Cloning and characterization of a farnesyl pyrophosphate synthase from Matricaria recutita L. and its upregulation by methyl jasmonate.

Matricaria recutita (L.), commonly known as chamomile, is one of the most valuable medicinal plants because it synthesizes a large number of pharmacologically active secondary metabolites known as α-bisabolol and chamazulene. Although the plant has been well characterized in terms of chemical constituents of essential oil as well as pharmacological properties, little is known about the genes responsible for biosynthesis of these compounds. In this study, we report a new full-length cDNA encoding farnesyl diphosphate synthase (FPS), a key enzyme in the pathway of biosynthesis of isoprenoids, from M. recutita. The cDNA of MrFPS comprises 1032 bp and encodes 343 amino acid residues with a calculated molecular mass of 39.4 kDa. The amino acid sequence homology and phylogenetic analysis indicated that MrFPS belongs to the plant FPS super-family and is closely related to FPS from the Asteraceae family. Expression of the MrFPS gene in Escherichia coli yielded FPS activity. Using real-time quantitative PCR, the expression pattern of the MrFPS gene was analyzed in different tissues of M. recutita as well as in response to methyl jasmonate. The expression analysis demonstrated that MrFPS expression varies in different tissues (with maximal expression in flowers and stems) and was significantly elevated in response to methyl jasmonate. This study will certainly enhance our understanding of the role of MrFPS in the biosynthesis and regulation of valuable secondary metabolites in M. recutita at a molecular level.

Molecular cloning and characterization of the full-length Hsp90 gene from Matricaria recutita.

Heat shock protein 90 (Hsp90) is one of the most abundant and conserved chaperone proteins and plays important roles in plant growth and responses to environmental stimuli. However, little is known regarding the sequence and function of Hsp90s in Matricaria recutita. In the present study, we cloned the full-length cDNA sequence of the hsp90 gene from this species. Using rapid amplification of cDNA ends technologies with 2 degenerate primers that were designed based on the hsp90 gene sequence from other members of Asteraceae, we isolated and characterized an Hsp90 homolog gene from M. recutita (Mr-Hsp90). The full-length Mr-hsp90 cDNA sequence, containing 2097 base pairs, encodes a protein of 698 amino acids. Based on amino acid sequence identity, Mr-Hsp90 showed high similarity to other cloned Hsp90 proteins. The Mr-Hsp90 protein was closely clustered with the Lactuca sativa in a phylogenetic tree. These results indicate that the cloned sequence of Mr-Hsp90 is a member of the Hsp90 family, which is reported for the first time in M. recutita. Next, we conducted a salt stress experiment to determine the protein's function under salt stress conditions. Survival of chamomile seedlings subjected to heat-shock pretreatment was significantly increased compared with groups that had not undergone heat-shock pretreatment in a salt stress environment. This indicates that Mr-Hsp90 plays an important role in the salt resistance of chamomile seedlings.

Patient-related constraints on get- and be-passive uses in English: evidence from paraphrasing.

In English, transitive events can be described in various ways. The main possibilities are active-voice and passive-voice, which are assumed to have distinct semantic and pragmatic functions. Within the passive, there are two further options, namely be-passive or get-passive. While these two forms are generally understood to differ, there is little agreement on precisely how and why. The passive Patient is frequently cited as playing a role, though again agreement on the specifics is rare. Here we present three paraphrasing experiments investigating Patient-related constraints on the selection of active vs. passive voice, and be- vs. get-passive, respectively. Participants either had to re-tell short stories in their own words (Experiments 1 and 2) or had to answer specific questions about the Patient in those short stories (Experiment 3). We found that a given Agent in a story promotes the use of active-voice, while a given Patient promotes be-passives specifically. Meanwhile, get-passive use increases when the Patient is marked as important. We argue that the three forms of transitive description are functionally and semantically distinct, and can be arranged along two dimensions: Patient Prominence and Patient Importance. We claim that active-voice has a near-complementary relationship with the be-passive, driven by which protagonist is given. Since both get and be are passive, they share the features of a Patient-subject and an optional Agent by-phrase; however, get specifically responds to a Patient being marked as important. Each of these descriptions has its own set of features that differentiate it from the others.