than the wild type (60.0 + 1.6 %). Higher and rapid germination was recorded
in the transgenic seeds than that in the wild type in presence of 150 mM NaCl. Germination in wild type seeds
was completely inhibited in presence of higher (200 mM) salt concentration whereas 9.0 + 1.6 % transgenic seeds
germinated successfully. Under polyethylene glycol (PEG 6000, 20 % w/v) induced osmotic stress, significantly
higher and rapid seed germination was observed in the transgenic than the wild type. The present study reports
salt and osmotic stress tolerance through constitutive over expression of Osmotin at seed germination in tomato.

Key words: Genetic engineering, Osmotin, Stress, Germination, Tolerance

Tomato (Solanum lycopersicum L.) is one of the most Materials and Methods
important vegetable crops cultivated worldwide. Sur-
The Osmotin transgenic tomato (cv. Pusa Ruby) was
vival, growth, productivity and the quality of produce
developed using Agrobacterium mediated genetic trans-
of the sensitive tomato cultivars are severely affected
formation and validated for improved cold tolerance at
on exposure to environmental stresses. Stress tolerance
germination, seedling and mature stages (Patade et al.,
being polygenic trait, a great emphasis is on molecular
2013, 2014, 2018). Seeds processed from the ripe fruits of
breeding and genetic engineering to develop stress tol-
transgenic (T8 generation) and wild type plants grown
erant cultivars. Function and mode of action of Osmotin
under containment conditions were used in the study.
has been well documented in improving plant defence
The transgenic and wild type seeds (30 per replicate)
under various stresses (Goel et al., 2010; Subramanyam
were placed for germination on filter papers saturated
et al., 2012; Krishna et al., 2019). Though several indepen-
with salt (NaCl; 100, 150, 200 mM) or polyethylene glycol
dent reports are available on improved stress tolerance
(PEG 6000; 15, 20 %; w/v) solution in Petri dishes. The
through over expression of Osmotin, scant information
filter papers in control group were saturated with dou-
is available on response of a particular transgenic line
ble distilled water. The Petri dishes were incubated at a
against the multiple stresses at germination stage. Plants
room temperature (25.0 ± 2.0 oC). Data on radical protru-
are more sensitive to the osmotic and ionic stress fac-
sion was recorded daily up to 15 days after placing the
tors at germination stage than the later developmental
seeds for germination. Percentage germination and rate
stages. Therefore, present study was designed to evalu-
of seed germination (Timson’s Index) were calculated
ate the effect of salt and osmotic stresses at germination
(Patade et al., 2018).
stage in the cold tolerant transgenic tomato expressing
Nicotiana tabacum Osmotin. The treatments and controls of the experiment were
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