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The importance of cassava as the fourth largest source of calories in the world requires that contributions of biotechnology to improving this crop, advances and current challenges, be periodically reviewed. Plant biotechnology offers a wide range of opportunities that can help cassava become a better crop for a constantly changing world. We therefore review the state of knowledge on the current use of biotechnology applied to cassava cultivars and its implications for breeding the crop into the future. The history of the development of the first transgenic cassava plant serves as the basis to explore molecular aspects of somatic embryogenesis and friable embryogenic callus production. We analyze complex plant-pathogen interactions to profit from such knowledge to help cassava fight bacterial diseases and look at candidate genes possibly involved in resistance to viruses and whiteflies—the two most important traits of cassava. The review also covers the analyses of main achievements in transgenic-mediated nutritional improvement and mass production of healthy plants by tissue culture and synthetic seeds. Finally, the perspectives of using genome editing and the challenges associated to climate change for further improving the crop are discussed. During the last 30 yr, great advances have been made in cassava using biotechnology, but they need to scale out of the proof of concept to the fields of cassava growers.

Auxotrophic strains of Agrobacterium tumefaciens can contribute to the development of more efficient transformation systems, especially for crops historically considered recalcitrant. Homologous recombination was used to derive methionine auxotrophs of two common A. tumefaciens strains, LBA4404 and EHA105. The EHA105 strains were more efficient for switchgrass transformation, while both the EHA105 and LBA4404 strains worked equally well for the rice control. Event quality, as measured by transgene copy number, was not affected by auxotrophy, but was higher for the LBA4404 strains than the EHA105 strains. Ultimately, the use of auxotrophs reduced bacterial overgrowth during co-cultivation and decreased the need for antibiotics.

Cassava bacterial blight (CBB) caused by Xanthomonas axonopodis pv. manihotis (Xam) is a prevalent disease in all regions, where cassava is cultivated. CBB is a foliar and vascular disease usually controlled through host resistance. Previous studies have found QTLs explaining resistance to several Xam strains. Interestingly, one QTL called XM5 that explained 13% of resistance to XamCIO136 was associated with a similar fragment of the rice Xa21-resistance gene called PCR250. In this study, we aimed to further identify and characterize this fragment and its role in resistance to CBB. Screening and hybridization of a BAC library using the molecular marker PCR250 as a probe led to the identification of a receptor-like kinase similar to Xa21 and were called RXam1 (Resistance to Xam 1). Here, we report the functional characterization of susceptible cassava plants overexpressing RXam1. Our results indicated that the overexpression of RXam1 leads to a reduction in bacterial growth of XamCIO136. This suggests that RXAM1 might be implicated in strain-specific resistance to XamCIO136.

A conventional breeding program was established to transfer the bacterial phytoene synthase transgene– crtB –from a transgenic, white-rooted cassava to yellow-rooted cassava plants carrying the endogenous phytoene synthase alleles named psy 2- y 1 and/or psy 2- y 2 . Combining endogenous phytoene synthase enzymes (PSYs) with CRTB in a single cassava plant would allow the molecular dissection of individual allele contributions to carotenoid synthesis and/or accumulation in cassava roots. The simultaneous expression of the crtB transgene and psy 2- y 2 in individuals planted in the field coincided with higher total, HPLC-quantified carotenoid content in roots, although the variability among replications (plants) precluded the detection of statistically significant differences. Nevertheless, the highest total carotenoid content in roots within a family coincided with one individual of the F1 progeny carrying both psy 2- y 2 and crtB genes. The results also indicated the presence of at least one more key gene–different from psy or crtB —which too is necessary for the synthesis and/or accumulation of Pro-Vitamin A carotenoids in cassava roots.

A conventional breeding program was established to transfer the bacterial phytoene synthase transgene–crtB–from a transgenic, white-rooted cassava to yellow-rooted cassava plants carrying the endogenous phytoene synthase alleles named psy2-y 1 and/or psy2-y 2. Combining endogenous phytoene synthase enzymes (PSYs) with CRTB in a single cassava plant would allow the molecular dissection of individual allele contributions to carotenoid synthesis and/or accumulation in cassava roots. The simultaneous expression of the crtB transgene and psy2-y 2 in individuals planted in the field coincided with higher total, HPLC-quantified carotenoid content in roots, although the variability among replications (plants) precluded the detection of statistically significant differences. Nevertheless, the highest total carotenoid content in roots within a family coincided with one individual of the F1 progeny carrying both psy2-y 2 and crtB genes. The results also indicated the presence of at least one more key gene–different from psy or crtB—which too is necessary for the synthesis and/or accumulation of Pro-Vitamin A carotenoids in cassava roots.

A conventional breeding program was established to transfer the bacterial phytoene synthase transgene-crtB-from a transgenic, white-rooted cassava to yellow-rooted cassava plants carrying the endogenous phytoene synthase alleles named psy2-y and/or psy2-y . Combining endogenous phytoene synthase enzymes (PSYs) with CRTB in a single cassava plant would allow the molecular dissection of individual allele contributions to carotenoid synthesis and/or accumulation in cassava roots. The simultaneous expression of the crtB transgene and psy2-y in individuals planted in the field coincided with higher total, HPLC-quantified carotenoid content in roots, although the variability among replications (plants) precluded the detection of statistically significant differences. Nevertheless, the highest total carotenoid content in roots within a family coincided with one individual of the F1 progeny carrying both psy2-y and crtB genes. The results also indicated the presence of at least one more key gene-different from psy or crtB-which too is necessary for the synthesis and/or accumulation of Pro-Vitamin A carotenoids in cassava roots.

Auxotrophic strains of Agrobacterium tumefaciens can contribute to the development of more efficient transformation systems, especially for crops historically considered recalcitrant. Homologous recombination was used to derive methionine auxotrophs of two common A. tumefaciens strains, LBA4404 and EHA105. The EHA105 strains were more efficient for switchgrass transformation, while both the EHA105 and LBA4404 strains worked equally well for the rice control. Event quality, as measured by transgene copy number, was not affected by auxotrophy, but was higher for the LBA4404 strains than the EHA105 strains. Ultimately, the use of auxotrophs reduced bacterial overgrowth during co-cultivation, with a concomitant decrease in the need for antibiotics. In turn, reduced overgrowth allowed longer co-cultivation periods, with a trend towards higher transformation frequencies for switchgrass, but not for rice.

Transformation of switchgrass (Panicum virgatum L.) is important for studies of gene function. New approaches of transformation are alternatives to improve efficiency of transformation. The objective was to test the potential of using Agrobacterium tumefaciens auxotrophic strains and morphogenic genes Baby Boom and Wuschel2 in genetic transformation of a lowland genotype (Performer 7). Data of transient and stable transformation were used to test the effectiveness in transformation of the Agrobacterium strains. The auxotrophic EHA105 showed high frequencies of transient transformation, which was confirmed in stable transformation (> 50% of hygromycin resistant calli). Direct transformation of in vitro plantlets with a morphogenic gene cassette reduced the time for generation of a transgenic plant, as a previous induction of embryogenic callus is not needed. Agrobacterium auxotrophs combined with morphogenic genes represents a new era of genetic transformation in switchgrass.