JES-01826; No of Pages 15

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X ) X X X

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1Q1 Perspective

2 Assessment of sunflower germplasm for

3 phytoremediation of lead-polluted soil and
4 production of seed oil and seed meal for human
5 and animal consumption

Q36Q2
Afsheen Zehra1,2, Zulfiqar Ali Sahito1, Wenbin Tong3, Lin Tang1, Yasir Hamid1,
7 Muhammad Bilal Khan1, Zarina Ali2, Beena Naqvi4, Xiaoe Yang1,⁎

8 1. Ministry of Education (MOE) Key Laboratory of Environmental

Remediation and Ecosystem Health, College of Environmental and Resources
9 Science, Zhejiang University, Hangzhou 310058, China
10 2. Department of Botany, Federal Urdu University of Arts, Science and
Technology, Karachi 75300, Pakistan
11 3. Technical Extension Station of Soil Fertilizer and Rural Energy,
Qujiang, Quzhou 324000, China
12 4. PCSIR Laboratories Complex, Karachi, Dr Salimuzzaman Road, Karachi
75280, Pakistan
13

1 6 A R T I C L E I N F O
A B S T R A C T

1272
1283
1294
25
26
Article history:
Received 5 December 2018
Revised 29 May 2019
Accepted 30 May 2019
Available online xxxx
Phytoremediation is a valuable technology for mitigating soil
contamination in agricultural lands, but phytoremediation without economic revenue
is unfeasible for land owners and farmers. The use of crops with high biomass
and bioenergy for phytoremediation is a unique strategy to derive supplementary
benefits along with
remediation activities. Sunflower (Helianthus annuus L.) is a high-biomass crop
that can
27 be
used for the phytoremediation of polluted lands with additional
advantages
2468
2479
3480
3491
3502
3513
3524
3535
3546
Keywords:
Heavy metal
Soil contamination
Metal removal efficiency (MRE) Germplasm (GP) Phytoextraction
Edible oil
Fatty acids (FA) Amino acids (AAs)
(biomass and oil). In this study, 40 germplasms of sunflower were screened in
field conditions for phytoremediation with the possibility for oil and meal
production. The study was carried out to the physiological maturity stage. All
studied germplasms mopped up substantial concentrations of Pb, with maximum amounts
in shoot > root > seed respectively. The phytoextraction efficiency of the
germplasm was assessed in
terms of the Transfer factor (TF), Metal removal efficiency (MRE) and Metal
extraction ratio (MER). Among all assessed criteria, GP.8585 was found to be
most appropriate for restoring moderately Pb-contaminated soil accompanied with
providing high biomass
and high yield production. The Pb content in the oil of GP.8585 was below the
Food
3557
safety standard of China, with 59.5% oleic acid and 32.1% linoleic acid. Moreover,
amino
38
acid analysis in meal illustrated significant differences among essential
and non-
39
essential amino acids. Glutamic acid was found in the highest percentage
(22.4%),
40
whereas cysteine in the lowest percentage (1.3%). Therefore,
its efficient

⁎ Corresponding author.
E-mail address: xeyang@zju.edu.cn. (X. Yang).

https://doi.org/10.1016/j.jes.2019.05.031
1001-0742 © 2019 The Research Center for Eco-Environmental Sciences, Chinese
Academy of Sciences. Published by Elsevier B.V.

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
https://doi.org/10.1016/j.jes.2019.05.031
2 J O U R N A L O F E N V
I R O N M E N T A L S C I E N C E S X X ( X X X X ) X X X

41
phytoextraction ability and good quality edible oil and meal production makes
GP.8585
42
the most convenient sunflower germplasm for phytoremediation of moderately Pb-
43
contaminated soil, with fringe benefits to farmers and landowners.
44 ©
2019 The Research Center for Eco-Environmental Sciences, Chinese Academy of
Sciences.
45
Published by Elsevier B.V.
58
59

61 Introduction

62 Contamination of farming land has been a worldwide

63 concern. Heavy metals (HMs), mainly lead (Pb), is the most
64 devastating pollutant that pose a severe menace to human
65 and animal health and cause soil pollution (Duruibe et al.,
66 2007; Vamerali et al., 2010; Malar et al., 2014). Contamination
67 of soil by lead occurs primarily through anthropogenic
68 activities such as irrigation with wastewater, and industrial
69 activity such as disposal of solid wastes including sewage
70 sludge and vehicular exhaust to the surroundings (Mielke et
71 al., 2010; Venkatachalam et al., 2016). Other anthropogenic
72 sources include mining and metallurgical industries and
73 manufacturing of batteries, sheet, ammunition, pipe, cable
74 sheeting, solder, paint, and trash incineration (Garcia-Alix et
75 al., 2013). Soil pollution with Pb is a serious problem in China,
76 where20% of overallarable farmlands have beencontaminated
77 with noxious metals and more than one-third have been
78 deteriorated and degraded due to unsustainable land-
79 management practices (Chen et al., 1999; Zhang et al., 2015;
80 Lei et al., 2016). Unnecessary accretion of lead into edible
81 fractions of plants may induce toxicity in the food chain and

Agro-production (i.e., oil and meal) from phytoremediation 113 is an important

avenue that facilitates not only the social 114 economy for sustainable
utilization of contaminated farm- 115 lands but also helps in meeting global
food demands (Cheng 116 et al., 2015). Recently, most of the arable lands
across the 117 world have been polluted with several toxic metals, including 118
lead (Pb) (Wei and Yang, 2010; Manikandan et al., 2015). Since 1Q149 cultivated
lands for food production are alarmingly concen- 120 trated, using moderately
contaminated soils for agro- 121 production is a good option to minimize food
shortage issues 122 (Sytar et al., 2015). Therefore, phytoremediation of
such 1Q253 neglected and abandoned lands for agro-production is receiv- 124 ing
worldwide attention (Lombi et al., 2001; Wong, 2004). 125
Sunflower is a profitable oil crop having high biomass, and 126 ranks second to
soybean among all oil seeds globally as a 127 source of vegetable oil (Lopez-
Beceiro et al., 2011). Sunflower 128 oil is composed of monounsaturated (Oleic
acid) and polyun- 129 saturated fats (Linoleic acid) and essential vitamins
(Vitamin 130
A, B and E) (Quiles et al., 2002). Its consumption helps in 131 maintaining
an ideal LDL/HDL (good cholesterol) ratio in the 132 body (http://www.webmd.com).
It is used in treatment of 133
acne, arthritis and hair damage (Alexander, 1998; Rele and 134
82 cause serious health problems in humans such as high blood
Mohile, 2003; Lambert et al., 2007).
135
83 pressure, joint and muscle pain, and difficulties with memory
84 and concentration (Tong et al., 2000; Gall et al., 2015; Zhong et
85 al., 2018). Lead poisoning also causes one of the most common
86 pediatric problems in China, and it was reported that one-third
87 of Chinese children are suffering from elevated serum lead
88 levelsthat causedelaysin development,difficultiesinlearning,
89 loss of appetite, weight loss, hearing loss, seizure and fatigue
90 (Huo et al., 2007; Bellinger, 2008; Wang et al., 2008). Strict
91 initiatives have been undertaken to control lead pollution
92 includingthebanningof leadingasoline,aleadingcauseof lead
93 pollution (Meyer et al., 2008; Venkatachalam et al., 2016).
94 The cleaning of Pb-contaminated soil is mandatory, and
95 traditional phytoremediation techniques are not very pro-
96 ductive, as they focus on special hyperaccumulator plants
97 that can accumulate high concentrations of phytotoxic
98 elements, (Peer et al., 2005) and harvested plants require
99 incineration treatments that cause secondary pollution in
100 the environment. Thus, conventional phytoremediation in-
101 volves cost year by year and owners of the contaminated
102 sites have no income during the remediation period, which
Sunflower meal is an excellent source of protein for 136 livestock feed
for ruminants and non-ruminants (https:// 137 www.sunflowernsa.com) and
contains high protein value, 138 with essential amino acids like leucine, valine
and isoleucine, 139 etc., which are well known for promotion of muscle growth 140
and development (Figueroa et al., 2003; González-Pérez and 141
Vereijken, 2007). Sunflower meal is considered to be the 1Q462 largest
by-product produced after the oil extraction process 143 and is consumed by
livestock for enhancing milk production 144 in cows and weight gains in young
calves and growing heifers 145 (Yildiz, 2015). The low-fiber and high-protein
meals can be 1Q476 utilized in poultry feed (non-ruminants) (Senkoylu and Dale,
147
1999; Torok et al., 2011). Therefore, the objective of this study 148 was to
screen potential sunflower germplasms grown under 149 natural environmental field
conditions for phytoremediation 150 of Pb-contaminated soil. Furthermore, the
main focus was on 151 (1) searching for an ideal sunflower germplasm with high
152 extraction ability of Pb; (2) high biomass for bioenergy (biofuel) 153
production; (3) safe products with economic value (oil and 154
meal within permissible limits of Pb concentration); (4) quality 155
103 has restricted the practical application of traditional
104 phytoremediation. In recent years, high biomass and eco-
105 nomically important crops like maize (Zea mays L.), sorghum
106 (Sorghum bicolor L.), Indian mustard (Brassica juncea L.), oilseed
107 rape (Brassica napus L.) and broad bean (Vicia faba L.) have
108 received increasing attention to remediate contaminated
109 soils as well as produce valuable biomass, through which
oil for human consumption; and (5) meal for animal feed.

1. Materials and methods

1.1. Study site and experimental design

156

1578

159
110 the practical application of phytoremediation may be pro-
111 moted (Diwan et al., 2008; Soudek et al., 2014; Wuana and
112 Okieimen, 2010; Tang et al., 2019).
The experimental site was located in farmland in eastern 160
Zhejiang province. The field climatic conditions throughout 161
the study were Avg temp: 10.5–29.2°C, Avg precipitation: 45%– 162

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
https://doi.org/10.1016/j.jes.2019.05.031
J O U R N A L O F E N V I R O N M E N T A L S C I E N C E S X X ( X X X
X ) X X X
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163 67%, whereas the soil was clay-like in texture. The land is
164 used for the agriculture of different crops, and before starting
1.5. Lead analysis in various plant tissues
217
165 the sunflower plantation, Indian mustard was grown in it. The
166 soil of the field was moderately contaminated with lead due
167 to frequent exposure to various fertilizers and pesticides in
168 the vicinity. The experiment was conducted in a split-split
169 plot design on fixed sites in March 2017. In this study, 40
170 sunflower germplasms were used. Each germplasm was
171 considered as the main plot in a randomized block design
172 method with four replicated blocks. The four replicates of
173 each germplasm were used as minor plots in the 40 m2 area.
174 The cultivated area was also protected from grazing.

175 1.2. Plant material and sampling

176 Forty different germplasms of sunflower were obtained from

177 the seed market of Hangzhou city in Zhejiang province and
178 used for the screening experiment in moderately Pb-polluted
179 farmland soil of Zhejiang province, China. Random soil
180 samples from the experimental site were collected at a
181 depth of 0–20 cm prior to plantation. After tilling of the field,
182 sunflower seeds were sown at a depth of 0.5 in. One healthy
Plants were harvested at the reproductive stage fom the field 218 and each plant
was than divided into various subsamples 219 including stem, leaf, root and seed.
Each sample (except seed) 220 was kept in a paper envelope for drying in an oven
at 65°C for 221
72 hr. The dry biomass of each sample was measured and the 222 samples were
milled into fine powder with a mixer. The 223 powdered samples were stored in
dark brown bottles for 224 further metal analysis. From each sample, 0.2 g was
digested 225 in two acids, i.e., HNO3:H2O2 in a 5:1 ratio (5 mL of 68%–70%
226
HNO3 and 1 mL of (30%) H2O2. Each sample was placed into a 227
test tube and 5 mL of nitric acid mixture was added into each 228 test tube. The
tubes were then put on an electric hot plate 229 until white fumes were observed
and the plant tissue was 230 completely digested. After digestion, each sample was
filtered 231 with a syringe micro-filter and the volume of solution was 232 made
up to 20 mL Metal analysis was performed using ICP-MS 233 (7500a, Agilent, USA).
For quality assurance, GBW(E) 100351 234 certified reference material for rice
flour chemical composi- 235 tion was used as the standard. The recovery of
spiked 236
standard was 90%–110% for the metals and was also digested 237
183 seedling was selected after seedling emergence for further
184 experiments. The plants from each replicated block were
185 harvested at the physiological maturity stage after 126 days.
186 Phenotypic characteristics such as plant height, biomass and
by a method similar to plant sample digestion.

1.6. Measurement of phytoremediation efficiency

238

239
187 grain yield were recorded for assessing high-accumulator
The phytoremediation efficiency of the sunflower germplasm 240
188 germplasms, which were used for further analysis. Plant
189 samples were than divided into root, stem, leaves and seeds.
was calculated as follows (Eqs. (1)–(3)):

Cmetal-shoot
241
190 The roots were thoroughly washed with fresh water and three
191 times with 0.02 mol/L EDTA, and then rinsed with Milli-Q
TF ¼
Cmetal-root
ð1Þ
192 water. The other plant parts were also washed carefully with
193 Milli-Q water and blotted dry with tissue paper, and the fresh
where TF is transfer factor, Cmetal-shoot (mg/kg) is the metal 2423
concentration in plant shoot, Cmetal-root (mg/kg) is the metal 244
194 weight was noted. Root and shoot parts of plants were oven-
concentration in plant root.
245
195 dried at 65°C for 72 hr to constant weight for plant biomass
196 and metal analysis.

MRE ¼

Csoil1
C

-Csoil2

100% ð2Þ

197 1.3. Soil sample preparation and chemical analysis

soil1

where, MRE is the metal removal efficiency, Csoil1 (mg/kg) is 2467

the soil Pb concentration before plantation; Csoil2 (mg/kg) is 248
198 The collected soil samples were air-dried at room temperature
the soil Pb concentration after harvesting.
249
199 for 60 day(s), ground into fine particles, and sieved (2 mm
Cplant

x Mplant
200 sieve). For assessment of physiochemical properties, the soil
201 samples were examined according to the Routine Analytical
MER ¼ C

soil x
Mrooted zone
100% ð3Þ
202 Methods of Agricultural Chemistry (Carter and Gregorich,
203 2008). Total metal content was determined by the nitric acid
204 digestion method described in (Ganzler et al., 1986) with some
205 modifications. 0.5 g soil samples were weighed and digested
206 with HNO3: HClO4: HF (5:1:1, V/V/V) in an oven at 180°C for
where, MER is metal extraction ratio, Cplant (mg/kg) is the 2501 metal
concentration in the harvested component of the plant 252 biomass; Mplant
(kg/m2) is the mass of harvestable above- 253 ground biomass produced in one
harvest; Csoil (mg/kg) is the 254 metal concentration in the soil volume; Mrooted
zone (kg) is the 255
207 12 hr. ICP-MS (7500a, Agilent, USA) was used for measuring
208 the Pb concentration of the digested solution. Chinese
209 standard reference soils (GSS6, GSS7 and GSS16) were used
210 for quality assurance of the studied field soil samples. The
mass of soil volume rooted by the species.

1.7. Quantification of seed oil content

256

257
211 recovery rates for lead (Pb) in the reference materials were
212 good (78%–118%).

213 1.4. Plant samples and measurements

214 Plant biomass, head diameter, number of seeds per plant,

215 1000 seeds' weight, and grain yield per unit area were
216 calculated after harvesting.
Oil extraction of all studied germplasms was conducted 258 according to a
standard method by using a Soxhlet apparatus 259 with hexane as solvent. 20 g of
powdered seed material was 260 loaded in a thimble and placed in the extraction
chamber. 261
This extraction chamber was assembled with a round-bottom 262 flask containing 200
mL of n-hexane. The system was allowed 263 to reflux for oil extraction for 12
h, then the hexane was 264 vaporized using a rotary evaporator. The sunflower oil
yield 265

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
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266 (Yoil, %) was calculated by Eq. (4):

W oil
where, Sblank is the control; Vtrit is the titration volume; 1.401 is 3145
the Kjeldahl nitrogen constant; 6.25 is the Jones factor for 316
Yoil ¼ W
100%
ð4Þ
converting nitrogen into protein; N is the normality of H2SO4; 317
seed

2678 where, Woil is the weight of extracted oil, and Wseed is the
Wsample
is the weight of sample.
318
269 weight of seeds used.
1.12. Determination of meal amino acids
319

270 1.8. Analysis of lead (Pb) content in oil

271 The Pb concentration in oil was analyzed through an acid

272 digestion method with slight modification (Cindric et al., 2007).
273 An aliquot (0.2 g) of extracted oil was added to 1 mL of 10%
274 diluted nitric acid. The mixture was shaken for 40 sec until the
275 layers were homogenized. The test tubes were incubated at 50°C
The amino acid composition in high-accumulator germplasm 320
seed meal was determined by the method described in 321
Ivanova et al. (2013). The acidic (5 mol/L HCl) hydrolyzed 322 samples were
used for the determination of amino acids using 323 a high-speed amino acid
analyzer (Hitachi, LA 8080 SAAYA, 324
Japan). The content of amino acid was measured in triplicate 325
as the ratio of each essential amino acid determined in the 326
276 in a shaking water bath for 2 hr. After centrifugation at
277 3000 rpm for 15 min, the lower aqueous layer was pipetted out
278 and was added to 25 mL of deionized water. The solution was
279 filtered and loaded into the auto-sampler of the ICP-MS system
isolated protein.

1.13. Measurement of mineral contents

327

328
280 for lead analysis (ICP-MS, 7500a, Agilent, USA).

281 1.9. Determination of oil composition (fatty acid analysis) by

282 GC-FID

283 Fatty acid analysis was performed by the method described in

Different minerals (Ca, K, Mg, and Na) were determined in all 329 studies of
germplasm seed meal. A 0.2 g sample was digested 330 with a HNO3:H2O2 (V/V)
solution by heating on a hot plate 331 until the solution turned colorless. The
digested solution was 332 cooled to room temperature and then diluted up to 20 mL
333 with deionized water. The mineral content analysis was 334
284 Nasopoulou et al. (2011) with some modifications. The extracted
285 oil from the screened high-accumulator germplasm was used in
286 the fatty acid analysis. The determination of n-hexane-
287 extracted oil quality was carried out by a gas chromatograph
performed by using ICP-MS (7500a, Agilent, USA).

1.14. Statistical analysis

335
336
288 (Agilent Technologies; Model: 7890A) equipped with HP-88 MS
289 column (30 m long × 0.32 mm; 0.25 film thickness). Nitrogen gas
290 was used as a carrier gas. Before quantification of the
291 components in the extracted oil, samples were filtered by
292 0.2 μm poly-tetra-fluoro-ethylene syringe micro-filters. The
The experimental data of Means comparison was statistically 337 analyzed by using
SPSS software version 20. A post hoc test 338 (Bonferroni, p < 0.05) was used for
determining the signifi- 339 cance levels of different studied parameters
(biomass, yield, 340 metal concentration, and metal accumulation) among differ- 341
293 GC/MSD CHEMSTATION software was used for data analysis.

294 1.10. Analysis of lead (Pb) in seed meal

295 The lead (Pb) concentration in meal samples was analyzed by

296 the acid digestion method. 0.2 g dried seed meal was digested
297 with HNO3:H2O2 (5:2, V/V) at 120°C for 4 hr. The digested
298 solution was diluted up to 15 mL and filtered using filter
ent germplasms.

2. Results

2.1. Soil characterization and metal analysis

342

3434

345

299 membranes. The meal Pb concentration was determined

300 using ICP-MS (7500a, Agilent, USA).

301 1.11. Quantification of protein content in seed meal

302 The total protein content in sunflower germplasm meal was

303 analyzed by the Kjeldahl standard method of nitrogen
304 determination described in Lynch and Barbano (1999) with
305 slight modifications. A 1 g meal sample from each germplasm
306 was placed in a Kjeldahl flask and digested with K2SO4:HgO:
The basic physical and chemical properties of the soil and 346
metal content in the field soil were analyzed in order to 347 determine the
contamination of soil and its removal after the 348 plantation of oil crop
sunflower. The following are the values 349 of soil-related parameters: pH:
5.55, CEC: 10.40 cmol/kg, 350 organic matter: 11.05%, total N: 3.17 g/kg,
total P: 81.74 g/kg, 351 total K: 247.91 g/kg, total Pb: 106.46 mg/kg. The
results showed 352 the presence of a moderate Pb concentration in soil at acidic
353 pH (Table 1). This level of Pb in farmland soil requires the best 354
management practice on an initial basis to minimize the 355
307 HCl (W/W/V) for 30 min until the solution became clear. The
308 digested solution was diluted with deionized water and 25 mL
probable increase of Pb toxicity to agriculture and humans.
356
309 Na2SO4 was added. The distillation process was carried out
2.2. Biomass of sunflower germplasm grown on Pb- 357
310 with 40% NaOH for 30 min; afterward, the distilled liquid was
311 titrated by using 0.1 mol/L H2SO4 until the blue liquid turned
contaminated soil under field conditions
358
312 to a purplish color. Sunflower meal crude protein content
313 (Ccrude protein, %) was calculated by Eq. (5):

ðS blank - V trit Þ 1 : 401 6 : 25 N

The biomass and grain yield of sunflower germplasms were 359
calculated at the physiological maturity stage, at which time 360 the plant
possesses its highest biomass. A significant differ- 361 ence (p < 0.05) was
observed in germplasm biomass and yield 362
Ccrude protein ¼
Wsample
ð5Þ
(Fig. 1).
363

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
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t1:1
t1:23
Table 1 – Field soil physiochemical properties ⁎.
The plant biomass ranged from 0.661 kg/m2 (GP. 18: S-9178) 364
to 0.235 kg/m2 (GP. 38: Ponum) with a 3-fold difference (mean 365
t1:4 Characters
Soil

t1:5 Type
Interceptisol
t1:6 Color
Red
t1:7 Texture
Clay
t1:8 pH
5.55 ± 0.07 t1:9 CEC (cmol/kg)
10.40 ± 0.03 t1:10 OM (%)
11.05 ± 0.03 t1:11 Total N (g/kg)
3.17 ± 0.09

value 0.426 kg/m2) from highest to lowest (Fig. 1a). The 366 sunflower
germplasms also showed significant differences 367 (p < 0.05) in seed
production. The maximum yield was 368 observed for GP.18: S-9178 (0.363
kg/m2) and minimum yield 369 was obtained for GP.38: Ponum (0.645 kg/m2); a 6-
fold differ- 370 ence (mean value 0.299 kg/m2) from maximum to minimum 371
grain yield was achieved in moderately Pb-contaminated soil 372
t1:12 Total P (g/kg)
81.74 ± 2.25
t1:13 Total K (g/kg)
247.91 ± 4.15
t1:14 Total Pb (mg/kg)
106.46 ± 4.43
(Fig. 1b).
373
2.3. Pb content in individual parts of sunflower germplasm 374
⁎ Soil pH measured in water: soil (1:1, m/V) mixture; Cation exchange capacity
(CEC) measured by using ammonium acetate
grown in Pb-contaminated soil under field conditions
375

t1:15
saturation method (Chapman, 1965); Organic matter (OM) was measured by using
the Walkley-Black method (Walkley and Black,
1934); Soil texture was analyzed by the hydrometer method; Total
N, P, K and Pb were measured by the acid digestion method.
Different parts of sunflower germplasm performed differen- 376 tially in terms of
Pb concentration and accumulation, with the 377 highest metal concentration and
accumulation in shoot, root 378
and seed respectively (Fig. 2).
379

Fig. 1 – Biomass (a) and grain yield (b) of sunflower germplasms grown on Pb-
contaminated soil under field conditions. Bars having the same letters represent
non-significant differences (p 0.05) among germplasms (Bonferroni test). Bars
indicate Mean ± S.E.

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
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6 J O U R N A L O F E N V
I R O N M E N T A L S C I E N C E S X X ( X X X X ) X X X

380 2.4. Concentration of Pb in individual parts of sunflower

381 germplasm

382 The lead (Pb) concentration at the reproductive stage in root

383 ranged from 56.984 mg/kg (GP.2: S-606) to 14.41 mg/kg (GP.39:
384 Agsun-5246) with a 4-fold difference (mean value, 42.57 mg/
kg) from highest to lowest metal concentration (Fig. 2 a1). In 385
shoot, the Pb concentration ranged from 98.234 mg/kg (GP.31: 386
DW-567) to 32.167 mg/kg (GP.36: NK-S-278), with a 3-fold 387 difference
(mean value, 66.067 mg/kg) from highest to lowest 388 concentration (Fig. 2 a2).
The Pb concentration in seeds ranged 389
from 0.840 mg/kg (GP.28: Huanong HN aijvren) to 0.288 mg/kg 390

Fig. 2 – Concentration (a) and accumulation (b) of Pb in individual parts of

sunflower germplasm grown on Pb-contaminated soil. Bars having similar letters
represent non-significant differences (p 0.05) between germplasms (Bonferroni
test). Bars indicate Mean ± S.E. (n = 4 replicates).

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
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J O U R N A L O F E N V I R O N M E N T A L S C I E N C E S X X ( X X X
X ) X X X
7

391 (GP.13: 988), with a 3-fold difference (mean value, 0.552 mg/kg)
392 from highest to lowest Pb concentration in seeds (Fig. 2 a3).

393 2.5. Accumulation of Pb in individual sunflower germplasms

394 The metal accumulation in plants can be obtained by

395 interaction of the plant biomass and metal concentration.
396 Individual parts of sunflower germplasm showed significant
397 differences in Pb accumulation, and the highest accumulation
percentage (Fig. 4 a1), whereas the total protein of sunflower 445 meal ranged
from 51.18% (GP.16: 988 F1) to 23.46% (GP.3: S-958F1) 446 with a 2-fold
difference (mean value, 27.12%) from highest to 447 lowest (Fig. 4 a2). The oil
Pb content ranged from 0.035 mg/kg 448 (GP.28: Huanong HN aijvren) to 0.012 mg/kg
(GP.13: 988) with a 3- 449 fold difference (mean value, 0.023 mg/kg) from highest
to lowest 450 concentration (Fig. 4 b1) and the meal Pb content in sunflower 451
germplasm ranged from 0.418 mg/kg (GP.25: Mei kui) to 452
0.057 mg/kg (GP.36: NK-S-278) with a 7-fold difference (mean 453
398 was observed in shoot, root and seed respectively (Fig. 2b). The
399 Pb accumulation in root ranged from 0.777 mg/plant dw (GP.2:
value, 0.361 mg/kg) from highest to lowest (Fig. 4 b2).
454
400 S-606) to 0.156 mg/plant dw (GP.3: S-958F1) with a 5-fold
2.8. Mineral composition of sunflower germplasm seed meal 455
401 difference (mean value, 0.621 mg/plant dw) from highest to
402 lowest accumulation in root (Fig. 2 b1). In shoot, the Pb
grown on Pb-contaminated soil under field conditions
456
403 accumulation ranged from 5.999 mg/plant dw (GP.7: Youwang
404 zhizun F1) to 0.997 mg/plant dw (GP.3: S-958F1) with a 6-fold
405 difference (mean value, 5.002 mg/plant dw) from highest to
406 lowest (Fig. 2 b2). In seeds the Pb accumulation ranged from
407 0.109 mg/plant dw (GP.20: 8898 F1) to 0.018 mg/plant dw
408 (GP.12: Huanong gaoyouwang), with a 6-fold difference
409 (mean value, 0.091 mg/plant dw) from highest to lowest
410 accumulation in seeds (Fig. 2 b3).

411 2.6. Transfer factor and Pb phytoextraction with sunflower

412 germplasms grown on Pb-contaminated soil under field
413 conditions
The nutritional quality of sunflower germplasm seed meal is 457 given in Table.
2. A great variation was observed in the meal 458 mineral contents of different
germplasms. Calcium (Ca) ranged 459 from 0.31% (GP.28: Huanong HN aijvren) to
0.08% (GP.13: 988) 460 with a 4-fold difference (mean value, 0.28%) from
highest to 461 lowest. Magnesium (Mg) ranged from 0.25%
(GP.22: 462
Youhuanghou F1) to 0.08% (GP.4: D-7676) with a 3-fold difference 463
(0.173%) from highest to lowest. Potassium (K) ranged from 464
1.40% (GP.25: Mei kui) to 0.33% (GP.13: 988) with a 4-fold 465
difference (mean value, 1.06%) from highest to lowest. Sodium 466 (Na) ranged from
0.008% (GP.24: JD-985) to 0.001% (GP.32: A-19) 467 with an 8-fold difference
(0.007%) from highest to lowest. The 468
average mineral content in sunflower genotypes was K >Ca > 469
414 Values of TF > 1 for plant species are considered adequate for
415 phytoextraction and phytostabilization of particular soil
Mg >Na (0.63, 0.17, 0.13, 0.003% respectively (Table 2)).
470
416 metals, whereas the metal extraction ratio is calculated to
2.9. Identification of Pb-accumulator sunflower germplasms 471
417 evaluate the metal cleanup from polluted soil in one harvest
418 of the crop. A significant difference (p < 0.05) for these factors
after screening in Pb-contaminated soil under field conditions
472
419 was observed in sunflower germplasm (Fig. 3).
420 At the physiological maturity stage, all studied
421 germplasms showed TF > 1 except GP.2: S-606 (0.633), and
422 the highest transfer factor of Pb was observed in GP.25: Mei kui
423 (6.04) (Fig. 3 a) with a 9-fold difference from highest to lowest
424 TF. The metal removal efficiency (MRE) of sunflower
425 germplasms ranged from 82.2% (GP.2: S-606) to 8.20% (GP.40:
426 No.4. youkui) with a 10-fold difference from highest to lowest
Forty different sunflower germplasms were used for 473 phytoremediation of
Pb-polluted soil, and different parameters 474 were considered (biomass, grain
yield, metal concentration and 475 accumulation in shoot, metal transfer, metal
removal efficiency 476 and metal extraction ratio) for screening of Pb high-
accumulator 477 germplasms. The best 15 germplasms were screened as per the 478
assigned parameters, with significant differences (p < 0.05) given 479
in Fig. 5, and only GP.8: 8585 was identified as a Pb accumulator 480
427 MRE (Fig. 3 b). The highest metal extraction ratio (MER) was
428 observed in GP.4: D-7676 (41.6%) and the lowest ratio was
after considering all these parameters.
481
429 found in GP.38: Ponum (1.13%), with a 38-fold difference from
430 highest to lowest metal extraction ratios for different sun-
2.10. Fatty acid composition in oil and amino acid profile in 482
meal of Pb-accumulator sunflower germplasm grown on Pb- 483
431 flower germplasms in Pb-contaminated soil (Fig. 3 c).
contaminated soil under field conditions
484

432 2.7. Total oil yield and protein content in seed meal and
433 concentration of Pb in oil and meal in sunflower germplasms
434 grown on Pb-contaminated soil under field conditions

435 The oil and meal of sunflower seeds have immense nutri-
436 tional value both for human consumption and livestock feed.
437 The Pb concentration in sunflower oil and meal was analyzed
438 to ensure their safety and healthy quality. Fig. 4 shows
439 significant differences (p < 0.05) in the total oil and protein
440 content as well as Pb concentration in the oil and meal of
The high biomass of sunflower is an important feature for 485 phytoremediation,
and its oil and meal have great nutritive 486 value for humans and animals. The
oil fatty acid and meal 487 amino acid composition of high accumulator GP.8:
8585 was 488 analyzed to determine the level of usefulness in the agriculture 489
products. A significant difference (p < 0.05) in fatty acid and 490 amino acid
composition was obtained in the high-accumulator 491 germplasm. The percentage of
oleic acid was the highest (59.5%) 492 compared to other FA (Table 3). The fatty
acid contents of GP.8: 493
8585 varied in the following order: oleic acid > linoleic acid > 494
441 sunflower germplasms.
palmitic acid > stearic acid > Gadoleic acid.
495
442 The oil percentage of sunflower germplasms ranged from
443 43.60% (GP.18: S-9178) to 3.40% (GP.38: Ponum) with a 13 13-fold
444 difference (mean value, 40.2%) from highest to lowest oil
Amino acid profiling revealed different concentrations of 496
essential and non essential amino acids in the solvent- 497
extracted meal of the high-accumulator germplasm (Table 498

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8 J O U R N A L O F E N V
I R O N M E N T A L S C I E N C E S X X ( X X X X ) X X X

Fig. 3 – (a) Translocation factor (TF), (b) metal removal efficiency (MRE) of Pb
and (c) Metal extraction ratio (MER) of sunflower germplasms grown on Pb-
contaminated soil. Bars having different letters represent significant differences
(p < 0.05) between germplasms (Bonferroni test). Bars indicate Mean ± S.E. (n =
4).

499 4). Among non-essential amino acids (NEAA), the glutamic

500 acid concentration was the highest (22.4%) and the histidine
501 concentration was lowest (2.41%). The pattern of NEAA was:
502 glutamic acid > aspartic acid > arginine > glycine > alanine >
503 serine > tyrosine > histidine, respectively (Table 4). Whereas,
504 among essential amino acids (EAA), leucine was found in the
505 highest percentage (7.3%) and the cysteine percentage was the
506 lowest (1.36%). The pattern of EAA was: leucine > phenylala-
507 nine > valine > isoleucine > threonine > lysine >methio-
508 nine >cysteine (Table 4).
present at trace levels (<1000 mg/kg), with relatively low 513 influence
upon soil (Alamgir, 2016). However, increasing 514 application of
fertilizers, pesticides, sewage sludge, organic 515 manure, industrial wastes,
etc., is causing them to increase at 516 alarming levels, contributing to the
contamination and 517 destruction of soil quality and fertility (Su et al.,
2014). At the 5Q188 optimal concentrations, metals are beneficial to some extent,
519 while higher concentrations tend to be harmful to the 520
environment and living organisms. In the last few decades, 521 scientists and
agriculturists have been involved in overcom- 522
ing the threat of soil metal pollution, and phytoremediation, 523

especially by growing high biomass crops, has been adopted 524

5190 3. Discussion

511 Heavy metals are naturally occurring substances in the soil as

512 a consequence of pedeogenic processes of parental materials
as a solution (Lv et al., 2018; Sytar et al., 2015). Cultivation of 525 high
biomass crops for phytoremediation of metal-polluted 526 soils is a viable
approach towards the identification of the 527 most suitable plant species for
this system. In our study, 528

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sunflower germplasm for phytoremediation of lead- polluted soil and production of
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J O U R N A L O F E N V I R O N M E N T A L S C I E N C E S X X ( X X X
X ) X X X
9

Fig. 4 – (a) Total oil and protein content and (b) Pb concentration in oil and meal
of sunflower germplasms grown on Pb- contaminated soil. Bars having different
letters represent significant differences (p ˂ 0.05) between germplasms
(Bonferroni test). Bars indicate Mean ± S.E. (n = 4).

529 different sunflower germplasms were grown in moderately

530 Pb-contaminated soil, and significant differences in biomass
531 production and yield were observed for all germplasms in
532 uniform soil and environmental conditions (Fig. 1).
533 Genotypic variations in biomass production and yield have
534 also been reported by several authors, who showed that plant
535 response in particular environments was correlated to spe-
536 cific genotypes (Alake and Ayo-Vaughan, 2015; Matsunami et
Effective phytoremediation comprises utilization of plant 542 biomass and
accumulation of metals in harvestable parts 543 (Chaturvedi et al., 2018).
Several plant species have been 544 documented as Pb-tolerant plant species,
including Phragmites 545 australis L., Leucaena leucocephala L., Sesbania rostrata
L (Pratas et 546
al., 2013; Shu et al., 2013; Zhuang et al., 2007; Yang et al., 2016). In
5Q4171Q123Q14
the present study, the concentration of Pb in sunflower 548
germplasm was evaluated, and the Pb concentration and 549
5Q379
al., 2013; Morris et al., 2016). The resistance of sunflower to
accumulation was found to vary in the order shoot > root > 550
538 heavy metals was also investigated by Madejón et al. (2003),
539 while low and moderate Pb-concentration soil was found to
540 stimulate the sunflower growth and biomass (Jadia and
seed respectively (Fig. 2). The uptake of metals is influenced by 551
several factors (soil pH, organic matter, cation exchange 552
capacity, plant species, cultivation and harvesting time), in 553
5Q4110
Fulekar, 2008; Roongtanakiat and Sanoh, 2015).
which the pH of soil regulates the solubility and sorption of 554

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sunflower germplasm for phytoremediation of lead- polluted soil and production of
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10 J O U R N A L O F E N V
I R O N M E N T A L S C I E N C E S X X ( X X X X ) X X X

t2:1
t2:23
Table 2 – Total mineral content in seed meal of different sunflower germplasms ⁎.
t2:4 Germplasm Ca (%)
Mg (%) K (%)
Na (%)
t2:5 1 0.131
± 0.015 0.116 ± 0.011
0.540 ± 0.056 0.004 ± 0.001 t2:6 2
0.115 ± 0.010 0.086 ± 0.008
0.416 ± 0.037 0.003 ± 0.001 t2:7 3
0.138 ± 0.016 0.106 ± 0.013
0.470 ± 0.054 0.002 ± 0.001 t2:8 4
0.155 ± 0.026 0.085 ± 0.014
0.487 ± 0.080 0.003 ± 0.001 t2:9 5
0.238 ± 0.021 0.164 ± 0.016
0.513 ± 0.046 0.003 ± 0.001 t2:10 6
0.163 ± 0.014 0.097 ± 0.008
0.450 ± 0.041 0.003 ± 0.001 t2:11 7
0.176 ± 0.013 0.131 ± 0.009
0.449 ± 0.034 0.004 ± 0.001 t2:12 8
0.188 ± 0.022 0.158 ± 0.020
0.529 ± 0.072 0.003 ± 0.001 t2:13 9
0.181 ± 0.029 0.092 ± 0.014
0.520 ± 0.084 0.004 ± 0.001 t2:14 10
0.193 ± 0.014 0.125 ± 0.010
0.492 ± 0.037 0.005 ± 0.001 t2:15 11
0.185 ± 0.029 0.127 ± 0.021
0.522 ± 0.084 0.003 ± 0.001 t2:16 12
0.140 ± 0.007 0.111 ± 0.006
0.386 ± 0.019 0.003 ± 0.001 t2:17 13
0.085 ± 0.006 0.085 ± 0.006
0.338 ± 0.028 0.002 ± 0.001 t2:18 14
0.158 ± 0.033 0.119 ± 0.025
0.357 ± 0.072 0.004 ± 0.001 t2:19 15
0.191 ± 0.027 0.126 ± 0.024
0.341 ± 0.049 0.002 ± 0.001 t2:20 16
0.164 ± 0.021 0.091 ± 0.010
0.383 ± 0.051 0.003 ± 0.001 t2:21 17
0.161 ± 0.017 0.094 ± 0.009
0.405 ± 0.033 0.003 ± 0.001 t2:22 18
0.204 ± 0.022 0.159 ± 0.016
0.749 ± 0.077 0.002 ± 0.001 t2:23 19
0.129 ± 0.017 0.116 ± 0.014
0.638 ± 0.080 0.003 ± 0.001 t2:24 20
0.215 ± 0.041 0.165 ± 0.032
0.988 ± 0.177 0.005 ± 0.002 t2:25 21
0.183 ±
0.014 0.165 ± 0.013
0.913 ± 0.069 0.005 ± 0.002 t2:26 22
0.297 ± 0.022 0.258 ± 0.017
1.00 ± 0.066 0.007 ± 0.001 t2:27 23
0.180 ± 0.031 0.103 ± 0.018
0.533 ± 0.098 0.003 ± 0.001 t2:28 24
0.226 ± 0.029 0.175 ± 0.020
0.660 ± 0.095 0.008 ± 0.003 t2:29 25
0.237 ± 0.048 0.213 ± 0.040
1.402 ± 0.255 0.006 ± 0.002 t2:30 26
0.150 ± 0.003 0.095 ± 0.007
0.859 ± 0.010 0.003 ± 0.001 t2:31 27
0.106 ± 0.013 0.120 ± 0.016
0.567 ± 0.079 0.002 ± 0.001 t2:32 28
0.313 ± 0.035 0.237 ± 0.030
1.297 ± 0.189 0.006 ± 0.002 t2:33 29
0.146 ± 0.005 0.106 ± 0.007
0.549 ± 0.029 0.003 ± 0.001 t2:34 30
0.231 ± 0.007 0.181 ± 0.021
1.034 ± 0.084 0.005 ± 0.002 t2:35 31
0.123 ± 0.003 0.103 ± 0.004
0.643 ± 0.016 0.002 ± 0.001 t2:36 32
0.182 ± 0.033 0.130 ± 0.022
0.548 ± 0.096 0.002 ± 0.001 t2:37 33
0.213 ± 0.006 0.183 ± 0.003
0.735 ± 0.014 0.004 ± 0.001 t2:38 34
0.197 ± 0.009 0.179 ± 0.012
0.836 ± 0.045 0.005 ± 0.001 t2:39 35
0.094 ± 0.007 0.200 ± 0.001
0.578 ± 0.033 0.003 ± 0.001 t2:40 36
0.193 ± 0.020 0.208 ± 0.022
0.795 ± 0.087 0.006 ± 0.001 t2:41 37
0.151 ± 0.012 0.168 ± 0.009
0.721 ± 0.098 0.004 ± 0.001 t2:42 38
0.148 ± 0.011 0.156 ± 0.010
0.664 ± 0.045 0.005 ± 0.002 t2:43 39
0.138 ± 0.004 0.144 ± 0.006
0.685 ± 0.015 0.004 ± 0.001
t2:44 40 0.086 ±
0.010 0.102 ± 0.011
0.451 ± 0.060 0.003 ± 0.001

t2:45

⁎ Values are Mean ± S.E. (n = 3 replicates).

555 metals, and low pH values provide a favorable environment to

556 plants for metal uptake (Aransiola et al., 2013). Our studied
557 sunflower germplasms showed the highest concentration of Pb
558 in shoot, and the concentration pattern was significantly
559 different (p < 0.05) between germplasms. Plant varietal differ-
560 ences in Pb accumulation in shoot have also been reported in
561 different genotypes of fibrous crops (Uddin et al., 2016). The
562 high accumulation of Pb in the harvestable parts of several
563 germplasms showed the easy excess of Pb to plants in Pb-
564 contaminated soil through binding with other ions during
germplasms showed TF > 1 except GP.2: S-606 (Fig. 3a), which 574 indicates that
all germplasms remediate the Pb-contaminated 575 soil by phytoextraction, while
GP.2: S-606 is involved in the 576 phytostabilization of Pb-contaminated soil.
The differences in 577
Transfer factor (TF) value might be affected by soil properties 578
and plant genotype (Adesodun et al., 2010; Pritsa et al., 2008). 579
The soil Pb concentration was measured before planting and 580 after harvesting
sunflower germplasm in field conditions, and 581 significant differences were
observed for MRE (p < 0.05) among 582
them (Fig. 3b). MRE indicates the potential of plants to remove 583
5Q6515
nutrient uptake (Bada and Kalejaiye, 2010). Several sunflower
Pb from soil, and 82.2% to 8.2% of Pb was eliminated from the 584
566 varieties showed permissible concentrations of Pb in the seed
567 portion consistent with their use for livestock feed (Chaney,
568 1999), whereas Pb was reported to exhibit the lowest concen-
569 tration in sunflower seeds in comparison to Cd and Zn
570 (Roongtanakiat and Sanoh, 2015).
soil; it translocated and accumulated in aboveground parts as 585
described by the TF values of the germplasm. Although GP.2: S- 586
606 showed TF < 1, its maximum MRE among all varieties 587 shows that GP.2: S-
606 removes the Pb from soil by retaining it 588 in roots, whereas other varieties
accomplished the removal by 589
571 The ratio of shoot Pb content and root Pb content was
transferring Pb from root to shoot.
590
572 measuredto evaluate thetranslocationof accumulatedPb from
573 rhizosphere parts to above-ground parts. All studied
The evaluation of plant extraction capacity is a basic 591
concern for phytoremediation of metals from polluted soils. 592

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sunflower germplasm for phytoremediation of lead- polluted soil and production of
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J O U R N A L O F E N V I R O N M E N T A L S C I E N C E S X X ( X X X
X ) X X X
11

Fig. 5 – Screening of Pb-accumulator sunflower germplasms in comparison with 15

best germplasms grown on Pb- contaminated soil. Different letters on bars indicate
significant differences (p 0.05) within germplasms (Bonferroni test). Bars
indicate Mean ± S.E.

593 Continuous harvesting of high biomass plants can reduce the

594 levels of soil heavy metals to legally defined values. Several
595 authors have investigated the phytoextraction potential of
596 certain plant species in controlled environmental conditions,
597 but data for phytoextraction under field conditions is still
lacking. Phytoextraction of metals has been assessed by 598 calculation of
different indices such as the bioconcentration 599 factor (BCF) and
bioaccumulation factor (BAF); however, 600
Mertens et al. (2005) proposed the use of the phytoextraction 601
capacity of plants in terms of the metal extraction ratio by 602

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sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
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I R O N M E N T A L S C I E N C E S X X ( X X X X ) X X X

t3:1
t3:23
Table 3 – Fatty acid (FA) profile of Pb-accumulator sunflower germplasm grown on
Pb-contaminated soil. ⁎
t3:4 Serial no. Fatty acid
Formula Retention time (min) Fatty acid
(%)

t3:5 1 Palmitic acid

(16:0) C16H32O2
16.46 6.14 ±
0.02c t3:6 2 Stearic acid
(18:0) C18H36O2
19.68 1.952 ±
0.01d t3:7 3 Oleic acid
(18:1) C18H34O2
20.19 59.522 ±
3.10a t3:8 4 Linoleic acid
(18:2) C18H32O2
23.85 32.199 ±
2.01b t3:9 5 Gadoleic acid
(20:1) C18H32O2
22.85 0.176 ±
0.001d

t3:10

⁎ Values having similar letters represent non-significant difference (P <0.05) of

FAs for germplasm. Values represents Mean ± S.E. (n = 3).

603 expressing the interaction of plant biomass and the unit

604 volume of soil to be cleaned in one harvest. The results of our
605 field study illustrate the metal extraction ratios (41.6%–1.1%)
606 of different sunflower germplasms, with significant differ-
607 ences (p < 0.05) among germplasms (Fig. 3c). The highest MER
608 indicates the best phytoextraction potential of a germplasm
609 in one harvest, and repeated harvesting can promptly clean
content in sunflower oil, which is essential for production 630
quality control as well as from the food safety point of view. 631
Significant differences in Pb concentrations in sunflower oil 632 were observed
among the germplasms, but the level of Pb 633 content in oil was below the food
safety standard in China 634 (GB-2762-2012). These results indicate that the
variation in oil 635
Pb content also depends upon the genotype and the environ- 636
610 up metal-polluted soil. Similar results were also found in six
mental conditions in which Pb is incorporated.
637
611 Chinese cabbage genotypes, whose phytoextraction was
612 evaluated by MER and showed the ability of the genotypes to
613 extract metal in one growth season (Liu et al., 2011).
614 The sunflower seed and oil production are strongly
615 seasonally dependent, and change in the crop cultivation
616 period markedly affects the seed and oil yield (Killi and
617 Altunbay, 2005). The planting season of our field experiment
618 was March and sunflower germplasm showed the best yield
619 components, although it varied among species (Fig. 1b and 4
620 a1). A great reduction in sunflower seed yield and oil content
621 with change in cultivation time was also reported by several
622 authors in previous studies (Barros et al., 2004; de la Vega and
623 Hall, 2002). Secondly, sunflower oil is generally consumed by
624 the human population due to its high nutritive value,
Sunflower seed meal, a byproduct obtained after oil extrac- 638 tion, is a rich
source of proteinand fiber and a valuable source of 639 feed for poultry and
livestock (Akande, 2011; Oliveira et al., 6Q4106
2016). The presence of metals has been reported in meal 641 products of
oilseed crops, and scientists believe that the end- 642 products of high-
accumulator crops should be disposed of 643 properly (Barbaroux et al., 2009;
Hussain et al., 2013). Yang et al. 644 (2016) assessed Pb and Cd in the oil and
meal of peanut and 645 oilseed rape grown on highly Pb-contaminated soil, and
found 646
<0.1 mg/kg Pb in oil and <5 mg/kg Pb in meal products of both 647 plant species.
Information related to metal content in sun- 648 flower meal is still lacking.
Our experimental site was moder- 649 ately Pb-contaminated, and the meal Pb
contents in the studied 650
sunflower germplasms were under the permissible limit of the 651
625 antioxidant properties and unsaturated fatty acids. Besides
Hygienic Standard for Feeds (GB13078-1991) (Yang et al., 2016).
652
626 this, metal ions catalyze the decomposition of hydroperoxides
627 in oil, which leads to the formation of undesirable substances
628 that ultimately alter the oxidative stability of edible oil
629 (Pehlivan et al., 2008). Therefore, we determined the Pb
The quality and quantity of protein are very important 653 aspects for animal
feed. The meal total protein contents in 654 different sunflower germplasms are
given in Fig. 4 b1, which 655 indicate significant differences in meal
nutritive values 656

t4:1
t4:23
Table 4 – Chromatograph and amino acid (AA) profile of high-accumulator sunflower
meal grown on Pb-contaminated soil ⁎.
t4:4 Serial no. Amino acid Formula
Retention time (min) Concentration (ng/20 μL) Amino acid (%)

t4:5 1 Aspartic acid C4H7NO4

4.960 1453.845
10.74 ± 0.03b t4:6 2 Threonine
C4H9NO3 5.607
597.657 4.41 ± 0.021d t4:7
3 Serine C3H7NO3
6.227 686.196
5.06 ± 0.015d t4:8 4 Glutamic acid
C5H9NO4 6.927
3032.72 22.40 ± 0.05a t4:9
5 Glycine C2H5NO2
9.913 805.158
5.94 ± 0.014d t4:10 6 Alanine
C3H7NO2 10.773
707.407 5.22 ± 0.011d t4:11
7 Cysteine C3H7NO2S
12.160 184.491
1.36 ± 0.01d t4:12 8 Valine
C5H11NO2 12.707
694.04 5.12 ± 0.02d t4:13
9 Methionine C2H11NO2S
13.900 276.126
2.03 ± 0.01d t4:14 11 Isoleucine
C6H13NO2 16.087
598.112 4.41 ± 0.01d t4:15
12 Leucine C6H13NO2
17.187 997.438
7.36 ± 0.02c t4:16 13 Tyrosine
C9H11NO3 18.560
355.373 2.62 ± 0.01d t4:17
14 Phenylalanine C9H11NO2
19.633 715.304
5.28 ± 0.02d t4:18 15 Lysine
C6H14N2O2 22.247
534.178 3.94 ± 0.01d t4:19
16 Histidine C6H9N3O2
24.520 326.783
2.41 ± 0.01d t4:20 17 Arginine
C6H14N4O2 28.633
1144.784 8.45 ± 0.03c t4:21
18 Proline C5H9NO2
7.52 426.526
3.15 ± 0.01d

t4:22

⁎ Values having similar letters represent non-significant differences (P ˂0.05) of

AAs for germplasm. Values represents Mean ± S.E. (n = 3).

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sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
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X ) X X X
13

657 among germplasms. Among non-essential amino acids

658 (NEAAs), glutamic acid was found to have the maximum
659 percentage (22.4%) and cysteine the minimum (2.6%), similar
660 to the findings of Ingale and Shrivastava (2011), who found
661 that the glutamic acid content was the highest in the two
662 sunflower germplasms LSF-8 and LSF-11 in a non-

Acknowledgments 71

This work was supported by Zhejiang Science and Technology 713

Bureau (No. 2018C02029), the National Natural Science Foun- 714
dation of China (Nos. 41721001 and 31872956), and the 715
663 contaminated medium; whereas, among essential amino
664 acids leucine had the highest percentage (7.3%) and cysteine
665 the lowest (1.3%) (Table 3). Leucine is involved in protein
666 synthesis and inhibits protein degradation in the catabolic
667 process. Increases in muscular protein synthesis in pigs and
Fundamental Research Funds for the Central Universities.

Appendix A. Supplementary data

7Q1169

7178
668 the meat yield in chickens have been observed after oral
Supplementary data to this article can be found online at 719
669 application of leucine (Erwan et al., 2009; Escobar et al., 2006).
670 Although the naturally occurring amino acid glutamate/
671 glutamic acid serves as an energy source and is a precursor
672 of glutathione, several scientists have stated that glutamate is
673 the main component of meat taste (Fujimura and Kadowaki,
https://doi.org/10.1016/j.jes.2019.05.031.

R E F E R E N C E S
720

721
6Q7417
2006; Kurihara, 2009). Watanabe et al. (2017) reported that low
Adesodun, J.K., Atayese, M.O., Agbaje, T., Adesodun, J.K., Atayese, 722
675 levels of lysine in poultry feed increase the free glutamic acid
676 content and enhance the taste of chicken meat. These
677 findings suggested that application of phytoremediation to
678 metal-contaminated soil must emphasize certain factors (soil,
679 environment, plant species, and cultivation period) with
680 economically feasible agriculture production, and screening
681 of different sunflower germplasms in moderately Pb-
682 contaminated soil provides a promising option for identifying
683 the most suitable cultivar for phytoremediation accompanied
M.O., Agbaje, T.A., et al., 2010. Phytoremediation potentials of sunflowers
(Tithonia diversifolia and Helianthus annuus) for metals in soils contaminated
with zinc and lead nitrates. Water. Air. Soil. Poll. 207 (4), 195–201.
Akande, K., 2011. Proximate and amino acid analyses of full-fat sunflower
(Helianthus annuus L.) seed meal. Sing. J. Sci. Res. 1 (2), 179–183.
Alake, C.O., Ayo-Vaughan, M.A., Ariyo, 2015. Selection criteria for grain yield
and stability in bambara groundnut (Vigna subter- ranean (L.) Verdc) landraces.
Acta. Agr. Sca 65 (5), 433–447.
724
725
726
727
728
729
730
731
732
733
684 with bio-agro production.
Alamgir, M., 2016. The effects of soil properties to the extent of soil 734

6856 4. Conclusions

687 In this study, sunflower GP.8: 8585 has been recognized as a

688 potential germplasm source for recovery of moderately lead-
689 polluted soil. Our results revealed that the said germplasm
690 grown in contaminated soil produced considerable amounts
691 of biomass, which correlated and was directly proportional to
692 lead uptake. Furthermore, GP.8: 8585 produced significant
contamination with metals. Environmental Remediation Technologies for Metal-
Contaminated Soils. Springer, Tokyo, Japan, pp. 1–19.
Alexander, J., 1998. Immunonutrition: the role of ω-3 fatty acids.
Nutrition. 14 (7–8), 627–633.
Aransiola, S.A., Ijah, U.J.J., Abioye, O.P., 2013. Phytoremediation of lead
polluted soil by Glycine max L. Appl. Environ. Soil Sci. 2013,
1–7.
Bada, B.S., Kalejaiye, S.T., 2010. Response of kenaf (Hibiscus Cannabinus L.)
grown in different soil textures and lead concentrations. Res. J. Agr. Bio. Sci.
6 (5), 659–664.
Barbaroux, R., Meunier, N., Mercier, G., Taillard, V., Morel, J.,
735
736
737
738
739
740
741
742
743
744
745
746
693 grain yield with oil lead concentrations below the standard
Simonnot, M., et al., 2009. Chemical leaching of nickel from the 747
694 limit for human consumption. Seed meal is also an important
695 component of agro-products, and analysis of the meal
seeds of the metal hyperaccumulator plant Alyssum murale. Hydrometallurgy 100 (2),
10–14.
748
749
696 showed high protein and nutrient values, with all the
Barros, J.F., de Carvalho, M., Basch, G., 2004. Response of sunflower 750
697 essential amino acids necessary for ruminant and non-
698 ruminant feed. Moreover, the metal content in seed meal
699 was also under the permissible limit. Therefore, the above-
700 mentioned sunflower germplasm can be used for
701 phytoremediation of lead-contaminated soils, along with
702 valuable agro-economic production to generate economic
703 revenue for farmers during the remediation process. More-
704 over, further research will focus on advanced and precise
705 agronomic practices for optimizing the field evaluation of
706 high-accumulator germplasms for a positive contribution
707 towards the environment and human welfare.
(Helianthus annuus L.) to sowing date and plant density under
Mediterranean conditions. Eur. J. Agr. 21 (3), 347–356. Bellinger, D.C., 2008. Very
low lead exposures and children's neurodevelopment. Cur. Opi. Pediatrics. 20 (2),
72–177.
Carter, M.R., Gregorich, E.G., 2008. Soil Sampling and Methods of
Analysis. 2nd Ed. CRC Press, Boca Raton FL.
Chaney, R.L., 1999. Improving metal hyperaccumulator wild plants to develop
commercial phytoextraction systems. Ap- proaches and progress. Phytoremediation of
Contaminated Soil and Water. CRC Press LLC, Boca Raton, FL, pp. 130–158.
Chaturvedi, R., Favas, P., Pratas, J., Varun, M., Paul, M.S., 2018.
Assessment of edibility and effect of arbuscular mycorrhizal fungi on Solanum
melongena L. grown under heavy metal (loid) contaminated soil. Ecotoxicol.
Environ. Saf. 148, 318–326.
Chen, H.M., Zheng, C.R., Tu, C., Zhu, Y.G., 1999. Heavy metal
pollution in soils in China: status and countermeasures.
751
752
753
754
755
756
757
758
759
760
761
762
763
764
765
766
7Q08918
Uncited reference
J. Human Environ. 28 (2), 130–134.
Cheng, S.F., Huang, C.Y., Lin, Y.C., Lin, S.C., Chen, K.L., 2015.
767
768
710 De Morais et al., 2016
Phytoremediation of lead using corn in contaminated
769

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
https://doi.org/10.1016/j.jes.2019.05.031
14 J O U R N A L O F E N V
I R O N M E N T A L S C I E N C E S X X ( X X X X ) X X X

770 agricultural land—an in situ study and benefit assessment.

771 Ecotoxicol. Environ. Saf. 111, 72–77.
772 Cindric, I.J., Zeiner, M., Steffan, I., 2007. Trace elemental charac-
773 terization of edible oils by ICP–AES and GFAAS. Microchem. J.
Kurihara, K., 2009. Glutamate: from discovery as a food flavor to role as a basic
taste (umami). The. Am. J. Clinical. Nutr. 90 (3),
719–722.
Lambert, E.V., Goedecke, J.H., Bluett, K., Heggie, K., Claassen, A.,
838
839
840
841
774 85 (1), 136–139.
Rae, D.E., et al., 2007. Conjugated linoleic acid versus high-oleic 842
775 De La Vega, A.J., Hall, A.J., 2002. Effects of planting date, genotype,
776 and their interactions on sunflower yield. Crop Sci. 42 (4),
777 1191–1201.
acid sunflower oil: effects on energy metabolism, glucose tolerance, blood lipids,
appetite and body composition in regularly exercising individuals. Brit. J. Nut. 97
(5), 1001–1011.
843
844
845
778 De Morais, O.V.R., De Arruda, A.M.V., Silva, L.N.S., De Souza, J.B.F.,
Lei, K., Giubilato, E., Critto, A., Pan, H., Lin, C., 2016. Contamination 846
779 De Queiroz, J.P.A.W., Da Silva, M.A., et al., 2016. Sunflower meal
780 as a nutritional and economically viable substitute for soybean
781 meal in diets for free-range laying hens. Ani. Fee. Sci. Tec. 220,
and human health risk of lead in soils around lead/zinc smelting areas in China.
Environ. Sci. Poll. Res. 23 (12),
13128–13136.
847
848
849
782 103–108.
Liu, W., Zhou, Q., Zhang, Z., Hua, T., 2011. Evaluation of cadmium 850
783 Diwan, H., Ahmad, A., Iqbal, M., 2008. Genotypic variation in the
784 phytoremediation potential of Indian mustard for chromium.
785 Environ. Manag. 41 (5), 734–741.
786 Duruibe, J.O., Ogwuegbu, M.O.C., Egwurugwu, J.N., 2007. Heavy
787 metal pollution and human biotoxic effects. Int. J. Phy. Sci. 2
788 (5), 112–118.
789 Erwan, E., Alimon, A., Sazili, A., Yaakub, H., Hilmi, M., 2009. Effect
790 of L-leucine supplementation on growth performance and
791 carcass characteristics of grower-broiler chickens fed low
792 protein diets. Am. J. Ani. Vet. Sci. 4 (4), 95–100.
793 Escobar, J., Frank, J.W., Suryawan, A., Nguyen, H.V., Kimball, S.R.,
phytoremediation potential in Chinese cabbage cultivars. J. Agr. Food. Chem. 59
(15), 8324–8330.
Lombi, E., Zhao, F., Dunham, S., McGrath, S.P., 2001.
Phytoremediation of heavy metal–contaminated soils. J. Environ. Qual. 30 (6), 1919–
1926.
Lopez-Beceiro, J., Artiaga, R., Gracia, C., Tarrio-Saavedra, J., Naya, S., Mier,
J.T., 2011. Comparison of olive, corn, soybean and sunflower oils by PDSC. J.
Ther. Anal. Calori 104 (1), 169–175.
Lv, S., Yang, B., Kou, Y., Zeng, J., Wang, R., Xiao, Y., et al., 2018.
Assessing the difference of tolerance and phytoremediation potential in mercury
contaminated soil of a non-food energy
851
852
853
854
855
856
857
858
859
860
861
794 Jefferson, L.S., et al., 2006. Regulation of cardiac and skeletal
crop, Helianthus tuberosus L. (Jerusalem artichoke). Peer. J 6, 2–18. 862
795 muscle protein synthesis by individual branched-chain amino
Lynch, J.M., Barbano, D.M., 1999. Kjeldahl nitrogen analysis as a
863
796 acids in neonatal pigs. Am. J. Phy. End. Met. 290 (4), 612–621.
reference method for protein determination in dairy products. 864
797 Figueroa, J., Lewis, A., Miller, P., Fischer, R., Diedrichsen, R.M.,
798 2003. Growth, carcass traits, and plasma amino acid concen-
799 trations of gilts fed low-protein diets supplemented with
800 amino acids including histidine, isoleucine, and valine. J. Ani.
801 Sci. 81 (6), 1529–1537.
J. AOAC Int. 82 (6), 1389–1398.
Madejón, P., Murillo, J.M., Marañón, T., Cabrera, F., Soriano, M.A.,
2003. Trace element and nutrient accumulation in sunflower plants two years after
the Aznalcóllar mine spill. Sci. Total Environ. 307 (1–3), 239–257.
865
866
867
868
869
802 Fujimura, S., Kadowaki, M., 2006. Improvement of meat taste by
Malar, S., Favas, P.J.C., Sahi, S.V., Venkatachalam, P., 2014. Effect of 870
803 dietary components. Bull. Fac. Agric. Nigata. Univ. 58, 151–153.
804 Gall, J.E., Boyd, R.S., Rajakaruna, N., 2015. Transfer of heavy metals
lead on phytotoxicity, growth, biochemical alterations and its role on genomic
template stability in Sesbania randiflora: a
871
872
805 through terrestrial food webs: a review. Environ. Mont. Assess.
potential plant for phytoremediation. Ecotoxicol. Environ. Saf. 873
806 187 (4), 201–214.
807 Ganzler, K., Salgó, A., Valkó, K., 1986. Microwave extraction: a
808 novel sample preparation method for chromatography.
809 J. Chromatogr 371, 299–306.
810 García-Alix, A., Jiménez-Espejo, F.J., Lozano, J.A., Jiménez-Moreno,
811 G., Martinez-ruiz, F., Sanjuan, L.G., et al., 2013. Anthropogenic
812 impact and lead pollution throughout the Holocene in South-
813 ern Iberia. Sci. Total Environ. 449, 451–460.
814 González-Pérez, S., Vereijken, J.M., 2007. Sunflower proteins:
815 overview of their physicochemical, structural and functional
816 properties. J. Sci. Food. Agri. 87 (12), 2173–2191.
817 Huo, X., Peng, L., Xu, X., Zheng, L., Qiu, B., Qi, Z., et al., 2007.
818 Elevated blood lead levels of children in Guiyu, an electronic
819 waste recycling town in China. Environ. Health. Per. 115 (7),
820 1113–1117.
108, 249–257.
Matsunami, M., Matsunami, T., Kon, K., Ogawa, A., Kodama, I., Kokubun, M., 2013.
Genotypic variation in nitrogen uptake during early growth among rice cultivars
under different soil moisture regimes. Plant. Prod. Sci. 16 (3), 238–246.
Mertens, J., Luyssaert, S., Verheyen, K., 2005. Use and abuse of trace metal
concentrations in plant tissue for biomonitoring and phytoextraction. Environ. Pol.
138 (1), 1–4.
Meyer, P.A., Brown, M.J., Falk, H., 2008. Global approach to reducing lead
exposure and poisoning. Mutation. Res. Funda- mental. Mol. Mec. Mut 659 (1–2),
166–175.
Mielke, H.W., Laidlaw, M.A., Gonzales, C., 2010. Lead (Pb) legacy from vehicle
traffic in eight California urbanized areas: continuing influence of lead dust on
children's health. Sci. Total Environ. 408 (19), 3965–3975.
874
875
876
877
878
879
880
881
882
883
884
885
886
887
888
821 Hussain, A., Alamzeb, S., Begum, S., 2013. Accumulation of heavy
Morris, G.P., Hu, Z., Grabowski, P.P., Borevitz, J.O., De Graaff, M.A., 889
822 metals in edible parts of vegetables irrigated with waste water
823 and their daily intake to adults and children, District Mardan,
824 Pakistan. Food Chem. 136 (3), 1515–1523.
825 Ingale, S., Shrivastava, S.J., 2011. Chemical studies of new varieties
826 of sunflower (Helianthus annuus) LSF-11 and LSF-8 seeds. Agri.
827 Bio. J. North Am. 2, 171–1181.
828 Ivanova, P., Chalova, V., Koleva, L., Pishtiyski, I., 2013. Amino acid
829 composition and solubility of proteins isolated from sunflower
830 meal produced in Bulgaria. Int. Food Res. J. 20 (6), 2995–3000.
831 Jadia, C.D., Fulekar, M.H., 2008. Phytoremediation: the application
832 of vermicompost to remove zinc, cadmium, copper, nickel and
833 lead by sunflower plant. Environ. Eng. Man. J. 7 (5), 547–558.
834 Killi, F., Altunbay, S.G., 2005. Seed yield, oil content and yield
835 components of confection and oilseed sunflower (Helianthus
836 annuus L.) cultivars planted in different dates. Int. J. Agri. Biol.
837 7, 21–24.
Miller, R., et al., 2016. Genotypic diversity effects on biomass production in
native perennial bioenergy cropping systems. Global Change Biol. Bioenergy. 8 (5),
1000–1014.
Nasopoulou, C., Stamatakis, G., Demopoulos, C.A., Zabetakis, I.,
2011. Effects of olive pomace and olive pomace oil on growth performance, fatty
acid composition and cardio protective properties of gilthead sea bream (Sparus
aurata) and sea bass (Dicentrarchus labrax). Food Chem. 129 (3), 1108–1113.
Peer, W.A., Freeman, J.L., Murphy, A.S., 2005. Phytoremediation and
Hyperaccumulator Plants, Molecular Biology of Metal Homeostasis and Detoxification.
Topics in Current Genetics. Springer, Berlin, Heidlberg, pp. 299–340.
Pehlivan, E., Arslan, G., Gode, F., Altun, T., Özcan, M.M., 2008.
Determination of some inorganic metals in edible vegetable oils by inductively
coupled plasma atomic emission spectros- copy (ICP-AES). Grasas Aceites 59 (3),
239–244.
890
891
892
893
894
895
896
897
898
899
900
901
902
903
904
905

Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
https://doi.org/10.1016/j.jes.2019.05.031
J O U R N A L O F E N V I R O N M E N T A L S C I E N C E S X X ( X X X
X ) X X X
15

906 Pratas, J., Favas, P.J., D'Souza, R., Varun, M., Paul, M.S., 2013.
907 Phytoremedial assessment of flora tolerant to heavy metals in
908 the contaminated soils of an abandoned Pb mine in Central
909 Portugal. Chemosphere. 90 (8), 2216–2225.
910 Pritsa, T.S., Fotiadis, E.A., Lolas, P.C., 2008. Corn tolerance to
911 atrazine and cadmium and sunflower to cadmium in soil and
912 hydroponic culture. Comm. Soil. Sci. Plant. Ana. 39 (7–8),
913 1168–1182.
914 Quiles, J.L., Ramırez-Tortosa, M.C., Gómez, J.A., Huertas, J.R.,
microbiotas in broiler chickens across various feeding trials. Appl. Env. Mic. 77
(17), 5868–5878.
Uddin, M.N., Wahid-Uz-Zaman, M., Rahman, M.M., Islam, M.S., Islam, M.S., 2016.
Phytoremediation potentiality of lead from contaminated soils by fibrous crop
varieties. Am. J. App. Sci. Res. 2 (5), 22–28.
Vamerali, T., Bandiera, M., Mosca, G., 2010. Field crops for phytoremediation of
metal-contaminated land. A review. Environ. Chem. Lett. 8 (1), 1–17.
948
949
950
951
952
953
954
955
956
915 Matiax, J., 2002. Role of vitamin E and phenolic compounds in
Venkatachalam, P., Jayaraj, M., Manikandan, R., Geetha, N., Rene, 957
916 the antioxidant capacity, measured by ESR, of virgin olive,
917 olive and sunflower oils after frying. Food Chem. 76 (4),
918 461–468.
919 Rele, A.S., Mohile, R., 2003. Effect of mineral oil, sunflower oil, and
920 coconut oil on prevention of hair damage. J. Cos. Sci 54 (2),
E.R., Sharma, N.C., et al., 2016. Zinc oxide nanoparticles (ZnONPs) alleviate
heavy metal induced toxicity in Leucaena leucocephala seedlings: a physiochemical
analysis. Plant Phys- iol. Biochem. 110, 59–69.
Wang, H.L., Chen, X.T., Yang, B., Ma, F.L., Wang, S., Tang, M.L., et
958
959
960
961
962
921 175–192.
al., 2008. Case–control study of blood lead levels and attention 963
922 Roongtanakiat, N., Sanoh, S., 2015. Comparative growth and
923 distribution of Zn, Cd and Pb in rice, vetiver and sunflower
924 grown in contaminated soils. Kasetsart J. (Nat. Sci.) 49 (5),
925 663–675.
deficit hyperactivity disorder in Chinese children. Environ. Health Perspect. 116
(10), 1401–1406.
Watanabe, G., Kobayashi, H., Shibata, M., Kubota, M., Kadowaki, M., Fujimora, S.,
2017. Reduction of dietary lysine increases
964
965
966
967
926 Senkoylu, N., Dale, N., 1999. Sunflower meal in poultry diets: a
free glutamate content in chicken meat and improves its taste. 968
927 review. World's Poultry Sci. J 55, 153–174.
Anim. Sci. J. 88 (2), 300–305.
969
928 Soudek, P., Petrová, Š., Vaňková, R., Song, J., 2014. Accumulation of
Wei, B., Yang, L., 2010. A review of heavy metal contaminations in 970
929 heavy metals using Sorghum sp. Chemosphere. 104, 15–24.
930 Su, C., Jiang, L.Q., Jun, W., 2014. A review on heavy metal
931 contamination in the soil worldwide: situation, impact and
932 remediation techniques. Env. Skeptics. Critics. 3 (2), 24–38.
933 Sytar, O., Brestic, M., Taran, N., Zivcak, M., 2015. Plants used for
934 biomonitoring and phytoremediation of trace elements in soil
urban soils, urban road dusts and agricultural soils from
China. Micro. J. 94 (2), 99–107.
Wong, J., 2004. Phytoremediation of contaminated soils. J. Nat.
Res. Life. Sci. Edu. 33, 51.
Wuana, R., Okieimen, F.E., 2010. Phytoremediation potential of maize (Zea mays
L.). a review. Afr. J. Gen. Agr. 6 (4), 275–287.
971
972
973
974
975
976
935 and water. Plant Metal Interaction: Emerging Remediation
Yang, Y., Li, H., Peng, L., Chen, Z., Zeng, Q., 2016. Assessment of Pb 977
936 Techniques. Elsevier, Amsterdam, pp. 61–384.
937 Tang, L., Hamid, Y., Zehra, A., Sahito, Z.A., He, Z., Hussain, B.,
and Cd in seed oils and meals and methodology of their extraction. Food Chem.
197, 482–488.
978
979
938 Yang, X., 2019. Characterization of fava bean (Vicia faba L.)
Yildiz, E., 2015. Comparison of rations for dairy cows with soybean 980
939 genotypes for phytoremediation of cadmium and lead co-
940 contaminated soils coupled with agro-production. Ecotoxicol.
941 Environ. Saf. 171, 190–198.
942 Tong, S., Schirnding, Y.E.V., Prapamontol, T., 2000. Environmental
943 lead exposure: a public health problem of global dimensions.
944 Bull. World Health Organ. 78 (9), 1068–1077.
945 Torok, V.A., Hughes, R.J., Mikkelsen, L.L., Perez-Maldonado, R.,
946 Balding, K., MacAlpine, R., et al., 2011. Identification and
947 characterization of potential performance-related gut

990
meal or with rapeseed meal in which the main source of protein is sunflower meal.
Bulg. J. Agr. Sci. 21 (3), 662–667.
Zhang, W., Chen, L., Zhang, R., Lin, K., 2015. Effects of decabromodiphenyl ether
on lead mobility and microbial toxicity in soil. Chemosphere 122, 99–104.
Zhong, T., Xue, D., Zhao, L., Zhang, X., 2018. Concentration of heavy metals in
vegetables and potential health risk assess- ment in China. Environ. Geochem.
Health 40 (1), 313–322.
981
982
983
984
985
986
987
988
989
Please cite this article as: A. Zehra, Z.A. Sahito, W. Tong, et al., Assessment of
sunflower germplasm for phytoremediation of lead- polluted soil and production of
seed ..., Journal of Environmental Sciences,
https://doi.org/10.1016/j.jes.2019.05.031