Synthetic promoters consisting of defined cis-acting elements link multiple signaling pathways to probenazole-inducible system.

Probenazole (3-allyloxy-1,2-benzisothiazole-1,1-dioxide, PBZ), the active component of Oryzemate, could induce systemic acquired resistance (SAR) in plants through the induction of salicylic acid (SA) biosynthesis. As a widely used chemical inducer, PBZ is a good prospect for establishing a new chemical-inducible system. We first designed artificially synthetic promoters with tandem copies of a single type of cis-element (SARE, JERE, GCC, GST1, HSRE, and W-box) that could mediate the expression of the ß-glucuronidase (GUS) reporter gene in plants upon PBZ treatment. Then we combined different types of elements in order to improve inducibility in the PBZ-inducible system. On the other hand, we were surprised to find that the cis-elements, which are responsive to jasmonic acid (JA) and ethylene, also responded to PBZ, implying that SA, JA, and ethylene pathways also would play important roles in PBZ's action. Further analysis demonstrated that PBZ also induced early events of innate immunity via a signaling pathway in which Ca(2+) influx and mitogen-activated protein kinase (MAPK) activity were involved. We constructed synthesized artificial promoters to establish a PBZ chemical-inducible system, and preliminarily explored SA, JA, ethylene, calcium, and MAPK signaling pathways via PBZ-inducible system, which could provide an insight for in-depth study.

Post-harvest induced production of salvianolic acids and significant promotion of antioxidant properties in roots of Salvia miltiorrhiza (Danshen).

Danshen, the dried roots of Salvia miltiorrhiza, is an extremely valued Traditional Chinese Medicine. Previously, we have demonstrated that salvianolic acid B (SaB), the important bioactive ingredient in this herb, was a post-harvest product. Here, we further reported that all salvianolic acids (SAs) in the roots were post-harvest products of the drying process. In addition, the results of various radical scavenging activity assays, including lipid peroxidation (1), DPPH (2), hydroxyl (3) and superoxide (4), were significantly increased along with the accumulation of total salvianolic acids in the process. The contents of chemical targets and antioxidant activities both reached the highest value under thermal treatment at 130 °C for 80 min. In this dehydration period, contents of SaB, and sum of nine SAs increased from 0.01% to 5.51%, and 0.20% to 6.61%; and IC50 of antioxidant activity decreased from 4.85 to 2.69 (1); 7.75 to 0.43 (2); 2.57 to 1.13 (3) and 17.25 to 1.10 mg/mL. These results further supported the hypothesis that the newly harvested plant roots were still physiologically active and the secondary metabolites might be produced due to dehydration stress after harvest. Our findings supplied an important and useful theoretical basis for promoting the quality of Danshen and other medicinal plant materials.

Regulation of leaf senescence and crop genetic improvement.

Leaf senescence can impact crop production by either changing photosynthesis duration, or by modifying the nutrient remobilization efficiency and harvest index. The doubling of the grain yield in major cereals in the last 50 years was primarily achieved through the extension of photosynthesis duration and the increase in crop biomass partitioning, two things that are intrinsically coupled with leaf senescence. In this review, we consider the functionality of a leaf as a function of leaf age, and divide a leaf's life into three phases: the functionality increasing phase at the early growth stage, the full functionality phase, and the senescence and functionality decreasing phase. A genetic framework is proposed to describe gene actions at various checkpoints to regulate leaf development and senescence. Four categories of genes contribute to crop production: those which regulate (I) the speed and transition of early leaf growth, (II) photosynthesis rate, (III) the onset and (IV) the progression of leaf senescence. Current advances in isolating and characterizing senescence regulatory genes are discussed in the leaf aging and crop production context. We argue that the breeding of crops with leaf senescence ideotypes should be an essential part of further crop genetic improvement.

Ectopic overexpression of AtHDG11 in tall fescue resulted in enhanced tolerance to drought and salt stress.

Tall fescue (Festuca arundinacea Schreb.) is a cool-season perennial grass, which has been conventionally grown in the temperate area. However, as a major type of cool-season turf grass, its growth has been extended to the sub-tropical climate or even to the transitional climate between the sub-tropical and the tropical, and, in some cases, to heavily salinized lands. The extended growth imposes a serious challenge to its tolerance to the abiotic stress, particularly to drought, salt and high temperature. Here, we report a successful introduction of Arabidopsis AtHDG11 into the tall fescue via Agrobacterium-mediated transformation. The ectopic overexpression of AtHDG11 under the control of CaMV 35S promoter with four enhancers resulted in significantly enhanced tolerance to drought and salt stress. No obvious adverse effects on growth and development were observed in the transgenic plants. The enhanced stress tolerance was associated with a more extensive root system, a lower level of malondialdehyde, a nearly normal Na(+)/K(+) ratio, a higher level of proline and a kinetically accelerated induction of SOD and CAT activities observed in the transgenic plants during drought and/or salt stress, indicating that an enhanced ROS scavenging capability might play a significant role in the acquired tolerance to the abiotic stress.

Analysis of probenazole-responsiveness of rice RPR1 upstream fragments.

RPR1 (rice probenazole-responsive) is a rice gene, the expression of which is responsive to probenazole (PBZ), a synthetic compound that may act as a plant defense activator. It has been shown that RPR1 gene may be involved in disease resistance responses. In this study, a series of amplified fragments from the rice RPR1 promoter region, including 2,416 bp, 1,574 bp, 819 bp, 568 bp and 208 bp fragments upstream to the ATG translation start site, were prepared and linked to the coding region of beta-glucuronidase (GUS) gene. Analysis of GUS gene transient expression in rice calli demonstrated that the 568 bp fragment was sufficient for probenazole responsiveness. Analysis of GUS gene stable expression in Arabidopsis thaliana indicated that the 2,416 bp and 1,574 bp fragments drove GUS expression only in shoot apical meristem and petiole. Identification of these PBZ-responsive fragments provides a basis on which PBZ-inducible gene regulatory systems can be constructed for experimental analysis of gene expression and for field application.

Repression of AtCLH1 expression results in a decrease in the ratio of chlorophyll a/b but doesnot affect the rate of chlorophyll degradation during leaf senescence.

To explore the possible regulatory role of chlorophyllase (Chlase) in chlorophyll (Chl) degradation during leaf senescence, RNAi Arabidopsis (Arabidopsis thaliana) plants were constructed to repress the expressions of AtCLH1 and/or AtCLH2. Transcript levels of AtCLH1 and/or AtCLH2 were dramatically lowered and Chlase activity was correspondingly inhibited, but the Chl degradation kinetics was not affected in the RNAi plants. Results of further analysis indicated that the Chl a/b ratio decreased in AtCLH1 RNAi lines, in comparison with the increasing Chl a/b ratio in the wide type during leaf senescence. In addition, an induced Chlase activity was consistently detected at the initial stage of senescence in all the plants examined. In contrast, transcript levels of both AtCLH1 and AtCLH2 decreased dramatically upon the initiation of senescence in both the wide-type and the RNAi plants. Interestingly, compared with the wide type, lower but still significant transcript levels of the RNAi targeted Chlase gene(s) were sustained during the whole period of dark incubation in all the three RNAi lines examined, indicating the functioning of some compensatively regulating mechanism. Based on these results, along with related reports, we conclude that Chlase might be required at the initial stage of leaf senescence, quite likely playing a role in converting Chl b to a.

Characterization of a novel developmentally retarded mutant (drm1) associated with the autonomous flowering pathway in Arabidopsis.

A developmentally retarded mutant (drm1) was identified from ethyl methanesulfonate (EMS)-mutagenized M2 seeds in Columbia (Col-0) genetic background. The drm1 flowers 109 d after sowing, with a whole life cycle of about 160 d. It also shows a pleiotropic phenotype, e.g., slow germination and lower germination rate, lower growth rate, curling leaves and abnormal floral organs. The drm1 mutation was a single recessive nuclear mutation, which was mapped to the bottom of chromosome 5 and located within a region of 20-30 kb around MXK3.1. There have been no mutants with similar phenotypes reported in the literature, suggesting that DRM1 is a novel flowering promoting locus. The findings that the drm1 flowered lately under all photoperiod conditions and its late flowering phenotype was significantly restored by vernalization treatment suggest that the drm1 is a typical late flowering mutant and most likely associated with the autonomous flowering pathway. The conclusion was further confirmed by the revelation that the transcript level of FLC was constantly upregulated in the drm1 at all the developmental phases examined, except for a very early stage. Moreover, the transcript levels of two other important repressors, EMF and TFL1, were also upregulated in the drm1, implying that the two repressors, along with FLC, seems to act in parallel pathways in the drm1 to regulate flowering as well as other aspects of floral development in a negatively additive way. This helps to explain why the drm1 exhibits a much more severe late-flowering phenotype than most late-flowering mutants reported. It also implies that the DRM1 might act upstream of these repressors.

Tetraenol, a novel sesquiterpenoid from the relict plant Tetraena mongolica in China.

A novel furansesquiterpenoid, tetraenol, was isolated from a relict shrub plant, Tetraena mongolica, collected from the northern desert of the Ningxia Hui Autonomous Region. The structure of the new compound was elucidated on the basis of spectroscopic analysis.

[Screening for lifespan-extension mutants with paraquat in Arabidopsis].

Genetic analyses of lifespan in model animals have revealed that extended lifespans are closely associated to increased resistance to oxidative stress. In the model plant Arabidopsis, late-flowering mutants are also found to be more tolerant to oxidative stress. However, Arabidopsis mutants with extended lifespans are poorly studied so far. In this study, a screening system for mutants with extended lifespans in Arabidopsis was established using paraquat, a potent herbicide that exacerbates O2-. radical production. The relationship between lifespan and resistance to oxidative stress was examined with one of the mutants, SFNA-9-4. Compared to that of wild type, the lifespan of SFNA-9-4 is significantly extended, and its resistance to oxidative stress is also significantly elevated. These results suggest that, as in Drosophila, paraquat can also be used to screen for mutants with extended lifespans in Arabidopsis.