Eucalyptus gunnii CCR and CAD2 promoters are active in lignifying cells during primary and secondary xylem formation in Arabidopsis thaliana

Cell-specific expression patterns of the Eucalyptus gunnii cinnamoyl coenzymeA reductase (EgCCR) and cinnamyl alcohol dehydrogenase (EgCAD2) promoters were analyzed by promoter-GUS histochemistry in the primary and secondary xylem tissues from floral stems and roots of Arabidopsis thaliana. Expression patterns indicated that the EgCCR and EgCAD2 genes were expressed in a coordinated manner in primary and secondary xylem tissues of the Arabidopsis floral stem and root. Both genes were expressed in all lignifying cells (vessel elements, xylem fibers and paratracheal parenchyma cells) of xylem tissues. The capacity for long-term monolignol production appeared to be related to the cell-specific developmental processes and biological roles of different cell types. Our results suggested that lignification of short-lived vessel elements was achieved by a two-step process involving (i) monolignol production by vessel elements prior to vessel programmed cell death and (ii) subsequent monolignol production by vessel-associated living paratracheal parenchyma cells following vessel element cell death. EgCCR and EgCAD2 gene expression patterns suggested that the process of xylem cell lignification was similar in both primary and secondary xylem tissues in Arabidopsis floral stems and roots.     

Structural analysis of subterranean organs in Zingiberaceae

Xylem structural characteristics are rarely used to separate tribes. This study aims to characterize the morphology of subterranean organs and determine both the occurrence and distribution of the vessel elements in ten species, belonging to Alpinieae and Zingibereae tribes. Species were from protected areas in the Brazilian Atlantic Rain Forest or from private cultivation areas. Roots and rhizomes processing followed both light and scanning electron microscopy procedures. According to morphological and anatomical characteristics, Euclidean distances among taxa were calculated, resulting in a dendrogram based on the unweighted pair group method with arithmetic mean (UPGMA) clustering. Despite our results indicating a similarity among all the species studied, they could be segregated by using qualitative and special metabolites characters in both the rhizome and the root. Vessel elements were found to be confined to the roots, presenting several basal characters in most of species, except for Alpinia purpurata and Etlingera species.     

白鲜根的发育解剖学研究

应用半薄切片、常规石蜡切片并结合离析法,对药用植物白鲜(Dictamnus dasycarpus Turcz.)根的发生发育过程进行了研究。结果表明:白鲜根的发生发育过程包括4个阶段,即原分生组织阶段、初生分生组织阶段、初生结构阶段以及次生结构阶段。原分生组织位于根冠内侧及初生分生组织之间,衍生细胞分化为初生分生组织。初生分生组织由原表皮、基本分生组织以及中柱原组成。原表皮分化为表皮,基本分生组织分化为皮层,中柱原分化为维管柱,共同组成根的初生结构;在初生结构中,部分表皮细胞外壁向外延伸形成根毛,皮层中分布有油细胞,内皮层有凯氏带,初生木质部为二原型或偶见三原型,外始式;根初生结构有髓或无。次生结构来源于原形成层起源的维管形成层的活动以及中柱鞘起源的木栓形成层的活动;白鲜次生韧皮部宽广,其中多年生根中可占根横切面积的85%,另外除基本组成分子外,还分布有油细胞;周皮发达,木栓层厚;初生皮层、次生木质部和次生韧皮部薄壁细胞中常充满丰富的淀粉粒。     

Effects of Inhibitory Concentrations of 3-Indolylacetic Acid and 3-Indolylacetonitrile on Cell Division and Tissue 2

Inhibitory concentrations of 3-indolylacetonitrile (IAN) cause,in cultured excised tomato roots, a marked decrease in the rateof cell division at the apical meristem but only a slight reductionin the lengths of mature exodermal and cortical cells. The reducedrate of cell division is associated with a decrease in the.number of meristematic cells at the root apex. By contrast,3-indolylacetic acid (IAA) causes marked reduction in the lengthsof mature cortical cells but does not markedly reduce cell-divisionrate at the apical meristem. Various lines of evidence indicate that both IAA and IAN causea relative increase in the number of longitudinal and a decreasein the number of transverse division walls in the meristematiczone of the root apex. Partial inhibition of the linear growth of excised tomato rootsby IAA and IAN is accompanied by increases in root and stelardiameters. These increases result from radial enlargement ofthe cortical cells and increase in the number of stelar cellsin the transverse section. The enlarged steles contain an increasednumber of lignified xylem elements, but only with the most inhibitoryconcentration of IAN (10^–4g./ml.) is there evidence ofthe development of secondary xylem. Both auxins increase significantlythe xylem vessel unit length.     

栓皮栎根系解剖结构、水力特性及碳、氮含量研究

为阐明栓皮栎根系随径级的变化规律,探究其细根的合理划分标准。以1年生栓皮栎幼苗为研究对象,将其根系分为1、1～2、2～3、3～4 mm四个径级,分别制作石蜡切片观察解剖结构,比较木质部水力特性,测定碳氮含量及其比值,并采用主成分法对根系进行分类。结果表明:(1)随着径级增加,栓皮栎根系周皮、韧皮部和形成层组织厚度增加而占径比降低,木质部直径及其占径比均增加。(2)直径2 mm以上的栓皮栎根系木质部平均最大和最小导管直径、根比导水率和栓塞脆弱性指数增加显著;而导管密度显著下降,导管面积与木质部面积之比变化不显著。(3)直径2 mm以上栓皮栎根系碳含量表现出显著增加,随着径级增加,根系氮含量下降、碳氮比升高。(4)主成分分析表明,13项根系结构和元素含量指标降维后,前2个主分量方差贡献率达62%,PCA双序轴显示栓皮栎根系可划分为2 mm以下的吸收根群和2 mm以上的运输根群。综上认为,以2 mm作为栓皮栎细根划分的标准兼顾了形态和功能的特点,更具有准确性。     

栽培太子参块根的发育解剖学研究

应用石蜡制片技术研究了栽培太子参纺锤状块根的发育过程。结果表明,栽培太子参的块根是由其不定根发育而成。太子参不定根的初生结构与次生结构的发育可分为4个阶段:原分生组织阶段、初生分生组织阶段、初生结构与次生结构阶段,类似一般草本双子叶植物根的发育。其特点是初生结构的皮层细胞大,仅3 ~4层,内皮层细胞具凯氏带;初生木质部多为三原型,少数为二原型、四原型与五原型。次生结构中次生木质部约占根面积80 %,主要为薄壁组织细胞,导管呈稀疏的放射状分布其中。由不定根发育成块根过程中,根据从根头至根尾不同距离的各组成部分的面积及细胞层数分析,从上向下其维管形成层活动强度不同,从而根的直径大小不同,使根发育成上粗下细的纺锤状肉质块根。高碘酸-Schiff反应显示,在成熟的块根中次生韧皮部的薄壁组织细胞和次生木质部射线间的木薄壁组织细胞内富含淀粉粒,在有些木薄壁组织细胞中还含有草酸钙簇晶。     

Age-related and origin-related control of the numbers of plasmodesmata in cell walls of developing Azolla roots

Plasmodesmata were counted in the longitudinal and transverse walls in developmental sequences of merophytes in roots of Azolla pinnata R.Br. The differences between certain categories of longitudinal wall were traced to factors that govern the surface area of the cell plates, the density of plasmodesmata (number per unit area of cell plate), and the amount by which each type of plate expands. No evidence for secondary augmentation of plasmodesmatal numbers after the cell-plate stage of development was found, but plasmodesmata are lost from the walls of sieve and xylem elements during their differentiation. Losses caused by cell separation occur in other tissues. The relatively high density of plasmodesmata in transverse walls is based not so much on a high density in the cell plates as on the relatively low expansion that these walls undergo. There appears to be a compensatory mechanism that relates initial plasmodesmatal density to the future expansion of the cell plate. The root shows determinate growth, the apical cell dividing about 55 times. Beginning at about the 35th division there is a progressive failure to maintain the plasmodesmatal frequencies that were developed in earlier cell divisions in the apical cell. The divisions that occur within the later-produced merophytes also show progressive diminution of plasmodesmatal numbers. The result is that the apex of the root, and particularly the apical cell, becomes more and more isolated symplastically, a phenomenon which could account for its limited lifespan and the determinate growth pattern of the root.     

An AGP epitope distinguishes a central metaxylem initial from other vascular initials in theArabidopsis root

Summary The developmental expression of an arabinogalactan protein (AGP) recognised by a monoclonal antibody, JIM 13, is described inArabidopsis roots. It is expressed in the single initial of the central metaxylem vessel that lies immediately above the four central cells of the quiescent centre. AGP expression spreads in a non-clona] fashion to neighbouring cell files as they mature. The AGP first appears in other pre-metaxylem elements and subsequently in the layer of parenchyma cells on either side of the xylem axis. Later, it spreads to the entire ring of eight endodermal cell files and to those pericycle cell files next to the xylem pole. It is postulated that this epitope reflects an underlying set of cell signalling events that may be involved in defining local positional values of cells important in establishing cellular patterns in the root.     

Halogenated Auxins Affect Microtubules and Root Elongation in Lactuca sativa

We studied the effect of 4,4,4-trifluoro-3-(indole-3-)butyric acid (TFIBA), a recently described root growth stimulator, and 5,6-dichloro-indole-3-acetic acid (DCIAA) on growth and microtubule (MT) organization in roots of Lactuca sativa L. DCIAA and indole-3-butyric acid (IBA) inhibited root elongation and depolymerized MTs in the cortex of the elongation zone, inhibited the elongation of stele cells, and promoted xylem maturation. Both auxins caused the plane of cell division to shift from anticlinal to periclinal. In contrast, TFIBA (100 micromolar) promoted elongation of primary roots by 40% and stimulated the elongation of lateral roots, even in the presence of IBA, the microtubular inhibitors oryzalin and taxol, or the auxin transport inhibitor naphthylphthalamic acid. However, TFIBA inhibited the formation of lateral root primordia. Immunostaining showed that TFIBA stabilized MTs orientation perpendicular to the root axis, doubled the cortical cell length, but delayed xylem maturation. The data indicate that the auxin-induced inhibition of elongation and swelling of roots results from reoriented phragmoplasts, the destabilization of MTs in elongating cells, and promotion of vessel formation. In contrast, TFIBA induced promotion of root elongation by enhancing cell length, prolonging transverse MT orientation, delaying cell and xylem maturation.     

Taproot promoters cause tissue specific gene expression within the storage root of sugar beet

The storage root (taproot) of sugar beet (Beta vulgaris L.) originates from hypocotyl and primary root and contains many different tissues such as central xylem, primary and secondary cambium, secondary xylem and phloem, and parenchyma. It was the aim of this work to characterize the promoters of three taproot-expressed genes with respect to their tissue specificity. To investigate this, promoters for the genes Tlp, His1-r, and Mll were cloned from sugar beet, linked to reporter genes and transformed into sugar beet and tobacco. Reporter gene expression analysis in transgenic sugar beet plants revealed that all three promoters are active in the storage root. Expression in storage root tissues is either restricted to the vascular zone (Tlp, His1-r) or is observed in the whole organ (Mll). The Mll gene is highly organ specific throughout different developmental stages of the sugar beet. In tobacco, the Tlp and Mll promoters drive reporter gene expression preferentially in hypocotyl and roots. The properties of the Mll promoter may be advantageous for the modification of sucrose metabolism in storage roots.     

Morpho-anatomical responses ofTrigonella foenum graecum Linn. to induced cadmium and lead stress

Effect of different concentrations of cadmium (0, 5,15, 30, 50 μg/g of soil) and lead (0,25, 50,100,200 μg/g of soil) on morphological and anatomical features ofTrigonella foenum graecum Linn, was studied at pre-flowering, flowering and post flowering stages. Morphological attributes, like number of leaves per plant, total leaf area of the plant and single leaf area, dry mass of stem, root and leaf, length of shoot, root and plant, size of stomata and stomatal pore, and the density of stomata on both epidermises were significantly reduced under metal stress at all the developmental stages. Trichome length on both epidermises increased while their density decreased under metal stress. Under cadmium stress, proportion of pith and vasculature decreased but cortex increased in the stem. Under lead stress, proportion of pith and vasculature increased but cortex decreased in the stem. In the root, proportion of vasculature and pith increased and cortex decreased in response to both cadmium and lead stresses. Dimensions of vessel element and xylem fibre in the wood of stem and root decreased under the cadmium and lead stresses. Decrease in density of vessel element in the stem and root with advancement of age was more pronounced in plants grown under cadmium and lead stresses.     

Root hydraulic conductivity and vessel structure modification with increasing soil depth of two oak species: Quercus pubescens and Quercus robur

There are numerous studies on water transport characteristics of trees from the base to tops, but only few deal with the variation in xylem conduit diameters from shallow to deep roots. This study compares variation in root conduit properties as a function of increasing soil depths for two oak species (Quercus pubescens Willd and Quercus robur L.) growing on two different plots. We measured root vessel characteristics at three soil depths including 0, 50 and 100 cm, and calculated the associated root-specific hydraulic conductivities. Vessel diameter and specific hydraulic conductivity increased with increasing soil depth from 0 to 50 cm, but did not change in the deeper soil layer in both species. We conclude that freeze–thaw events in upper soil layer limit vessel diameters and thus hydraulic conductivity of roots.     

The role of hormones and gradients in the initiation of cortex proliferation and nodule formation in Pisum sativum L.

Summary The effect of exogenous phytohormones on proliferation of the root cortex, and their relation to the division factors from Rhizobium which participate in the initiation of root nodules, were studied using explants of root-cortex tissue from 7-day-old, sterile pea plants. The explants were cultured for 7 days on a synthetic nutrient medium supplemented with auxin, or auxin and cytokinin. With only auxin present in the medium, ca. 10% of the explants showed cell proliferation. With both auxin and cytokinin this percentage was much higher (ca. 80%). The active explants showed proliferation patterns which were similar to or could be derived from a pattern with three predominant meristematic areas in the inner cortex opposite the three xylem radii of the excised central cylinder. These proliferation patterns were similar to the initial proliferative stages in root-nodule formation in seedling intact roots. From this restricted division response of the explants to the hormones, a hypothesis of endogenous division factors is proposed. To test this hypothesis, extractions of root tissue were performed. The addition of a crude alcoholic extract from the central cylinder or the cortex to the medium resulted in cell divisions throughout the cortex. The results are interpreted as evidence for the presence of a transverse gradient system of (an) unknown cell-division factor(s) in the root cortex which may control the induction of cell divisions in nodule initiation brought about by the release of auxin and cytokinin from Rhizobium.     

桔梗根的发育解剖学研究

以桔梗(Platycodon grandiflorum A.DC)根为材料,运用石蜡切片和半薄切片法对其根的发育过程及结构进行解剖学观察,并对不同年限根的结构进行了比较。结果表明:桔梗根的结构发育过程包括原生分生组织、初生分生组织、初生生长和次生生长4个阶段。其原生分生组织由3群原始细胞组成,表现出典型分生组织的细胞学特征;初生分生组织包括根冠原、表皮原、皮层原和中柱原;初生结构由表皮、皮层和中柱组成,其中皮层薄壁细胞占主要地位,初生木质部为二原型;次生生长主要依靠维管形成层和木栓形成层的活动来完成,其次生结构从外到内由周皮和次生维管组织组成,次生维管组织占主导地位,其中以薄壁细胞为主,维管分子少量,分散在薄壁组织中。不同年限的根的结构基本相同,但它们在主根长度和直径、周皮厚度、木质部与韧皮部面积之比等方面存在差异。     

灰叶胡杨根蘖繁殖的形态解剖学特征

利用常规石蜡切片法对灰叶胡杨(Populus pruinosa)根蘖繁殖特性进行形态解剖学研究。结果表明: 灰叶胡杨横走侧根由周皮、次生维管组织和四原型的初生木质部构成, 具有次生维管组织中维管射线、次生韧皮薄壁组织发达的结构特征。灰叶胡杨的根蘖繁殖源于横走侧根上不定芽的发生及生长发育。不定芽起源于横走侧根的木栓形成层, 木栓形成层经细胞分裂活动形成不定芽原基, 不定芽原基细胞分裂和生长分化形成在横走侧根表面可观察到的不定芽, 进而生长发育为根蘖苗。不定芽的发生具有同步或非同步的时间特征和单点或多点聚集的空间分布特点, 在生长发育过程中其基部可以产生新的不定芽。不定芽发生、分布和生长特点是根蘖苗大小不一、密集丛生的内在原因, 表明灰叶胡杨具有较强的根蘖繁殖能力。     

勃氏甜龙竹不同部位根的解剖结构比较

该研究选择勃氏甜龙竹7月份(雨季)和11月份(干季)地面以下10 cm处土壤中秆基处生的根,秆下部节内(地上部分3~4节)和秆上部枝条基部在空气中自然生的根,以及7月份枝条高空压条产生的伸长阶段和成熟阶段的根作为实验材料,采用树脂切片和徒手切片法,对不同部位根的根尖和根毛区进行横切,甲苯胺蓝O和苏丹红7B染色,研究不同起源根的形态解剖结构,为竹子根解剖学研究提供新的理论信息。结果显示:(1)不同来源根的皮层宽度占根直径的比例、周缘纤维组织宽度及其细胞层数、内皮层细胞壁厚度均随生长时间延长而增加。(2)所有起源根的外皮层和内皮层细胞壁均有显著的木栓质沉积,但环内皮层木栓化程度存在显著差别。(3)测量不同来源根的后生木质部导管数目和直径发现,7月份采集的根导管数目更多、直径更大。(4)枝条基部自然生根的根尖与其他起源部位的根相比更为钝平,但根冠面积较小;秆下部节内自然生根非常硬,所以未能成功观察其根尖结构特点。研究认为,勃氏甜龙竹4种不同起源根的基本结构一致,但在各自的具体结构上还存在显著差别,可能与其生长的环境以及发育状况密切相关。     

Exogenous Auxin and N-1 -Naphthylphthalamic Acid Effects on Populus deltoides Xylogenesis

A method was developed for evaluating the empirical alterationof xylem vessel differentiation in the central leaf trace ofPopulus deltoides, a species that exhibits helical phyllotaxis.Effects of experimental treatments for a period of six plastochronswere evaluated by vessel parameter ratios = 2.P_T/ (P_T+1 + P_T–1),where P was either vessel number or mean transverse vessel areameasured at mid-intern ode at Leaf Plastochron Indices of T– 1, T, and T + 1. Excising leaf laminae reduced vesselnumber and mean vessel area in the associated central leaf traceby 50% and 70%, respectively, compared to unexcised laminaecontrols. Replacing excised laminae with a concentration seriesof exogenous indoleacetic acid (IAA) resulted in a 5% increaseper log mol m^–3 of IAA in the number of vessels differentiatingin the associated central leaf traces compared to excised controls.Mean vessel areas within these leaf traces were 50% of thoseof intact leaf traces. No significant effects of different concentrationsof exogenously applied IAA on mean vessel area could be demonstrated.A lanolin paste ring of N-1 -naphthylphthalamic acid (NPA),an auxin transport inhibitor, around the petioles of intactleaves reduced the number of differentiating vessels by 7% andmean vessel area by 29% per log mol m^–3 of NPA comparedto central leaf traces of leaves ringed with plain lanolin paste.The results suggest that NPA treatments may be used to distinguishexperimentally, at least in part, the cell division from thecell enlargement phases of primary xylogenesis within centralleaf traces of P. deltoides stems. Key words: Auxin transport, Vessel area, Vessel number     

Vessel-diameter distribution in roots of grapevines (Vitis vinifera L. cv. Shiraz)

The distribution frequency patterns of diameter of xylem vessels and percentage of total predicted axial conductances were studied in 190-day and 212-day-old main roots of grapevine (Vitis vinifera L. cv. Shiraz) grown under well-watered and stressed conditions. The protoxylem were the first to mature and were responsible for most of the theoretical conductance in root segments between the tip and 2.5 cm from the tip. Some large xylem vessels retained cross walls and protoplasm up to 22.5 cm from the tip. Statistical tests using the Kolmogorov-Smirnov two sample test showed that the pattern of distribution frequency of xylem vessels classified in different diameter classes varied with distance from the root tip. The distribution frequency of xylem vessels was similar in both well-watered and stressed plants from the tip up to 15 cm from the tip. At distances further from the tip the distribution frequency of xylem vessels of well-watered plants was significantly different from that of stressed plants, with the former having more larger vessels than the latter. The pattern of vessel distribution frequency was different from that of percent total axial conductance (K_h) predicted with fewer large vessels carrying most of the axial flow.     

Expression of transferred genes during hairy root development in pea

Root border cell development and expression of reporter genes were evaluated in transgenic pea hairy roots. Successful induction of hairy roots in pea is conditioned by bacterial strain and plant genotype, as well as by developmental and environmental factors. Morphological changes sometimes occur when hairy roots are transferred from infected plants to tissue culture media, but such changes are confined to specific clones. Expression of reporter genes under the control of promoters from bean (Phaseolus vulgaris L.) stress genes encoding phenylalanine ammonia lyase and chalcone synthase were evaluated. Expression patterns vary between hairy roots taken directly from infected plants, and those grown in culture; most hairy roots taken from infected plants exhibit expression throughout all tissues, whereas expression in cultured hairy roots is most often localized to specific tissues. Patterns of expression that occur during different stages of hairy root development are very similar to those observed in transgenic plants expressing the same fusion genes. Border cell separation and release in hairy roots is normal, and expression of glucuronidase in border cells of some transgenic roots resulted in development of bright blue single cells. Cultured hairy roots should provide a very useful model for studying the effect of defined changes in root border cells on microbial associations with roots of this important legume.Abbreviations YEM yeast extract-mannitol - GUS glucuronidase - PAL phenylalanine ammonium lyase - CHS chalcone syntase     

Relationships among xylem transport, biomechanics and storage in stems and roots of nine Rhamnaceae species of the California chaparral

Here, hypotheses about stem and root xylem structure and function were assessed by analyzing xylem in nine chaparral Rhamnaceae species. Traits characterizing xylem transport efficiency and safety, mechanical strength and storage were analyzed using linear regression, principal components analysis and phylogenetic independent contrasts (PICs). Stems showed a strong, positive correlation between xylem mechanical strength (xylem density and modulus of rupture) and xylem transport safety (resistance to cavitation and estimated vessel implosion resistance), and this was supported by PICs. Like stems, greater root cavitation resistance was correlated with greater vessel implosion resistance; however, unlike stems, root cavitation resistance was not correlated with xylem density and modulus of rupture. Also different from stems, roots displayed a trade-off between xylem transport safety from cavitation and xylem transport efficiency. Both stems and roots showed a trade-off between xylem transport safety and xylem storage of water and nutrients, respectively. Stems and roots differ in xylem structural and functional relationships, associated with differences in their local environment (air vs soil) and their primary functions.