Alpha-picolinic acid, a fungal toxin and mammal apoptosis-inducing agent, elicits hypersensitive-like response and enhances disease resistance in rice

Alpha-picolinic acid (PA), a metabolite of tryptophan and an inducer of apoptosis in the animal cell, has been reported to be a toxin produced by some of plant fungal pathogens and used in screening for disease resistant mutants. Here, we report that PA is an efficient apoptosis agent triggering cell death of hypersensitive-like response in planta. Confirmed by Fluorescence Activated Cell Sorter (FACS), rice suspension cells and leaves exhibited programmed cell death induced by PA. The PA-induced cell death was associated with the accumulation of reactive oxygen species that could be blocked by diphenylene iodonium chloride, indicating that the generation of reactive oxygen species was NADPHoxidase dependent. We also demonstrated the induction of rice defense-related genes and subsequent resistant enhancement by PA against the rice blast fungus Magnaporthe grisea. Hence, it was concluded that the PA-stimulated defense response likely involves the onset of the hypersensitive response in rice, which also provides a simple eliciting tool for studying apoptosis in the plant cell.     

Enhancement of Indole-3-Acetic Acid Photodegradation by Vitamin B6

Dear Editor, The plant hormone indole-3-acetic acid （IAA） has long been used in plant culture media for practical applications and sci- entific inquiries. The use of IAA is complicated by the fact that IAA is a photo-labile compound. In Murashige and Skoog （MS） plant media （Murashige and Skoog, 1962）, the concen- trations of salts and mineral nutrients are known to hasten the photodegradation of IAA under white light （Dunlap and Robacker, 1988）. This degradation can be virtually eliminated by the use of a yellow-colored light filter that removes UV, violet, and some of the blue wavelengths from the incident light （Stasinopoulos and Hangarter, 1990）. However, the use of yellow light clearly affects the quality of light that the plants under study receive. In addition to applications in plants, IAA has been used in human health applications.     

Huma should be Linum usitatissimum,not Sesamum indicum—With special reference to the source of confusion of names for traditional Chinese medicine and the written time and author of Shên Nung Pên Ts’ao Ching

The confusion of Huma (胡麻) in Chinese history was discussed after an introduction of Chih Wu Ming Shi T’u K’ao of the Qing Dynasty and its author—Wu Qijun’s contribution to Chinese botany. The morphological characters and distribution of Huma and Jusheng (巨胜) documented in ancient Chinese literature were comparatively studied. Simultaneously, other questions about the Chinese medicament and medicine were inspected in the backgrounds of historical development and social stratum differentiation. We concluded that the earliest recorded Huma in Chinese literature should be Linum usitatissimum. The Chinese name for this plant has been used by the folks until well into modern times. Jusheng (巨胜) should be Sesamum indicum. The reason for the confusion of these two names was also explored. It was further inferred that the confusion of names for traditional Chinese medicine (TCM) originated in the similarity of their property and function while the difference of their morphology and distribution was not taken seriously. We also concluded that Shên Nung Pên Ts’ao Ching was forged by TAO Hongjing (陶弘景) in his Collected Commentaries on Pên Ts’ao Ching but attributed to an ancient author, Shennong (神农). In this book TCM was classified by their property and function, which became the root cause of the confusion of Chinese names for TCM from then on. Some suggestions were also given for the development of Chinese materia medica in the future.     

Highly efficient uptake of ultrafine mesoporous silica nanoparticles with excellent biocompatibility by Liriodendron hybrid suspension cells

The characteristics of the interactions co-cultures of ultrafine mesoporous silica nanoparticles (MSNs) and the Liriodendron hybrid suspension cells were systematically investigated using laser scanning confocal microscope (LSCM) and scanning electron microscopy (SEM). Using fluorescein isothiocyanate (FITC) labeling, the LSCM observations demonstrated that MSNs (size, 5-15nm) with attached FITC molecules efficiently penetrated walled plant cells through endocytic pathways, but free FITC could not enter the intact plant cells. The SEM measurements indicated that MSNs readily aggregated on the surface of intact plant cells, and also directly confirmed that MSNs could enter intact plant cells; this was achieved by determining the amount of silicon present. After 24 h of incubation with 1.0mg mL-1 of MSNs, the viability of the plant cells was analyzed using fluorescein diacetate staining; the results showed that these cells retained high viability, and no cell death was observed. Interestingly, after the incubation with MSNs, the Liriodendron hybrid suspension cells retained the capability for plant regeneration via somatic embryogenesis. Our results indicate that ultrafine MSNs hold considerable potential as nano-carriers of extracellular molecules, and can be used to investigate in vitro gene-delivery in plant cells.     

Recycling Isolation of Plant DNA,A Novel Method

DNA is one of the most basic and essential genetic materials in the field of molecular biology.To date,isolation of sufficient and good-quality DNA is still a challenge for many plant species,though various DNA extraction methods have been published.In the present paper,a recycling DNA extraction method was proposed.The key step of this method was that a single plant tissue sample was recycled for DNA extraction for up to four times,and correspondingly four DNA precipitations(termed as the 1st,2nd,3rd and 4th DNA sample, respectively) were conducted.This recycling step was integrated into the conventional CTAB DNA extraction method to establish a recycling CTAB method.This modified CTAB method was tested in eight plant species,wheat,sorghum,barley,corn,rice,Brachypodium distachyon,Miscanthus sinensis and tung tree.The results showed that high-yield and good-quality DNA samples could be obtained by using this new method in all the eight plant species.The DNA samples were good templates for PCR amplification of both ISSR and SSR markers.The recycling method can be used in multiple plant species and can be integrated with multiple conventional DNA isolation methods,and thus is an effective and universal DNA isolation method.     

TRANSGENIC PLANTS EXPRESSING BACILLUS THURINGIENSIS DELTA-ENDOTOXINS

Commercial varieties of transgenic Bacillus thuringiensis (Bt) plants have been developed in many countries to control target pests. Initially, the expression of native Bt genes in plants was low due to mRNA instability, improper splicing, and post-translation modifications. Subsequently, modifications of the native Bt genes greatly enhanced expression levels. This is a review of the developments that made modem high-expression transgenic Bt plants possible, with an emphasis on the reasons for the low-level expression of native Bt genes in plant systems, and the techniques that have been used to improve plant expression of Bt toxin genes.     

Genome-wide Expression Profiling in Seedlings of the Arabidopsis Mutant uro that is Defective in the Secondary Cell Wall Formation

Plant secondary growth is of tremendous importance, not only for plant growth and development but also for economic usefulness. Secondary tissues such as xylem and phloem are the conducting tissues in plant vascular systems, essentially for water and nutrient transport, respectively. On the other hand, products of plant secondary growth are important raw materials and renewable sources of energy. Although advances have been recently made towards describing molecular mechanisms that regulate secondary growth, the genetic control for this process is not yet fully understood. Secondary cell wall formation in plants shares some common mechanisms with other plant secondary growth processes. Thus, studies on the secondary cell wall formation using Arabidopsis may help to understand the regulatory mechanisms for plant secondary growth. We previously reported phenotypic characterizations of an Arabidopsis semi-dominant mutant, upright rosette （uro）, which is defective in secondary cell wall growth and has an unusually soft stem. Here, we show that lignification in the secondary cell wall in uro is aberrant by analyzing hypocotyl and stem. We also show genome-wide expression profiles of uro seedlings, using the Affymetrix GeneChip that contains approximately 24 000 Arabidopsis genes. Genes identified with altered expression levels include those that function in plant hormone biosynthesis and signaling, cell division and plant secondary tissue growth. These results provide useful information for further characterizations of the regulatory network in plant secondary cell wall formation.     

Disruption of Cortical Microtubules by Overexpression of Green Fluorescent Protein-Tagged α-Tubulin 6 Causes a Marked Reduction in Cell Wall Synthesis

It has been known that the transverse orientation of cortical microtubules （MTs） along the elongation axis is essential for normal cell morphogenesis, but whether cortical MTs are essential for normal cell wall synthesis is still not clear. In the present study, we have investigated whether cortical MTs affect cell wall synthesis by direct alteration of the cortical MT organization in Arabidopsis thaliana. Disruption of the cortical MT organization by expression of an excess amount of green fluorescent protein-tagged a-tubulin 6 （GFP-TUA6） in transgenic Arabidopsis plants was found to cause a marked reduction in cell wall thickness and a de- crease in the cell wall sugars glucose and xylose. Concomitantly, the stem strength of the GFP-TUA6 overexpressors was markedly reduced compared with the wild type. In addition, expression of excess GFP- TUA6 results in an alteration in cell morphogenesis and a severe effect on plant growth and development. Together, these results suggest that the proper organization of cortical MTs is essential for the normal synthesis of plant cell walls.     

Effects of Nutrient Limitation on Pigments in Thalassiosira weissflogii and Prorocentrum donghaiense

The response of Prorocentrum donghaiense and Thalassiosira weissfiogii pigments under nitrate （N） and phosphate （P） limitation were studied using HPLC and in vivo fluorescence protocols in batch cultures. For P. donghaiense, the pigment ratio was kept stable under different nutrient conditions from the results of HPLC. For T. weissflogii, there was a lower ratio of chlorophyllide to Chl a during the exponential phase, but the reverse during the stationary phase. Different members of the phytoplankton had different pigments response mechanisms under nutrient limitation. From the results of in vivo fluorescence, the ratio of peridinin to Chl a for P. donghaiense increased in nutrient-free culture, while it was kept stable for nutrient-limited cultures during the exponential phase. For T. weissflogii, the ratio of fucoxanthin to Chl a for each culture increased during the exponential phase, but the ratio under N limitation was apparently lower than that for P limitation during the stationary phase. The results indicate that both pigment ratios from HPLC and in vivo fluorescence of To weissflogii were changed greatly under different nutrient conditions, which suggests that both ratios could be used as indicators of algal physiological status in different nutrient conditions.     

Clues to the signals for chloroplast photo-relocation from the lifetimes of accumulation and avoidance responses

Chloroplast photo-relocation movement is crucial for plant survival; however, the mechanism of this phenomenon is still poorly understood. Especially, the signal that goes from photoreceptor to chloroplast is unknown, although the photoreceptors(phototropin 1 and 2) have been identified and an actin structure(chloroplast actin filaments) has been characterized that is specific for chloroplast movement. Here,in gametophytes of the fern Adiantum capillus-veneris,gametophores of the moss Physcomiterella patens, and leaves of the seed plant Arabidopsis thaliana, we sought to characterize the signaling system by measuring the lifetime of the induced response. Chloroplast movements were induced by microbeam irradiation with high-intensity blue light and recorded. The lifetime of the avoidance state was measured as a lag time between switching off the beam and the loss of avoidance behavior, and that of the accumulation state was measured as the duration of accumulation behavior following the extinction of the beam. The lifetime for the avoidance response state is approximately 3–4 min and that for the accumulation response is 19–28 min. These data suggest that the two responses are based on distinct signals.     

Investigation of Combining Plant Genotypic Values and Molecular Marker Information for Constructing Core Subsets

In the present study, a strategy was proposed for constructing plant core subsets by clusters based on the combination of continuous data for genotypic values and discrete data for molecular marker InformaUon. A mixed linear model approach was used to predict genotyplc values for eliminating the environment effect. The ＂mixed genetic distance＂ was designed to solve the difficult problem of combining continuous and discrete data to construct a core subset by cluster. Four commonly used genetic distances for continuous data （Euclidean distance, standardized Euclidean distance, city block distance, and Mahalanobls distance） were used to assess the validity of the conUnuous data part of the mixed genetic distance; three commonly used genetic distances for discrete data （cosine distance, correlaUon distance, and Jaccard distance） were used to assess the validity of the discrete data part of the mixed genetic distance, A rice germplasm group with eight quantitative traits and information for 60 molecular markers was used to evaluate the validity of the new strategy. The results suggest that the validity of both parts of the mixed geneUc distance are equal to or higher than the common geneUc distance. The core subset constructed on the basis of a combination of data for genotyplc values and molecular marker information was more representative than that constructed on the basis of data from genotypic values or molecular marker informaUon alone. Moreover, the strategy of using combined data was able to treat dominant marker informaUon and could combine any other continuous data and discrete data together to perform cluster to construct a plant core subset.     

Effects of hunger and tannic acid on food intake and foraging behavior in Microtus fortis

The influence of hunger and plant secondary compounds on food selection and foraging behavior in Microtus fortis were measured in this study. The three selected kinds of tannic acid food were measured with the cafeteria method. The voles were first offered with food ad libitum for a period of 4 days. Food intake was recorded daily for calculation of average food intake by each vole. The calculated average food intake was multiplied by a hunger index (0, 25, 50, 75 or 100). Measurements were made over 4 consecutive nights for 2–4 feeding bouts per animal per night. The results indicated that hunger increased food intake, but had no significant effects on food selection. 0% tannic acid food was the most preferred food, and the intake of 6% tannic acid food was the least. Ingestion rate and bite size of voles increased with severity of hunger, but the feeding frequency was not significantly affected by hunger, and the time of feeding bout increased slowly. When the amount of food that voles have been offered is 25% more than that when they freely access to the food, the time of the feeding bout of hungry voles was significantly increased. These results suggested that voles increased their food intake mainly by increasing bite size when they were in hunger. The changes in foraging behavior indicated that herbivores increased their bite sizes and food intake rate to satisfy their nutritional demands rather than prolonged their foraging time and decreased the time for defending or reproductive activities to increase the amount of food intake.     

Use of the modified viral satellite DNA vector to silence mineral nutrition-related genes in plants: silencing of the tomato ferric chelate reductase gene, FRO1, as an example

Virus-induced gene silencing (VIGS) is potentially an attractive reverse-genetics tool for studies of plant gene function, but whether it is effective in silencing mineral nutritional-related genes in roots has not been demonstrated. Here we report on an efficient VIGS system that functions in tomato roots using a modified viral satellite DNA (DNAmβ) associated with Tomato yellow leaf curl China virus (TYLCCNV). A cDNA fragment of the ferric chelate reductase gene (FRO1) from tomato was inserted into the DNAmβ vector. Tomato roots agro-inoculated with DNAmβ carrying both a fragment of FRO1 and TYLCCNV used as a helper virus exhibited a significant reduction at the FRO1 mRNA level. As a consequence, ferric chelate reductase activity, as determined by visualization of the pink FeBPDS3 complex was significantly decreased. Our results clearly demonstrated that VIGS system can be employed to investigate gene function associated with plant nutrient uptake in roots.     

Epigenetic reprogramming of embryos derived from sperm frozen at −20°C

Cryopreservation of spermatozoa is a strategy that has been used to conserve the sperm of animal species and animal strains that are valuable for biomedical research. A simple method for preserving spermatozoa after application of intracytoplasmic sperm injection (ICSI) is much needed. It has been shown previously that spermatozoa frozen at 20°C can activate oocytes and support full-term embryo development. However, epigenetic reprogramming could be affected by the environment and by the in vitro manipulation of gametes. Here, we investigated embryo epigenetic reprogramming including DNA methylation and histone modification, in embryos derived from sperm preserved at 20°C without cryoprotectants. The results showed that although both fertilization and embryo developmental competence were decreased, the dynamic epigenetic reprogramming of embryos derived from frozen sperm was similar to the reprogramming of embryos derived from fresh sperm. The results reported in this study indicate that sperm frozen without cryoprotectant is epigenetically safe for ICSI.     

Chlorophyll in desiccated seeds of a euhalophyte, Suaeda physophora, and its significancy in plant adaptation to salinity during germination

The seed cotyledons of a euhalophyte, Suaeda physophora, were found to be dark green. The pigment extracted from the cotyledons was proved to be chlorophyll for the absorption spectra curve of extracts the same as that for leaves. Photosynthetic oxygen-exchanging could be detected after the seeds were moistened for 6 h. Microstructure of organelles in cotyledons of ungerminated seeds was detected by transmission electron microscopy. Histochemical pigmentation was used to investigate the degree of damage on the membrane of radicles. A xerophyte, Haloxylon persicum, whose cotyledons of desiccated seeds also contain chlorophyll was used as a comparative species. The results showed that S. physophora maintained the ultra-structure of chloroplasts, the integrity of plasma membranes of radicles kept much better than that of H. persicum, which showed the great adaptability to salinity of the euhalophyte even at the seed-germination stage. Seeds were incubated in 0 and 700 mmol/L NaCl for 10 days in darkness at 20 ℃ , then ungerminated seeds in NaCl solution were transferred to deionised water and reincubated for another 8 days and was recorded the germination recovery. Dry seeds moistened with deionised water germinated gradually in the dark and germination was maximal after 3 or 4 d. Seeds at 700 mmol/L NaCl for 10 d and transferred back to deionised water germinated abruptly, most seeds germinated in one day and photosynthesis was also detected. It is concluded that recovery germination of S. physophora ensured the seeds could germinate rapidly after salinity declines, for example under the mild but transitory favorable edaphic condition in early spring. Photosynthesis function in seeds might also promoted radicle growth and seedling establishment.     

Target organ-specific inactivation of drug metabolizing enzymes in kidney of hamsters treated with estradiol

Chronic treatment of hamsters with estradiol for several months has previously been shown to decrease the specific content of cytochrome P450 in the kidney, a target of hormonal carcinogenesis, but not in liver. The reason for this decrease in metabolic enzyme activity is unknown and has been examined in this investigation. We now report that the decrease in specific content of renal cytochrome P450 by 73% in response to estradiol was not affected by co-treatment with tamoxifen for 1 month. The subcutaneous infusion of 250 μg/day estradiol for 7 days lowered renal cytochrome P450 by 71% from control values and was therefore used for further mechanistic studies. This treatment decreased renal activities of estradiol 2- or 4-hydroxylase by 77 to 80%, of 7-ethoxycoumarin-O-deethylase by 66% of control values, respectively, and completely eliminated aryl hydrocarbon hydroxylase activities, whereas liver enzymes remained unaffected. After 7 days of infusion of estradiol, fluorescent products of lipid peroxidation were more than doubled in hamster kidney but remained unchanged in liver. The possibility of enzyme destruction by binding of estradiol 2,3-quinone to metabolizing enzymes was investigatedin vitro. In the presence of 2-hydroxyestradiol, cumene hydroperoxide, and microsomes, conditions known to favor the oxidation of the steroid to quinone, the binding of catechol estrogen metabolite to microsomal protein increased 60 fold over control values in the absence of cofactor. Purified rat liver cytochrome P450c also oxidized 2-hydroxyestradiol to 2,3-estradiol quinone. The rate of oxidation was linear for the first 2–3 min, but thereafter decreased with time. Under these incubation conditions, irreversible binding of catechol estrogen metabolite to cytochrome P450c increased for the first 2–3 min and then remained at this plateau level. It was concluded that enzyme destruction by a reactive estrogen metabolite or by lipid peroxides may be a major reason for the organ-specific decrease in cytochrome P450 enzymes in kidneys of estrogen-treated hamsters.     

Prasinoxanthin-constaining Prasinophyceae Discovered in Jiaozhou Bay, China

The class Prasinophyceae （Chlorophyta） contains some photosynthetic eukaryotic ultraplankton species characterized by containing prasinoxanthin. The existence and abundance of these organisms can be estimated by the diagnostic pigment. We detected the unique pigments of prasinoxanthin-containing Prasinophyceae in Jiaozhou Bay, China using high performance liquid chromatography （HPLC）. This was the first finding of this kind in Chinese seas. Using the ratio of prasinoxanthin to chlorophyll a, the abundance of prasinoxanthin-containing Prasinophyceae has been calculated. The average contribution of prasinoxanthin-containing Prasinophyceae to the chlorophyll a pool was 8.5% and 17.0% in May and August 2004 in Jiaozhou Bay, and the maximums were 25.9% and 36.3%. Size fractionated pigment analysis suggested that more than 80% of prasinoxanthin were in the fraction of 2-20 μm. According to the results of pigment and morphological analysis, the possible genera of prasinoxanthin-containing Prasinophyceae and the reasons for causing this high abundant phytoplankton in Jiaozhou Bay were discussed. This kind of phytoplankton can not be discovered in traditional biological investigation, but its contribution to the coastal ecosystem is significant enough to be studied further.     

咖啡种子中的内生真菌

Green coffee seeds from Colombia, Guatemala, India, Kenya, Papua New Guinea, Puerto Rico and Vietnam were sampled for the presence of fungal endophytes. Stions of surface sterilized seeds were plated on yeast malt agar, and fungal growth was isolated for subsequent DNA extraction and sequencing. Several fungal genera were isolated, including Acremonium, Aspergillus, Eurotium, Fusarium, Gibberella, Penicillium, Pseudozyma and an undescribed clavicipitaceous species. The biological activities that these fungi might be playing in coffee seeds remain unknown, but in other plants some of the genera isolated have been reported to protect against plant pathogens.     

An Effective Method for Raising Antisera Against β-defensins： Double-copy Protein Expression of mBinlb in E. coli

Binlb is a rat epididymis specific β-defensin which may have fertility related functions in addition to its antimicrobial activity. β-defensins are cysteine-rich cationic antimicrobial peptides that have their important implications in innate and adaptive immunity. Though considerable numbers of new β-defensins have been discovered, few corresponding antibodies have been reported. The small peptide with special structure and antimicrobial nature of β-defensins make them very difficult to express in prokaryotic system. Here we adopted a double-copy protein expression scheme based on which not only the mBin 1 b protein was successfully expressed but also the immunity of the antigen was enhanced. The validity of the antisera was verified by using Western blotting and immunohistochemical analyses. It will be a useful tool for deeply investigating the roles of Bin 1b and also provide a simple but effective method in raising antisera against other members of the β-defensin gene family.