Wounding coordinately induces cell wall protein, cell cycle and pectin methyl esterase genes involved in tuber closing layer and wound periderm development

Little is known about the coordinate induction of genes that may be involved in agriculturally important wound-healing events. In this study, wound-healing events were determined together with wound-induced expression profiles of selected cell cycle, cell wall protein, and pectin methyl esterase genes using two diverse potato genotypes and two harvests (NDTX4271-5R and Russet Burbank tubers; 2008 and 2009 harvests). By 5 d after wounding, the closing layer and a nascent phellogen had formed. Phellogen cell divisions generated phellem layers until cessation of cell division at 28 d after wounding for both genotypes and harvests. Cell cycle genes encoding epidermal growth factor binding protein (StEBP), cyclin-dependent kinase B (StCDKB) and cyclin-dependent kinase regulatory subunit (StCKS1At) were induced by 1 d after wounding; these expressions coordinated with related phellogen formation and the induction and cessation of phellem cell formation. Genes encoding the structural cell wall proteins extensin (StExt1) and extensin-like (StExtlk) were dramatically up-regulated by 1-5 d after wounding, suggesting involvement with closing layer and later phellem cell layer formation. Wounding up-regulated pectin methyl esterase genes (StPME and StPrePME); StPME expression increased during closing layer and phellem cell formation, whereas maximum expression of StPrePME occurred at 5-14 d after wounding, implicating involvement in later modifications for closing layer and phellem cell formation. The coordinate induction and expression profile of StTLRP, a gene encoding a cell wall strengthening "tyrosine-and lysine-rich protein," suggested a role in the formation of the closing layer followed by phellem cell generation and maturation. Collectively, the genes monitored were wound-inducible and their expression profiles markedly coordinated with closing layer formation and the index for phellogen layer meristematic activity during wound periderm development; results were more influenced by harvest than genotype. Importantly, StTLRP was the only gene examined that may be involved in phellogen cell wall thickening after cessation of phellogen cell division.     

Studies on Endosperm Development and Deposition of Storage Reserves in Coix lacryma-jobi

The transition from free nuclear to cellular endosperm of Coix lacryma-jobi was eompleted 2 days after pollination. By 3 days after pollination the central cell was filled with endosperm cells. At first all cells of endosperm underwent division, later cell division was limited mainly in the peripheral region. 10 days after pollination the epidermal layer ceased its periclinal division and became the aleurone layer. Cell division persisted in the subepidermal 'cambium-like layers until the caryopsis nearly matured. Ceils of the inner region of endosperm became enlarged. Several layers of transfer cells were formed at the basal part of the endosperm. Starch grains appeared in endosperm cells on the 9th day after pollination. 10 days after pollination, lipid bodies occurred in the aleurone layer and the underlying layers. 13 and 15 days after pollination, the small vacuoles of aleurone cells contained protein and 20 days after pollenation they became aleurone grains. By 15 days after pollination pro tein bodies were formed in starch endosperm. Storage reserve deposition continued until the grain ripened. A correlation between endosperm and emoryo development was also observed.     

Expression of DNA methyltransferases is involved in Quercus suber cork quality

Cork oak (Quercus suber) is an important Portuguese species, mainly due to the economic value of the cork it produces. Cork results from phellogen, a meristematic tissue, which can locally produce lenticels or have discontinuities, originating “defects”: pores and nail inclusions that are detrimental to cork industrial use. Epigenetic processes control plant development and its deregulation can lead to altered phenotypes; therefore, the study of epigenetic players in the phellogen is important to understand the emergence of cork's defects. DNA methyltransferases (DNMTs) and one protein associated to MET1 (DMAP1) were characterized in Q. suber, and their gene expression was analyzed in phellogen and contiguous differentiating cell layers of trees producing high and low quality cork, after the evaluation of their defects by physical and image analysis methods. All classes of DNMTs (MET, DRM, and CMT) with the respective canonical motifs were identified in Q. suber. The expression analyses of these genes showed that QsDRM2 was the most active methyltransferases in the cells analyzed, and that all the genes were differentially expressed in trees with distinct cork quality, with a tendency for higher expression levels in low quality producers. Interestingly, the global methylation level was higher in cells with low expression of DNA methyltransferases. A positive and significant correlation was obtained between QsDMAP1 gene expression and the percentage of cork defects. This work provides the first evidence that cork quality in Q. suber is likely influenced by epigenetic mechanisms.     

薏苡胚乳发育及营养物质积累的研究

薏苡 ( Coix lacryma- jobi)授粉后 2 d,游离核胚乳已转变为细胞胚乳。授粉后 3d,中央细胞被胚乳细胞充满。起初 ,全部胚乳细胞均进行分裂 ,一定时期后 ,细胞分裂主要发生在胚乳周边区。授粉后 1 0 d,表皮停止平周分裂变为糊粉层 ,内方的数层形成层状细胞行平周分裂直到颖果接近成熟。胚乳内部生长则依赖于细胞体积扩大。胚乳基部 (颖果基部的胚乳 )形成了数层传递细胞。授粉后 9d,淀粉积累。授粉后 1 0 d,糊粉层及其内方数层细胞产生了脂体 ,后者的脂体以后又消失。授粉后 1 3、1 5 d,糊粉层细胞的液泡积累蛋白质。授粉后 2 0 d,液泡变为糊粉粒。授粉后 1 5 d淀粉胚乳细胞产生蛋白质体 ,营养物质积累持续到颖果成熟。还观察了胚和胚乳发育的对应关系。     

The Importance of Phellogen Cells and their Structural Characteristics in Susceptibility and Resistance to Excoriation in Immature and Mature Potato Tuber (Solanum tuberosum L.) Periderm

Potato tuber (Solanum tuberosum L.) periderm maturation is animportant physiological process that directly affects the susceptibilityand development of resistance to costly excoriation (skinning-typewounds) at harvest. The objectives of this research were toidentify the specific types of cells and the cellular changesassociated with susceptibility and resistance to tuber excoriationin immature and mature tubers respectively. Epifluorescent microscopicexamination of immature tuber periderm (phellem, phellogen andphelloderm cells) from several genetically diverse cultivarshas shown that the cellular damage resulting from excoriationoccurs within the phellogen (cork cambium), a meristematic layerof cells that gives rise to neighbouring phellem and phellodermcells. Tuber excoriation is the result of the fracture of radialphellogen cell walls linking the skin (phellem) to the phelloderm.As the tuber periderm matures, phellogen cells become inactiveand the radial walls of these cells become more resistant tofracture; resistance to excoriation develops. Ultrastructuralstudies of immature tuber periderm show that radial walls ofactive phellogen cells are thin and fragile. During peridermmaturation, both radial and tangential phellogen cell wallsthicken as they strengthen and become resistant to fracture,thereby providing resistance to excoriation. These results refuteprevious theories of the physiological changes responsible forthe onset of resistance to tuber skinning injury. The combinedresults establish a paradigm whereby the thickening and strengtheningof tuber phellogen cell walls upon periderm maturation are thedeterminant for resistance to tuber excoriation. Copyright 2001Annals of Botany Company Cambium, meristematic, periderm, phellem, phelloderm, phellogen, potato, skinning, Solanum tuberosum L., 0tuber     

miRNA profiling in leaf and cork tissues of Quercus suber reveals novel miRNAs and tissue-specific expression patterns

The differentiation of cork (phellem) cells from the phellogen (cork cambium) is a secondary growth process observed in the cork oak tree conferring a unique ability to produce a thick layer of cork. At present, the molecular regulators of phellem differentiation are unknown. The previously documented involvement of microRNAs (miRNAs) in the regulation of developmental processes, including secondary growth, motivated the search for these regulators in cork oak tissues. We performed deep sequencing of the small RNA fraction obtained from cork oak leaves and differentiating phellem. RNA sequences with lengths of 19–25 nt derived from the two libraries were analysed, leading to the identification of 41 families of conserved miRNAs, of which the most abundant were miR167, miR165/166, miR396 and miR159. Thirty novel miRNA candidates were also unveiled, 11 of which were unique to leaves and 13 to phellem. Northern blot detection of a set of conserved and novel miRNAs confirmed their differential expression profile. Prediction and analysis of putative miRNA target genes provided clues regarding processes taking place in leaf and phellem tissues, but further experimental work will be needed for functional characterization. In conclusion, we here provide a first characterization of the miRNA population in a Fagacea species, and the comparative analysis of miRNA expression in leaf and phellem libraries represents an important step to uncovering specific regulatory networks controlling phellem differentiation.     

Phytotoxic material from associations betweenErwinia amylovora and pear tissue culture: Possible role in necrotic symptomatology of fireblight disease

Pear cell suspension cultures (PCSC) inoculated with virulent strains of the fireblight bacterium,Erwinia amylovora, exhibited massive necrosis within 7 days, whereas avirulentE. amylovora strains and other enterobacteria generally elicited very slight or no necrotic reactions. These results were generally repeated when pear seedlings (1–2 months old) were inoculated with these same bacterial strains. Fractions derived from massively necrotized PCSC were tested for biological activity against healthy PCSC and pear seedling cuttings. Of these fractions, the dialyzable, 70%-ethanol-soluble material—remaining after precipitation by ethanol of high-molecular-weight polysaccharides (“amylovorin”)—exclusively inhibited growth of pear callus, plasmolyzed and disintegrated pear cells in PCSC, and caused some browning of pear callus. Although cuttings of young pear seedlings placed in solutions containing this dialyzable, ethanol-soluble material generally became massively necrotized within 3 days, some residual methodological problems with this bioassay procedure must be solved. These observations suggest that some relatively small molecule(s) formed in PCSC inoculated with virulentE. amylovora can exert antagonistic biological activity against pear tissue and may play a role in the symptomatology characteristic of the fireblight disease.     

NaCl处理对空心莲子草营养器官解剖结构的影响

利用光学显微技术,对不同浓度NaCl处理后的空心莲子草的营养器官进行形态解剖学研究。结果表明:不同浓度的NaCl处理下生长的空心莲子草解剖结构与对照相比发生了显著变化,这些变化表现为:随着盐浓度的逐渐增加,叶片面积变小而厚度增加;茎的横向生长受抑制;角质膜进一步增厚;营养器官的通气组织进一步发达,根、茎、叶中的胞间隙或孔隙紧密相连,使植物体上下贯通进行气体交换;根中的木栓细胞层数增加。     

Formation of Interfamilial Cell Symplast During Cell Co-Culture in Vitro

Two cell lines, white tobacco (Nicotiana tabacum) cell line and green carrot (Daucus carota) cell line, each with very distinct cellular structure markers by which the two cell lines could be identified at levels of callus cells, with light microscopy and electron microscopy, were established and used for interfamilial cell co-culture in which callus cells were well mixed and finely dispersed and treated with K+ hypotonic solution. Variegated interfamilial chimeral calli were observed after 10 to 15 days of co-culture. An isolation layer was formed and became thickened at the interface between the two attached unrelated callus cells and the heteroplastic cell wall complex was gradually established. Then the isolation layer became thinned and disappeared and plasmodesmata were formed secondarily within the thinned region and the interfamilial cell symplast was established. The wall in the region with isolation layer was about twice as thick as the fused cell wall of the symplastic. The developmental process of the interfamilial cell symplastic connection was discussed and it was suggested that (1) the nonspecific formation of isolation layer as initial adhesion between the two attached unrelated cells was the prerequisit for symplastic connection de novo formation, and (2) the specific cell recognition leading to disappearance or thickening to lignification or suberization of isolation layer and formation of plasmodesmata started within the isolation layer.     

Histological study of in vitro development of adventitious buds on leaf explants of oriental beech (Fagus orientalis lipsky)

Summary Adventitious shoots were induced on the proximal portion of leaves excised from Fagus orientalis shoot cultures derived from a 2-mo.-old or a 4-yr-old seedling. Up to 90% of the explants formed between 13 and 19 buds after culture on Woody Plant Medium containing 2.9 μM indole-3-acetic acid and 4.5 μM thidiazuron. Adventitious buds developed mostly on petiole stub callus, but also on the midvein. The histological events leading to shoot organogenesis were examined. Some shoots developed directly from subepidermis or epidermis, but most originated indirectly from cell file proliferation produced by periclinally dividing cells subadjacent to the epidermis. Some cells in the outermost layers of these files became meristematic and divided extensively, resulting in the formation of meristemoids after 16 d of culture. Dedifferentiation into meristematic cells, which exhibited a large, prominent nucleus, densely-stained cytoplasm, and a high nucleus-to-cell area ratio, was generally associated with anticlinal divisions in cells previously originated by periclinal division. Subepidermal cell proliferation occurred mainly in the abaxial surface of the explant, which initially formed a diffuse cambium and later evolved to a phellogenic cambium. Some meristemoids were also differentiated by lenticel phellogen. Organized cell divisions in meristemoids gave rise to bud primordia that emerged from the explant surface and differentiated a protoderm. The progressive structural differentiation of the apical meristem, leaf primordia, and procambial strands led, after about 28 d of culture, to shoots with vascular connections with treachery elements previously differentiated in adjacent tissues.     

离体细胞共培养中科间细胞共质体的形成

离体培养下选出的绿色胡萝卜（Ｄａｕｃｕｓ ｃａｒｏｔａ）细胞系和白色普通烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ ）细胞系,各自具有独特的细胞结构标志,在愈伤组织、光镜和电镜３ 个水平上均可区分。对两个细胞系进行分散、混合、Ｋ＋ 液低渗处理后在固体培养基上共培养,１０—１５ ｄ 后可观察到两种细胞的镶嵌生长。光镜和电镜下均观察到烟草细胞和胡萝卜细胞之间隔离层的存在与消失。在隔离层消失的区域可见到异种细胞间次生胞间连丝的形成,从而将独立的两个共质体连成一个统一的共质体。对科间细胞共质联系的建立过程进行了讨论,认为细胞接触后首先非特异粘连——以隔离层形成并适度加厚为标志,然后特异的细胞识别在隔离层中启动,从而导致隔离层或消失而重新建立共质联系或加厚、木质化、木栓化     

缺铁和矫治缺铁对梨树叶片结构的影响

通过对不同缺铁黄化程度的酥梨叶片进行解剖结构分析和矫治缺铁后叶片解剖结构的恢复情况的研究,结果表明,缺铁黄化使叶片厚度,叶肉厚度,栅栏组织厚度,栅栏组织／海绵组织的比值均明显下降;栅栏组织细胞排列疏松,细胞变短甚至断裂成块状,叶绿体数量明显减少甚至无,并出现解体细胞;栅栏组织与海绵组织界限模糊,海绵组织细胞排列变密;维管束的导管口径变小,维管束鞘和韧皮部的细胞番红染色加重。进行缺铁黄化矫治后,叶片结构恢复正常。这些结构的变化,可作为梨树缺铁诊断和矫治的指标。     

Morphogenetic determination of the dermoepidermal interface during tail regeneration in Rana catesbeiana.

During regeneration of the amputated tadpole tail, reconstruction of the epithelial basal lamina and basement lamella occurs only after the other major morphogenetic processes are well established. At 4 days after tail transection of the bullfrog tadpole, electron microscopy of the internal surface of the basal cell layer of the blastemal epithelium reveals it to be relatively free of extracellular matrix. By 11 days a basal lamina of distinct regularity has formed, and the first rodlets and fibers signaling the replacement of the collagenous basement lamella are identified. At 15 days the basal cells of the epithelium start to exhibit specialization of their internal cell surfaces: Hemidesmosomes and associated tonofilaments appear, and the adepidermal globular layer is formed. Orthogonal packing of collagen plies begins by 19 days after transection, the number of layers exceeding 22 in the latter stages of regeneration.     

库尔勒香梨果实石细胞发育及其对果实肉质影响初探

采用植物解剖学方法对库尔勒香梨果实石细胞形成过程和分布状况进行观察研究,结果表明,库尔勒香梨果实中石细胞从化后15d开始形成,随着时间的推移石细胞的直径和密度逐渐增大,花后110d左右直径达到最大值,花后50d左右密度达到高峰．之后随着果实的增大与成熟度的提高,石细胞团的密度表现下降的趋势。石细胞的分布特点是石细胞团以近果心和近果皮处分布密度高,直径大,中层果肉中石细胞团密度低,直径小。与其它品种相比,成熟库尔勒香梨的行细胞团密度最低,含有较大石细胞的含量相对较少,肉质较细。     

Regeneration of Tissues in Wounded Stems: A Quantitative Study

When a dicotyledonous stem is wounded by longitudinally splittinga young internode into halves, cells near the cut surface proliferateto form a callus within which vascular tissues differentiateand tend to restore a vascular cylinder in each half. Threephases of regeneration after wounding were identified and quantifiedin stems of three Solanaceous species. (1) In an initial ‘lag’phase, lasting about 2 d, neither cell division nor enlargementwere detected, but mitotic figures were observed within about300 µm of the cut surface. (2) Throughout a second, ‘division’phase, from about days 2–10, cell division and enlargementoccurred. Both were initiated mainly in the two cell layersnearest the surface. A mass of callus formed, with new cellwalls mostly parallel to the surface. Cell enlargement laggedbehind cell division for the first few days, so that mean radialcell diameter decreased until day 6, thereafter remaining almostconstant at 30–40 µm. Towards the end of this phase,mitoses ceased within the callus except in the positions ofthe future vascular and cork cambia, where radial cell diameterfell towards a constant 15–20 µm. (3) During a third,‘differentiation’ phase, cell division was restrictedto the cambial zones, and derivatives differentiated into cork,phloem or xylem according to position. The rate of increasein cell number per transect was 1.5–2.0 cells d^–1,of which more than half was xylem. Capsicum annuum L., sweet pepper, Lycopersicon esculentum Mill., tomato, cambium, cell division, differentiation, regeneration, wounding of stems, xylem     

Ultrastructure of Ascodichaena rugosa on beech bark

Ascodichaena rugosa Butin is a corkinhabiting fungus, found frequently on the bark of Fagus sylvatica L. The hyphae of the fungus are distributed solely in the phellem cells, stopping their growth in the last-formed cork cell layer. The cell to cell invasion is effected by penetration hyphae, causing no extensive dissolution of the cork wall. Electron microscopical observations revealed fine structural details of the fruit bodies and of the intracellular hyphae. Of special interest were the finger-like hyaline hyphae in the last-formed layer of cork cells, which are interpreted as haustoria on the basis of the fine structure both of hyphae and host cells. This situation is considered as reflecting a parasitic relationship of Ascodichaena to beech bark. The activity of the fungus led also to the increased production of cork cells, perhaps related to the nutrient supply of the fungus.     

The development of the tuber in seedlings of five species of Dioscorea from Nigeria

Tubers in all five species develop from the hypocotyl region of the seedlingS. A perivascular cambium arises cutting off mainly starch-storing parenchyma and collateral vascular bundles to the inside. A phellogen gives rise to cork on the outside. Between the two cambial layers there may or may not be layers of parenchyma, not storing starch but containing raphides. The vascular bundles consist of xylem with vessels, scalariform tracheids and parenchyma; and phloem, with sieve tubes and parenchyma.     

银杏雌性生殖器官发育过程的显微观察

对银杏(Ginkgo biloba)雌性生殖器官的发育过程进行了连续显微观察.结果表明:功能大孢子经过大约1个月的分裂形成约5000个游离核后开始细胞化.授粉后约45天近珠孔端两侧各产生1个颈卵器母细胞.授粉后约50天.颈卵器母细胞平周分裂形成初生颈细胞和中央细胞.授粉后约55天,初生颈细胞垂周分裂形成2个扁平状次生颈细胞,之后次生颈细胞体积逐渐增大并突入颈卵器腔.授粉后约130天,2个次生颈细胞斜向分裂形成4个颈细胞,中央细胞不均等分裂形成腹沟细胞和卵细胞.套细胞起源于颈卵器母细胞的周围细胞,授粉后70天至受精作用发生前,套细胞内不断积累营养物质,且套细胞与中央细胞间的细胞壁以及套细胞之间角隅处的细胞壁均出现明显增厚现象.在受精及胚胎早期发育过程中,套细胞内营养物质逐渐消失,细胞逐渐解体.授粉后55天,2个颈卵器之间的一些细胞向上突起形成帐篷柱,之后帐篷柱体积逐渐增加,并突入颈卵器腔.自授粉后120天至受精前帐篷柱细胞内开始积累大量营养物质,随后这些营养物质在受精过程中被逐渐消耗.到了原胚游离核后期,帐篷柱的顶端细胞发生变形并解体.