Cell lineage analysis of maize bundle sheath and mesophyll cells

Maize leaves are divided into repeated longitudinal units consisting of vascular tissue, bundle sheath (BS), and mesophyll (M) cells. We have carried out a cell lineage analysis of these cell types using six spontaneous striping mutants of maize. We show that certain cell division patterns are preferentially utilized, but not required, to form the characteristic arrangement of cell types. Our data suggest that early in development a central cell layer is formed, most frequently by periclinal divisions in the adaxial subepidermal layer of the leaf primordium. Lateral and intermediate veins are initiated in this central layer, most often by divisions which contribute daughter cells to both the procambium and the ground meristem. These divisions generate "half vein" units which comprise half of the bundle sheath cells around a vein and a single adjacent M cell. We show that intermediate veins are multiclonal both in this transverse direction and along their lengths. BS cells are more closely related to M cells in the middle layer of the leaf than to those in the upper and lower subepidermal layers. An examination of sector boundaries has shown that photosynthetic differentiation in M cells is affected by the phenotype of neighboring BS cells.     

A Transmission and Cryo-Scanning Electron Microscopy Study of the Formation of Aerenchyma (Cortical Gas-Filled Space) in Adventitious Roots of Rice (Oryza sativa)

The initiation and development ofaerenchyma in adventitiousroots of rice (Oryza sativa) was studied in tissuc up to approximately36-h-old using transmission electron microscopy (TEM) and cryoscanningelectron microscopy (cryo-SEM). Aerenchyma resulting from selective cortical cell collapse isa naturally occurring feature of rice roots. Evidence of somecortical cell disruption was noticeable by TEM in cells thatwere approximately 6-h-old; it became more advanced as the cellsaged. At 12 h, early stages of cell wall breakdown and lossof cell turgidity were seen. Complete collapse of columns ofcells had occurred by 24 h. In tissue that was 36-h-old, thesenescent cytoplasm of many remaining cells began to disperse.The visual evidence suggests that cell collapse was the resultof autolysis. This pattern of cortical degeneration in ricewas dissimilar to that reported elsewhere for Zea roots grownin an oxygen depleted environment. Cryo-SEM revealed the occurrence of small structures withinthe cortex with the external appearance of miniature, intactcells which are not preserved during conventional SEM preparativeprocedures. Key words: Rice (Oryza sativa L.), roots, aerenchyma, ultrastructure     

云南野生稻叶茎根组织结构特性的比较研究

采用徒手切片法对云南3种野生稻和栽培稻的叶片、茎秆及根的组织结构进行比较研究,以明确野生稻的内部结构,为进一步揭示其结构与云南野生稻的生长势旺盛、营养吸收能力强、抗某些病虫害能力强的关系奠定基础。结果表明,(1)云南野生稻与栽培稻叶片主脉、茎秆及根的组织结构差异显著,其中景洪疣粒野生稻、景洪药用野生稻与栽培稻的差异最明显。(2)在叶主脉结构中,景洪疣粒野生稻无气腔结构,维管束数量少、面积小;景洪药用野生稻、3个普通野生稻材料存在多个维管束和气腔结构,维管束、束内导管直径及气腔面积较栽培稻大,而栽培稻中的气腔均为2个。(3)在茎秆结构方面,景洪疣粒野生稻茎秆最小,维管束数量最少,其茎壁内的维管束排列方式与栽培稻不同;景洪药用野生稻和普通野生稻茎秆及茎壁较栽培稻粗厚,维管束数量也较栽培稻多,普通野生稻的茎壁中有通气组织。(4)在根的组织结构中,3种野生稻的导管数量较多,导管直径及中柱面积较栽培稻大,外皮层出现了具有凯氏带功能的凯氏点等。     

The Effect of Salinity upon Photosynthesis in Rice (Oryza sativa L.): Gas Exchange by Individual Leaves in relation to their Salt Content

Yeo, A. R., Caporn, S. J. M.and Flowers, T. J. 1985. The effectof salinity upon photosynthesis in rice (Oryza sativa L.): Gasexchange by individual leaves in relation to their salt content.—J.exp. Bot. 36: 1240–1248. The effect of salinity upon net photosynthesis and transpirationby individual leaves of rice has been investigated by gas exchangemeasurements in seedlings at the five to six leaf stage. Salinitydid not, initially, reduce net photosynthesis in the whole plantbut only in the older leaves in which sodium accumulated. Analysisof the course of events in leaf four following salinizationof the medium showed that net photosynthesis was inversely correlatedwith the sodium concentration in the leaf tissue. There wasno evidence of a threshold effect; net photosynthesis declinedlinearly with increasing leaf sodium concentration and was reducedby 50% at only 05 mmol sodium per gram dry weight. The relationshipbetween transpiration rate and leaf sodium concentration closelyparalleled that for photosynthesis; there was no effect of leafsodium concentration on the carbon dioxide concentration inthe intercellular spaces, showing that sodium accumulation inthe leaf affected stomatal aperture and carbon dioxide fixationsimultaneously. Photosynthesis was reduced by half at a sodiumconcentration in the leaf which did not reduce the concentrationof chlorophyll. The nature of the effect of salinity upon leafgas exchange is discussed. Key words: Salinity, rice, Oryza sativa L., photosynthesis, apoplastic salt load     

Inhibition of Gibberellin-Induced Elongation Growth of Rice by Feruloyl Oligosaccharides

The biological activity of cell wall-derived feruloyl oligosaccharideswas investigated using a modified micro-drop bioassay. A feruloylarabinoxylan trisaccharide (FAXX) and a feruloyl arabinoxylantetrasaccharide (FAXXX) were found to inhibit the gibberellin-inducedelongation of dwarf rice (Oryza sativa L., cv, Tan-ginbozu)that had been treated with uniconazole (S-3307), an inhibitorof the biosynthesis of gibberellins. An arabinoxylan trisaccharide(AXX) was ineffective. The growth-inhibitory effecf of feruloyloligosaccharides depended on their feruloyl and glycosyl moieties.The amount of esterified diferulic acid residues in the cellwalls of the second leaf sheath of rice seedlings that had beentreated with FAXX was almost same as that of controls. Feruloyloligosaccharides did not inhibit the incorporation of [¹⁴C]leucineinto acid-precipitable proteins by suspension-cultured maizecells, whereas cinnamic acid and its derivatives strongly inhibitedsuch incorporation. (Received May 10, 1995; Accepted August 23, 1995)     

The Use of 14C-Ethane Diol as a Quantitative Tracer for the Transpirational Volume Flow of Water and an Investigation of the Effects of Salinity upon Transpiration, Net Sodium Accumulation and Endogenous ABA in Individual Leaves of Oryza sativa L.

Yeo, A. R., Yeo, M. E., Caporn, S. J. M., Lachno, D. R. andFlowers, T. J. 1985. The use of ¹⁴C-ethane diol as a quantitativetracer for the transpirational volume flow of water and an investigationof the effects of salinity upon transpiration, net sodium accumulationand endogenous ABA in individual leaves of Oryza sativa L.—J.exp. Bot. 36: 1099–1109. Oryza sativa L. (rice) seedlings growing in saline conditionsexhibit pronounced gradients in leaf sodium concentration whichis always higher in the older leaves than the younger ones.Individual leaf transpiration rates have been investigated todiscover whether movement of sodium in the transpiration streamis able to explain these profiles from leaf to leaf. The useof ¹⁴C labelled ethane diol to estimate transpiration was evaluatedby direct comparison with values obtained by gas exchange measurements.Ethane diol uptake was linearly related to the transpirationalvolume flow and accurately predicted leaf to leaf gradientsin transpiration rate in saline and non-saline conditions. ¹⁴C-ethanediol and ²²NaCl were used to compare the fluxes of water andsodium into different leaves. The youngest leaf showed the highesttranspiration rate but the lowest Na accumulation in salineconditions; conversely, the older leaves showed the lower transpirationrates but the greater accumulation of Na. The apparent concentrationof Na in the xylem stream was 44 times lower into the youngerleaf 4 than into the older leaf 1. Exposure to NaCl (50 molm^–3) for 24 h elicited an increase in endogenous ABA inthe oldest leaf only, but no significant changes occurred inthe younger leaves. Key words: —Salinity, rice, Oryza sativa L., transpiration, volume flow, abscisic acid     

Coleoptile Senescence in Rice (Oryza sativa L.)

We investigated the cellular events associated with cell deathin the coleoptile of rice plants (Oryza sativa L.). Seeds germinatedunder submergence produced coleoptiles that were more elongatedthan those grown under aerobic conditions. Transfer of seedlingsto aerobic conditions was associated with coleoptile opening(i.e. splitting) due to death of specific cells in the sideof the organ. Another type of cell death occurred in the formationof lysigenous aerenchyma. Senescence of the coleoptile was alsonoted, during which discolouration of the chlorophyll and tissuebrowning were apparent. DNA fragmentation was observed by deoxynucleotidyltransferase-mediateddUTP nick end labelling (TUNEL) assay, and further confirmedby the appearance of oligonucleosomal DNA ladders in senescentcoleoptile cells. Two nucleases (Nuc-a and Nuc-b) were detectedby in-gel-assay from proteins isolated from coleoptiles. Nuc-a,commonly observed in three cell death phases required eitherCa²⁺or Mg²⁺, whereas Nuc-b which appeared during senescencerequired both Ca²⁺and Mg²⁺. Both nucleases were strongly inhibitedby Zn²⁺. Copyright 2000 Annals of Botany Company Aerenchyma, rice, cell death, coleoptile, fragmentation, nuclease, Oryza sativa, senescence, split, submergence, TUNEL     

Distribution of Endogenous Gibberellins in Vegetative Shoots of Rice

Levels of endogenous gibberellins in rice seedlings (Oryza sativaL., cv. Nipponbare) were compared between young and old leavesat the 4- and 5-leaf stages. The levels of GA₁, GA₁₉ and GA₅₃were higher in the youngest leaf than in older leaves at the5-leaf stage, but they did not differ significantly betweenthe leaf sheath and the leaf blade. At the 4-leaf stage, thelevel of GA₁, was highest in the third leaf sheaths which containedyoung elongating tissues. These results indicate that gibberellinsare synthesized in young vegetative tissues to promote theirelongation growth. The levels of GA₁ in the youngest leaf sheathsof two cultivars of dwarf rice, Tan-ginbozu and Waito-C, wereapproximately 10% of that in the normal rice at the 5-leaf stage.This result could explain the retardation of shoot elongationin these dwarf cultivars. (Received February 15, 1995; Accepted June 1, 1995)     

The Vascular Pattern of Italian Ryegrass (Lolium multiflorum Lam.): 3. The Leaf Trace System, and Tiller Insertion, in the Adult

The leaf trace system in the region of congested internodesat the base of Lolium multiflorum is described. A typical major trace in a leaf consists of a collateral bundlehaving a double bundle sheath and incorporating a certain amountof sclerenchyma. As such a leaf trace is followed down intothe stem it increases in diameter, loses the inner (mestome)bundle sheath, and the xylem becomes associated with xylem transfercells. Lower down, the bundle diameter is reduced although nowit has become amphivasal. The internal xylem only is still associatedwith transfer cells. The proximal portions of the bundle aremuch reduced, transfer cells, mestome sheath, and sclerenchymaare lacking and the now insignificant bundle merges with a lowerleaf trace or some other vascular tissue. Such a bundle in thestem may be in direct contact via bridges with other leaf traces,with the nodal plexus, and with the peripheral plexus that surroundsthe inner leaf trace system. In the base of a typical young plant, approximately one-halfof all leaf traces, including all the median veins, join bundlesfrom the next oldest leaf. Approximately one-third join thenodal plexus, and the remainder variously join bundles fromthe same or next but one oldest leaf to join the peripheralplexus. The differentiation of tiller insertions into the pre-existingmain stem system is highly variable. In a very young tillera number of traces were seen to terminate before the main systemwas reached suggesting basipetal differentiation. The actualconnections made by the tiller traces may occur with any nearbyleaf trace, the nodal plexus, or with the peripheral plexus.Later differentiating leaf traces in a tiller join leaf tracesof the tiller itself. Occasional bundles from secondary tillers by-pass the vasculartissue of the primary tiller to join directly with that of theparent plant. Vascular connections between parent and tiller,although very variable, appear to be totally comprehensive froma functional standpoint.     

Anatomical and Chemical Alterations but not Photosynthetic Dynamics and Apoplastic Transport Changes are Involved in the Brittleness Culm Mutation of Rice

Brittleness culm is an important agronomic trait that has a potential usefulness in agricultural activity as animal forage although the developmental mechanism is not clear yet. In the present study, the anatomical and chemical characteristics as well as some ecophysiological features in the brittleness culm mutation of rice (Oryza sativa L.) were investigated. Compared with the wild type (WT), the brittleness culm mutant (bcm) exhibited higher culm vascular bundle distance and lower culm wall thickness, leaf interveinal distance and leaf thickness. Ratio of bundle sheath cell/whole bundle and areas of whole vascular bundles and bundle sheath of leaves were reduced while ratios of xylem and phloem to whole bundles were elevated in bcm. The Fourier transform infrared (FTIR) microspectroscopy analysis and further histochemical and physiological measurements revealed that the different contents and depositions of cell wall components such as pectins, lignin, suberin and cellulose all participated in the mutation of brittleness. However, the mutant presented no significant changes in leaf photosynthetic dynamics and apoplastic transport ability. These results strongly indicate that the alterations in anatomical and chemical characteristics, rather than changes in major ecophysiological features such as photosynthesis and apoplastic transport were involved in the brittleness mutation of rice.     

不同水稻品种叶片显微结构和超微结构的比较研究

以江苏省常规粳稻品种武育粳3号、武运粳7号和日本优质粳稻品种关东194为材料,利用光学显微镜和透射电子显微镜,研究了抽穗期叶片主脉的显微结构和超微结构。结果表明武育粳3号和武运粳7号叶片主脉的维管束数目和气腔数目均较多,叶绿体基粒片层排列紧密,中间无空隙。而关东194叶片主脉的维管束数目和气腔数目较少,叶绿体基粒片层排列疏松,中间有空隙。武育粳3号和武运粳7号的这种结构特点可能与其高产特性有关。     

Abscisic Acid Accumulation in Detached Leaves of Rice (Oryza sativa L.) in Response to Water Stress: A Correlation with Leaf Size

When water stress was imposed on detached leaves of two rice(Oryza sativa L.) cultivars, more ABA per unit fresh weightaccumulated in IR20, a small-leaved cultivar, than in 63–83,a large-leaved cultivar; the difference being up to threefold.In an F₂ population of a cross between the two cultivars ABAaccumulation was found to be significantly negatively correlatedwith leaf fresh weight. This correlation persisted in the F₃generation. Such a correlation was not evident, however, whena number of rice cultivars, which varied widely in leaf size,were examined. The difference in ABA accumulation between IR20 and 63–83was not accounted for by different spatial patterns of waterloss or ABA accumulation within a leaf, and cultivar differencesin ABA content were maintained both across, and at various positionsalong the leaf. No major differences in leaf anatomy were observed between thetwo cultivars. Differences found in leaf water relations characteristicswere few and generally minor. It therefore seems unlikely thatthese properties account for the difference between the cultivarsin the ability to accumulate ABA or for the correlation withleaf size. Oryza sativa L, rice, water stress, abscisic acid, leaf size     

Aerenchyma (Gas-space) Formation in Adventitious Roots of Rice (Oryza sativa L.) is not Controlled by Ethylene or Small Partial Pressures of Oxygen

Jackson, M. B., Fenning, T. M., and Jenkins, W. 1985 Aerenchyma(gas-space) formation in adventitious roots of rice (Oryza sativaL.) is not controlled by ethylene or small partial pressuresof oxygen.—J. exp. Bot. 36: 1566–1572. The extent of gas-filled voids (aerenchyma) within the cortexof adventitious roots of vegetative rice plants (Oryza sativaL. cv. RB3) was estimated microscopically from transverse sectionswith the aid of a computer-linked digitizer drawing board. Gas-spacewas detectable in 1-d-old tissue and increased in extent withage. After 7 d, approximately 70% of the cortex had degeneratedto form aerenchyma. The extent of the voids in 1-4-d-old tissuewas not increased by stagnant, poorly-aerated external environmentscharacterized by sub-ambient oxygen partial pressures and accumulationsof carbon dioxide and ethylene. Treatment with small oxygenpartial pressures, or with carbon dioxide or ethylene appliedin vigorously stirred nutrient solution also failed to promotethe formation of cortical gas-space. Furthermore, ethylene productionby rice roots was slowed by small oxygen partial pressures typicalof stagnant conditions. Silver nitrate, an inhibitor of ethylene action, did not retardgas-space formation; similarly when endogenous ethylene productionwas inhibited by the application of aminoethoxyvinylglycine(A VG), aerenchyma development continued unabated. Cobalt chloride,another presumed inhibitor of ethylene biosynthesis, did notimpair formation of the gas in rice roots nor did it decreasethe extent of aerenchyma even if A VG was supplied simultaneously.These results contrast with those obtained earlier using rootsof Zea mays L. We conclude that in rice, aerenchyma forms speedily even inwell-aerated environments as an integral part of ordinary rootdevelopment There seems to be little or no requirement for ethyleneas a stimulus in stagnant root-environments where aerenchymais likely to increase the probability of survival. Key words: Rice (Oryza sativa L.), ethylene, flooding, aeration, aerenchyma, environmental stress     

Organ and cellular localization of asparagine synthetase in rice plants

DNA gel blot analysis suggested that asparagine synthetase (AS; EC 6.3.5.4) occurred as a single gene in rice. A fusion protein consisting of 17 kDa tagged-region from pET32a(+) expression plasmid and 42 kDa N-terminal region of rice AS was first expressed in Escherichia coli. The resulting polypeptide was purified and a mono-specific antibody for rice AS was prepared after affinity-purification with the antigen. Immunoblotting revealed a high content of AS protein in the leaf sheath at the second position from the fully expanded top leaf and in grains at the middle stage of ripening. Accumulation of mRNA for AS was also observed in these organs. During the ripening of the spikelets, the AS protein contents increased during the first 21 days after flowering, then declined rapidly. Immunolocalization analysis revealed signals for AS protein in the companion cells of vascular bundles of leaf sheath and phloem-parenchyma cells, nucellar projection, and nucellar epidermis of dorsal vascular bundles of grains.     

基于主成分分析水稻植株CH4排放估算模型

估算稻田甲烷（CH4）排放量是开展稻田甲烷排放研究的重要内容之一．通过观测南方红黄壤稻田不同水稻品种甲烷排放通量,测定了16个早稻、20个晚稻品种的植株节间组织的数量特征．选取株高、茎秆长度、茎秆维管束面积／茎壁横切面积、茎壁横切面积／节间横切面积、叶鞘横切面积／节间横切面积、气腔面积／茎壁横切面积、维管束总面积／茎壁横切面积等相关因子进行了主成分分析,建立基于水稻植株的CHa排放估算模型,早、晚稻估算模型相关系数分别为0．827、0．853．同时构造了综合评价函数,得出了水稻品种CH4排放综合分值,与实测结果相比较,吻合度较高．利用估算模型进行模拟,比较模拟值与实测值,相对误差较小,证明模型具有有效性和可行性,为估算水稻CH4排放提供参考依据,为评价水稻品种CH4排放高低提供经验参考．     

Characterization of a xyloglucan endotransglucosylase gene that is up-regulated by gibberellin in rice

Xyloglucan endotransglucosylases/hydrolases (XTHs) that mediate cleavage and rejoining of the beta (1-4)-xyloglucans of the primary cell wall are considered to play an important role in the construction and restructuring of xyloglucan cross-links. A novel rice (Oryza sativa) XTH-related gene, OsXTH8, was cloned and characterized after being identified by cDNA microarray analysis of gibberellin-induced changes in gene expression in rice seedlings. OsXTH8 was a single copy gene; its full-length cDNA was 1,298 bp encoding a predicted protein of 290 amino acids. Phylogenetic analysis revealed that OsXTH8 falls outside of the three established subfamilies of XTH-related genes. OsXTH8 was preferentially expressed in rice leaf sheath in response to gibberellic acid. In situ hybridization and OsXTH8 promoter GUS fusion analysis revealed that OsXTH8 was highly expressed in vascular bundles of leaf sheath and young nodal roots where the cells are actively undergoing elongation and differentiation. OsXTH8 gene expression was up-regulated by gibberellic acid and there was very little effect of other hormones. In two genetic mutants of rice with abnormal height, the expression of OsXTH8 positively correlated with the height of the mutants. Transgenic rice expressing an RNAi construct of OsXTH8 exhibited repressed growth. These results indicate that OsXTH8 is differentially expressed in rice leaf sheath in relation to gibberellin and potentially involved in cell elongation processes.     

Biochemical Basis of Different Sensitivity to Methotrexate in Daucus carota and Oryza sativa Cell Cultures

The biochemical basis of the different sensitivity to methotrexateof Daucus carota and Oryza sativa cell cultures has been investigated.Carrot cells have a dihydrofolate reductase activity about tentimes higher than rice cells. In addition, they show a loweruptake rate of the inhibitor. No relevant differences have beenfound in the K_m value for the dihydrofolate of the two enzymesand in the degree of inhibition of their activity by methotrexate. Key words: Dihydrofolate reductase, Methotrexate resistance, Plant cell suspension cultures, Oryza sativa, Daucus carota     

Influence of Osmotic Adjustment on Leaf Rolling and Tissue Death in Rice (Oryza sativa L.)

Osmotic adjustment, measured by the lowering of the osmotic potential at full turgor, and its influence on leaf rolling and leaf death was assessed in the lowland rice (Oryza sativa L.) cultivar IR36 in both the greenhouse and field. The degree of osmotic adjustment varied with the degree and duration of stress, but was usually 0.5 to 0.6 megapascal (maximally 0.8 to 0.9 megapascal) under severe stress conditions. In leaves in which osmotic adjustment was 0.5 to 0.6 megapascal, leaf rolling and leaf death occurred at lower leaf water potentials in adjusted than in nonadjusted leaves. We conclude that osmotic adjustment aids in the drought resistance of rice by delaying leaf rolling, thereby maintaining gas exchange, and by delaying leaf death.     

Potentiality of leaf sheath cells for regeneration of rice (Oryza sativa L.) plants

Summary Regeneration of rice plantlets (Oryza sativa L.) from calli originated from leaf sheath cells was made possible. This was possible in tissues initially grown in media containing 2.4-D (2.4-dichlorophenoxyacetic acid) at low temperature and illumination. The slow growing tissues were subsequently subjected to growth conditions at an elevated temperature and higher illumination with addition of kinetin and IAA and without 2.4-D. The suitability of leaf sheath cells for protoplast technology is indicated by this success.     

Deposition pattern of hydrogen peroxide in the leaf sheaths of rice under salt stress

Deposition pattern of hydrogen peroxide (H₂O₂) under salt stress (100 mM NaCl) was examined cytochemically in rice (Oryza sativa L. cv. Pokkali) through the reaction of H₂O₂ with cerium chloride (CeCl₃) to produce electron dense precipitates of cerium perhydroxide. The distribution pattern of cerium perhydroxide precipitates in leaf sheath was considerably different from other parts of rice under salinity stress. Cerium perhydroxide precipitates were mainly accumulated on the tonoplast of leaf sheath under salinity, although they were localized on the cell wall and plasma membrane in all other tissues such as leaf blade and root.