Expression of CT-wpr, screened by cDNA-AFLP approach, associated with hydroxysafflor yellow A in Carthamus tinctorius L.

Hydroxysafflor yellow A (HSYA) is the main active compound in petals of Carthamus tinctorius L. In this study, we used cDNA-amplified fragment length polymorphism (cDNA-AFLP), coupled with bulked segregant analysis (BSA), to screen differentially expressed genes in two cDNA pools, and obtained 2560 reproducible bands ranging from 1500 bp to 100 bp with 64 AFLP primer combinations. Two TDFs (TDF-4 and TDF-11) were identified with low recombination and successfully sequenced. The genetic distance of the linkage map from the HSya locus was 9.9 cM and 2.9 cM, respectively. The full-length cDNA sequence of TDF-11 gene, designated as CT-wpr (GenBank Accession No. GU227360) was cloned. Nucleotide sequence alignment revealed that CT-wpr shared 66% identity with Populus trichocarpa. Semi-quantitative RT-PCR analysis revealed that the mRNA of the two TDFs only expressed in the HSYA-absent line. Possible functions of CT-wpr are also discussed.     

Gene expression changes in response to drought stress in <Emphasis Type="Italic">Citrullus colocynthis</Emphasis>

Citrullus colocynthis (L.) Schrad, closely related to watermelon, is a member of the Cucurbitaceae family. This plant is a drought-tolerant species with a deep root system, widely distributed in the Sahara-Arabian deserts in Africa and the Mediterranean region. cDNA amplified fragment length polymorphism (cDNA-AFLP) was used to study differential gene expression in roots of seedlings in response to a 20% polyethylene glycol-(PEG8000) induced drought stress treatment. Eighteen genes which show similarity to known function genes were confirmed by quantitative relative (RQ) real-time RT-PCR to be differentially regulated. These genes are involved in various abiotic and biotic stress and developmental responses. Dynamic changes with tissue-specific pattern were detected between 0 and 48 h of PEG treatment. In general, the highest induction levels in roots occurred earlier than in shoots, because the highest expression was detected in roots following 4 and 12 h, in shoots following 12 and 48 h of drought. These drought-responsive genes were also affected by the plant hormones abscisic acid (ABA), salicylic acid (SA), or jasmonic acid (JA), indicating an extensive cross-talk between drought and plant hormones. Collectively, these results will be useful to explore the functions of these multiple signal-inducible genes for unveiling the relationship and crosstalk between different signaling pathways.     

A sulphur deficiency-induced gene, sdi1, involved in the utilization of stored sulphate pools under sulphur-limiting conditions has potential as a diagnostic indicator of sulphur nutritional status

A sulphate deficiency-induced gene, sdi1 , has been identified by cDNA-amplified fragment length polymorphism (AFLP) analysis utilizing field-grown, nutrient-deficient wheat ( Triticum aestivum var. Hereward). The expression of sdi1 was specifically induced in leaf and root tissues in response to sulphate deficiency, but was not induced by nitrogen, phosphorus, potassium or magnesium deficiency. Expression was also shown to increase in plant tissues as the external sulphate concentration in hydroponically grown plants was reduced from 1.0 to 0.0 m m . On this basis, sdi1 gene expression has potential as a sensitive indicator of sulphur nutritional status in wheat. Genome-walking techniques were used to clone the 2.7-kb region upstream of sdi1 from genomic DNA, revealing several cis -element motifs previously identified as being associated with sulphur responses in plants. The Arabidopsis thaliana gene most highly homologous to sdi1 is At5g48850, which was also demonstrated to be induced by sulphur deficiency, an observation confirmed by the analysis of microarray data available in the public domain. The expression of Atsdi1 was induced more rapidly than previously characterized sulphur-responsive genes in the period immediately following the transfer of plants to sulphur-deficient medium. Atsdi1 T-DNA 'knockout' mutants were shown to maintain higher tissue sulphate concentrations than wild-type plants under sulphur-limiting conditions, indicating a role in the utilization of stored sulphate under sulphur-deficient conditions. The structural features of the sdi1 gene and its application in the genetic determination of the sulphur nutritional status of wheat crops are discussed.