Establishment of bispyribac selection protocols for <i>Agrobacterium tumefaciens</i>- and <i>Agrobacterium rhizogenes</i>-mediated transformation of the oil seed plant <i>Jatropha curcas</i> L.

Abstract

Cultivation of the biofuel plant Jatropha (Jatropha curcas L.) has spread around the world because of its drought resistance, high seed oil content, and adaptability to di erent environmental conditions. Because of these attributes, Jatropha has the potential to be one of the main resources for next-generation biodiesel fuel. To improve the productivity of Jatropha biomass, it is important to understand the molecular functions of key Jatropha genes, and to modify various agronomic traits of Jatropha via molecular breeding. A reliable and e cient protocol for genetic transformation of Jatropha is a prerequisite for molecular biology research and breeding on this plant. Here, we developed a system in which the herbicide bispyribac sodium salt, which inhibits acetolactate synthase, was used as the selection agent, and a two-point-mutated acetolactate synthase gene (mALS) was used to confer resistance upon transformants. Application of this system signi cantly improved the e ciency of Agrobacterium tumefaciens-mediated stable transformation of the high-yielding elite Jatropha population, IP-2P. e bispyribac-mALS system was also successfully applied in the Agrobacterium rhizogenes-mediated hairy roots system, which allowed integration of a foreign gene and expression in Jatropha root tissues within 2 weeks. e new protocols described here are powerful tools not only for functional studies on endogenous genes, but also for the molecular breeding of Jatropha to develop elite varieties.

Agrobacterium tumefaciens-mediated transformation of Jatropha curcas L. with a polyhydroxyalkanoate gene (phaC)

“…Rooting induction of transformed J. curcas was rather difficult to be obtained, but nevertheless was successfully obtained when using liquid half-strenght MS with 4.9 µM IBA (Figure 3e). Similar finding was also reported by Toppo et al (2012) and Kajikawa et al (2012) that the use of indole butiric acid (IBA) as a growth factor gave a better result in the induction of root than indole acetic acid (IAA) as an endogenous auxin. Auxin induces root fomation by breaking root apical dominance through cytokinins.…”

“…Kajikawa et al (2012) succesfully transformed J. curcas using A. tumefaciens and found the shoot frequency of cotyledon explant was higher than hypocotyl explant. Pan et al (2010) and Khemkladngoen et al (2011) also succesfully transformed J. curcas using A. tumefaciens with cotyledon explant.…”

Improving the transformation efficiency of Cucurbita species: factors and strategy for practical application

“…Kanamycin was used in this experiment as a selective agent. But in other plant species, for example Pisum sativum, Cucumis sativus, Jatropha curcus and Brassica rapa, some herbicides such as glyphosate, bialaphos, and bispyribac are more effective for producing transgenic plants (Bean et al 1997;Kajikawa et al 2012;Konagaya et al 2013;Vengadesan et al 2005). An efficient transformation system for other Cucurbita species may be established by optimization of the factors described above.…”

Introduksi Gen Sitrat Sintase Pseudomonas aeruginosa ke Nicotiana tabacum dan Jatropha curcas untuk Meningkatkan Toleransi terhadap Cekaman Aluminium

“…Penelitian Pan et al (2010) hanya mendapat satu planlet dari 120 tunas in vitro. Kajikawa et al (2012) Plasmid A1 A2 A8 A11 M22 M24 M13 M6 P1 P7 P13 marker TL …”