Scope and Limitations

The study will focus on the use of the Brassica juncea plant wherein the morphological
and physiological effects of salicylic acid (SA) and ascorbic acid (AsA) will be measured under
salt stress. The treatment application for SA and AsA will be exogenous through foliar
application. The parameters which will be measured in the study are the morphology of the
leaves, stem, and roots; the chlorophyll content; and biochemical analysis of secondary
metabolite, specifically, flavonoid, present in the leaves.
The plant treatment replicates will be monitored inside of the greenhouse and other
laboratory works will be conducted at the SEC B Biology Laboratories at the Ateneo de Manila
University.

Hypothesis
The plants subjected to salicylic acid and ascorbic acid treatment will have higher
resistance to salt stress than the plants which will not undergo SA and AsA treatment. The treated
plants will have better morphology and may have a higher photosynthetic rate. Overall, plant
treatments with salicylic acid under salt stress will be healthier compared to the plants which will
not be treated with SA and AsA.
Ascorbic Acid (AsA)
L- ascorbic acid (AsA) in plants, the main source of vitamin C for humans, is an essential
compound with important roles as an antioxidant, a modulator of plant development through
hormone signaling, and a cofactor for enzymes involved in photosynthesis, hormone
biosynthesis, and the regeneration of antioxidants (Pastori, et. al 2003; Gallie 2012). AsA is
important in plants such that it has a protective role against reactive oxygen species that are

formed from photosynthetic and respiratory processes (Pastori, et. al 2003; Gallie 2013). It also
protects the plant from reactive oxygen species by like hydroxyl radical formed from hydrogen
peroxide occurring in photosynthetic and respiratory processes by transferring of a single
electron (Zhang, 2013; Ismail, et. al 2015).
In most mammals, with exception of humans, few primates, bats, and guinea pigs,
synthesize AsA by converting D-glucuronic acid generated from D-glucose to L-gulonic acid and
with the help of aldono-lactonase convert it to gulono-1,4-lactone. Gulono-1, 4-lactone is
converted to L-ascorbic acid by gulono-1, 4-lactone oxidase. (Valpuesta and Botella 2004; Gallie
2012).
In plants, AsA synthesis has plenty of pathways in which Smirnoff- Wheeler pathway
could be considered as the primary AsA biosynthetic pathway as the other pathways have not
compensated for the Arabidopsis mutated with vtc2 vtc5 mutant (Gallie 2013).
AsA under environmental stress has been known to regulate the plant functions under
abiotic and biotic stress (Gallie, 2012). Thus, effects of AsA on plant growth and development
had been further studied by applying environmental stresses such as water deficiency and salt
stress on different plant specimens. AsA is the main factor in tolerating salinity which causes
oxidative stress through enhancing production of reactive oxygen species (Hameed, et. al 2015).
Naz, et. al (2016) had found out that cucumber (Cucumis sativus L) under water deficit
condition, when applied with AsA on the leaves, would have improved shoot fresh and dry
weight, relative membrane permeability, proline, and glycine betaine contents. On another
research conducted on drought stress, the role of AsA in seed priming of wheat was observed in
two independent experiments that resulted in finding out that hydropriming was still substantially

higher from the osmopriming of AsA (Farooq, et. al 2013). But significantly, osmopriming of
AsA had improved the morphology of roots, leaves, and its dry weight (Farooq, et. al 2012).
Tuna, et al (2013) used non-enzymatic antioxidative compounds (ascorbic acid, thiamine
HCl, and -carotene) to find its effects on salt stressed corn plants. The non-enzymatic
antioxidative compounds were sprayed on the plant once a week for the whole duration of the
experiment but salt treatment started 25 days after sowing. The physiological and biochemical
measurements were collected. The shoot and root dry weights increased in ascorbic acid
treatments. The activities of superoxide dismutase increased with the ascorbic acid application
while peroxidase increased with the -carotene application. Overall, ascorbic acid was more
effective in protecting the maize plants from salt stress if compared to other non-enzymatic
antioxidative compounds.
A subtropical halophyte, Limonium stocksii, was treated with 0, 300mM, and 600 mM
NaCl for 30 days with the exogenous application of 20 mM AsA and distilled water (Hameed, et.
al 2015). The sap osmolality, AsA concentrations, and AsA-dependent antioxidant activities
increased at increasing level of salinity and AsA treated plants under salinity improved in both
growth and AsA-dependent antioxidant activities compared to water treated plants (Hameed, et.
al 2015).

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