Frontiers in Life Science

ISSN: 2155-3769 (Print) 2155-3777 (Online) Journal homepage: https://www.tandfonline.com/loi/tfls20

Development of halophytes as energy feedstock

by applying genetic manipulations

Neelma Munir, Zainul Abideen & Nadia Sharif

To cite this article: Neelma Munir, Zainul Abideen & Nadia Sharif (2019): Development of
halophytes as energy feedstock by applying genetic manipulations, Frontiers in Life Science, DOI:
10.1080/21553769.2019.1595745

To link to this article: https://doi.org/10.1080/21553769.2019.1595745

© 2019 The Author(s). Published by Informa

UK Limited, trading as Taylor & Francis
Group

Published online: 27 Mar 2019.

Submit your article to this journal

Article views: 94

View Crossmark data

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=tfls20
FRONTIERS IN LIFE SCIENCE
https://doi.org/10.1080/21553769.2019.1595745

Development of halophytes as energy feedstock by applying genetic

manipulations
Neelma Munira , Zainul Abideenb and Nadia Sharifc
a Department of Biotechnology, Lahore College for Women University, Lahore, Pakistan; b Institute of Sustainable Halophyte Utilization,
University of Karachi, Karachi, Pakistan; c Department of Biotechnology, Women University, Mardan, Pakistan

ABSTRACT ARTICLE HISTORY

Decreasing arable land and fresh water resources, and increasing soil salinization and production of Received 19 April 2018
energy from food crops pose a threat to plant productivity and caused several environmental prob- Accepted 6 March 2019
lems. Using plant species which can grow on saline degraded soils for energy production can be a KEYWORDS
sustainable approach because they do not compete for the agricultural lands and availability of fresh Bioenergy; desalination;
water. These plants can be cultivated with seawater without compromising their biomass for com- drought; genetic
mercializing purposes such as lignocellulosic content and seed yields and could strongly benefit as manipulation; halophytes;
energy plants. Optimizing biomass production and its economic conversion to the end product are, lignocellulose; salinity
however, of paramount importance. The biomass of halophytes can be improved through studying
unexplored aspects of these plants related to genetic manipulations. Here, we recommended current
advances and highlight the key genetic approaches required in halophytes for biofuel production.
Genetic approaches might lead to desired alterations in the composition of halophytic lignocellulosic
biomass, including higher quantity of cellulose and hemicelluloses and lower lignin. In summary, the
genetic manipulations in halophytes might lead to improvement in biomass compositions hence can
be used as a feedstock for the production of biofuel on nutrient poor degraded soils.

Abbreviations energy resources are necessary to improve energy effi-

ciency and to drop energy demand by offering more
EIA Energy Information Agency
opportunity for daily energy consumption.
IPCC Intergovernmental Panel on Climate Change

Creeping energy crisis Shortage of fresh water

The world is experiencing the energy crisis because of
consumption of the limited fossil fuel assets (Amaro
et al. 2012). The utilization of fossil fuel as a primary
energy source is currently perceived to be unsustain-
able in light of exhausting assets and environmental
pollution (Khan et al. 2009). According to Energy
Information Agency (EIA), the world energy require-
ments are expected to be raised 60% more than today
till 2030. If this trend continues then the world overall
fossil oil stores will be depleted in less than 45 years
(Ahmad et al. 2011) that might cause a shortage of
energy supply for economic activity. Thusly to unravel
these essential issues the advancements in renewable
Fresh water provisions are running lower throughout
the world and this water emergency would worsen
as the population is increasing. Most of the world’s
Availability of fresh water is limited because they are
mainly located in the polar icecaps and underground
stream frameworks which could only supply through
wells and springs. In addition, irregular precipitation,
periodic rise in temperature and change in land cli-
mate interactions also presented imbalance in water
availability. Global climate models predict many dry
periods in future that might experience greater water
stress as these regions become warmer and drier (Ward
et al. 2016). Uneven rain timing, decreased access

CONTACT Neelma Munir neelma.munir@yahoo.com

© 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
2 N. MUNIR ET AL.
to water supplies, winds and low relative humidity
caused innate dry periods over a extended duration
in many countries that affects millions of people of
the world (Sun et al. 2006; Grainger 2013). The grow-
ing aridity and freshwater demands build a serious
need to link scientific research with optimal water
management that will assist to allocate water more effi-
ciently in extremes water scarce areas. (Salinas et al.
2016). One solution is to use saline water in sub-
tropical habitats for agricultural purposes instead of
fresh water. Use of saline water impairs more than 45
million hectares of irrigated land (Munns and Tester
2008). Fresh water can be generated by using largely
available saline water through the process of desalina-
tion. Numerous nuclear powered desalination plants
are present; though the cost is higher especially for
poor developing countries in the world. It will cre-
ate the transporting cost to transfer huge amounts of
desalinated seawater in to the plant (Hinrichsen and
Tacio 2002). Zhoua and Tolb (2005) estimated that
the cost of desalination is equal to the cost required
to transport it over 1600 km or raise the water up to
2000 km. Somewhat high and far places have higher
desalination costs while other places cost is higher
due to transport rather than desalination. Desalinated
water might be an answer for some water-push dis-
tricts, however not designed for spots that are poorer,
somewhere down in the inside of a landmass, or at
elevated rise. Tragically, that incorporates a portion of
the areas with greatest water issues. Another poten-
tial issue with desalination is the creation of salt water,
which can be a noteworthy reason for marine contami-
nation when drained once again into the sea at elevated
temperature (Zhou and Tol 2005). Limitation of water
or presence of salty water both can decrease biomass
of economic important and might cause shortage
in food supply.
Food security
Lack of affordable nutrition by conventional channels
has been a serious issue of experience malnutrition
and starvation on daily basis especially in third world
countries. Many scientists recommended that energy
made from edible plants for transport is not appropri-
ate (Nouairi et al. 2006).
Food crops such as sugarcane, wheat, maize and
corn are the most efficient energy crops in current
scenario (Cherubini et al. 2009). These edible crops
that are grown and processed for bioenergy will have
huge impact on food supply and demand system. This
is particularly fact that the cultivation of these dedi-
cated food crops for the purpose of bioenergy rather
than food purposes only. The utilization of prime
agricultural lands which was patents to feed the peo-
ples in previous decades sharply boasting the food
prices up to the limit which can harm life of common
people (Colombo and Onorati 2013). Growing food
crops for bioenergy instead of those for food prod-
ucts really invented the huge contention between food
and fuel. In order to represent and provides relax-
ation to extensive utilization for food-based feedstock
crops to biofuel raises emerging technology that con-
vert non-edible grasses and forest products. These are
termed as second generation biofuel feedstocks plant
productivity changes are still held to baseline levels.
These second generation feedstocks for biofuel still
can pop up the land compactions against food crops
can harm food prices and food security (Flora et al.
2012).
Now the whole world is searching for more sustain-
able and environmental friendly crops for the biofuels
otherwise the image of fuels production from plants
can get bad views in the world regarding environmen-
tal and food shortage issues. The compatible solution
for all the food curse issues may the plants which
cannot compete to use the cultivated lands for their
survival. Those plants which can grow and complete
their life events on marginal soil, saline soil and other
then the prime agricultural lands. On the basis of these
plants, we may solve dilemma of food vs fuel.
Halophytic crops for a saline agriculture and
bioenergy
A consistent supply of reliable feedstock regarding
quantity and quality of plant biomass is required for
the monetary viability of the biofuel industry. The
plants specific to saline environments and can survive
and complete their life cycles, no less than, 200 mM
NaCl are called halophytes (Grigore et al. 2014). How-
ever, many can develop at salt concentration signifi-
cantly higher than that of seawater (Rabie and Alma-
dini 2005). Salt resistance plants cover around 0.25% of
angiosperm species that represent approximately 600
taxa of plants consist of genera and families (Fita et al.
2015). Halophytes are highlighted as wild plants but
huge numbers of halophytes can possibly be changed
 FRONTIERS IN LIFE SCIENCE 3

into valuable ‘new crops’, after extensive a domestica- Table 1. Ligno-cellulosic contents of halophytic biomass (% dry
tion procedure. They are already salt resistant plants – weight).
which is the most essential and the most difficult qual- Species Cellulose Hemicellulose Lignin
ity to introduce and manipulate- it ought to be moder- Grass halophytesa
1 Aeluropus lagopoides 26.67 29.33 7.67
ately simple to practice particular breeding programs 2 Cenchrus ciliaris 22.67 23.17 7
to quickly enhance the required agronomic attributes 3 Chloris baraeta 25.33 23 8.33
4 D. bipinnata 26.67 24.68 6.67
of the most encouraging halophytic taxa. For spe- 5 D. annulatum 19 24.33 7
cific development conditions, it might be important 6 Eleusine indica 22 29.67 7
7 H. mucronatum 37 28.67 5
to choose the best genotypes, by killing or possibly 8 Lasiurus scindicus 24.67 29.67 6
9 Panicum turgidum 28 27.97 6
decreasing the substance of dangerous mixes or hostile 10 Paspalum paspaloides 20.33 32 2.33
to supplements, by expanding yields, or by enhancing 11 P. karka 26 29 10.33
12 S. ioclados 15.33 30.67 2
showcasing attributes (time span of usability, market 13 Urochondra setulosa 25.33 25 6.33
accessibility, consistency of the item in size, shad- 14 Typha domingensis 26.33 38.67 4.67
ing, taste, and so forth), and to modify general farm- Non grass halophytesa
1 A. javanica 15.67 13.33 6.33
ing techniques to specific crops (Baram and Bourrier 2 A. indicum 11.33 13 7
2011). Individuals of halophytes have been collected 3 Calotropis procera 12.33 11 5
4 Ipomea pescaprea 12.67 17 5.33
from nature for hundreds of years. Sometimes they 5 Salsola imbricata 9 18.33 2.67
are pickled or cooked, or their leaves are regularly 6 Salvadora persica 22 13.33 7
7 Suaeda fruticosa 8.67 21 4.67
eaten as raw vegetables and salad, or sold in local mar- 8 Sueda monoica 10.67 11.33 2.33
9 Tamarix indica 12.17 24.67 3.33
kets (Nyankanga et al. 2012). The conventional use
Conventional Speciesb
as sustenance of these species will make them all the 1 Panicum virgatum 45 31.4 12
more effectively worthy by the overall population, with 2 Popolar 40 23 20
3 Cynodon dactylon 25 35.7 6.4
the goal that they are fitting possibility to be tamed a Abideen et al. (2011).
and changed into verdant vegetable products for saline b Karp and Shield (2008), Reshamwala et al. (1995).

agribusiness. Halophytes are exceptionally nutritious

alongside eatable trademark; for the most part they are
rich in basic supplements – minerals, vitamins, amino
acids, as well as unsaturated fats, protein and antiox-
idants (Fita et al. 2015). These plants can grow under
seawater and many of them are cultivated and domes-
ticated as vegetable crops due to their salt resistance
ability. For instance Salicornia and Sarcocornia are two
suitable halophytes attracted due to their utilization in
food purposes (Fita et al. 2015). Numerous halophytes
can be potential oilseed plant to their chemical com-
position (Abideen et al. 2015). For instance, Salicornia
bigelovii is fascinating because of its seed production,
approximately two tons per hectare every year, which
are similar to those of oilseed harvests of conventional
species such as soybean. The chemical composition of
these halophytes are acceptable in term of their seed
total protein (30%), oil contents (30%), and the oil con-
tains of these plants composed of a high percentage
of polyunsaturated fatty acids (70% of linoleic acid)
so that it can be considered as promising edible oil.
Chenopodium quinoa also need to be focused regard-
ing food nutrient (Ladeiro 2012). Seeds of S. salsa
can be used as a raw material to produce fatty acid
methyl esters from their high oil contents, about 97%
of S. salsa oil can be processed to fatty acid methyl
esters (Piloto-Rodríguez et al. 2014). Substantial scale
advancement and modern production of halophytes
could be received as another agribusiness alternative
by using reasonable areas along coastal areas. There
is a requirement for the efficient investigation of halo-
phytes to screen out high value industrial species and
their rearing for desirable plant attributes Many halo-
phytic species including Phragmites karka, Halopyrum
mucronatum, Panicum antidotale and Desmostachya
bipinnata has been reported for promising lignocellu-
losic content (Table 1) (Panta et al. 2014).
Halophytes are reported as a sustainable source of
bioethanol production in many studies. The produc-
tion of ethanol from plant biomass depends on cellu-
lose, hemicelluose and lignin content of a potential fast
growing species. Presence of higher lignin (30–40%)
in energy feedstock ensure resistant to their sequen-
tial conversion from dry biomass to fermentable
sugars and in to ethanol. It was reported earlier
studies that five promising plants such as Phrag-
mites karka, Panicum antidotale, Desmostchya bipin-
nata, Halophyrum mucronatum and Typha domengesis
4 N. MUNIR ET AL.
Figure 1. Some halophytes that can be a potential candidate for biofuel production.
(Figure 1) showed exceptionally optimal ratios of cel-
lulose, hemicellouse and lignin and in some cases
better than our food crops. There are also some
plant species those showed lower cellulose and hemi-
cellulose content such as Dichanthium annulatum,
Sporobolus ioclados, Aerva javanica and Arthrocne-
mum indicum (Tables 1 and 2). A recent study
assessed Juncus maritimus a salt marsh plant that
can be used for producing lignocellulosic biomass,
because its total carbohydrate content can reach up
to 73%, with cellulose and hemicellulose represent-
ing approximately 41% and 31%, respectively, of the
lignocellulose biomass (Smichi et al. 2016). Tamarix
aphylla plants were irrigated from domestic sewage
(EC approximately 3 dS/m−1 ) to different salinity lev-
els or with brine (EC approximately 7–10 dS/m−1 ),
produced 52 and 26 t/ha, respectively, of organic
biomass. Tamarix was selected as a biofuel plants
for its higher cellulose and low hemicellulose and
polyphenol contents, properties particularly suitable
for bioethanol production, because the species of yeast
commonly used for fermentation prefer C6 − sugars
to C5 − sugars (Calvin 1980; Santi et al. 2015). The
variation in lignocellulosic contents and oil yields may
be due to the harvesting stage, different plant species,
cultivation time, climate change, ripening stage of
plant, and/or extraction method of chemical analysis.
Considerable genetic variability exists among halo-
phytic genotypes (Llanes et al. 2011) and genotypes of
some populations may perform better in suboptimal
conditions (Maron et al. 2004). Now a day’s mod-
ern genetic manipulation tools are also in operation
to enhance biomass of salt and the drought resistance
plants to produce higher biomass which can be useful
for bioenergy.
Genetic manipulation of biosynthetic pathways
of halophytes to enhance bioethanol production
It is essential to outline the most suitable approach to
convert plant complex polysaccharides into the sug-
ars and their conversion to ethanol by fermentation
for biofuel production. If cell wall degrading enzymes
(hemicellulase and cellulase) production can be induced
inside the growing cell through bioengineering of the
crop, then the reliance on microbial bioreactors for
 FRONTIERS IN LIFE SCIENCE 5

Table 2. Oil content, Saponification number (SN) Iodine value (IV) suitable genes to modify synthesis of sugars, lignin and
Cetane number (CN) of fatty acid methylesters of some halophytes lipids in plants..
seeds oils.
Finding an operative method for tissue culturing
Name of species Oil content % SN IV CN
and transformation of water hyacinth cultivars is the
Alhagi marorum 21.9 202.31 97.31 51.38
Allenrolfea occidentialis 14 193.44 121.36 47.21
initial step. Tissue culture specifies the strategies and
Arthrocnemum macrostachyum 25 205.66 115.5 46.86 procedures accessible for cultivating a huge number
Atriplex bunge 15.8 194.89 75.29 57.35
Atriplex rosea 12.9 194.05 115.5 49.33 of cells in controlled and axenic environment. Cell
Cressa cretica 23.3 203.5 114.27 47.41 totipotency that is to regenerate intact generally as
Halogeton glomeratus 24.7 199.82 103.47 50.31
H. mucronatum 22.7 202.95 124.63 45.15 fertile plants, that depends on the expression of devel-
Haloxylon stocksii 23.3 203.27 121.71 45.77 opmental genes is the basic principle of tissue cul-
Kochia scoparia 9.7 200.56 138.69 42.31
Kosteletzkya virginica 17.54 205.99 113.14 47.37 turing. The last are typically heritable, in light of the
Sacrobatus vermiculatus 17.5 199.22 139.84 42.23 tissue culture conditions. Hence, numerous permu-
S. bigelovii 29.7 200.86 154.32 38.78
Salicornia brachiata 22.4 129.89 29.7 81.67 tations of plant growth regulators are needed to be
S. europaea 30 202.55 155.96 38.19 evaluated for plant organogenesis and regeneration
Salicornia fruticosa 25.98 204.38 93.68 51.96
Sarcocornia ambigua 13 187.19 106.66 51.49 (Hassanein and Soltan 2000). In order to use the tissue
Suaeda fruticosa 25.98 204.38 93.68 51.96
Suaeda aralocaspica 29.92 198.7 146.59 40.82
culture as a baseline for genetic transformation, this is
Suaeda salsa 25.99 171.1 133.9 48.1 a crucial pre-requisite. In biolistic gene delivery and
Sueda torreyana 25.25 202.02 154.77 38.49
Agrobacterium tumefaciens-mediation tissue culturing
Source: Abideen et al. (2015).
techniques this is a key component needed (Bhatia
et al. 2004). Mass multiplication of plants and micro-
propagation is a major application of tissue cultur-
enzymes will be reduced and the production costs ing. It offers another method for acquiring particular
could be efficient (Balan 2014). Cost optimization can pathogen free plants from meristem culture, heredi-
be accomplished by engineering of lignin biosyn- tary control, and the generation of haploid plants. In
thetic pathway in plants. These measures can substan- the propagation of various plant species the Micro-
tially reduce the cost of pretreatments but the plants propagation has been applied broadly. The compo-
those already have lower lignin may be more cost nents that are responsible for the success of a tissue
effective in hydrolysis of biomass. The yield of cellu- culture system includes composition of the growth
losic biofuel can be enhance by up regulation of cellu- media, plant species, availability of utilizable carbon,
lose biosynthesis pathway enzymes to support higher growth regulators, the axenic culture conditions and
polysaccharide production (Bita and Gerats 2013). It callus induction response. Some additional factors are
is reported earlier that adaptable polymerization is carbon source, trace elements, plant growth regulators,
possible in plants and mono-lignins can be substi- inorganic salts and organic compounds (Bhattacharya
tuted by polyphenols (Wymelenberg et al. 2006). These and Kumar 2010). Tissue culture of halophytes includ-
above describe procedure may not suffer the advance- ing S. bigelovii (Lee et al. 1992), Leymus chinensis T,
ment procedure of plants, but will increase extrac- Suaeda maritima has been explored recently. There are
tion of important saccharides which is critical for the many halophytes that show potential in biofuel pro-
energy production from the plants. Similar approach duction but still needs careful studies to improve their
can be applied to water hyacinth and many other yield by genetic transformation.
bioenergy feedstocks which is composed of lower
lignin content and might enhance extraction of sac-
Genetic transformation can enhance oil content
charides (Lukuyu et al. 2014). Vanden Wymelenberg
of halophyte
et al. (2006) revealed a substantial number of genes
involved in breakdown of lignin from the fungus Knockout and overexpression strategies have been
Phanerochaetechrysosporium genome. Accordingly, applied lately to clarify the genes role in lipid synthesis
expression of such genes in water hyacinth may also and accumulation in order to enhance the Arabidop-
reduce the cost of biofuel. The utilization of transgenes sis thaliana, rapeseed (Brassica napus) and soy bean
could support already existing breeding methods to (Glycine max), plants lipid contents. These above
enhance biofuel production through accessibility of described studies might improve quality and quantity
6 N. MUNIR ET AL.
triacylglycerols of seed oil and other plant organs.
Ohlrogge and Jaworski (1997) that seed might be pre-
programmed to produce the particulate amount of
fatty acids (Ohlrogge and Jaworski 1997); substan-
tial efforts have been made to make the expression of
pathways of fatty acid synthesizing enzymes.
The initial step in synthesis of fatty acid in many
organisms is catalyzed by acetyl-CoA carboxylase
(ACCase) is the exchange of acetyl-coenzyme A (CoA)
to malonyl-CoA. Nevertheless, significant efforts were
made to enhance lipid content in a range of sys-
tems by utilizing ACCase overexpression which seems
somehow disappointing. Insignificant increase in seed
lipid content 6% (384 mg g−1 and 408 mg g−1 dry
weight for wild-type and transgenic ACCase rapeseed
lines, respectively) was noted by the overexpression of
ACCase in the oleaginous seeds of B. napus. In lipid
Figure 2. Mechanism pathways for drought tolerance in B. napus.
biosynthesis the ACCase levels are a restricting stride
for the most part in cells that ordinarily don’t store
a lot of lipid (Figure 2). Another endeavor to expand
the lipid production involving unsaturated fat amal-
gamation through 3-ketoacyl-acyl-transporter protein
synthase III (KASIII) was made but was not much
effective (Dehesh et al. 2001). Many fascinating results
have been performed through the overexpression of
genes required in TAG assembly. A 40% increase in
lipid content was achieved through the overexpres-
sion of cytosolic yeast, glycerol-3-phosphate dehydro-
genase (G3PDH), in the seeds of B. napus (Vigeolas
et al. 2007). Synthesis of glycerol-3-phosphate which is
needed for TAG formation is catalyzed by the G3 PDH.
Overall seed oil production thus is also dependent
somewhat on genes involved in TAG assembly. This
finding was further strengthening by studies on the

correlation of TAG assembly genes overexpression
and increase in seed oil content. Studies (Jain et al.
2000; Jako et al. 2001; Taylor et al. 2002; Lardiza-
bal et al. 2008) reported significant increase in plant
lipid productivity by overexpressing the TAG assembly
genes (lysophosphatidic acid acyltransferase, glycerol-
3-phosphate acyltransferase or diacylglycerol acyl-
transferase). Due to the fact that enzymes seem to
be good candidates for overexpression strategies to
increase storage lipid contents, an attempt has also
been made to use directed evolution for increasing the
efficiency of DAGAT enzymes (Siloto et al. 2009).
Genetic manipulation in halophytes for the biofuel
productivity can be introduced for two purposes (a)
to increase lipids contents (b) to improve oil quality.
Recently researchers have observed that high levels of
NaCl enhance membrane lipids’ unsaturated fatty
acid in halophytes. α-linolenic acid 18:3 is the main
fatty acid enhance under NaCl stress in halophytes.
Stress resistance ability of transgenic tobacco plant
at water deficit and salinity was improved through x-
3 desaturases (Sui and Han 2014; Zhang et al. 2005),
which infers that the drought and salt resistance of
plant is dependent on the unsaturated fatty acids
(Berberich et al. 1998; Mikami and Murata 2003).
This concept was further strengthen by the lower
of the x-3 and x-6 desaturase activity in Synechocys-
tis mutants (Allakhverdiev et al. 2001). In another
study an increased tolerance to low temperature and
salt stress was observed when x-6 desaturase sun-
flower gene were transformed in yeast cells. Three
sorts of halophytes were noticed which increment their
resilience to salt stress through expanding or keep-
ing up their unsaturated fats composition. Possible
clarification about role of higher unsaturated fatty
acid is linked with the membrane (Na+ or K+) ion
channels and Na+/H+ antiporter frameworks stabil-
ity under stress situations. Higher quantity of unsat-
urated fatty acids robust the membrane fluidity, and
triggers the activity of Na+/H+ antiporter and H+-
ATPase to protect the photosynthesis apparatus and
ultimately increase carboxylation efficiency. It was also
reported that with the change in membrane fluid-
ity can activate certain membrane bound enzymes
which can alter physiological responses of plants under
suboptimal conditions (Zhang et al. 2012). Accord-
ing to Sui and Han (Sui and Han 2014) to enhance
membrane fluidity for the ion compartments, it’s prob-
able that euhalophytes for example, S. salsa required
FRONTIERS IN LIFE SCIENCE 7
additional un-saturated fatty acids. It is also pragmatic
that at lower temperature and light intensity favors the
synthesis of unsaturated fatty acids protect photosyn-
thesis apparatus by reducing the photoinhibition and
photodamage of PSII and PSI (Sui et al. 2007). Plant
seedlings of Solanum tuberosum L. was transformed
with the desA encoding gene 12 acyl-lipid desaturase
in to another organism cyanobacterium Synechocys-
tissp. PCC 6803. In this situation sequence of genes
was translationally fuzed with the sequence of the
reporter gene encoding to thermostable lichenase to
analyze the efficiency of this gene expression in plant.
Interestingly the lichenase retained the thermostability
and its activity within the hybrid protein observed by
the comparison of hybrid and native gene expression
however another enzyme named as desaturase started
inserting the double bond in fatty acid chains to amend
their composition in membrane lipids. (Maali-Amiri
et al. 2007). The shoots enclosed higher linoleic acid
(39–73%) and linolenic acid (12–41%) in most trans-
formed plants. In comparison to wild-type plants, the
total absolute unsaturated FAs content was (20–42%)
higher in transformants. When severely cooled to
−7°C wild-type plants increased their membrane lipid
substantially by 25% whereas in under such condi-
tions the membrane lipid peroxidation rate was not
increased in transformed plants. The reason for the
higher resistance to lower temperatures and the
oxidative injury could have induced by hypothermia
(Maali-Amiri et al. 2007).
Gas–liquid chromatography (GLC) technique was
used for the oil compositions (qualitative and quanti-
tative) of esterified fatty acids (FAs) in the total fat con-
tent of halophytic plants such as Salicornia europaea,
Artemisia lerchiana and Suaedaaltissima collected in
their natural environments. GLC results showed that
the vegetative tissues of these halophytes contained
16 very-long-chain FAs among total 24 FA species.
Around four very-long-chain FAs groups were C20,
C21, C22, and C23, each including saturated, mono-,
and di-unsaturated components; C24 and C25 FAs
were also present. The concentration of VLCFAs in
the total FAs comprised 4–64%. In vegetative organs
of higher plants not subjected to genetic transforma-
tion, such a high VLCFA content was found for the
first time. Saturated and even-numbered components
predominated among the VLCFAs, and the roots
exceeded several fold the above-ground organs in the
total VLCFA content. Possible pathways of VLCFA
8 N. MUNIR ET AL.
biosynthesis in plants, VLCFA content in the vegeta-
tive tissues, and the physiological role of membrane
lipid FA composition in the plant salt metabolism are
discussed (Ivanova et al. 2009).
The study by Ramani et al. (2004) focused on
sulfolipids role and salt resistance mechanisms of
halophytes including Sesuvium portulacastrum, Aster
tripolium L., members of Compositae and L., Aizoa
ceae families, and glycophyte A. thaliana (L.) Heynh,
Brassicaceae. The sulfolipid contents of Sesuvium and
Aster increased significantly at 517 mM or 864 mM
salt stress conditions. At up to 100 mM NaCl changes
in sulfolipid contents were not observed in Arabidop-
sis. Whereas with an increase in NaCl concentra-
tion of Aster modified the fatty acid profile of sulfo-
quinovosyldiacyl glycerol. The presence of 16:0/18:3
and 18:3/18:3 molecules was confirmed through sul-
folipids quantification performed by LC-MS from
Aster and Sesuvium.
The sulfolipid content improved substantially in the
presence of NaCl in Crithmum maritimum halophyte.
Plants treated with salinity were similar in fatty acid
composition of sulfolipids, except for linolenic acids
and linoleic composition.
There was a significant decline in sulfolipids con-
tent under NaCl-treated plants in the unsaturated fatty
acid (C18:3) composition as compared to the con-
trol plants, whereas the percentage of unsaturated
fatty acids (C18:2) increase analogously (Ben Hamed
et al. 2005). In a conclusion from the above studies it
is found that the sulfolipds are an important aspect
of strategy in salt tolerance of halophytes. Nouairi
et al. (2006) carried out the experiments on young
small-sized hydroponically grown S. portulacastrum
and aseptically germinated seeds of Mesembryanthe-
mum crystallinum. They exposed these halophytes to
0, 50, 100 and 200 μM of cadmium concentration
for four weeks. The effect of cadmium on fatty acid
composition and leaf lipid contents of S. portulacas-
trum and M. crystallinum were studied recently. It was
found that the total lipids (TL) contents as well as
lipid fractions such as neutral lipids (NL) galactolipids
(GL) and phospholipids (PL) reduced more in M.
crystallinum as compared to S. portulacastrum at 200
μM cadmium concentration. Additionally there was
no noteworthy changes in the aggregate unsaturated
fat composition of S. portulacastrum leaves. How-
ever, an increase in the di-unsaturated fatty acid
(C18:2) and decrease of the tri-unsaturated fatty acid
(C18:3) was observed in M. crystallinum leaves. These
changes in fatty acid composition interestingly block
the sugar transport to leaves and conversion path-
ways but resulted in higher oil production and accu-
mulation in these plants (Zhai et al. 2017). Another
future option would be to dissolve lignocellulose mate-
rial from halophytes in saline ionic liquids, which
are well established as alternative and ‘green’ sol-
vents to be used in the pre-treatment of the walls
of plant cells prior to enzymatic hydrolysis (Gunny
et al. 2014). Lignocellulosic biomass from halophytes
for ethanol production proved advantageous both in
term of high net productivity and low maintenance
costs (Fooladvand and Fazelinasab 2014). Consider-
ing the advantages of second-generation biofuels, it is
recommended that biofuel production be increased up
to 10–20 EJ a year by 2050 (Searle and Malins 2015)
and the share of biofuels in the transport sector be
increased from 3% to 8% worldwide between 2013 and
2035.
Conclusions
Halophytes have a strong potential of biofuel pro-
duction as there are multiple feedstocks which could
be exploited industrially at lower cost. Exploration of
halophytes to produce biofuel can reduce the depen-
dence on conventional fuels in the world and can
discover some environmental benefits. However, bio-
fuel productions from some halophytes stocks need
special attention to improve their biomass for higher
yields by using genetic manipulations. A suitable com-
position of lignocellulosic biomass (higher cellulose
and hemi cellulose but lower lignin content) induce
through genetic manipulations manifest the poten-
tial of halophytes as a compatible biofuel candidate to
existing edible feedstock. These plant not only avoids
competition with water and land meant for production
of edible crops because of its ability to grow in soils
with salinity but will also help in CO2 sequestration
and reclaiming degraded lands.
Disclosure statement
No potential conflict of interest was reported by the authors.
References
Abideen Z, Ansari R, Khan MA. 2011. Halophytes: potential
source of ligno-cellulosic biomass for ethanol production.
Biomass Bioenergy. 35(5):1818–1822.

Abideen Z, Qasim M, Rizvi RF, Gul B, Ansari R, Khan MA.
2015. Oilseed halophytes: a potential source of biodiesel
using saline degraded lands. Biofuels. 6(5–6):241–248.
Ahmad AL, Yasin NHM, Derek CJC, Lim JK. 2011. Microal-
gae as a sustainable energy source for biodiesel production:
a review. Renew Sust Energ Rev. 15(1):584–593.
Allakhverdiev SI, Kinoshita M, Inaba M, Suzuki I, Murata N.
2001. Unsaturated fatty acids in membrane lipids protect the
photosynthetic machinery against salt-induced damage in
synechococcus. Plant Physiol. 125(4):1842–1853.
Amaro HM, Macedo ÂC, Malcata FX. 2012. Microalgae:
an alternative as sustainable source of biofuels? Energy.
44(1):158–166.
Balan V. 2014. Current challenges in commercially produc-
ing biofuels from lignocellulosic biomass, ISRN. Biotechnol.
2014:31.
Baram M., Bourrier M. 2011. Governing risk in GM agricul-
ture: An introduction. In: Baram M., Bourrier M., editors.
Governing risk in GM agriculture. New York: Cambridge
University Press; p. 1–12.
Ben Hamed K, Ben Youssef N, Ranieri A, Zarrouk M, Abdelly
C. 2005. Changes in content and fatty acid profiles of total
lipids and sulfolipids in the halophyte Crithmum maritimum
under salt stress. J Plant Physiol. 162(5):599–602.
Berberich T, Harada M, Sugawara K, Kodama H, Iba K, Kusano
T. 1998. Two maize genes encoding omega-3 fatty acid desat-
urase and their differential expression to temperature. Plant
Mol Biol. 36(2):297–306.
Bhatia P, Ashwath N, Senaratna T, Midmore D. 2004. Tissue
culture studies of tomato (Lycopersicon esculentum). Plant
Cell Tissue Organ Cul. 78(1):1–21.
Bhattacharya A, Kumar P. 2010. Water hyacinth as a poten-
tial biofuel crop. Electron J Environ Agric Food Chem.
9(1):112–122.
Bita C, Gerats T. 2013. Plant tolerance to high temperature in
a changing environment: scientific fundamentals and pro-
duction of heat stress-tolerant crops. Front Plant Sci. 4:
273.
Calvin M. 1980. Hydrocarbons from plants: analytical methods
and observations. Naturwissenschaften. 67(11):525–533.
Cherubini F, Bird ND, Cowie A, Jungmeier G, Schlamadinger
B, Woess-Gallasch S. 2009. Energy- and greenhouse gas-
based LCA of biofuel and bioenergy systems: key issues,
ranges and recommendations. Resour Conserv Recycl.
53(8):434–447.
Colombo L, Onorati A. 2013. Food. Riots and rights. London
(UK): IIED.
Dehesh K, Tai H, Edwards P, Byrne J, Jaworski JG. 2001. Over-
expression of 3-ketoacyl-acyl-carrier protein synthase IIIs
in plants reduces the rate of lipid synthesis. Plant Physiol.
125(2):1103–1114.
Fita A, Rodríguez-Burruezo A, Boscaiu M, Prohens J, Vicente
O. 2015. Breeding and domesticating crops adapted to
drought and salinity: a new paradigm for increasing food
production. Front Plant Sci. 6:978.
Flora CB, Bain C, Call C. 2012. Sustainability standards and
their implications for agroecology. In: Campbell W., López
FRONTIERS IN LIFE SCIENCE 9
Ortíz S., editors. Integrating agriculture, conservation and
ecotourism: societal influences. Issues in agroecology –
present status and future prospectus. Dordrecht: Springer;
p. 163–225.
Fooladvand Z, Fazelinasab B. 2014. Evaluate the potential halo-
phyte plants to produce biofuels. Eur J Biotechnol Biosci.
2:1–3.
Grainger A. 2013. Controlling tropical deforestation. London:
Routledge/CRC Press.
Grigore MN, Ivanescu L, Toma C. 2014. Halophytes: An inte-
grative anatomical study. New York, NY: Springer.
Gunny AA, Arbain D, Edwin Gumba R, Jong BC, Jamal P. 2014.
Potential halophilic cellulases for in situ enzymatic sacchari-
fication of ionic liquids pretreated lignocelluloses. Bioresour
Technol. 155:177–181.
Hassanein A, Soltan D. 2000. Solanum nigrum is a model sys-
tem in plant tissue and protoplast cultures. Biol Plantarum.
43(4):501–509.
Hinrichsen D, Tacio H. 2002. The coming freshwater crisis is
already here, the linkages between population and water.
Washington (DC): Woodrow Wilson International Center
for Scholars, 1–26.
Ivanova T, Myasoedov N, Pchelkin V, Tsydendambaev V,
Vereshchagin A. 2009. Increased content of very-long-chain
fatty acids in the lipids of halophyte vegetative organs, Rus-
sian. J Plant Physiol. 56(6):787–794.
Jain RK, Coffey M, Lai K, Kumar A, MacKenzie SL. 2000.
Enhancement of seed oil content by expression of glycerol-
3-phosphate acyltransferase genes. Biochem Soc T. 28(6):
958–961.
Jako C, Kumar A, Wei Y, Zou J, Barton DL, Giblin EM, Cov-
ello PS, Taylor DC. 2001. Seed-specific over-expression of an
Arabidopsis cDNA encoding a diacylglycerol acyltransferase
enhances seed oil content and seed weight. Plant Physiol.
126(2):861–874.
Karp A, Shield I. 2008. Bioenergy from plants and the sustain-
able yield challenge. New Phytolog. 179(1):15–32.
Khan SA, Rashmi, Hussain MZ, Prasad S, Banerjee UC. 2009.
Prospects of biodiesel production from microalgae in India.
Renew Sust Energ Rev. 13(9):2361–2372.
Ladeiro B. 2012. Saline agriculture in the 21st century: using
salt contaminated resources to cope food requirements. J
Botany. 2012:1–7.
Lardizabal K, Effertz R, Levering C, Mai J, Pedroso MC, Jury T,
Aasen E, Gruys K, Bennett K. 2008. Expression of Umbelop-
sis ramanniana DGAT2A in seed increases oil in soybean.
Plant Physiol. 148(1):89–96.
Lee CW, Glenn EP, O’Leary JW. 1992. In vitro propaga-
tion of Salicornia bigelovii by shoot-tip cultures. Hort Sci.
27(5):472–472.
Llanes A, Bonercarrere V, Capdevielle F, Vidal S, Luna V. 2011.
Genetic diversity in a natural population of the halophytic
legume Prosopis strombulifera revealed by AFLP finger-
printing. B Soc Argent Bot. 46(3–4):305–312.
Lukuyu B, Okike I, Duncan AJ, Beveridge M, Blummel M.
2014. Use of cassava in livestock and aquaculture feeding
programs. ILRI Discussion Paper 25. Nairobi, Kenya: ILRI.
10 N. MUNIR ET AL.
Maali-Amiri R, Goldenkova-Pavlova IV, Yur’eva NO, Pchelkin
VP, Tsydendambaev VD, Vereshchagin AG, Deryabin AN,
Trunova TI, Los DA, Nosov AM. 2007. Lipid fatty acid
composition of potato plants transformed with the 12-
desaturase gene from cyanobacterium, Russian. J Plant Phys-
iol. 54(5):600–606.
Maron JL, Vilà M, Bommarco R, Elmendorf S, Beardsley P.
2004. Rapid evolution of an invasive plant. Ecol Monogr.
74(2):261–280.
Mikami K, Murata N. 2003. Membrane fluidity and the percep-
tion of environmental signals in cyanobacteria and plants.
Prog Lipid Res. 42(6):527–543.
Munns R, Tester M. 2008. Mechanisms of salinity tolerance.
Annu Rev Plant Biol. 59:651–681.
Nouairi I, Ghnaya T, Youssef NB, Zarrouk M, Ghorbel MH.
2006. Changes in content and fatty acid profiles of total lipids
of two halophytes: Sesuvium portulacastrum and Mesem-
bryanthemum crystallinum under cadmium stress. J Plant
Physiol. 163(11):1198–1202.
Nyankanga RO, Onwonga RN, Wekesa FS, Nakimbugwe D,
Masinde D, Mugisha J. 2012. Effect of inorganic and organic
fertilizers on the performance and profitability of grain ama-
ranth (Amaranthus caudatus L.) in Western Kenya. J Agricul
Sci. 4(1):223.
Ohlrogge JB, Jaworski JG. 1997. Regulation of fatty acid syn-
thesis annual review of plant physiology and plant molec-
ular biology. Annu Rev Plant Physiol Plant Mol Biol. 48:
109–136.
Panta S, Flowers T, Lane P, Doyle R, Haros G, Shabala S. 2014.
Halophyte agriculture: success stories. Environ Exper Bot.
107(Supplement C):71–83.
Piloto-Rodríguez R, Melo EA, Goyos-Pérez L, Verhelst S. 2014.
Conversion of by-products from the vegetable oil industry
into biodiesel and its use in internal combustion engines: a
review. Braz J Chem Eng. 31(2):287–301.
Rabie G, Almadini A. 2005. Role of bioinoculants in devel-
opment of salt-tolerance of Vicia faba plants under salinity
stress. Afr J Biotechnol. 4(3):210.
Ramani B, Zorn H, Papenbrock J. 2004. Quantification and fatty
acid profiles of sulfolipids in two halophytes and a glycophyte
grown under different salt concentrations. Z Naturforsch C.
59(11–12):835–842.
Reshamwala S, Shawky BT, Dale BE. 1995. Ethanol produc-
tion from enzymatic hydrolysates of AFEX-treated coastal
bermudagrass and switchgrass. Appl Biochem Biotechnol.
51–52(1):43–55.
Salinas CX, Gironás J, Pinto M. 2016. Water security as a chal-
lenge for the sustainability of La Serena-Coquimbo conurba-
tion in northern Chile: global perspectives and adaptation.
Mitig Adapt Strat Gl. 21(8):1235–1246.
Santi G, Muzzini VG, Galli E, Proietti S, Moscatello S, Bat-
tistelli A. 2015. Mycelial growth and enzymatic activities
of white-rot fungi on anaerobic digestates from industrial
biogas plants. Environ Engin Manag J. 14(7):1713–1719.
Searle S, Malins C. 2015. A reassessment of global bioenergy
potential in 2050. GCB Bioen. 7(2):328–336.
Siloto RM, Truksa M, Brownfield D, Good AG, Weselake RJ.
2009. Directed evolution of acyl-CoA:diacylglycerol acyl-
transferase: development and characterization of Brassica
napus DGAT1 mutagenized libraries. Plant Physiol Bioch.
47(6):456–461.
Smichi N, Messaoudi Y, Moujahed N, Gargouri M. 2016.
Ethanol production from halophyte Juncus maritimus using
freezing and thawing biomass pretreatment. Renew Energ.
85:1357–1361.
Sui N, Han G. 2014. Increases of unsaturated fatty acids in
membrane lipids protects photosystem II from photoinhi-
bition under salinity in different halophytes. J Agricul Sci.
6(12):251.
Sui N, Li M, Zhao S-J, Li F, Liang H, Meng Q-W. 2007. Over-
expression of glycerol-3-phosphate acyltransferase gene
improves chilling tolerance in tomato. Planta. 226(5):1097–
1108.
Sun Y, Solomon S, Dai A, Portmann RW. 2006. How often does
it rain? J Climate. 19(6):916–934.
Taylor DC, Katavic V, Zou J, MacKenzie SL, Keller WA, An
J, Friesen W, Barton DL, Pedersen KK, Giblin EM. 2002.
Field testing of transgenic rapeseed cv. Hero transformed
with a yeast sn-2 acyltransferase results in increased oil con-
tent, erucic acid content and seed yield. Molecul Breed.
8(4):317–322.
Vigeolas H, Waldeck P, Zank T, Geigenberger P. 2007. Increas-
ing seed oil content in oil-seed rape (Brassica napus L.) by
over-expression of a yeast glycerol-3-phosphate dehydroge-
nase under the control of a seed-specific promoter. Plant
Biotechnol J. 5(3):431–441.
Ward RD, Friess DA, Day RH, MacKenzie RA. 2016. Impacts of
climate change on mangrove ecosystems: a region by region
overview. Ecosyst Health Sustainabil. 2(4):e01211.
Wymelenberg AV, Minges P, Sabat G, Martinez D, Aerts A,
Salamov A, Grigoriev I, Shapiro H, Putnam N, Belinky
P. 2006. Computational analysis of the Phanerochaete
chrysosporium v2.0 genome database and mass spectrom-
etry identification of peptides in ligninolytic cultures reveal
complex mixtures of secreted proteins. Fungal Genet Biol.
43(5):343–356.
Zhai Z, Liu H, Xu C, Shanklin J. 2017. Sugar potentiation of
fatty acid and triacylglycerol accumulation. Plant Physiol.
175(2):696–707.
Zhang M, Barg R, Yin M, Gueta-Dahan Y, Leikin-Frenkel A,
Salts Y, Shabtai S, Ben-Hayyim G. 2005. Modulated fatty
acid desaturation via overexpression of two distinct ω-3
desaturases differentially alters tolerance to various abi-
otic stresses in transgenic tobacco cells and plants. Plant J.
44(3):361–371.
Zhang J, Liu H, Sun J, Li B, Zhu Q, Chen S, Zhang H. 2012. Ara-
bidopsis fatty acid desaturase FAD2 is required for salt tol-
erance during seed germination and early seedling growth.
PloS One. 7(1):e30355.
Zhou Y, Tol RSJ. 2005. Evaluating the costs of desali-
nation and water transport. Water Resour Res. 41(3).
doi:10.1029/2004WR003749.