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

CAATINGACAATINGA

� 11% brazilian territory;

� 70% Notheastern Region;

� 1300 counties; � 1300 counties;

� 63% of northeastern population.

CAATINGACAATINGA

�Exclusively brazilian;

�Rich in species and endemism;

�Highly degraded

� less than 50% original area

�Less studied ecossystem

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

Dantas, B.F. Dantas, B.F.

Rainy season Dry season

Kiill, L.H.P. Kiill, L.H.P.

CAATINGACAATINGAWATER RESORUCES

� São Francisco River

CAATINGACAATINGA

WATER RESORUCES

� Brackish groundwater

CAATINGACAATINGAWATER RESORUCES

� Fish farming wastewater

� Ponds filled with brackish groundwater

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??

Caatinga Caatinga nativenative forestforest speciesspecies

Pereiro (Aspidosperma pyrifolium Mart.)Apocynaceae

•Ornamental tree;

•Bark- farmaceutical;

•Seeds- craftwork

Photos: J.E.B. Silva; R.C. Ribeiro

Angico (Anadenanthera macrocarpa (Benth.) Brenan)Mimosaceae

•Widely distributed in brazillan territory;

•Farmaceutical,

•Ornamental,

•Wood for coal and construction;

•Forage

Catingueira-verdadeira (Poincianella pyramidalis [Tul.] L.P.Queiroz)Leguminosae

Endemic from Caatinga

Pioneer species

Forage

Mulungu (Erythrina velutina Willd.)Leguminosae

Beautiful tree

Ornamental, farmaceutical, craftwork

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

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

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.

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

-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

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.

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®

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)

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

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)

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

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

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

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

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

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.

ACKNOWLEDGEMENTSACKNOWLEDGEMENTS

ACKNOWLEDGEMENTSACKNOWLEDGEMENTS

Merci!