Aerenchyma Formation In Roots Research Articles

Demonstration of osmotically dependent promotion of aerenchyma formation at different levels in the primary roots of rice using a ‘sandwich’ method and X-ray computed tomography

The effect of environmental factors on the regulation of aerenchyma formation in rice roots has been discussed for a long time, because aerenchyma is constitutively formed under aerated conditions. To elucidate this problem, a unique method has been developed that enables sensitive detection of differences in the development of aerenchyma under two different environmental conditions. The method is tested to determine whether aerenchyma development in rice roots is affected by osmotic stress. To examine aerenchyma formation both with and without mannitol treatment in the same root, germinating rice (Oryza sativa) caryopses were sandwiched between two agar slabs, one of which contained 270 mm of mannitol. The roots were grown touching both slabs and were thereby exposed unilaterally to osmotic stress. As a non-invasive approach, refraction contrast X-ray computed tomography (CT) using a third-generation synchrotron facility, SPring-8 (Super photon ring 8 GeV, Japan Synchrotron Radiation Research Institute), was used to visualize the three-dimensional (3-D) intact structure of aerenchyma and its formation in situ in rice roots. The effects of unilateral mannitol treatment on the development of aerenchyma were quantitatively examined using conventional light microscopy. Structural continuity of aerenchyma was clearly visualized in 3-D in the primary root of rice and in situ using X-ray CT. Light microscopy and X-ray CT showed that the development of aerenchyma was promoted on the mannitol-treated side of the root. Detailed light microscopic analysis of cross-sections cut along the root axis from the tip to the basal region demonstrated that aerenchyma developed significantly closer to the root tip on the mannitol-treated side of the root. Continuity of the aerenchyma along the rice root axis was morphologically demonstrated using X-ray CT. By using this 'sandwich' method it was shown that mannitol promoted aerenchyma formation in the primary roots of rice.

Drought-Induced Root Aerenchyma Formation Restricts Water Uptake in Rice Seedlings Supplied with Nitrate

Previous studies demonstrated that ammonium nutrition results in higher water uptake rate than does nitrate nutrition under water stress, and thus enhances the tolerance of rice plants to water stress. However, the process by which water uptake is related to nitrogen form under water stress remains unknown. A hydroponic experiment with simulated water stress induced by polyethylene glycol (PEG6000) was conducted in a greenhouse to study the relationship between root aerenchyma formation and water uptake rate, such as xylem sap flow rate and hydraulic conductance, in two different rice cultivars (cv. 'Shanyou 63' hybrid indica and cv. 'Yangdao 6' indica, China). The results showed that root aerenchyma tissue increased in water-stressed plants of both cultivars fed by nitrate. No significant difference was found in root hydraulic conductivity and/or xylem sap flow rate between the two rice cultivars fed by ammonium regardless of water status, whereas these parameters decreased significantly in water-stressed plants fed by nitrate. It was concluded that aerenchyma that formed in the root cortex impeded the radial transport of water in the root cylinder and decreased water uptake in water-stressed rice plants fed by nitrate. Water transport occurred mainly through Hg-sensitive water channels in rice roots supplied with ammonium.

Screening of candidate genes associated with constitutive aerenchyma formation in adventitious roots of the teosinte Zea nicaraguensis

Zea nicaraguensis (teosinte), a wild relative of maize (Zea mays ssp. mays), constitutively forms aerenchyma, which contributes to plant waterlogging tolerance, in the root cortex in drained soil, whereas maize (inbred line Mi29) does not. One with highest logarithm of odds (LOD) among quantitative trait loci (QTLs) that control constitutive aerenchyma formation in Z. nicaraguensis is Qaer1.05-6 on chromosome 1. Here, we attempted to identify genes in Qaer1.05-6 by comparing cDNA libraries from Mi29, Z. nicaraguensis and a hybrid (BC4F1 #62) carrying Qaer1.05-6. We first confirmed that constitutive aerenchyma formation was apparently observed in the order Z. nicaraguensis > BC4F1 #62 > Mi29. Contigs were assembled from cDNAs pooled from the three lines. We identified 1,868 contigs in the region on chromosome 1 that contained Qaer1.05-6. These contigs were screened for contigs that were predominantly composed of cDNAs from BC4F1 #62 and Z. nicaraguensis (no more than 10% of cDNAs from Mi29). Twenty-one such contigs were found and the genes they encoded were identified. In a real-time reverse transcription-polymerase chain reaction (RT- PCR) analysis, expression of six of these genes in BC4F1 #62 was at least double that in Mi29, making them candidates for genes associated with constitutive aerenchyma formation.

Lysigenous aerenchyma formation involves non-apoptotic programmed cell death in rice (Oryza sativa L.) roots

In waterlogged soil, deficiency of oxygen triggers development of aerenchyma in roots which facilitates gas diffusion between roots and the aerial environment. However, in contrast to other monocots, roots of rice (Oryza sativa L.) constitutively form aerenchyma even in aerobic conditions. The formation of cortical aerenchyma in roots is thought to occur by either lysigeny or schizogeny. Schizogenous aerenchyma is developed without cortical cell death. However, lysigenous gas-spaces are formed as a consequence of senescence of specific cells in primary cortex followed by their death due to autolysis. In the last stage of aerenchyma formation, a 'spoked wheel' arrangement is observed in the cortical region of root. Ultrastructural studies show that cell death is constitutive and no characteristic cell structural differentiation takes place in the dying cells with respect to surrounding cells. Cell collapse initiation occurs in the center of the cortical tissues which are characterized by shorter with radically enlarged diameter. Then, cell death proceeds by acidification of cytoplasm followed by rupturing of plasma membrane, loss of cellular contents and cell wall degradation, while cells nuclei remain intact. Dying cells releases a signal through symplast which initiates cell death in neighboring cells. During early stages, middle lamella-degenerating enzymes are synthesized in the rough endoplasmic reticulum which are transported through dictyosome and discharged through plasmalemma beneath the cell wall. In rice several features of root aerenchyma formation are analogous to a gene regulated developmental process called programmed cell death (PCD), for instance, specific cortical cell death, obligate production of aerenchyma under environmental stresses and early changes in nuclear structure which includes clumping of chromatin, fragmentation, disruption of nuclear membrane and apparent engulfment by the vacuole. These processes are followed by crenulation of plasma membrane, formation of electron-lucent regions in the cytoplasm, tonoplast disintegration, organellar swelling and disruption, loss of cytoplasmic contents, and collapse of cell. Many processes in lysing cells are structural features of apoptosis, but certain characteristics of apoptosis i.e., pycnosis of the nucleus, plasma membrane blebbing, and apoptotic bodies formation are still lacking and thus classified as non-apoptotic PCD. This review article, describes most recent observations alike to PCD involved in aerenchyma formation and their systematic distributions in rice roots.

Comparative spatiotemporal analysis of root aerenchyma formation processes in maize due to sulphate, nitrate or phosphate deprivation

Nitrate (N), phosphate (P) or sulphate (S) deprivation causes aerenchyma formation in maize (Zea mays L.) nodal roots. The exact mechanisms that trigger the formation of aerenchyma under these circumstances are unclear. We have compared aerenchyma distribution across the nodal roots of first whorl (just emerging in 10-day-old seedlings), which were subject to S, N or P deprivation over a period of 10days in connection with oxygen consumption, ATP concentration, cellulase and polygalacturonase activity in the whole root. The effect of deprivation on aerenchyma formation was examined using light and electron microscopy, along with in situ detection of calcium and of reactive oxygen species (ROS) by fluorescence microscopy. Aerenchyma was not found in the root base regardless of the deprivation. Programmed cell death (PCD) was observed near the root tip, either within the first two days (-N) or a few days later (-S, -P) of the treatment. Roots at day6 under all three nutrient-deprived conditions showed signs of PCD 1cm behind the cap, whereas only N-deprived root cells 0.5cm behind the cap showed severe ultrastructural alterations, due to advanced PCD. The lower ATP concentration and the higher oxygen consumptions observed at day2 in N-, P- and S-deprived roots compared to the control indicated that PCD may be triggered by perturbations in energy status of the root. The peaks of cellulase activity located between days3 (-N) and 6 (-P), along with the respective alterations in polygalacturonase activity, indicated a coordination which preceded aerenchyma formation. ROS and calcium seemed to contribute to PCD initiation, with ROS possessing dual roles as signals and eliminators. All the examined parameters presented both common features and characteristic variations among the deprivations.

Lysigenous aerenchyma formation in maize root is confined to cortical cells by regulation of genes related to generation and scavenging of reactive oxygen species

To adapt to waterlogging, maize (Zea mays) forms lysigenous aerenchyma in root cortex as a result of ethylene-promoted programmed cell death (PCD). Respiratory burst oxidase homolog (RBOH) gene encodes a homolog of gp91phox in NADPH oxidase, and has a role in the generation of reactive oxygen species (ROS). Recently, we found that, during aerenchyma formation, RBOH was up-regulated in all maize root tissues examined, whereas an ROS scavenging-related metallothionein (MT) gene was down-regulated specifically in cortical cells. Together, these changes should lead to high accumulations of ROS in root cortex, thereby inducing PCD for aerenchyma formation. As further evidence of the involvement of ROS in root aerenchyma formation, the PCD was inhibited by diphenyleneiodonium (DPI), an NADPH oxidase inhibitor. Based on these results, we propose a model of cortical cell-specific PCD for root aerenchyma formation.

Constraints of root response to waterlogging in Alisma triviale

To understand the economics of root aerenchyma formation in wetland plants, we investigated in detail the response of Alisma triviale to waterlogging. We hypothesized costs being associated with development of a large root air space. In three out-door pot experiments, seedlings (1 experiment) and mature plants (2 experiments) were grown under waterlogged and drained conditions for up to 2 months. Waterlogging promoted growth, and was associated with increased root porosity and decreased root density (fresh mass per volume). The increased formation of aerenchyma was associated with a higher root dry matter content for a given root density. Despite improved growth and earlier flowering, the waterlogged plants also showed signs of being constrained by the anoxic substrate, such as shallower roots, and a higher leaf dry matter content. The formation of aerenchyma was associated with costs, such as increased root dry matter content and reduced metaxylem vessel diameter. The faster growth of the seedlings under the waterlogged conditions, despite some signs of being stressed, was possibly a result of decreased requirements to allocate biomass below ground. In mature plants the increased aerenchyma allowed deeper root penetration, and ameliorated the effects of anoxia, reducing the differences in plant traits between the treatments.

The effect of nitrate nitrogen and salicylic acid on aerenchyma formation in Typha angustifolia grown in mesocosms

Macrophytes play a significant role in the functioning of hydroecosystems. Their activities include changes at the physiological and anatomical levels toward the action of various pollutants. This study was intended to reveal some features in aerenchyma formation in roots of Typha angustifolia exposed to various concentrations of nitrate nitrogen and salicylic acid. It was found that the resistance of T. angustifolia to nitrate nitrogen is mediated by redistribution of aerenchyma formation in the root system. The action of salicylic acids results in decreasing influence of nitrate nitrogen in two types of adventitious roots. Features of aeren- chyma formation in the soilborne roots and aquatic ones were different.

Contrasting dynamics of radial O2-loss barrier induction and aerenchyma formation in rice roots of two lengths.

Many wetland species form aerenchyma and a barrier to radial O(2) loss (ROL) in roots. These features enhance internal O(2) diffusion to the root apex. Barrier formation in rice is induced by growth in stagnant solution, but knowledge of the dynamics of barrier induction and early anatomical changes was lacking. ROL barrier induction in short and long roots of rice (Oryza sativa L. 'Nipponbare') was assessed using cylindrical root-sleeving O(2) electrodes and methylene blue indicator dye for O(2) leakage. Aerenchyma formation was also monitored in root cross-sections. Microstructure of hypodermal/exodermal layers was observed by transmission electron microscopy (TEM). In stagnant medium, barrier to ROL formation commenced in long adventitious roots within a few hours and the barrier was well formed within 24 h. By contrast, barrier formation took longer than 48 h in short roots. The timing of enhancement of aerenchyma formation was the same in short and long roots. Comparison of ROL data and subsequent methylene blue staining determined the apparent ROL threshold for the dye method, and the dye method confirmed that barrier induction was faster for long roots than for short roots. Barrier formation might be related to deposition of new electron-dense materials in the cell walls at the peripheral side of the exodermis. Histochemical staining indicated suberin depositions were enhanced prior to increases in lignin. As root length affected formation of the barrier to ROL, but not aerenchyma, these two acclimations are differentially regulated in roots of rice. Moreover, ROL barrier induction occurred before histochemically detectable changes in putative suberin and lignin deposits could be seen, whereas TEM showed deposition of new electron-dense materials in exodermal cell walls, so structural changes required for barrier functioning appear to be more subtle than previously described.

Construction of intraspecific linkage maps, detection of a chromosome inversion, and mapping of QTL for constitutive root aerenchyma formation in the teosinte Zea nicaraguensis

The teosinte Zea nicaraguensis, a wild relative of maize, possesses a flooding tolerance-related trait: the formation of constitutive root aerenchyma under drained (non-flooded) soil conditions. A previous study suggested that the degree of constitutive aerenchyma formation varies within Z. nicaraguensis. The objectives of this study were to construct linkage maps, to determine the marker order in a region of chromosome 4 in which recombination between maize and Z. nicaraguensis is suppressed, and to identify quantitative trait loci (QTL) controlling constitutive root aerenchyma formation in two segregating populations of Z. nicaraguensis. A total of 236 simple sequence repeat (SSR) markers were screened for polymorphism in an S1 population of Z. nicaraguensis. Seventy-one polymorphic SSR markers were assigned to 10 chromosomes, and a linkage map was constructed covering 793.5 cM. In the S1 map, a paracentric inversion was detected on the long arm of chromosome 4; this rearrangement was confirmed in an S1 linkage map of a different Z. nicaraguensis accession. Composite interval mapping analysis in 96 S1 plants revealed QTL for aerenchyma formation on chromosomes 1 (bins 1.06–1.07) and 7 (bin 7.01), explaining 17 and 12% of the total phenotypic variance, respectively. The QTL on chromosome 1 was verified by using 156 S2 plants. Near-isogenic lines exhibiting the presence or absence of the aerenchyma QTL have been developed that should be useful for genetic and physiological analyses of root aerenchyma formation.

PROGRAMMED CELL DEATH AND POSTHARVEST DETERIORATION OF HORTICULTURAL PRODUCE

Programmed cell death (PCD) is a process where cells or tissues are broken down in an orderly and predictable manner, whereby nutrients are re-used by other cells, tissues or plant parts. The process of (petal) senescence shows many similarities to autophagic PCD in animal cells including a massive breakdown of protein, DNA and RNA, the formation of autophagic vacuoles for the breakdown of cytoplasm and organelles therein and, the eventual rupture of these vacuoles that kills the cell. Common storage disorders such as scald, internal browning, core breakdown and senescent breakdown in fruit and formation of a variety of storage-related problems in vegetables that are induced by severe conditions such as low temperature, low oxygen and increased carbon dioxide concentrations, are accompanied by death and sometimes disappearance of cells. This type of cell death shows similarities to autophagic cell death during aerenchyma formation in flooded roots. This paper discusses different types of cell death in relation to flower petal senescence and storage disorders in fruit and vegetables.

Avaliação da porosidade e placa férrica de raízes de arroz cultivado em hipoxia

A alta difusividade do oxigênio em diversos materiais dificulta a criação e, ou, manutenção de um ambiente livre de O2. As técnicas utilizadas são pouco eficientes na exclusão do O2 e, portanto, não expressam a condição do solo alagado. Os objetivos deste trabalho foram desenvolver um método para obtenção de raízes em condição de hipoxia e avaliar a placa férrica e a formação de aerênquima no arroz. Foi criada uma condição de hipoxia semelhante à do solo alagado em tanques de 50 L, explorando a capacidade de difusão do O2 por meio do vinil em contato com solo reduzido. Cada tanque preenchido com solo (Gleissolo háplico) recebeu cinco sacos de vinil e foi mantido alagado. Plantas dos genótipos IRGA 423 e IRGA 424 previamente cultivadas em campo foram coletadas, tendo as raízes cortadas junto ao colo, lavadas e um terço da lâmina foliar removido. Cada saco de vinil recebeu 12 plantas de cada genótipo e solução nutritiva. Após sete dias, as novas raízes adventícias formadas foram utilizadas na determinação da porosidade e a da placa férrica em segmentos de 0-2, 2-4 e 4-6 cm a partir da ponta da raiz. As raízes foram colocadas em contato com a solução de um solo em processo de redução por 4 h e a placa férrica determinada após a extração do Fe com HCl 0,5 mol L-1. A porosidade foi determinada pela aplicação de ciclos de vácuo, com auxílio de seringas. A diferença de peso antes e depois do tratamento com vácuo e entrada de água foi assumida como sendo a estimativa da magnitude do aerênquima ao longo da raiz. A eficácia do método foi testada com a produção de raízes adventícias em saco de vinil com aeração e hipoxia. A porosidade nas raízes foi maior em ambiente hipóxico, comparado à aeração. A porosidade foi maior na proximidade da base da planta e, à medida que a porosidade aumentou, houve aumento do conteúdo de Fe na superfície das raízes, indicando que a placa férrica pode servir como estimativa da formação de aerênquima no arroz. O método de obtenção de raízes foi eficiente em promover a eliminação de O2 do saco de vinil para estudar a formação do aerênquima.

Hypoxia induced non-apoptotic cellular changes during aerenchyma formation in rice (Oryza sativa L.) roots.

The stress of low oxygen concentrations in a waterlogged environment is minimized in some plants that produce aerenchyma, a tissue characterized by prominent intercellular spaces. It is produced by the predictable collapse of root cortex cells, indicating a programmed cell death (PCD) and facilitates gas diffusion between root and the aerial environment. The objective of this study was to characterize the cellular changes take place during aerenchyma formation in root of rice that accompany PCD. Scanning electron microscopy and transmission electron microscopy were used for cellular analysis of roots. Aerenchyma development was observed in both aerobic and flooded conditions. Structural changes in membranes and organelles were examined during development of root cortex cells to compare with previous examples of PCD. There was an initial collapse which started at a specific position in the mid cortex, indicating loss of turgor, and the cytoplasm became more electron dense. These cells were distinct in shape from those located towards the periphery. Mitochondria and endoplasmic reticulum appeared normal at this early stage though the tonoplast lost its integrity. Subsequently it underwent further degeneration while the plasmalemma retracted from the cell wall followed by death of neighboring cells followed a radial path. However, pycnosis of the nucleus, blebbing of plasma membrane and production of apoptotic bodies were not found which in turn indicated nonapoptotic PCD during aerenchyma formation in rice.

Evaluation of native warm-season grass cultivars for riparian zones

Aerenchyma are air-filled spaces in the root cortex that help maintain oxygen supply to roots in saturated soils. We examined aerenchyma formation in roots and its effect on the suitability of 26 native warm-season grass cultivars from six species for use in riparian areas. In a greenhouse study, all cultivars exhibited extensive aerenchyma formation in well-drained soil, and percent aerenchyma was greater in the well-drained control than in the saturated treatment (89% versus 71%, respectively). However, the presence of extensive aerenchyma development did not guarantee good root growth under anaerobic conditions. Suitable plant materials for inclusion in riparian areas were found among four of the six warm-season species examined, although a wide range in suitability was observed within species where multiple cultivars were tested. Nine cultivars representing a range of responses were further evaluated at four field locations subjected to flooding and soil saturation. Red River prairie cordgrass (<i>Spartina pectinata</i> L.) provided superior performance at all locations. The worst performing cultivars in the greenhouse study also performed poorly in the field. However, not all cultivars that performed well in the greenhouse also performed well in the field. Osage Indiangrass (<i>Sorghastrum nutans</i> L.) was the third ranked cultivar for flooding tolerance in the greenhouse but had acceptable performance at only one of the four field sites. Conversely, Meadowcrest eastern gamagrass (<i>Tripsacum dactyloides</i> L.) exhibited only moderate flooding tolerance in the pot study but had among the lowest mortality, highest vigor, and highest biomass at three of the four locations. Controlled environment studies were useful for eliminating unacceptable cultivars, but field studies were necessary to identify suitable material for riparian areas.

Effects of Ethephon on Aerenchyma Formation in Rice Roots

The effects of ethephon on the constitutive aerenchyma formation in roots were studied with a rice variety Yangdao 6 as material. The number of air spaces formed by disintegrated cells in mediopellis increased significantly with the rising ethephon concentrations, whereas superoxide dismutase (SOD) activity showed downward trends. Compared with the control, the expression levels of xyloglucan endotransglycosylase ( XET) gene were markedly higher both at the apical 10 mm and distal parts of roots in 100 mg/L ethephon treated plants. The accumulation of XET was supposed to be associated with the aerenchyma development. Furthermore, earlier cortical cell death was observed under the ethephon treatments, and most of nuclei of cells at 4 mm from the root apex disintegrated with many Golgi apparatus, mitochondria and membrane- bound vesicles around the cell wall.

Glomus intraradices induces changes in root system architecture of rice independently of common symbiosis signaling

Arbuscular mycorrhizal fungi colonize the roots of most monocotyledons and dicotyledons despite their different root architecture and cell patterning. Among the cereal hosts of arbuscular mycorrhizal fungi, Oryza sativa (rice) possesses a peculiar root system composed of three different types of roots: crown roots; large lateral roots; and fine lateral roots. Characteristic is the constitutive formation of aerenchyma in crown roots and large lateral roots and the absence of cortex from fine lateral roots. Here, we assessed the distribution of colonization by Glomus intraradices within this root system and determined its effect on root system architecture. Large lateral roots are preferentially colonized, and fine lateral roots are immune to arbuscular mycorrhizal colonization. Fungal preference for large lateral roots also occurred in sym mutants that block colonization of the root beyond rhizodermal penetration. Initiation of large lateral roots is significantly induced by G. intraradices colonization and does not require a functional common symbiosis signaling pathway from which some components are known to be needed for symbiosis-mediated lateral root induction in Medicago truncatula. Our results suggest variation of symbiotic properties among the different rice root-types and induction of the preferred tissue by arbuscular mycorrhizal fungi. Furthermore, signaling for arbuscular mycorrhizal-elicited alterations of the root system differs between rice and M. truncatula.

Flooding tolerance of Tabebuia cassinoides: Metabolic, morphological and growth responses

Tabebuia cassinoides (Lam.) DC (Bignoniaceae) is a tree species that occurs in swampy areas of the coastal “restinga” in SE Brazil (a coastal sandy plains scrub and forest formation). To elucidate possible adaptive strategies that enable this species to occupy areas subjected to seasonal or perennial waterlogging, metabolic, morphological and growth responses of plants under flooding conditions were studied. The root system of T. cassinoides plants presented elevated amounts of ethanol (10.6 μmol g −1 fresh wt) only in the first 5 d of soil water saturation. The two-fold increase in ethanol production under flooding was corroborated by an increase in ADH activity in the same period. Lactic acid concentrations did not change significantly during four months of flooding treatment. The decrease of alcoholic fermentation under hypoxia was associated with the appearing of new roots. The induction of aerenchyma formation in roots developed under flooding conditions, allowed oxygen transport from the shoot to these organs, thus maintaining an aerobic respiration. We conclude that this characteristic and the capacity to oxidize the rhizosphere are probably responsible for the survival and growth of plants while flooded and for their success in an environment, which restricts the presence of the majority of competing tree species.

Proteomics Analysis of Rice Lesion Mimic Mutant (spl1) Reveals Tightly Localized Probenazole-Induced Protein (PBZ1) in Cells Undergoing Programmed Cell Death

Numerous reports have predicted/hypothesized a role for probenazole-induced protein (PBZ1) as a molecular marker in rice self-defense mechanism. However, the precise function of PBZ1 remains unknown. In the present study, we examined PBZ1 as a putative cell death marker in rice. For this, we focused our attention on a rice lesion mimic mutant (LMM), spotted leaf 1 ( spl1), which has been used to study the programmed cell death (PCD) phenomenon during lesion development in leaf. Using two-dimensional gel electrophoresis (2-DGE), 18 colloidal Coomassie brilliant blue stained protein spots were found to be differentially expressed in the leaves of spl1 mutant. After analysis of these spots by MALDI-TOF-MS, we identified the PBZ1 protein to be highly inducible in spl1. On the basis of these results, we proceeded to verify whether PBZ1 is highly expressed in the tissues undergoing PCD in rice. To do so, we performed immunoblot analysis and immunolocalization and used transgenic lines carrying the PBZ1 promoter fused with GFP. Results demonstrated that the expression levels and localizations of PBZ1 dramatically coincided with tissues undergoing PCD, namely, during leaf senescence, root aerenchyma formation, coleoptiles senescence, root cap, and seed aleurone layer. Furthermore, localization of the PBZ1 protein was also tightly correlated with TUNEL signal in the seed aleurone layer. As DNA fragmentation is a hallmark of PCD, this result clearly indicates a role for PBZ1 in rice tissues undergoing PCD. In conclusion, our results provide strong support for the hypothesis that PBZ1 is a molecular marker in rice defense response, and can serve as a novel potential marker for cell death/PCD in rice.

Utilizing Chromosome Segment Substitution Lines (CSSLs) for Evaluation of Root Responses to Transient Moisture Stresses in Rice

Drought and waterlogging that occur sequentially under field conditions are important abiotic stresses affecting plant growth and development. The ability to maintain the root system development during the contrasting moisture stresses may be one of the key traits for plant adaptation. This study aimed to identify the key root traits that contributed to the above ability by comparatively examining the effects of the two moisture stresses in succession on root system development. The chromosome segment substitution lines (CSSLs) from the crosses between the japonica rice cultivar Nipponbare and indica rice cultivar Kasalath were used for precise comparison of root system development. The rice seedlings were grown by hydroponics under a continuously well-aerated condition for 14 days (non-stressed), a drought condition for 7 days followed by an oxygen (O2)-deficient (stagnant) condition for 7 days (drought-to-stagnant, D-S), or a stagnant condition for 7 days followed by drought condition for 7 days (stagnant-to-drought, S-D). CSSL43 and 47 did not show any significant differences in growth from Nipponbare under the non-stressed condition, but exhibited greater lateral root production under the stresses. Lateral root production was most closely related to faster seminal root elongation mediated by higher aerenchyma formation in the D-S condition, and to more branching of lateral roots on the seminal root axis in the S-D condition. The D-S condition severely affected lateral root production due to reduced seminal root elongation and aerenchyma formation. These results confirmed the fact that those root traits previously identified using different cultivars greatly contribute to plant adaptation. Oxygen deficiency preceded by drought (D-S) was more stressful to roots than drought preceded by O2 deficiency (S-D), because drought reduced root aerenchyma formation during the subsequent stagnant condition.

Verification of QTL controlling root aerenchyma formation in a maize * teosinte "Zea nicaraguensis" advanced backcross population

Root aerenchyma formation is considered to be one of the most important characteristics for adapting to flooding. Using an F2 population of maize B64 × teosinte Zea nicaraguensis, we earlier found that, in Z. nicaraguensis, quantitative trait loci (QTLs) in two regions of chromosome 1 (Qaer1.02-3 and Qaer1.07) are associated with aerenchyma formation under non-flooding conditions. The objective of this study was to determine the utility of these (and any other) aerenchyma QTLs in marker-assisted maize breeding. For this study, an elite maize Mi29 × teosinte Z. nicaraguensis 214 BC2F1 individual population was used for advanced backcross-QTL mapping (AB-QTL). An SSR-based map was constructed using 94 markers, covering 852.7 cM for the ten chromosomes. Mapping revealed that a QTL for aerenchyma formation under non-flooding conditions exhibiting a larger effect was located on chromosome 1 (bin 1.06), the position of which was close to the previously identified Qaer1.07. Also, two QTLs with minor effects were mapped to a different position on chromosomes 1 (bin 1.11) and 5 (bin 5.09). With a few additional backcrosses, it will be possible to obtain near-isogenic lines possessing aerenchyma-forming genes. These materials may be useful in breeding flooding-tolerant maize hybrids.