Abstract
<pre><strong><span lang="EN-US">Background. </span></strong><em><span lang="EN-US">Jatropha curcas</span></em><span lang="EN-US"> L. is a crop whose oil can be converted to biodiesel. However, there is lack of varieties with high yields that limiting commercial production of the crop. An alternative is to obtain <em>in vitro </em>plants from unpollinated female gametophytes is known as gynogenesis which can provide an alternative to </span><span lang="EN-US">produce </span><span lang="EN-US">varieties with increased seed production.<strong> Objective</strong>. The objective was to establish a protocol for obtaining <em>in vitro</em> plants from unfertilized ovules of <em>Jatropha curcas</em> L. ovules. <strong>Methodology</strong>. The ovules were extracted from unfertilized inflorescences pretreated for 0, 1, 3 and 7 days at 2 to 5 ° C. Three combinations of plant growth regulator treatments were applied to culture media for induction of gynogenic calli. Gynogenic calli were cultured on MS basal medium with BAP and IBA for development of green, friable. The </span><span lang="EN-US">gynogenic </span><span lang="EN-US">calli from the development treatment </span><span lang="EN-US">were transferred to experiments to determine a regeneration treatment. </span><span lang="EN-US">The gynogenic plantlets were transferred to different treatments for root development. <strong>Results</strong>. The results show that pretreatment of inflorescences at 2 to 5°C had no significant effect on the number of ovules that formed friable white calli. Induction of gynogenic friable white calli occurred in the dark conditions, on half-strength </span><span lang="EN-US">Murashige and Skoog basal medium (</span><span lang="EN-US">½</span><span lang="EN-US"> MS) </span><span lang="EN-US">supplemented with two combinations of growth regulators: (i) 5.37 µM naphthalene acetic acid (NAA) combined with 6.65 µM 6-benzylaminopurine (BAP) and, (ii)</span><span lang="EN-US">8.05 µM NAA with 22.09 µM BAP. Development of green, friable, gynogenic calli under light–dark conditions was possible under treatment with complete Murashige and Skoog (1962) (MS) basal medium with 6.66 µM BAP and 4.9 µM indole-butyric acid (IBA).</span><span lang="EN-US">Friable green callus formed gynogenic embryos on MS basal medium supplemented with </span><span lang="EN-US">22.09 </span><span lang="EN-US">µM</span><span lang="EN-US">BAP and </span><span lang="EN-US">3.40 </span><span lang="EN-US">µM</span><span lang="EN-US"> paclobutrazol (PBZ), embryos regeneration occurred in photoperiod conditions</span><span lang="EN-US">. </span><span lang="EN-US">Embryos were able to develop and convert to plantlets in the treatment on MS basal medium containing 2.22 µM BAP and 0.28 µM of indole-3-acetic acid (IAA).</span><span lang="EN-US"> Root development of </span><span lang="EN-US">plantlets</span><span lang="EN-US"> occurred in </span><span lang="EN-US">½</span><span lang="EN-US"> MS basal medium with 18.65 µM IBA. <strong>Implications</strong>. A protocol of <em>in vitro</em> gynogenesis in <em>Jatropha curcas</em> would contribute to the improvement of its cultivation, reducing the time required for the generation of pure lines that would allow us to obtain varieties with increased seed production. <strong>Conclusions. </strong>The<strong> </strong></span><span lang="EN-US">work presented here describes a reproducible protocol to produce plants <em>in vitro</em> from unfertilized ovules of <em>Jatropha curcas</em> L. T</span><span lang="EN-US">his methodology will facilitate to obtain homozygotic lines with significant reduction in the time required by conventional methods. </span><strong></strong></pre>
Highlights
A protocol of in vitro gynogenesis in Jatropha curcas would contribute to the improvement of its cultivation, reducing the time required for the generation of pure lines that would allow us to obtain varieties with increased seed production, given that the current lack of varieties with high yields is limiting commercial production of the crop
Calluses were cultured on MS basal medium with bencilaminopurina 6.65 μM (BAP) and indole-butyric acid (IBA)
The experiment for this stage was set up with a 4 × 4 factorial design with two factors: a cytokinin (BAP) and an auxin (IBA). Both factors were evaluated at four levels, BAP at 2.22, 4.44, 6.66, and 8.88 μM and IBA at 2.45, 4.9, 7.35, and 9.9 μM, resulting in a total of 16 treatments (Table 1, DT1DT16)
Summary
Jatropha curcas is a promising energy crop whose oil is suitable for biodiesel production (Reale et al, 2012). A protocol of in vitro gynogenesis in Jatropha curcas would contribute to the improvement of its cultivation, reducing the time required for the generation of pure lines that would allow us to obtain varieties with increased seed production, given that the current lack of varieties with high yields is limiting commercial production of the crop. This is the first report of gynogenesis in Jatropha curcas L
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