Download - Apresentação em Paris, julho de 2013
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UNEB PPHI
Germinative metabolism of CaatingaGerminative metabolism of Caatingasavannah forest species in biosaline
agriculture
Barbara F. Dantas,Renata C. Ribeiro, Janete R. Matias, Gherman G.L. Araújo
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CAATINGACAATINGA
� 11% brazilian territory;
� 70% Notheastern Region;
� 1300 counties; � 1300 counties;
� 63% of northeastern population.
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CAATINGACAATINGA
�Exclusively brazilian;
�Rich in species and endemism;
�Highly degraded
� less than 50% original area
�Less studied ecossystem
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CAATINGACAATINGA� Semi-arid climate
� Low and poorly distributed rainfall
(250 to 800 mm/ year)
� 2012- 107,2 mm
� Rainy season – november- april;� Rainy season – november- april;
� Dry season – may- october;
� Temperatures – 12oC a 40oC;
� Today- min 20oC- max 26oC
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Dantas, B.F. Dantas, B.F.
Rainy season Dry season
Kiill, L.H.P. Kiill, L.H.P.
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CAATINGACAATINGAWATER RESORUCES
� São Francisco River
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CAATINGACAATINGA
WATER RESORUCES
� Brackish groundwater
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CAATINGACAATINGAWATER RESORUCES
� Fish farming wastewater
� Ponds filled with brackish groundwater
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CanCan caatinga caatinga forestforest speciesspecies germinategerminate andand growgrow
in in thesethese saline saline conditionsconditions????
WhatWhat is is thethe seedseed metabolismmetabolism comparedcompared withwith no no WhatWhat is is thethe seedseed metabolismmetabolism comparedcompared withwith no no
stress stress conditioncondition??
CanCan seedlingsseedlings bebe producedproduced in in biosalinebiosaline
agricultureagriculture??
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Caatinga Caatinga nativenative forestforest speciesspecies
Pereiro (Aspidosperma pyrifolium Mart.)Apocynaceae
•Ornamental tree;
•Bark- farmaceutical;
•Seeds- craftwork
Photos: J.E.B. Silva; R.C. Ribeiro
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Angico (Anadenanthera macrocarpa (Benth.) Brenan)Mimosaceae
•Widely distributed in brazillan territory;
•Farmaceutical,
•Ornamental,
•Wood for coal and construction;
•Forage
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Catingueira-verdadeira (Poincianella pyramidalis [Tul.] L.P.Queiroz)Leguminosae
Endemic from Caatinga
Pioneer species
Forage
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Mulungu (Erythrina velutina Willd.)Leguminosae
Beautiful tree
Ornamental, farmaceutical, craftwork
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Aroeira (Myracrodruon urundeuva (Fr. All.))Anacardiaceae
•Big tree
•Great wood for construction and furniture
•Bark- Farmaceutical, tannin
•Widely distributed in Brazil
•Slow growth
•Endangered species•Endangered species
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Evaluate the possibility of production of
Caatinga tree seedlings in biosaline agriculture,
through the study of the germinative
ObjectiveObjective
through the study of the germinative
metabolism of the seeds subjected to high
electrical conductivities (EC) due to the use of
salted and/or biosaline water
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Material and methodsMaterial and methodsSpeciesSpecies studiedstudied
Anadenanthera macrocarpa (Benth.) Brenan,
Aspidosperma pyrifolium (Mart.),
Erythrina velutina (Willd.),
Myracrodruon urundeuva (Fr. All.)Myracrodruon urundeuva (Fr. All.)
Poincianella pyramidalis (Tul.) L.P. Queiroz.
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Material and methodsMaterial and methodsSeedSeed harvestharvest
10 mother plants
September to November 2012
Jutaí, Lagoa Grande-PE
(8o38’45,23” S, 40o14’07,32” W)
Juremal, Juazeiro-BA
(9o43’51,12” S, 40o21’02,52” W)
Sun-dried, processed and
stored in a dry chamber at
10 oC
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-Biosaline water- collected from ponds
Material and methodsMaterial and methodsSaltSalt stressstress screenningscreenning
Germination in NaCl solutions with EC 0-18 dS.m-1
Germination in Germination in biosalinebiosaline waterwater
-Biosaline water- collected from ponds
supplied with brackish groundwater and
used for raising Thai tilapia
Photo: RIBEIRO, R.C , 2012
-Concentrations of BW: 0, 33,
50, 67, 100% in tap water
4, 09 dSm-1
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Material and methodsMaterial and methodsGerminationGermination teststests
�Paper rolls soaked in solutions (NaCl or BW), 2,5x paper weight
�BOD germinators at 25°C / 12-h photoperiod for 15 days
Germination vGermination variables accessedariables accessedGermination vGermination variables accessedariables accessed
�Germination percentage- G%
�Germination mean time (Labouriau, 1983) – GMT
�Germination speed index (Maguire, 1962)- GSI
�Germination uniformity coefficient (Heydecker, 1973)- GUC
Shoots and roots of germinated seeds were freezed until biochemical
assays.
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Material and methodsMaterial and methodsGerminativeGerminative metabolismmetabolism assaysassays
�Total soluble carbohydrates (TSC) –Morris (1948); Yemm &Willis
(1954);
�Reducing sugars (RS) – Miller (1959);
�Total soluble protein (TSP) – Bradford (1976);�Total soluble protein (TSP) – Bradford (1976);
�Proline content- Bates (1973);
�Ascorbate peroxidase (APX, EC 1.11.1.11) - Nakano &Asada (1981);
�Catalase (CAT,EC 1.11.1.6 )- Azevedo et al. (1998)
�Glutathione-S-transferase (GST, EC 2.5.1.18)- GST assay kit Sigma®
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SaltSalt stress stress screenningscreenning
GerminationGermination
Myracrodruon urundeuva
0
10
20
30
40
50
60
70
80
90
100
0
10
20
30
40
50
60
70
80
90
100
Erythrina velutina Poincianella pyramidalis
Ger
min
ati
on
(%
)
Silva et al. 2008
0
0 2 4 6 8 10 12 14 16 18
0
0 2 4 6 8 10 12 14 16 18 20 22
0
10
20
30
40
50
60
70
80
90
100
0 2 4 6 8 10 12 14 16 18
Aspidosperma pyrifolium
0
10
20
30
40
50
60
70
80
90
100
0 2 4 6 8 10 12 14 16 18 20
Anadenanthera macrocarpa
Ger
min
ati
on
(%
)
Electrical conductivity (dS.m-1)
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0
1
2
3
4
5
6
7
0
1
2
3
4
5
6
7
Erythrina velutina Poincianella pyramidalis
Ger
min
ati
on
mea
n t
ime
(day
s)
SaltSalt stress stress screenningscreenning
GerminationGermination kineticskinetics
Myracrodruon urundeuva
0
1
2
3
4
5
6
7
0 2 4 6 8 10 12 14 16 181
2
3
4
5
6
7
0 2 4 6 8 10 12 14 16 18 20
Aspidosperma pyrifolium Anadenanthera macrocarpa
Electrical conductivity (dS.m-1)
0
0 2 4 6 8 10 12 14 16 18
0
0 2 4 6 8 10 12 14 16 18 20 22
Ger
min
ati
on
mea
n t
ime
(day
s)
Silva et al. 2008
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Germination in Germination in biosalinebiosaline water concentrationswater concentrations
0
10
20
30
40
50
60
70
80
90
100
0 33 50 67 100
Germ
ina
tio
n (
%)
a
0
1
2
3
4
5
6
7
0 33 50 67 100
Germ
ina
tio
n m
ea
n t
ime
b
Aspidosperma pyrifoliumAnadenanthera macrocarpa
Erythrina velutina Myracrodruon urundeuva
0
10
20
30
40
50
60
70
80
90
100
0 20 40 60 80 100
Ge
rm
ina
tio
n (
%)
0
1
2
3
4
5
6
7
0 20 40 60 80 100
Ge
rm
ina
tio
n m
ea
n t
ime
(d
ay
s)
80
90
100a
6
7
Ger
min
ati
on
mea
n t
ime
(da
ys)
b
90
100a
7b
Poincianella pyramidalis
0
10
20
30
40
50
60
70
80
0 20 40 60 80 100
Ger
min
ati
on
(%
)
0
1
2
3
4
5
0 20 40 60 80 100
Ger
min
ati
on
mea
n t
ime
(da
ys)
0
10
20
30
40
50
60
70
80
90
0 20 40 60 80 100
Germ
ina
tio
n (
%)
0
1
2
3
4
5
6
0 20 40 60 80 100
Germ
ina
tio
n m
ea
n t
ime
0
10
20
30
40
50
60
70
80
90
100
0 33 50 67 100
Ger
min
ati
on
(%
)
a
0
1
2
3
4
5
6
7
0 33 50 67 100
Ger
min
ati
on
mea
n t
ime
b
Biosaline water (% in H2O distilled)
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Seedlings Seedlings biomoleculesbiomolecules-- EC variationEC variation
Anadenanthera macrocarpa- 4 days
0
200
400
600
800
1000
1200
1400
1600
TP
(µ
g/g
-1)
Shoot Roota
0
10
20
30
40
50
60
70
80
RS
(µ
mo
l.g-1
)
bTotal proteins
Reducing sugars
Different biochemical responses
Erythrina velutina- 4 days
Electrical conductivity (dSm-1)
0
0 2 4 6 8 10 12 14 16 18
0
0 2 4 6 8 10 12 14 16 18
0
5
10
15
20
25
30
35
0 2 4 6 8 10 12
TP
((µ
mol
g-1
)
a
0
20
40
60
80
100
120
140
0 2 4 6 8 10 12
RS
(µm
ol
g-1
)
b
c
Total proteinsReducing sugars
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Seed reserve degradationSeed reserve degradation-- BSW concentrationBSW concentration
Aspidosperma pyrifolium – 4 day seedlings
Total proteins Reducing sugars
No visible pattern to
biochemical responses
Biosaline water (% in H2O distilled)
0
2000
4000
6000
8000
10000
12000
0 33 50 67 100
TP
(µg
.g-1
)
leaves hypocotyl-roota
0
20
40
60
80
100
120
140
160
180
0 33 50 67 100
TS
S(µ
mo
l.g
-1)
b
Total proteins Reducing sugars
Myracrodruon urundeuva
biochemical responses
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ProlineProline contentcontent-- EC variationEC variationAnadenanthera macrocarpa
10
20
30
40
50
60
70
Pro
lin
e (
mm
ol
.g -
1)
d
Aspidosperma pyrifolium
12
0
300
600
900
1200
1500
1800
2100
2400
2700
0 2 4 6 8 10 12
Pro
lin
e (µ
g g
-1)
d
0
0 2 4 6 8 10 12 14 16 18
Erythrina velutina
Electrical conductivity (dSm-1)
Different biochemical responses
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AOX enzymes AOX enzymes -- EC variationEC variation
0
500
1000
1500
2000
2500
3000
3500
4000
4500
5000
Asc
orb
ato
per
oxid
ase
(η
mo
l AsA
.min
-1.m
g p
rote
ína
-1)
a
0
10
20
30
40
50
60
70
80
90
100
Cata
laase
(µ
mol
H2O
2.m
in-1
.mg p
rote
ína
-1)
Cot
Axis
b
Erythrina velutina
0
0 2 4 6 80
0 2 4 6 8
0.0
0.5
1.0
1.5
2.0
2.5
3.0
3.5
4.0
4.5
5.0
5.5
6.0
0 2 4 6 8
Glu
tati
on
a-S
-tra
nfe
ra
se
(µm
ol C
DN
B.m
in-1
.mg
p
ro
teín
a-1
)
c
0.0
0.2
0.4
0.6
0.8
1.0
1.2
1.4
1.6
1.8
2.0
0 2 4 6 8
Gu
aia
col
Per
ox
idase
(A
BS
.min
-1.m
g p
rote
ína
-1)
d
Electrical conductivity (dS.m-1)4 dSm-1���� treshold for stress response
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ConsiderationsConsiderations
� High germination in pure biosaline water;
� Analysis BSW � may explain the results
High content of organic matter, Ca 2 +, Mg 2 +High content of organic matter, Ca 2 +, Mg 2 +
Other variables of the germination process � to understand the
germination metabolism in this situation
� Seedling production � great ecological benefit
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ConclusionsConclusionsHigh salt tolerance with different mechanisms of tolerance to brackish water.
Biossaline Agriculture maybe an alternative, viable and sustainable for the production sustainable for the production of seedlings of this species.
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ACKNOWLEDGEMENTSACKNOWLEDGEMENTS
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ACKNOWLEDGEMENTSACKNOWLEDGEMENTS
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Merci!