睡莲叶脐着生胎芽与叶片不同部位碳水化合物代谢的关系

为了解睡莲叶脐胎芽的发育机理,以广热带亚属胎生睡莲‘玛格丽特’和‘鲁比’为材料,非胎生睡莲‘粉星’为对照,采用石蜡切片技术观察叶脐胎芽发育4个时期的形态变化,并比较胎生与非胎生睡莲叶片不同部位碳水化合物代谢的差异性。结果表明: 叶片展开后,胎生睡莲叶脐表皮以下细胞不断分裂和生长,形成排列紧密的细胞群,并逐渐向上凸起呈球形,非胎生睡莲叶脐则无任何变化。随着叶片的不断发育,除蔗糖和酶活性以外,各生理指标在胎生睡莲叶片中的含量均表现为先升后降,显著高于非胎生睡莲;从不同部位来看,碳水化合物含量总体表现为叶片>叶脐>叶柄(淀粉含量除外,其叶脐高于叶片和叶柄);不同品种蔗糖代谢酶活性表现为蔗糖合成酶(SS)和酸性转化酶(AI)活性高于蔗糖磷酸合成酶(SPS)和中性转化酶(NI)活性,胎生睡莲不同组织中的SPS和NI活性均显著高于非胎生睡莲,但SS和AI活性在胎生睡莲中并未表现出明显的品种优势;AI活性在品种间差异大,NI活性在品种间差异较小,且在不同组织中均处于较低水平。相关性分析表明,‘鲁比’叶片的蔗糖含量与SPS和AI呈极显著正相关,与NI呈显著正相关,且蔗糖含量的积累主要增加了睡莲叶片的SS和NI活性,有利于促进胎芽的形成。     

风信子试管苗芽体培养中的全息现象初探

风信子试管苗,取其芽体及切块在离体培养时存在全息现象,主要表现为,在风信子芽体的不同位置的切段上。不定芽发生的数量和频率呈梯度变化;在风信子芽体的不同大小的切段上,循着切段越小不定芽的分化数越少这一规律变化,这些现象都遵循了生物全息律。适当的激素浓度对全息胚的表现具有一定的促进作用。     

Selection<Emphasis Type="Italic">of nptll</Emphasis> transgenic sweetpotato plants using G<Subscript>418</Subscript> and paromomycin

We have used two aminoglycosides, G₄₁₈ and paromomycin, to develop a reliable selection system fornptll transgenic sweet-potato (Ipomoea batatas (L.) Lam.). Embryogenic calli derived from shoot apical meristems were bombarded with gold particles coated with pCAMBIA2301, which contained thenptll andgusA genes. When compared on a kill curve that was based on calli proliferation and cell viability, G₄₁₃-selection proved to be more efficient and had fewer escapes than kanamycin. These bombarded expiants were then selected on G₄₁₈-containing media. The total time required from bombardment to plant establishment in soil was seven to nine months. Multiple copies of the transgene were integrated into the sweetpotato genome. Northern analysis confirmed transgene expression in the regenerated plants, and a paromomycin assay demonstrated that thenptll gene was functionally expressed in transformed sweetpotato. These molecular analyses and assays all showed that selection with G₄₁₈ and paromomycin is reliable. So far, we have produced 69 transgenic events with this system, at a transformation frequency of approx. 1.1%. That efficiency is based on the number of transgenic plants obtained and the amount of calli bombarded. Thus, this selection method that combines G₄₁₈ with paromomycin is now available for selectingnptll transgenic sweetpotato.     

Apical sorting by galectin-3-dependent glycoprotein clustering

Epithelial cells are characterized by their polarized organization based on an apical membrane that is separated from the basolateral membrane domain by tight junctions. Maintenance of this morphology is guaranteed by highly specific sorting machinery that separates lipids and proteins into different carrier populations for the apical or basolateral cell surface. Lipid-raft-independent apical carrier vesicles harbour the beta-galactoside-binding lectin galectin-3, which interacts directly with apical cargo in a glycan-dependent manner. These glycoproteins are mistargeted to the basolateral membrane in galectin-3-depleted cells, dedicating a central role to this lectin in raft-independent sorting as apical receptor. Here, we demonstrate that high-molecular-weight clusters are exclusively formed in the presence of galectin-3. Their stability is sensitive to increased carbohydrate concentrations, and cluster formation as well as apical sorting are perturbed in glycosylation-deficient Madin-Darby canine kidney (MDCK) II cells. Together, our data suggest that glycoprotein cross-linking by galectin-3 is required for apical sorting of non-raft-associated cargo.     

Functional genomics and the biosynthesis of artemisinin

Artemisinin, a sesquiterpene lactone endoperoxide derived from the glandular secretory trichomes (GSTs) of Artemisia annua, provides the basis for the most effective treatments of malaria. The biology and biochemistry of GSTs of the Asteraceae and their biosynthesis of isoprenoids is reviewed. Recent efforts to understand the biosynthesis of artemisinin in A. annua GSTs are discussed in detail. This includes the development in the authors' laboratory of an expressed sequence tag (EST) approach to identifying the relevant biosynthetic genes using isolated GST as a source of mRNA. This has lead to the isolation of a cDNA encoding CYP71AV1, a multifunctional cytochrome P450 which catalyzes multiple oxidations of the sesquiterpene intermediate amorpha-4,11-diene to artemisinic acid. Further biochemical and molecular genetic work is required to elucidate the precise route from artemisinic alcohol to artemisinin and to engineer more efficient low cost production of artemisinin-based antimalarial drugs.     

A comparative analysis of the <Emphasis Type="Italic">Arabidopsis</Emphasis> mutant <Emphasis Type="Italic">amp1-1</Emphasis> and a novel weak <Emphasis Type="Italic">amp1</Emphasis> allele reveals new functions of the AMP1 protein

Ethylene and gibberellins have a synergistic stimulatory effect on hypocotyl elongation of light-grown Arabidopsis thaliana (L.) Heynh. seedlings. A screen for mutants with decreased response to these hormones led to the isolation of a novel allele (amp1-7) of the ALTERED MERISTEM PROGRAM (AMP) 1 locus. The amp1-7 allele contains a missense mutation causing a phenotype, which is weaker than that of the amp1-1 mutant that carries a nonsense mutation. The mutant phenotype prompted the hypothesis that AMP1 is involved in ethylene and GA signalling pathways or in a parallel pathway-controlling cell and hypocotyl elongation and cellular organization. Amp1 mutants contain higher zeatin concentrations causing enlargement of the apical meristem, which was confirmed by cytokinin application to wild type seedlings. Light grown amp1 seedlings have shorter hypocotyls than wild type; however, application of cytokinins promotes hypocotyl elongation of both Col-0 and amp1. We suggest that in amp1 mutants either zeatin overproduction or its action is strictly localized. Nelson J. M. Saibo and Wim H. Vriezen contributed equally to this work.     

慢性根尖周炎应用氢氧化钙根管内封药的临床疗效观察

目的:观察慢性根尖周炎患牙用氢氧化钙根管内封药的临床疗效。方法:选择100例慢性根尖周病变患者随机分为实验组和对照组各50例;观察组用氢氧化钙做根管内封药;对照组用甲醛甲酚做根管内封药,观察术后1周的临床及X线片表现,并分析评定临床疗效。结果:术后1周有效率比较,观察组为96%,对照组为86%;两组间疗效差异具有统计学意义。结论:氢氧化钙是较理想的根管内消毒剂。     

Antifungal triazole derivative triadimefon induces ectopic maxillary cartilage by altering the morphogenesis of the first branchial arch

BACKGROUND: The triazole derivative, triadimefon (FON), induces branchial arch abnormalities in post-implantation rat embryos cultured in vitro, and cranio-facial malformations in mouse fetuses. Ectopic maxillary cartilage has been also described as a typical FON-related malformation. This work studies the morphogenesis of the ectopic cartilage in rat embryos and fetuses exposed in vivo to FON during the early postimplantation period. METHODS: Pregnant rats were treated with 0, 250, and 500 mg/kg FON on Day 9.5 of pregnancy (D9.5) and sacrificed at term (D20), during the early fetal period (D17) or at different embryogenetic periods (D10, D11, D12). The skeleton was examined after stain of bone and cartilage or of cartilage alone respectively at term or at D17. The neural crest cell (NCC) migration and compaction was investigated at D10 and D11 and the cranial nerve organization described at D12. RESULTS: Triadimefon is teratogenic in rats under the chosen experimental conditions. The malformations were at the level of the cranio-facial and axial skeleton at term and of the hindbrain nerves in embryos. A NCC abnormal migration and compaction was observed at the level of the first branchial arch: in FON-exposed embryos NCC were detected at the level of both maxillary and mandibular processes, whereas control embryos showed the immunostained tissue only at the level of the mandibular bud. CONCLUSIONS: The pathogenic pathway, proposed to explain the ectopic cartilage, is the displacement of part of the NCC-derived tissues at the maxillary region of the first branchial arch.     

Genotypic variation in phenological plasticity: Reciprocal common gardens reveal adaptive responses to warmer springs but not to fall frost

Species faced with rapidly shifting environments must be able to move, adapt, or acclimate in order to survive. One mechanism to meet this challenge is phenotypic plasticity: altering phenotype in response to environmental change. Here, we investigated the magnitude, direction, and consequences of changes in two key phenology traits (fall bud set and spring bud flush) in a widespread riparian tree species, Populus fremontii. Using replicated genotypes from 16 populations from throughout the species’ thermal range, and reciprocal common gardens at hot, warm, and cool sites, we identified four major findings: (a) There are significant genetic (G), environmental (E), and GxE components of variation for both traits across three common gardens; (b) The magnitude of phenotypic plasticity is correlated with provenance climate, where trees from hotter, southern populations exhibited up to four times greater plasticity compared to the northern, frost‐adapted populations; (c) Phenological mismatches are correlated with higher mortality as the transfer distances between provenance and garden increase; and (d) The relationship between plasticity and survival depends not only on the magnitude and direction of environmental transfer, but also on the type of environmental stress (i.e., heat or freezing), and how particular traits have evolved in response to that stress. Trees transferred to warmer climates generally showed small to moderate shifts in an adaptive direction, a hopeful result for climate change. Trees experiencing cooler climates exhibited large, non‐adaptive changes, suggesting smaller transfer distances for assisted migration. This study is especially important as it deconstructs trait responses to environmental cues that are rapidly changing (e.g., temperature and spring onset) and those that are fixed (photoperiod), and that vary across the species’ range. Understanding the magnitude and adaptive nature of phenotypic plasticity of multiple traits responding to multiple environmental cues is key to guiding restoration management decisions as climate continues to change.