Isolation of a Tomato Protease that May Be Involved in Proteolysis of 1-Aminocyclopropane-l-Carboxylate Synthase

1-aminocyclopropane- 1-carboxylate （ACC） synthase is a principal enzyme that catalyses the committed step in phytohormone ethylene biosynthesis. Previous evidence indicates that the hypervariable C-terminus of ACC synthase is most likely to be processed proteolytically in vivo. However, the protease responsible has not been identified thus far. In the present study, we detected proteolytic activity against ACC synthase （LeACS2） in tomato （Lycopersicon esculentum Mill.） fruit extract based on a newly established in vitro assay system. Purification of the protease through DEAE, gel filtration and MonoQ chromatography resulted in considerable enrichment of a 64-kDa protein species. Subsequent biochemical analysis of the purified tomato protease revealed that the optimal conditions for its proteolytic activity were at pH 8.0 and at 37 ~C. In addition, the protease activity was blocked completely by the metalloprotease inhibitor 1,10-phenanthroline. The present study represents the first report on the isolation of an ACC synthase- processing protease from plant tissues.     

Large-Scale Analyses of Glycosylation in Cellulases

Cellulases are important glycosyl hydrolases (GHs) that hydrolyze cellulose polymers into smaller oligosaccharides by breaking the cellulose β (1→4) bonds,and they are widely used to produce cellulosic ethanol from the plant biomass.N-linked and O-linked glycosylations were proposed to impact the catalytic efficiency,cellulose binding affinity and the stability of cellulases based on observations of individual cellulases.As far as we know,there has not been any systematic analysis of the distributions of N-...     

The Mechanism of Yield Increasing Bacteria and Plant Ecological Engineering

Yield-Increasing-Bacteria (YIB) increased crop yields by two ways: 1) Ecological effect. The YIB produced the effect for rapid reproduction and resistance to extreme environmental conditions and the function of regulating the microflora on plant surfaces(including rhizosphere and phyllosphere).The improved microflora promoted plant growth and inhibited the plant pathogens and deleterioui rhizobzcteria(DRB.II) physiological effect.YIB produced growth rogulating substances,such as gibberellin and zeatin which increased the growth and yield of crops Based on related research in the past 20 years and current expleration of YIB,the concept of Plant Ecological Engineering was elucidated,i.e.changing the microflora artificially in and on plants so as to promote the exprossion of fine characteristies of plants and the prduction potantial, restraning the activity of plant pathogens and DRB,thus making the crop healthy.     

Two new triterpenoid saponins from Gymnema sylvestre

Two new oleanane-type triterpenoid saponins, gymnemoside-W1 and W2, together with seven known compounds were isolated from the leaves of Gymnema sylvestre R. Br. By means of spectral and chemical analysis, the structures of the new compounds were elucidated as 16 β-hydroxyl olean-12-en-3-O-[β-D-glucopyranosyl (1→6)-β.D-glucopyranosyl]-28-O-β-D-glucopyranoside(1) and 16 β,21 β,28-trihydroxyl-olean-12-ene-3-O-glucoronopyranoside (2). The EtOHIH2O extracts of this plant were shown to be able to inhibit glucose absorption in rats.     

特异切割番茄 1-氨基环丙烷羧酸合成酶mRNA的核酶基因序列的合成与克隆

根据番茄1-氨基环丙烷羧酸合成酶(ACC合成酶)基因cDNA序列, 按照锤头状核酶作用模式, 设计合成长度分别为40mer和48mer的一对引物,经体外退火、延伸、连接,插入质粒pGEM-3Zf(+)的KpnI和BamHI位点之间,经菌落原位杂交、双酶切、聚丙烯酰胺凝胶电泳和序列分析证明,人工合成了特异切割ACC合成酶mRNA第667-669位点GUC序列的核酶基因序列。 Abstract:In order to interrupting the ACC synthase activity in tomato,a hammerhead ribozyme targeting to cleave GUC triplet at the position 667~699 of ACC synthase mRNA was established and integrated into KpnI and BanHI site of vector pGEM-3Zf(+).Four positive clones were gotten by in situ hybridization.The recombinatant plasmids were digested with restriction endonuclease KpnI and BanHI.By polyacrylamide gels electrophoresis and sequence analysis,the size and the sequence of one of the digested product was the same as those of what we expected.These results showed that the ribozyme gene sequence has been obtained.     

Transformation of carnation with genes related to ethylene production and perception: towards generation of potted carnations with a longer display time

Potted carnation (Dianthus caryophyllus L. cv. Lillipot) plants were transformed with cDNAs for carnation 1-aminocyclopropane-1-carboxylate (ACC) synthase (DC-ACS1, s/aACS transgenes) or ACC oxidase (DC-ACO1, s/aACO transgenes) in sense or antisense orientation or mutated carnation ethylene receptor cDNA (DC-ERS2′) by Agrobacterium-mediated gene transfer. The presence of acetosyringone at 100 μM in media for shoot culture prior to leaf explant preparation and preculture of Agrobacterium in addition to the conventional method of addition to media for infection and coculture, and the use of water instead of nutrient media for infection and coculture increased the transformation efficiency to 4.0% compared to the 0.1% obtained by the conventional method. PCR analysis as well as Southern blot analysis confirmed the integration of the ethylene-related transgenes. Leaflet segments of cultured shoots of some lines transformed with s/aACO transgenes had less activity to convert ACC to ethylene than that of the non-transformed control plant, indicating that the integrated s/aACO transgenes reduced the expression of endogenous ACC oxidase gene (DC-ACO1) in the cultured shoots.     

BOTANICAL JOURNAL OF SOUTH CHINA CONTENTS Ⅰ-Ⅱ

No.I 1992,!New and noteworthy species of Myrsinaceae from China and Vietnam……………………………Hu Ofiming(1)A new Medidnal species of C_.hirita(G粼riaoeae)from Guangxi……Huan${~"fftngchun and Chert XitLxlang(14)Qvantitative studies on the seed plant flora of china I.Computer programs for the smtigical analysis of the seed plant flora·……·……·……-…·………………·……·……一Fu Dezhi,Li yi and Xue Yanhong(22)Studies On Epiphyllous Liverworts in China(V)一Ep.phyllous liverwo…     

Proteomics Study of COI1-regulated Proteins in Arabidopsis Flower

Jasmonates(JAs) are a new class of plant hormone that regulate expression of diverse genes to mediate various plant responses.The Arabidopsis F-box protein COI1 is required for plant defense and male fertility in JA signal pathway.To further investigate the regulatory role of COI1 in male fertility,we compared the proteomics profiles of Arabidopsis wild type(WT) flowers with coi1-1 mutan male-sterile flowers using two-dimensional difference gel electrophoresis coupled with matrix-assisted laser desoption/ionization-time-of-flight mass spectrometry.Sixteen proteins with potential function in specific biological processes such as metabolism processes and defense/stress responses were differentially expressed in WT and coi1-1 mutant flowers.Verification on a phi class glutathione transferase AtGSTF9,one out of these 16 identified proteins,revealed that the expression of AtGSTF9 was severely downregulated in flowers of coi1-1 mutant compared with that of WT.Further function analyses of these genes would provide new insights into the molecular basis of COI1-regulated male fertility.     

Chemical Constituents of Schisandra rubriflora Rehd. et Wils.

Schisandra rubriflora Rehd. et Wils. is a traditional Chinese medicine. To search for new and bioactive components from traditional Chinese medicines and provide scientific evidence for taxonomy, the chemical constituents of the plant were investigated by various column chromatography methods (silica gel, Sephadex LH-20, and RP-18). From the aerial parts of S. rubriflora, three new megastigmane glycosides,namely (3S, 5R, 6S, 9R)-megastigmane-3, 9-diol 3-O-[α-L-arabionfuranosyl-(1→6)-β-D-glucopyranoside](1), 7-megastigmene-3-ol-9-one 3-O-[α-L-arabionfuranosyl-(1→6)-β-D-glucopyranoside] (2), and megastigmane-3α, 4β, 9ξ-tfiol 3-O-β-D-glucopyranoside (3), along with 14 known compounds, were isolated.The structures of the new compounds were elucidated by a combination of spectroscopic and chemical methods.     

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.     

Prevalence of 1-aminocyclopropane-1-carboxylate deaminase in Rhizobium spp

This is the first report documenting the presence of 1-aminocyclopropane-1-carboxylate (ACC) deaminase in Rhizobium. This enzyme, previously found in free-living bacteria, yeast and fungi, degrades ACC, the immediate precursor of ethylene in higher plants. Thirteen different rhizobial strains were examined by Southern hybridization, Western blots and ACC deaminase enzyme assay. Five of them tested positive for ACC deaminase. Induction of the expression of ACC deaminase was examined in one of the positively tested strains, Rhizobium leguminosarum bv. viciae 128C53K. This rhizobial ACC deaminase had a trace basal level of expression without ACC, but could be induced by a concentration of ACC as low as 1 μM. The more ACC added to this Rhizobium the higher the expression level of the ACC deaminase. This revised version was published online in June 2006 with corrections to the Cover Date.     

Collections-based systematics in the new age of discovery: Celebrating the legacy and life of Professor Wen-Tsai Wang

Professor Wen-Tsai Wang (王文采, June 5, 1926–November 16, 2022) was an academician of the Chinese Academy of Sciences (CAS) and a legendary plant taxonomist at the Institute of Botany of CAS (Fig.1). Herein, we organize a virtual special issue in Journal of Systematics and Evolution (JSE) to celebrate the legacy and life of Professor Wang, who was a leading plant taxonomist in China and made important contributions toward advancing the understanding of the flora of China, the biogeography of eastern Asia, and biodiversity research in the vast Hengduan Mountains. He served as the Editor-in-Chief of Acta Phytotaxonomica Sinica (now JSE) for 6 years from 1982 to 1988, and trained several generations of plant taxonomists in China (Li,2001).     

Variation in the Agronomic and Morphological Traits of Iranian Chickpea Accessions

Landraces of chickpea (Cicer arietinum L.) in Iran have not been adequately characterized for their agronomic and morphological traits. Such characterization would be helpful in the development of improved cultivars, so in this study 362 chickpea accessions, collected from the major chickpea growing areas of Iran, were evaluated to determine their phenotypic diversity. High coefficients of variation (CVs) were recorded in pods/branch, seeds/pod, yield/plant, seeds/plant, pods/plant and branches/plant. Using principal component (PC) analysis, the first four PCs with eigenvalues more than 1 contributed 84.10% of the variability among accessions, whereas PC5 to PC10 were less than unity. PC1 was positively related to days to first maturity, days to 50% flowering and days to 50% maturity. The characters with the greatest weight on PC2 were seeds/plant and yield/plant, whereas PC3 was mainly related to pods/plant, seeds/pod and 100seed weight, and PC4 was positively related to pods/branch and negatively to branches/plant. The germplasm was grouped into four clusters using cluster analysis. Each cluster had some specific characteristics of its own and the cluster I was clearly separated from clusters II, III and IV. These accessions are an important resource for the establishment of a core collection of chickpeas in the world.     

Physical mapping of three fruit ripening genes：Endopolygalacturonase,ACC oxidase and ACC synthase from apple（Malus x domestica） in an apple rootstock A106（Malus sieboldii

The apple rootstock,A106(Malus sieboldii),had 17 bivalents in pollen mother cells at meiotic metaphase 1,and 17 chromosomes in a haploid pollen cell.Karyotypes were prepared from root-tip cells with 2n=34 chromosomes,Seven out of 82 karyotypes(8.5%) showed one pari of satellites at the end of the short arm of chromosome 3.C-bands were shown on 6 pairs of chromosomes 2,4,6,8,14,and 16 near the telomeric regions of short arms.Probes for three ripening-related genes from Malus x domestica:endopolygalacturonase(EPG,0.6kb),ACC oxidase(1.2kb),and ACC synthase(2kb)were hybridized in situ to metaphase chromosomes of A106.Hybridization sites for the EPG gene were observed on the long arm of chromosome 14 in 15 out of 16 replicate spreads and proximal to the centromere of chromosomes 6 and 11.For the ACC oxidase gene,hylridization sites were observed in the telomeric region of the short arm of chromosomes 5 and 11 in 87% and 81% of 16 spreads respectively,proxiaml to the centromere of chromosome 1 in 81% of the spreads,and on the long arm of chromosome 13 in 50% of the spreads. Physical mapping of three fruit ripening genes in an apple rootstock A106.Twenty five spreads were studied for the ACC synthase gene and hybridization sites were observed in the telomeric region of the short arm of chromosome 12 in 96% of the spreads.chromosomes 9 and 10 in 76% of the spreads,and chromosome 17 in 56% of the spreads.     

Simulated N and S deposition affected soil chemistry and understory plant communities in a boreal forest in western Canada

Aims We conducted a simulated nitrogen (N) and sulfur (S) deposition experiment from 2006 to 2012 to answer the following questions: (i) does chronic N and S deposition decrease cation concentrations in the soil and foliage of understory plant species, and (ii) does chronic N and S deposition decrease plant diversity and alter species composition of the understory plant community in a boreal forest in western Canada where intensifying industrial activities are increasing N and S deposition. Methods Our field site was a mixedwood boreal forest stand located ~100 km southeast of Fort McMurray, Alberta, Canada. The experiment involved a 2 × 2 factorial design, with two levels each of N (0 and 30 kg N ha^-1 yr^-1; applied as NH₄NO₃) and S addition (0 and 30 kg S ha^-1 yr^-1; applied as Na₂SO₄). Four blocks were established in July 2006, each with four plots of 20 × 20 m randomly assigned to the treatments. Soil and understory vegetation were sampled and cover (%) of individual species of herb (height ≤ 0.5 m) and shrub (height 0.5–1 m) layers was determined in August 2012. Important findings Seven years after the treatments began, N addition increased dissolved organic carbon and N in the mineral soil (P < 0.05), whereas S addition decreased exchangeable cations (P < 0.05) in the forest floor. In the shrub layer, species evenness, and overall diversity were decreased by N addition (P < 0.05) due to increases in abundance of nitrophilous species and S addition (P < 0.01) due to decreased cation concentrations in soils. Total shrub cover decreased with S addition (P < 0.10). Nitrogen and S addition affected neither species richness nor evenness in the herb layer. However, permutational multivariate analysis of variance and non-metric multidimensional scaling analyses (based on plant cover) indicated that the effect of N and S addition on understory plant species composition in the both shrub and herb layers was species-specific. Addition of N decreased foliar phosphorus and potassium concentrations in some species, suggesting potential risk of N-meditated nutrient imbalance in those species. Our results indicate that long-term elevated levels of N and S deposition can negatively impact plant nutrition and decrease the diversity of the understory plant community in boreal forests in northern Alberta, Canada. However, considering that the current N and S deposition rates in northern Alberta are much lower than the rates used in this study, N and S deposition should not negatively affect plant diversity in the near future.     

Plant villins:Versatile actin regulatory proteins

Regulation of actin dynamics is a central theme in cell biology that is important for different aspects of cell physiology.Villin, a member of the villin/gelsolin/fragmin superfamily of proteins, is an important regulator of actin. Villins contain six gelsolin homology domains(G1–G6) and an extra headpiece domain. In contrast to their mammalian counterparts, plant villins are expressed widely, implying that plant villins play a more general role in regulating actin dynamics. Some plant villins have a de fined role in modifying actin dynamics in the pollen Invitube; most of their in vivo activities remain to be ascertained.Recently, our understanding of the functions and mechanisms of action for plant villins has progressed rapidly, primarily due to the advent of Arabidopsis thaliana genetic approaches and imaging capabilities that can visualize actin dynamics at the single filament level in vitro and in living plant cells. In this review,we focus on discussing the biochemical activities and modes of regulation of plant villins. Here, we present current understanding of the functions of plant villins. Finally, we highlight some of the key unanswered questions regarding the functions and regulation of plant villins for future research.     

DEAP1 encodes a fasciclin-like arabinogalactan protein required for male fertility in rice

Arabinogalactan proteins(AGPs) are widely distributed in plant cells. Fasciclin-like AGPs(FLAs)belong to a subclass of AGPs that play important roles in plant growth and development. However,little is known about the biological functions of rice FLA. Herein, we report the identification of a male-sterile mutant of DEFECTIVE EXINE AND APERTURE PATTERNING1(DEAP1) in rice. The deap1 mutant anthers produced aberrant pollen grains with defective exine formation and a flattened aperture annulus and ex...     

Regulatory mechanisms of host plant defense responses to arbuscular mycorrhiza

Arbuscular mycorrhizal (AM) fungi colonize the roots of over 80% of terrestrial plant species, forming mutually beneficial symbioses. During the colonization process, symbiotic partners recognize each other, and undergo observable morphological and physiological changes; indicating that symbiosis formation involves multiple factors that are finely regulated. Sometimes host plants generate a transient, weak, defense response. This response and its down-regulation play a very important role in the development of AM symbioses. Although AM fungi can infect a wide range of host root tissues, which host defense may play a crucial role is hypothesized from the fact that hyphal expansion is only observed in the root cortex.
We discuss five defense mechanisms. (1) The degradation of exogenous elicitors. The host’s weak defense response may be due to the degradation of the exogenous elicitor chitin, or the prevention of release of an endogenous inductor from the plant cell wall. (2) The inactivation of defense signal molecules. Some defense signal molecules such as hydrogen peroxidase, salicylic acid (SA), and jasmonic acid (JA), are inactivated in host plants. This helps to avoid the turn-on of defense-related genes and facilitate mycorrhizal formation. (3) The regulation of plant hormones and plant photosynthates. Plant hormone levels and plant photosynthate metabolism both change during AM colonization. These mechanisms need further exploration. (4) Changes in levels of phosphorous (P), and (iso)flavonoids. High P levels can induce some defense genes to express hydrogen peroxidase, chitinase, and glucanase. These gene products can repress colonization by AM fungi. The plant defense response regulatory effect for different (iso)flavonoids varies, and their levels are regulated by P. (5) The suppressed expression of symbiotic genes. Some symbiosis-related genes inhibit plant defense responses, but it is still unclear which mechanisms underlie gene regulation. We provide here a theoretical basis for research into AM symbiosis that may promote study of host plant resistance and the mechanisms of symbiosis formation.
We provide a deeper insight into the signal transduction pathways of mycorrhization that will aid understanding and analysis of plant defense mechanisms in the AM context. The on-going development of genome sequencing technology will contribute greatly to the detailed study of symbiosis-related genes, and pathogenesis-related protein genes. These related genes may be induced to express corresponding proteins, be repressed, postpone expression or even shutdown, or both may work together to form symbioses. Elucidation of these features will help us understand the roles that plant defenses play in mycorrhizal formation; providing an unprecedented opportunity for research into mycorrhizal molecular biology and the interaction of symbiotic partners, and allowing the underlying mechanisms to be gradually uncovered.