Tolerance of subzero winter cold in kudzu (Pueraria montana var. lobata).

The use of species distribution as a climate proxy for ecological forecasting is thought to be acceptable for invasive species. Kudzu (Pueraria montana var. lobata) is an important invasive whose northern distribution appears to be limited by winter survival; however, kudzu's cold tolerance thresholds are uncertain. Here, we used biogeographic evidence to hypothesize that exposure to - 20 °C is lethal for kudzu and thus determines its northern distribution limit. We evaluated this hypothesis using survival tests and electrolyte leakage to determine relative conductivity, a measure of cell damage, on 14 populations from eastern North America. Relative conductivity above 36% was lethal. Temperatures causing this damage averaged - 19.6 °C for northern and - 14.4 °C for southern populations, indicating kudzu acclimates to winter cold. To assess this, we measured relative conductivity of above- and belowground stems, and roots collected throughout the winter at a kudzu population in southern Ontario, Canada. Consistent with acclimation, the cold tolerance threshold of aboveground stems at the coldest time of year was - 26 °C, while stems insulated from cold extremes survived to - 17 °C-colder than the survival limits indicated by kudzu's biogeographic distribution. While these results do not rule out alternative cold limitations, they indicate kudzu can survive winters north of its current distribution. For kudzu, biogeography is not a proxy for climatic tolerance and continued northward migration is possible. Efforts to limit its spread are therefore prudent. These results demonstrate that physiological constraints inform predictions of climate-related changes in species distribution and should be considered where possible.

Substrate promiscuity of a rosmarinic acid synthase from lavender (Lavandula angustifolia L.).

One of the most common types of modification of secondary metabolites is the acylation of oxygen- and nitrogen-containing substrates to produce esters and amides, respectively. Among the known acyltransferases, the members of the plant BAHD family are capable of acylating a wide variety of substrates. Two full-length acyltransferase cDNAs (LaAT1 and 2) were isolated from lavender flowers (Lavandula angustifolia L.) by reverse transcriptase-PCR using degenerate primers based on BAHD sequences. Recombinant LaAT1 exhibited a broad substrate tolerance accepting (hydroxy)cinnamoyl-CoAs as acyl donors and not only tyramine, tryptamine, phenylethylamine and anthranilic acid but also shikimic acid and 4-hydroxyphenyllactic acid as acceptors. Thus, LaLT1 forms esters and amides like its phylogenetic neighbors. In planta LaAT1 might be involved in the biosynthesis of rosmarinic acid, the ester of caffeic acid and 3,4-dihydroxyphenyllactic acid, a major constituent of lavender flowers. LaAT2 is one of three members of clade VI with unknown function.

Cinnamate metabolism in ripening fruit. Characterization of a UDP-glucose:cinnamate glucosyltransferase from strawberry.

Strawberry (Fragaria x ananassa) fruit accumulate (hydroxy)cinnamoyl glucose (Glc) esters, which may serve as the biogenetic precursors of diverse secondary metabolites, such as the flavor constituents methyl cinnamate and ethyl cinnamate. Here, we report on the isolation of a cDNA encoding a UDP-Glc:cinnamate glucosyltransferase (Fragaria x ananassa glucosyltransferase 2 [FaGT2]) from ripe strawberry cv Elsanta that catalyzes the formation of 1-O-acyl-Glc esters of cinnamic acid, benzoic acid, and their derivatives in vitro. Quantitative real-time PCR analysis indicated that FaGT2 transcripts accumulate to high levels during strawberry fruit ripening and to lower levels in flowers. The levels in fruits positively correlated with the in planta concentration of cinnamoyl, p-coumaroyl, and caffeoyl Glc. In the leaf, high amounts of Glc esters were detected, but FaGT2 mRNA was not observed. The expression of FaGT2 is negatively regulated by auxin, induced by oxidative stress, and by hydroxycinnamic acids. Although FaGT2 glucosylates a number of aromatic acids in vitro, quantitative analysis in transgenic lines containing an antisense construct of FaGT2 under the control of the constitutive 35S cauliflower mosaic virus promoter demonstrated that the enzyme is only involved in the formation of cinnamoyl Glc and p-coumaroyl Glc during ripening.

Up- and down-regulation of Fragaria x ananassa O-methyltransferase: impacts on furanone and phenylpropanoid metabolism.

A complex mixture of hundreds of substances determines strawberry (Fragaria x ananassa) aroma, but only approximately 15 volatiles are considered as key flavour compounds. Of these, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) is regarded as the most important, but it is methylated further by FaOMT (Fragaria x ananassa O-methyltransferase) to 2,5-dimethyl-4-methoxy-3(2H)-furanone (DMMF) during the ripening process. It is shown here that transformation of strawberry with the FaOMT sequence in sense and antisense orientation, under the control of the cauliflower mosaic virus 35S promoter, resulted in a near total loss of DMMF, whereas the levels of the other volatiles remained unchanged. FaOMT repression also affected the ratio of feruloyl 1-O-beta-D-glucose and caffeoyl 1-O-beta-D-glucose, indicating a dual function of the enzyme in planta. Thus, FaOMT is involved in at least two different biochemical pathways in ripe strawberry fruit.