442
rate of more than 80%. Later, Rodrigues (2019) also working with a clonal minigarden of this species observed that the best time to collect 5 cm mini- cuttings was
when the shoots were no more than 40 cm long. Other authors were able to clonally
propagate 15-year-old trees of this species by rooting 15-cm-long cuttings. They
found that the best time of extraction was in spring, achieving a 52% survival of the
cuttings with a rooting capacity of 30%, with application of indole butyric acid
3000 ppm (Ovando et al. 2013).
Using micropropagation techniques and from in vitro germinated H. impetiginosus seedlings, Larraburu (2014) adjusted a protocol which produced 83% of in vitro
rooted plants, and the subsequent acclimatization allowed a survival of 50% of the
obtained plants. Duarte et al. (2016) germinated in vitro H. heptaphyllus seeds and
used different parts of the seedling to regenerate shoots. The rooting of the sprouts
obtained in this way was achieved in a temporary immersion medium which showed
a good response during acclimatization.
In vitro experiments on vegetative propagation of H. impetiginosus were conducted using nodal segments cultivated in both Murashige and Skoog salts with
Gamborg vitamins (MSG) and Woody Plant Medium (WPM), with different concentrations of 6-benzylaminopurine (BA) and indole butyric acid (IBA).
Morphogenic responses were differentially affected by salt compositions and their
interactions with plant growth regulators in each micropropagation stage. According
to response surface analysis, the optimum multiplication rate with 1 μM IBA ranged
from 16.7 to 21.3 μM BA in WPM, and the inhibitors of endogenous auxins could
increase multiplication rates. A pulse with 50 μM IBA in MSG produced 83% rooting with 3.2 roots per shoot and higher fresh and dry weights of shoots and roots. In
the acclimatization stage, 50% of plants survived after 1 year (Larraburu et al. 2012).
Finally, in 2016 a Clonal Seed Orchard was installed in Famaillá Experimental
Station of INTA (27° 0′ 40.11″ S; 65° 22′ 41.88″ W; 380 m asl), including 160
clones from nine provenances (those of Table 16.2). The design of the CSO was in
randomize complete blocks with six repetitions of single tree plots with a distance
between plants of 5x5 m. The CSO is currently starting to produce seeds. In the near
future, those seeds will be used to establish a network of trials in order to evaluate
each mother-tree in adaptative and productive traits.
16.4 Anadenanthera colubrina var. cebil
16.4.1 Phenotypic and Genetic Studies: Development
of Specific nuSSRs and Spatial Distribution
of Phenotypic and Molecular Variability
In Argentina, genetic diversity studies of A. colubrina var. cebil have considered the
species as a model for population genetics studies of long-live species with disjunct
distribution. Initially, these studies were performed using four universal
L. F. Fornes et al.
rate of more than 80%. Later, Rodrigues (2019) also working with a clonal minigarden of this species observed that the best time to collect 5 cm mini- cuttings was
when the shoots were no more than 40 cm long. Other authors were able to clonally
propagate 15-year-old trees of this species by rooting 15-cm-long cuttings. They
found that the best time of extraction was in spring, achieving a 52% survival of the
cuttings with a rooting capacity of 30%, with application of indole butyric acid
3000 ppm (Ovando et al. 2013).
Using micropropagation techniques and from in vitro germinated H. impetiginosus seedlings, Larraburu (2014) adjusted a protocol which produced 83% of in vitro
rooted plants, and the subsequent acclimatization allowed a survival of 50% of the
obtained plants. Duarte et al. (2016) germinated in vitro H. heptaphyllus seeds and
used different parts of the seedling to regenerate shoots. The rooting of the sprouts
obtained in this way was achieved in a temporary immersion medium which showed
a good response during acclimatization.
In vitro experiments on vegetative propagation of H. impetiginosus were conducted using nodal segments cultivated in both Murashige and Skoog salts with
Gamborg vitamins (MSG) and Woody Plant Medium (WPM), with different concentrations of 6-benzylaminopurine (BA) and indole butyric acid (IBA).
Morphogenic responses were differentially affected by salt compositions and their
interactions with plant growth regulators in each micropropagation stage. According
to response surface analysis, the optimum multiplication rate with 1 μM IBA ranged
from 16.7 to 21.3 μM BA in WPM, and the inhibitors of endogenous auxins could
increase multiplication rates. A pulse with 50 μM IBA in MSG produced 83% rooting with 3.2 roots per shoot and higher fresh and dry weights of shoots and roots. In
the acclimatization stage, 50% of plants survived after 1 year (Larraburu et al. 2012).
Finally, in 2016 a Clonal Seed Orchard was installed in Famaillá Experimental
Station of INTA (27° 0′ 40.11″ S; 65° 22′ 41.88″ W; 380 m asl), including 160
clones from nine provenances (those of Table 16.2). The design of the CSO was in
randomize complete blocks with six repetitions of single tree plots with a distance
between plants of 5x5 m. The CSO is currently starting to produce seeds. In the near
future, those seeds will be used to establish a network of trials in order to evaluate
each mother-tree in adaptative and productive traits.
16.4 Anadenanthera colubrina var. cebil
16.4.1 Phenotypic and Genetic Studies: Development
of Specific nuSSRs and Spatial Distribution
of Phenotypic and Molecular Variability
In Argentina, genetic diversity studies of A. colubrina var. cebil have considered the
species as a model for population genetics studies of long-live species with disjunct
distribution. Initially, these studies were performed using four universal
L. F. Fornes et al.