A Putative Calcium-Permeable Cyclic Nucleotide-Gated Channel, CNGC18, Regulates Polarized Pollen Tube Growth

A tip-focused Ca^2＋ gradient is tightly coupled to polarized pollen tube growth, and tip-localized influxes of extracellular Ca^2＋ are required for this process. However the molecular identity and regulation of the potential Ca^2＋ channels remains elusive. The present study has implicated CNGC18 （cyclic nucleotide-gated channel 18） in polarized pollen tube growth, because its overexpression induced wider and shorter pollen tubes. Moreover, CNGC18 overexpression induced depolarization of pollen tube growth was suppressed by lower extracellular calcium （[Ca^2＋]ex）. CNGC18-yellow fluorescence protein （YFP） was preferentially localized to the apparent post-Golgi vesicles and the plasma membrane （PM） in the apex of pollen tubes. The PM localization was affected by tip-localized ROP1 signaling. Expression of wild type ROP1 or an active form of ROP1 enhanced CNGC18-YFP localization to the apical region of the PM, whereas expression of RopGAP1 （a ROP1 deactivator） blocked the PM localization. These results support a role for PM-Iocalized CNGC18 in the regulation of polarized pollen tube growth through its potential function in the modulation of calcium influxes.     

Evidence for Inbreeding and Genetic Differentiation among Geographic Populations of the Saprophytic Mushroom Trogia venenata from Southwestern China

During the past 40 years, more than 400 Sudden Unexplained Deaths (SUDs) have occurred in Yunnan, southwestern China. Epidemiological and toxicological analyses suggested that a newly discovered mushroom called Trogia venenata was the leading culprit for SUDs. At present, relatively little is known about the genetics and natural history of this mushroom. In this study, we analyzed the sequence variation at four DNA fragments among 232 fruiting bodies of T. venenata collected from seven locations. Our ITS sequence analyses confirmed that all the isolates belonged to the same species. The widespread presence of sequence heterozygosity within many strains at each of three protein-coding genes suggested that the fruiting bodies were diploid, dikaryotic or heterokaryotic. Within individual geographic populations, we found significant deviations of genotype frequencies from Hardy-Weinberg expectations, with the overall observed heterozygosity lower than that expected under random mating, consistent with prevalent inbreeding within local populations. The geographic populations were overall genetically differentiated. Interestingly, while a positive correlation was found between population genetic distance and geographic distance, there was little correlation between genetic distance and barium concentration difference for the geographic populations. Our results suggest frequent inbreeding, geographic structuring, and limited gene flow among geographic populations of T. venenata from southwestern China.     

Insights into protective effects of medium additives on animal cells under fluid stresses: the hydrophobic interactions

Animal cells in suspension culture can suffer severe mechanical damage from bursting gas bubbles or other hydrodynamic force sources. Certain chemical additives in the culture media, particularly some surface-active chemicals, can effectively protect animal cells against such damage. Previously we proposed that the protective effect is associated with the adsorption of the additives in the cell membrane through hydrophobic binding of the surface-active molecules to the membrane. Adsorption of the additives to the cell membrane may lead to decreased hydrophobicity of the cell surface, thus eliminating cell adhesion to bubbles and reducing cell damage from bursting bubbles. In this study, we measured the hydrophobicity of two insect cell lines based on cell adhesion to hydrocarbon phase and its influence by surface-active chemicals, Pluronic F68, a methylcellulose and a polyethylene glycol. The experimental results showed strong support for the aforecited cell protection mechanism.     

Plasminogen Deficiency Causes Reduced Corticospinal Axonal Plasticity and Functional Recovery after Stroke in Mice

Tissue plasminogen activator (tPA) has been implicated in neurite outgrowth and neurological recovery post stroke. tPA converts the zymogen plasminogen (Plg) into plasmin. In this study, using plasminogen knockout (Plg^-/-) mice and their Plg-native littermates (Plg^+/+), we investigated the role of Plg in axonal remodeling and neurological recovery after stroke. Plg^+/+ and Plg^-/- mice (n = 10/group) were subjected to permanent intraluminal monofilament middle cerebral artery occlusion (MCAo). A foot-fault test and a single pellet reaching test were performed prior to and on day 3 after stroke, and weekly thereafter to monitor functional deficit and recovery. Biotinylated dextran amine (BDA) was injected into the left motor cortex to anterogradely label the corticospinal tract (CST). Animals were euthanized 4 weeks after stroke. Neurite outgrowth was also measured in primary cultured cortical neurons harvested from Plg^+/+ and Plg^-/- embryos. In Plg^+/+ mice, the motor functional deficiency after stroke progressively recovered with time. In contrast, recovery in Plg^-/- mice was significantly impaired compared to Plg^+/+ mice (p<0.01). BDA-positive axonal density of the CST originating from the contralesional cortex in the denervated side of the cervical gray matter was significantly reduced in Plg^-/- mice compared with Plg^+/+ mice (p<0.05). The behavioral outcome was highly correlated with the midline-crossing CST axonal density (R²>0.82, p<0.01). Plg^-/- neurons exhibited significantly reduced neurite outgrowth. Our data suggest that plasminogen-dependent proteolysis has a beneficial effect during neurological recovery after stroke, at least in part, by promoting axonal remodeling in the denervated spinal cord.     

Integrated regulation of the photosynthetic gene family from Arabidopsis thaliana in transformed tobacco cells

Summary Expression of the three chlorophyll a/b binding protein (cab) genes of Arabidopsis thaliana was studied in transformed tobacco tissues. For each cab gene, approximately 1000 bp of the promoter region plus a portion of the structural gene was inserted into a promoter-expression vector such that a translational fusion between the cab gene and the promoter-less chloramphenicol acetyltransferase (cat) gene was formed. The constructed molecules were introduced into either cultured tobacco cells or tobacco leaves and the promoter activity was monitored as chloramphenicol acetyltransferase activity. The light-grown tissues exhibited 1.5- to 60-fold greater promoter activity than did dark-grown tissues. Expression of the cab promoters was tissue specific: activities were much stronger in green leaves than other tissues. The cab promoters were almost equally active in transformed calli or shoots derived from leaves. However, in cultured tobacco cells, one promoter was two to three times stronger than the other two. The chimeric gene fusion, cab-cat, segregated in the F1 generation as a dominant Mendelian trait.     

Three distinct regulatory elements comprise the upstream promoter region of the nopaline synthase gene

Summary Fine deletion mutants were generated in the upstream control region of the nopaline synthase (nos) promoter to define the position and role of upstream regulatory elements. The results indicated that the 8 bp sequence (CAGAAACC) at -106/-113 and its inverted repeat (GGTTTCTG) at -140/-147 are important for promoter function. The downstream element appears more important than the upstream element since deletion of the former reduced promoter activity more significantly than deletion of the latter. Deletion of the element alone, however, did not abolish promoter function, whereas, deletion of the 10 bp potential Z-DNA-forming (Z) element located between the repeat elements nullified promoter activity. Therefore, it appears that the Z element is an essential upstream regulator and the repeated elements are upstream modulators of the nos promoter. These elements are functionally distinct since alteration of stereospecificity or insertion of short oligonucleotides between the elements did not significantly influence promoter activity. These regulatory elements were unable to function from 200 bp upstream of the CCAAT-TATA box region.     

猕猴桃属美味猕猴桃和毛花猕猴桃种间杂交的胚胎学和胚援救

本文报道美味猕猴桃(Actintdia delictosa cv.Hayward)(6x)和毛花猕猴桃(A.eria-ntha)(2x)杂交当代种子的胚胎学分析和胚援救的结果:1.根据杂交种子外部形态和胚胎发育程度,区分为正常的和败育的两类,其比率接近1:1。正常种子的胚发育分化正常,且含有适量胚乳。败育种子中,除少数不含胚和胚乳外,绝大多数种子的胚发育终止于球形、心形和早子叶阶段,胚体畸形。胚乳处于消失或完全解体。2.在被试8组培养基中,最适合胚萌发和生长的是:MS+IAA0.5ppm+GA_30.5ppm,MS+2ip2ppm+IAA0.5ppm+GA_30.5ppm和 MS+2ip2ppm+GA_30.5ppm。适于实生苗生长的培养基为 MS+GA_30.5ppm 和 MS 培养基。在 MS+BAP2ppm 和 MS+IAA0.5ppm+GA_30.5ppm 培养基上,一些正常种子和少数不正常种子胚的下胚轴直接形成了许多不定芽,从而诱导产生了更多的杂种植株。但是在MS十BAP2ppm+GA_30.5ppm,MS+BAP2ppm+IAA0.5ppm 和 MS+2ip2ppm+GA_30.5ppm培养基上,虽然胚产生了愈伤组织,但都未分化出器官。杂种实生苗根尖细胞近半数的含有近4倍性染色体数(4x=116),约半数为其他倍性。     

虫草成熟子囊壳、子囊与子囊孢子的电镜观察

虫草子囊壳壁为拟薄壁组织,基部与子座菌丝相联,顶端有一孔口,其内无缘丝,中心腔内无侧丝,基部着生的子囊垂直平行排列。子囊顶端中央是乳状突起,围以隆起的环状膜质边缘。成熟时,其顶端乳状突起膨大,膜质边缘也随之翻卷,中央有一小孔。同一子囊壳内子囊顶端发育阶段不同,说明子囊的形成不同步。子囊内含有两条平行的具横隔的子囊孢子。虫草子囊顶端形态结构即不象虫草属模式种蛹虫草,也不象头状虫草,说明子囊顶端的形态结构的种间差异。