The discovery of the CRISPR/Cas genome-editing system has revolutionized our understanding of the plant genome. CRISPR/Cas has been used for over a decade to modify plant genomes for the study of specific genes and biosynthetic pathways as well as to speed up breeding in many plant species, including both model and non-model crops. Although the CRISPR/Cas system is very efficient for genome editing, many bottlenecks and challenges slow down further improvement and applications. In this review we discuss the challenges that can occur during tissue culture, transformation, regeneration, and mutant detection. We also review the opportunities provided by new CRISPR platforms and specific applications related to gene regulation, abiotic and biotic stress response improvement, and de novo domestication of plants.

The phytopathogenic bacterium Erwina carotovora spp. which infects potato plants causes severe losses in agriculture. No protective means or resistance traits usable for plant breeding are known. Introduction of a new resistance gene into potato by gene technology leads to a reduced susceptibility of the transgenic plants towards Erwinia carotovora atroseptica infection. Bacteriophage T4 lysozyme is the most active member of a class of bacteriolytic enzymes also detected in several plant species. Secretion of the foreign T4 lysozyme into the intercellular spaces of transgenic potato plants effects a resistance against the phytopathogenic bacterium already at low expression levels.

Summary: The expression of single rol genes of the T_L‐DNA of Agrobacterium rhizogenes strain A4 in transgenic tobacco (Nicotiana tabacum L.) and potato (Solanum tuberosum L.) plants alters the internal concentrations of, and the sensitivity to, several plant hormones. The levels of immunoreactive cytokinins, abscisic acid, gibberellins and indole‐3‐acetic acid were analysed in tissues of the apical shoots, stems, leaves, roots and undifferentiated callus tissue. The addition of the dominant and morphogenetically active rolA, rolB, or rolC genes resulted in alterations in the content of several hormones. rolC overexpression in particular led to an up to fourfold increase in the content of isopentenyladenosine, dihydrozeatin riboside and trans‐zeatin riboside‐type cytokinins in potato plants. This increase correlated well with different levels of expression of the rolC gene in different transgenic plants. Furthermore it was shown that the dwarfism of P_35s‐rolC transgenic tobacco and potato plants is correlated with a 28–60% reduction of gibberellic acid A₁ concentration in apical shoots. Exogenous addition of gibberellic acid completely restored stem elongation in P_35s‐_rolC transgenic plants. Apical shoots of dwarf rolA transgenic tobacco plants also contained 22% less gibberellic acid A₁ than control plants, but growth cannot be restored completely by exogenously added gibberellic acid. Similarly, the sensitivity of transgenic tobacco seedlings or callus tissues towards different phytohormone concentrations can be altered by the expression of single rol genes. The overexpression of the rolC gene in seedlings led to an altered response to auxins, cytokinins, abscisic acid, gibberellic acid and the ethylene precursor 1‐aminocyclopropane‐carboxylic acid. The overexpression of the rolB gene in tobacco calli led to necrosis at lower auxin concentrations than in the wild‐type, while other parameters of auxin action, like the induction of cell growth, remained unchanged.

The long juvenile period of trees delays the breeding of new varieties. Flowering begins within 5–10 years in most cultivated forest trees under intensive management, but can take up to 40 years in some species and environmental conditions. To accelerate the breeding process several agrotechnical and biotechnical methods have been developed. Knowledge about genes controlling flower initiation in model plants like Arabidopsis thaliana, and identification of homologous genes in trees, have led to new possibilities for early‐flower induction. Overexpression of MADS‐box and other floral regulatory genes resulted in early flowering in some tree species and/or varieties. However, these methods have not yet been shown to enable the production of fertile, viable or normal gametes and progeny; developmental research towards these ends is therefore of high priority. A breeding scheme has been developed to use early flowering trees for the introduction of genes from wild species that would allow several backcrosses to occur in only a few years, and to produce at the end a non‐transgenic improved variety. Research to develop practical early flowering methods could lead to several new methods for breeding and biotechnology.