A point mutation in the kinase domain of CRK10 leads to xylem vessel collapse and activation of defence responses in Arabidopsis.

Cysteine-rich receptor-like kinases (CRKs) are a large family of plasma membrane-bound receptors ubiquitous in higher plants. However, despite their prominence, their biological roles have remained largely elusive so far. In this study we report the characterization of an Arabidopsis mutant named crk10-A397T in which alanine 397 has been replaced by a threonine in the αC helix of the kinase domain of CRK10, known to be a crucial regulatory module in mammalian kinases. The crk10-A397T mutant is a dwarf that displays collapsed xylem vessels in the root and hypocotyl, whereas the vasculature of the inflorescence develops normally. In situ phosphorylation assays with His-tagged wild type and crk10-A397T versions of the CRK10 kinase domain revealed that both alleles are active kinases capable of autophosphorylation, with the newly introduced threonine acting as an additional phosphorylation site in crk10-A397T. Transcriptomic analysis of wild type and crk10-A397T mutant hypocotyls revealed that biotic and abiotic stress-responsive genes are constitutively up-regulated in the mutant, and a root-infection assay with the vascular pathogen Fusarium oxysporum demonstrated that the mutant has enhanced resistance to this pathogen compared with wild type plants. Taken together our results suggest that crk10-A397T is a gain-of-function allele of CRK10, the first such mutant to have been identified for a CRK in Arabidopsis.

Delivering sustainable crop protection systems via the seed: exploiting natural constitutive and inducible defence pathways.

To reduce the need for seasonal inputs, crop protection will have to be delivered via the seed and other planting material. Plant secondary metabolism can be harnessed for this purpose by new breeding technologies, genetic modification and companion cropping, the latter already on-farm in sub-Saharan Africa. Secondary metabolites offer the prospect of pest management as robust as that provided by current pesticides, for which many lead compounds were, or are currently deployed as, natural products. Evidence of success and promise is given for pest management in industrial and developing agriculture. Additionally, opportunities for solving wider problems of sustainable crop protection, and also production, are discussed.

The Arabidopsis F-box/Kelch-repeat protein At2g44130 is upregulated in giant cells and promotes nematode susceptibility.

We report that the F-box/Kelch-repeat protein At2g44130 is specifically induced by the root-knot nematode Meloidogyne incognita during the initial stages of the initiation and maintenance of the feeding site. In addition, we show that the expression of this gene promotes susceptibility of infection because knocking down the F-box gene (At2g44130) drastically reduces nematode attraction to and infection of roots. In contrast, F-box overexpressing (OE) lines had a hypersusceptible phenotype, with an increase of 34% in nematode attraction and 67% in nematode infection when grown in soil. This hypersusceptibility might be the result of an increased attraction of the second-stage juveniles toward root exudates of the F-box OE, which would suggest that the blend of compounds in the root exudates of the OE line was somewhat different from the ones present in the root exudates of the wild type and the F-box knockout and tilling lines. Although the function of the F-box/Kelch-repeat protein (At2g44130) is not known, we postulate that its activation by nematode effectors released during the infection process leads to the formation of SCF((At2g44130)) (Skp1-Cullin1-F-box protein) complexes, which are involved in facilitating successful infection by the nematode through targeting specific proteins for degradation.

The transcriptome of cis-jasmone-induced resistance in Arabidopsis thaliana and its role in indirect defence.

cis-jasmone (CJ) is a plant-derived chemical that enhances direct and indirect plant defence against herbivorous insects. To study the signalling pathway behind this defence response, we performed microarray-based transcriptome analysis of CJ-treated Arabidopsis plants. CJ influenced a different set of genes from the structurally related oxylipin methyl jasmonate (MeJA), suggesting that CJ triggers a distinct signalling pathway. CJ is postulated to be biosynthetically derived from jasmonic acid, which can boost its own production through transcriptional up-regulation of the octadecanoid biosynthesis genes LOX2, AOS and OPR3. However, no effect on these genes was detected by treatment with CJ. Furthermore, CJ-responsive genes were not affected by mutations in COI1 or JAR1, which are critical signalling components in MeJA response pathway. Conversely, a significant proportion of CJ-inducible genes required the three transcription factors TGA2, TGA5 and TGA6, as well as the GRAS regulatory protein SCARECROW-like 14 (SCL14), indicating regulation by a different pathway from the classical MeJA response. Moreover, the biological importance was demonstrated in that mutations in TGA2, 5, 6, SCL14 and the CJ-inducible gene CYP81D11 blocked CJ-induced attraction of the aphid parasitoid Aphidius ervi, demonstrating that these components play a key role in CJ-induced indirect defence. Collectively, our results identify CJ as a member of the jasmonates that controls indirect plant defence through a distinct signalling pathway.

Natural variation in responsiveness of Arabidopsis thaliana to methyl jasmonate is developmentally regulated.

A number of Arabidopsis thaliana (L.) Heynh ecotypes were assayed for their responses to methyl jasmonate in order to determine any natural variation in response to this volatile signal. We observed that the regulation of methyl jasmonate-induced expression of the vegetative storage proteins VSP1 and VSP2 is linked to the developmental stage of the plants. In two ecotypes investigated further, Gr-3 and Col-0, it was observed that the VSP1/2 genes became non-responsive to methyl jasmonate stimulation as the plants progressed to bolt formation and flowering. However, the onset of when this transcriptional inactivation occurred differed between the two ecotypes, with Col-0 displaying still high levels of transcript at the onset of flowering whereas Gr-3 showed no induction of VSP1/2 transcription at the same developmental stage. To our knowledge, this is the first time that such a pattern of regulation has been described for a methyl jasmonate-regulated gene. Moreover, in an F(2) population of a cross between these two ecotypes, the trait for 'VSP1/2 methyl jasmonate non-responsiveness' segregated among individuals, indicating the feasibility of mapping the genetic components of this response.

cis-Jasmone induces Arabidopsis genes that affect the chemical ecology of multitrophic interactions with aphids and their parasitoids.

It is of adaptive value for a plant to prepare its defenses when a threat is detected, and certain plant volatiles associated with insect damage, such as cis-jasmone (CJ), are known to switch-on defense metabolism. We used aphid and aphid parasitoid responses to Arabidopsis thaliana as a model system for studying gene expression and defense chemistry and its impact at different trophic levels. Differential responses to volatiles of induced Arabidopsis occurred for specialist and generalist insects: the generalist aphid, Myzus persicae, was repelled, whereas the specialist, Lipaphis erysimi, was attracted; the generalist aphid parasitoid Aphidius ervi was attracted, but the specialist parasitoid Diaeretiella rapae was not affected. A. ervi also spent longer foraging on induced plants than on untreated ones. Transcriptomic analyses of CJ-induced Arabidopsis plants revealed that a limited number of genes, including a gene for a cytochrome P450, CYP81D11, were strongly up-regulated in the treated plants. We examined transgenic Arabidopsis lines constitutively overexpressing this gene in bioassays and found insect responses similar to those obtained for wild-type plants induced with CJ, indicating the importance of this gene in the CJ-activated defense response. Genes involved in glucosinolate biosynthesis and catabolism are unaffected by CJ and, because these genes relate to interactions with herbivores and parasitoids specific to this family of plants (Brassicaceae), this finding may explain the differences in behavioral response of specialist and generalist insects.

Developments in aspects of ecological phytochemistry: the role of cis-jasmone in inducible defence systems in plants.

The challenges and opportunities for protecting agricultural production of food and other materials will be met through exploiting the induction of defence pathways in plants to control pests, diseases and weeds. These approaches will involve processes that can be activated by application of natural products, patented in terms of this use, to "switch on" defence pathways. Already, a number of secondary metabolite defence compounds are known for which the pathways are conveniently clustered genomically, e.g. the benzoxazinoids (hydroxamic acids) and the avenacins. For the former, it is shown that the small molecular weight lipophilic activator cis-jasmone can induce production of these compounds and certain genes within the pathway. Numerous groups around the world work on inducible defence systems. The science is rapidly expanding and involves studying the interacting components of defence pathways and the switching mechanisms activated by small molecular weight lipophilic compounds. Examples are described of how plant breeding can exploit these systems and how heterologous gene expression will eventually give rise to a new range of GM crops for food and energy, without the need for external application of synthetic pesticides.