Shoot Base Research Articles

Sodium nitroprusside promotes multiplication and regeneration of Malus hupehensis in vitro plantlets

The effects of sodium nitroprusside (SNP) on the multiplication, regeneration and rooting of Malushupehensis Rehd. var. pinyiensis Jiang in tissue culture have been investigated. The results showed that the multiplication of plantlets was promoted significantly by applying 20 μM SNP to the Murashige and Skoog (MS) medium containing 2.0 μM 6-benzylaminopurine (BA) and 1.0 μM zeatin (ZT). Multiplication of plantlets from the 1st subculture was more sensitive to SNP than that from the 4th or 7th subculture. The differentiation and regeneration of adventitious shoots from leaves or cotyledons increased significantly when 20–30 μM SNP was supplied to the medium MS containing 25 μM BA, 2.5 μM α-naphthaleneacetic acid (NAA) and 2.5 μM ZT. Adventitious shoots regeneration frequency from cotyledons was higher than that from leaves at the presence of SNP. The rooting of plantlets was promoted by SNP significantly and the best result for rooting was achieved in the half-strength MS medium containing 75 μM SNP. In addition, adventitious roots without callus distributed at the base of shoots when SNP was supplied.

How does burrowing by the isopod Limnoria agrostisa (Crustacea: Limnoriidae) affect the leaf canopy of the southern Australian seagrass Amphibolis griffithii?

In south-western Australia, the isopod Limnoria agrostisa commonly burrows into leaf clusters and immature shoots of Amphibolis griffithii. The isopod also burrows into the sheath and rhizomes of Posidonia species. In A. griffithii, the isopod consumes new tissue within the sheath, damaging or destroying the meristem. This results in malformation of new leaves or destruction of whole leaf clusters with the potential to reduce the photosynthetic area of a shoot. The isopod has been found in all but one meadow of A. griffithii examined over 1,000 km of the Western Australian coastline. It was present throughout the year and showed little variation in abundance. Young were produced year round, but were more abundant in the summer months. Females, 3.5 mm in length or larger, produced 2–5 young that were brooded within the leaf cluster or base of an immature shoot. Within a meadow, 40–70% of shoots and 10–20% of leaf clusters were damaged by isopods. Seasonal trends were not consistent, but damage appeared to be higher in summer when isopod abundance was higher. Approximately 40% of clusters were destroyed by isopod damage. Isopods attack shoots of all ages, but damage was often located on apical clusters. There was no evidence that isopod damage initiated branching or leaf cluster formation. Estimations of clusters damaged or destroyed may be conservative, as only those clusters remaining on a shoot could be counted, and cluster loss could not be quantified. Examination of defoliated stems on upright shoots and horizontal rhizomes indicated that many were destroyed by isopods. The effect of L. agrostisa although substantial appears to be a feature of healthy seagrasses throughout southern Australia.

Quantitation of endogenous levels of IAA, IAAsp and IBA in micro-propagated shoots of hybrid chestnut pre-treated with IBA

Endogenous levels of indole-3-acetic acid (IAA), indole-3-acetylaspartic acid (IAAsp) and indole-3-butyric acid (IBA) were measured during the first 8 d of in vitro rooting of rootstock from the chestnut ‘M3’ hybrid by high performance liquid chromatography (HPLC). Rooting was induced either by dipping the basal ends of the shoots into a 4.92-mM IBA solution for 1 min or by sub-culturing the shoots on solid rooting medium supplemented with 14.8-μM IBA for 5 d. For root development, the induced shoots were transferred to auxin-free solid medium. Auxins were measured in the apical and basal parts of the shoots by means of HPLC. Endogenous levels of IAA and IAAsp were found to be greater in IBA-treated shoots than in control shoots. In extracts of the basal parts of the shoots, the concentration of free IAA showed a significant peak 2 d after either root inductive method and a subsequent gradual decrease for the remainder of the time course. The concentration of IAAsp peaked at day 6 in extracts of the basal parts of shoots induced with 14.8-μM IBA for 5 d, whereas shoots induced by dipping showed an initial increase until day 2 and then remained stable. In extracts from basal shoot portions induced by dipping, IBA concentration showed a transient peak at day 1 and a plateau between day 2 and 4, in contrast to the profile of shoots induced on auxin-containing medium, which showed a significant reduction between 4 and 6 d after transferred to auxin-free medium. All quantified auxins remained at a relatively low level, virtually constant, in extracts from apical shoot portions, as well as in extracts from control non-rooting shoots. In conclusion, the natural auxin IAA is the signal responsible for root induction, although it is driven by exogenous IBA independently of the adding conditions.

Leaf heliotropism in Styrax camporum Pohl from the Brazilian cerrado: distinct gas exchange and leaf structure, but similar leaf temperature and water relations

Styrax camporum is a common shrub species from the cerrado regions of São Paulo State, Brazil. Although its leaves do not have a morphologically delimited pulvinus, the apical leaves are oriented parallel to the sun (paraheliotropic leaves), most notably around noon. Horizontal leaves at the base of shoots are oriented perpendicular to the sun's rays (diaheliotropic leaves). The ecophysiological significance of this is not fully understood. We investigated how paraheliotropism influences daily CO2 assimilation (A) and other gas exchange rates, water relations, leaf temperature (Tl), and how these relate to leaf structure as assessed by anatomical leaf descriptions. Paraheliotropic leaves had greater A and stomatal conductance (g s), which led to greater transpiration rates (E) during most of the day. However, no categorical data proved that these responses were responsible for the lower Tl for vertical leaves, which only occurred at 1100 h and 1600 h. There was no indication that Tl influenced A, E, or g s. However, diaheliotropic leaves did not assimilate less carbon because of a smaller stomatal aperture. Both leaf types had satisfactory stomatal apertures and the same g s range, resulting in similar intercellular CO2 levels. Paraheliotropic leaves had higher A rates under increased irradiance, suggesting an avoidance of excessive irradiance. Both leaf types had compact spongy parenchyma, representing an increased photosynthetic capacity per unit leaf area. Paraheliotropism in S. camporum does not seem to be related to leaf temperature control, even in the stressful Brazilian cerrado habitat.

Histopathology comparison and phenylalanine ammonia lyase (PAL) gene expressions in Fusarium wilt infected watermelons

Fusarium wilt disease of watermelon (Citrullus lanatus (Thunb.) Matsum & Nakai), caused by Fusarium oxysporum f. sp. niveum (FON), is one of the limiting factors of worldwide watermelon production. In this study, a Fusarium wilt resistant watermelon JSB, which was derived from a spontaneous mutation of the susceptible Sugar Baby (SB), was used to investigate histopathology. The number and diameter of xylem vessels in the root (10 mm below the shoot base) of resistant JSB plants were significantly higher than those in susceptible SB plants. At 9 days post inoculation (dpi), using the plate assay on Nash-PCNB media, FON could be recovered from 86% of the roots in the symptomless plants of both watermelon lines, and from 55% and 64% of the stem segments (5 mm above the shoot base) in resistant and susceptible plants, respectively. In paraffin and free-hand tissue sections, at 8, 13, and 35 dpi, the xylem of roots and stems close to the soil surface in resistant watermelon JSB plants was also colonised by FON, but to a much lower percentage than the susceptible SB ones. No colonisation below the middle of stems was observed in the resistant JSB plants. The susceptible plants grown in infested soil were all dead by 35 dpi, while the resistant plants remained healthy. These observations suggest that reducing FON colonisation in the vascular systems of the host may contribute to the resistance in JSB. Furthermore, the higher expression of the phenylalanine ammonia lyase (PAL) gene in JSB induced by FON and the effects of PAL inhibitor on the resistance of JSB suggested that PAL is involved in resistance of watermelon to Fusarium wilt pathogen.

An improved micropropagation of Eclipta alba by in vitro priming with chlorocholine chloride

An efficient method of micropropagation for Eclipta alba from young nodal axils of shoot tip explants has been developed by giving special attention to ‘priming’ in vitro plantlets in view of increasing their hardening ability after transplantation ex vitro. Among 3 cytokinins—BAP, kinetin and TDZ, BAP was found most effective in inducing and proliferating adventitious shoots. The highest frequency of responding explants (100%) and maximum number of shoots (23.0) per explant were obtained after 60 days culture on MS medium containing 8.8 μM BAP. Cent percent shoots developed roots directly from shoot base when transferred to growth regulator-free MS medium. For priming E. alba microshoots, 6.3 μM of chlorocholine chloride (CCC) was found most effective. The major changes observed in 30 days old treated shoots were, production of increased number of root, elevation of chlorophyll level in leaves and increase in plant biomass. Furthermore, arrested undesirable shoot elongation made the plants sturdier and more suitable for acclimatization. The primed micropropagated E. alba plants were healthy and survived by higher frequency (100%) in soil in comparison to the non-treated plants (84% survival).

SHOOT REGENERATION AND GENETIC TRANSFORMATION OF REGENERATIVE CALLUS CULTURE IN DIANTHUS HYBRIDA 'TELSTAR SCARLET'

Regenerative callus was established and was successfully applied to Agrobacterium-mediated genetic transformation. The callus was isolated from the base of adventitious shoots on cotyledon explants of Dianthus hybrida cv. 'Telstar Scarlet' cultured on MS media supplemented with 1 mg L -1 TDZ and 0.1 mg L -1 NAA. This callus culture was successfully applied to Agrobacterium-mediated genetic transformation. Transformed shoots were obtained from inoculated calli selected on MS medium containing 1.0 mg L -1 TDZ, 0.1 mg L -1 NAA, 20 g L -1 sucrose, 2 g L -1 gellan gum and 20-80 mg L -1 G-418 within 20 weeks from inoculation. All transformants grew, flowered normally and produced seeds when crossed with other Dianthus species.

Prophylactively parking sodium in the plant

Prophylactively parking sodium in the plant

Water relations of woody perennial plant species

<p style="text-align: justify;"><strong>Aims</strong>: To describe the relation of various water status measures of woody perennial plants (predawn and midday stem and leaf water potential), to indices of physiological activity such as leaf conductance, vegetative growth and fruit growth and composition.</p><p style="text-align: justify;"><strong>Methods and results</strong>: Almonds were exposed to three levels of irrigation over three years, and midday stem water potential (SWP) and leaf conductance, collected at approximately weekly intervals, is reported for the third year of the study. A strong linear increase in both leaf conductance and trunk growth occurred with increasing SWP, and this relation was consistent both within and between treatments. A similarly positive linear relation was found between SWP and fruit size in pear, with a negative relation between SWP and fruit soluble solids and fruit color. In grapevine, SWP was found to be uniform across all lower canopy positions tested (trunk, cordon and near the base of current year shoots) and positively correlated to early season shoot growth even before irrigation treatments were applied. Midday SWP was found to be more sensitive than midday leaf water potential (LWP) for detecting treatment differences over the course of the season, but both were well correlated to average seasonal leaf conductance within and between irrigation treatments. Predawn SWP and LWP were not as well correlated to average seasonal leaf conductance, but the most important factor determining midday leaf conductance was wind speed, indicating that grape leaf stomatal responses are quite sensitive to this environmental factor.</p><p style="text-align: justify;"><strong>Conclusion</strong>: In a wide variety of woody crop species midday stem water potential (SWP) has been found to be a valuable tool for quantifying the degree of water stress experienced by the plant, and for understanding the physiological responses of the plant to water limited conditions. In grapevine, SWP detected irrigation differences over 1 month sooner than midday leaf water potential when the number of vines used and the number of samples taken were the same for both methods, and SWP had a higher correlation to leaf conductance than predawn leaf or stem water potential.</p><p style="text-align: justify;"><strong>Significance and impact of study</strong>: SWP as a standard method for quantifying water stress in grapevine and other crops will aid research in the development of reliable management practices to improve crop productivity and quality.</p>

Dinâmica da dormência e brotação de gemas de vimeiro no Planalto Sul Catarinense

O objetivo deste trabalho foi avaliar a dinâmica e a heterogeneidade de dormência de gemas, e avaliar o efeito de diferentes épocas de colheita de ramos na brotação do vimeiro, no Planalto Sul Catarinense. Ramos de ano foram colhidos nos meses de abril a agosto de 2005, nos municípios catarinenses de Lages, Bocaina do Sul e Urupema. A dormência de gemas foi avaliada em estacas retiradas da base (0 a 12 cm), centro (12 a 24 cm) e ápice (24 a 36 cm) de ramos com comprimento superior a 150 cm. Foram utilizadas estacas com e sem redução para uma única gema. As brotações das plantas em campo também foram avaliadas. A dormência mais intensa de gemas de ramos de ano ocorreu em maio, quando a intensidade de dormência e a freqüência de gemas dormentes decresceram do ápice para a base do segmento basal dos ramos. A colheita do vime em abril induziu a brotação antes do inverno e prejudicou a brotação primaveril.

Common reed produces starch granules at the shoot base in response to salt stress

Common reed (Phragmites australis) is a well known salt-tolerant plant and it is suggested that reeds recover Na(+) in the xylem sap of the shoot base (basal part of the shoot), store it temporarily in the shoot base, release it into the phloem sap, and then retranslocate it to the roots. To investigate whether Na(+) is retained in the shoot base of reeds, confocal laser scanning microscope (CLSM) observations were conducted using an intracellular Na(+)-specific fluorescent probe. The CLSM observations revealed that reeds produced a large number of the starch granules at the shoot base when salt-stressed, and that the fluorescence indicating the location of intracellular free Na(+) was observed in the same position as the starch granules. The Na content of starch granules was considerably greater than that of the shoot base, whereas the potassium (K) contents of the granules was only slightly greater than that of the shoot base. Reeds produced Na(+)-binding starch granules in the parenchyma cells of the shoot base when salt-stressed; these starch granules may decrease intracellular free Na(+). It is proposed that the site-specific production of Na(+)-binding starch granules constitutes a novel salt tolerance mechanism.

Assessment of somaclonal variation in Dieffenbachia plants regenerated through indirect shoot organogenesis

The occurrence of somaclonal variation among regenerants derived through indirect shoot organogenesis from leaf explants of three Dieffenbachia cultivars Camouflage, Camille and Star Bright was evaluated. Three types of somaclonal variants (SV1, SV2, and SV3) were identified from regenerated plants of cv. Camouflage, one type from cv. Camille, but none from cv. Star Bright. The three variants had novel and distinct foliar variegation patterns compared to cv. Camouflage parental plants. Additionally, SV1 was taller with a larger canopy and longer leaves than parental plants and SV2. SV2 and SV3 did not produce basal shoots (single stem) but basal shoot numbers between SV1 and parental plants were similar ranging from three to four. The variant type identified from regenerated cv. Camille had lanceolate leaves compared to the oblong leaves of the parent. This variant type also grew taller and had a larger canopy than parental plants. The rates of somaclonal variation were up to 40.4% among regenerated cv. Camouflage plants and 2.6% for regenerated cv. Camille. The duration of callus culture had no effect on somaclonal variation rates of cv. Camouflage as the rates between plants regenerated from 8 months to 16 months of callus culture were similar. The phenotypes of the identified variants were stable as verified by their progenies after cutting propagation. This study demonstrated the potential for new cultivar development by selecting callus-derived somaclonal variants of Dieffenbachia.

Shoot regeneration from organogenic callus of sweet cherry (Prunus avium L.)

A complete method to regenerate adventitious shoots and to produce field-ready trees from three commercial cultivars of sweet cherry (Prunus avium L.) is described. The effects of explant types, pre-treatments, basal media, and phloroglucinol on cultivars Bing, Sweetheart, and Lapins were investigated. Callus developed on four explant types: apical shoot tips isolated from orchard trees; and punctured shoot tips, stem sections, and shoot bases of in vitro shoot cultures. Callus formed on Bing (5%), Sweetheart (8%), and Lapins (20%) shoot tips from orchard trees after 4 months on Murashige and Skoog medium (MS) at half-strength with 3 μM benzylaminopurine (BA). In vitro-derived explants formed callus after 3 months on Woody Plant Medium with 3 μM BA (W3B): punctured shoot tips (Sweetheart and Lapins 67%), stem sections (Sweetheart 31%, Lapins 27%), and shoot bases (Sweetheart 10%, Lapins 17%). Pre-treatment of shoot cultures on MS with 3 μM BA and 1 mM phloroglucinol increased callus formation three-fold on shoot base explants. Callus was separated from parental explants and maintained on MS with 3 μM BA. Shooting was induced by transferring callus to W3B. At 2 weeks, shoot development approached 100%. By 4 weeks, 7–17 shoots had formed on each explant. Callus was maintained for 1.5 years with no decrease in shoot production. Shoots were grafted onto Mazzard (P. avium) rootstocks with 54% (Sweetheart), 57% (Lapins), and 21% (Bing) success after 5 weeks.

Leaf exchange in a Mediterranean shrub: water, nutrient, non-structural carbohydrate and osmolyte dynamics

Leaf exchange is an abrupt phenological event that drastically modifies the morphology and physiology of the aerial portion of the plant. We examined if water and osmolyte differences between old leaves and new organs trigger leaf exchange, and whether the differences are closely linked to the resource resorption process in senescing leaves. We monitored concentrations of osmolyte, water, non-structural carbohydrate, nitrogen and potassium in senescing leaves and in emerging new leaves and inflorescences of a Mediterranean leaf exchanger (Cistus laurifolius L.) growing in NE Spain. Old leaves rehydrated markedly during most of the senescence process, which co-occurred with the extension of new shoots, suggesting the lack of a clear-cut switch in water supply from old to new organs. The accumulation of osmolytes in the early stage of leaf senescence might account for this rehydration. Osmolyte dynamics in old leaves depended largely on the progression of resource resorption from senescing organs but were mostly unrelated to water content during late senescence. We conclude that dehydration of old leaves is not a prerequisite for the triggering of leaf exchange. The finding that most nutrients and carbohydrates accumulated in new organs before senescing leaves massively exported resources, and the absence of relevant differences between the dynamics of old leaves at the base of inflorescences and those at the base of vegetative shoots, indicate that the nutrient and carbohydrate demands of new organs do not trigger leaf exchange.

Geographic variation in seaweed induced responses to herbivory

Recent work on European (Swedish and English) populations of Ascophyllum nodosum has detected inducible defenses in response to snail grazing that include a decrease in algal palat- ability and an increase in phlorotannin concentration, a class of potential chemical defenses. How- ever, tests of induced responses in other A. nodosum populations are lacking and we thus have a lim- ited understanding of how widespread such responses are in fucoid algae. We exposed a North American (Massachusetts) A. nodosum population to Littorina obtusata, the same snail species used in previous studies, and tested for the presence of inducible responses during 2 experiments. In con- trast to European populations, the North American population of A. nodosum did not develop any such responses. Exposure to either direct grazing or just waterborne cues associated with grazing for 26 d failed to suppress the palatability of artificial foods made from seaweed tissues exposed to these treatments. In addition, exposure to grazer cues did not increase seaweed phlorotannin levels com- pared to controls. The lack of induced responses in the North American population cannot be attrib- uted to tissue type or potential seasonal variation in inducibility because each of our 2 independent experiments tested for responses in 2 tissue types (apical tips and basal shoots) in both the winter and spring. Furthermore, consistent with European work, the North American population lacked obvious grazing damage at the time of collection, suggesting that the differences we observed were unrelated to recent history with grazers. Thus, our work and that of others indicates that A. nodosum displays considerable inter-population variability in its response to a single herbivore species.

Microarray gene expression profiling of developmental transitions in Sitka spruce (Picea sitchensis) apical shoots

The apical shoot drives the yearly new stem growth of conifer trees, is the primary site for the establishment of chemical and physical defences, and is important in establishing subsequent perennial growth. This organ presents an interesting developmental system, with growth and development progressing from a meristematic tip through development of a primary vascular system, to a base with fully differentiated and lignified secondary xylem on the inside and bark tissue with constitutive defence structures such as resin, polyphenolic phloem parenchyma cells, and sclereids on the outside. A spruce (Picea spp.) microarray containing approximately 16.7K unique cDNAs was used to study transcript profiles that characterize the developmental transition in apical shoots of Sitka spruce (Picea sitchensis) from their vegetative tips to their woody bases. Along with genes involved in cell-wall modification and lignin biosynthesis, a number of differentially regulated genes encoding protein kinases and transcription factors with base-preferred expression patterns were identified, which could play roles in the formation of woody tissues inside the apical shoot, as well as in regulating other developmental transitions associated with organ maturation. Preferential expression of known conifer defence genes, genes encoding defence-related proteins, and genes encoding regulatory proteins was observed at the apical shoot tip and in the green bark tissues at the apical shoot base, suggesting a commitment to constitutive defence in the apical shoot that is co-ordinated with rapid development of secondary xylem.

Amelioration of detrimental effects of waterlogging by foliar nutrient sprays in barley.

Six barley (Hordeum vulgare L.) cultivars contrasting in their waterlogging tolerance were subjected to waterlogging for 2 weeks under glasshouse conditions. The adverse effects of waterlogging were significantly alleviated by the foliar spray of nutrients (quarter- or full-strength Hoagland solution) in all cultivars. Beneficial effects of nutrient sprays included improved shoot and root growth and reduced leaf senescence. Chlorophyll content was increased, as was net CO2 assimilation, photochemical efficiency of PSII, and adventitious root production compared with waterlogged plants with no added foliar nutrients. Auxin was found accumulated at the shoot base in waterlogged plants, and the spray of foliar nutrients significantly increased this accumulation in waterlogged plants after 14 days of treatment. Foliar application of 1-NAA also promoted the production of adventitious roots. The highest concentration of auxin was measured in the stem region between 1.2 and 1.6 cm above the shoot-root junction. Foliar nutrient application also improved nitrogen and potassium content in both shoot and root, but calcium content was hardly affected. It is concluded that the improvement of waterlogged plant growth by foliar nutrient sprays could be related to both the improved plant nutrition and increased auxin accumulation in the shoot base, responsible for development of adventitious roots.

Water Uptake by Older Roots: Evidence From Desert Succulents

Desert succulents are renowned for their rapid recovery of shoot function after periods of drought as a result of shoot succulence, root–shoot interactions, and key root properties. Near the base of the shoot, the proximal (generally older) roots appear to play a major role in the rapid uptake and delivery of water, especially after a period of soil drying when the rest of the root system has a lower hydraulic conductance. In all of the cacti, agaves, and yuccas examined to date, hydraulic conductance for the proximal root zone is unexpectedly high. Substantial water uptake by older roots of cacti is associated with much higher axial (xylem) hydraulic conductance in proximal than in distal root regions. For agaves and yuccas, proximal roots have other anatomical and physiological traits that enhance water uptake near the shoot base. Despite suberization (waxiness) of external root tissues in the proximal region, internal cell layers appear more characteristic of young root regions with living cortical cells and a high proportion of unsuberized passage cells in the endodermis. Both features may be related to the existence of a contractile root zone possessed by all members of the Agavaceae surveyed to date. Passage cells in the endodermis and aquaporins (protein water channels) in the metabolically active cortex expedite radial water transport. The radial swelling that accompanies longitudinal root contraction may also help maintain root-to-soil contact that would otherwise be diminished in dry soil. Modifications that increase axial and radial hydraulic conductance in the proximal root region suggest that water uptake by older roots is critical to the success of desert succulents.

Foliar and shoot allometry of pollarded black locust, Robinia pseudoacacia L.

Browse of multipurpose tree species such as black locust could be used to broaden grazing options, but the temporal distribution of foliage has not been adequately studied. Our objective was to determine effects of harvest date, P fertilization (0 and 600 kg ha 1 yr 1 ), and pollard height (shoots clipped at 5-, 50-, and 100cm above ground) on foliar and shoot allometry of black locust. The experiment was conducted on a naturally regenerated 2-yr-old black locust stand (15,000 trees ha 1 ). Basal shoot diameter and foliar mass were measured monthly in June to October 2002 and 2003. Foliar and shoot dry mass (Y) was estimated from basal shoot diameter (D) by the function Y = aD b , with regression explaining ‡95% of variance. Allometry of foliar mass was affected by harvest date, increasing at a greater rate with D in September than in June or July, but not by P fertilization or pollard height. Foliar mass was predicted best by monthspecific equations for the June to October growth interval. Allometry of shoot dry mass was unaffected by harvest date, P fertilization, or pollard height. These equations could be used as a first approximation of foliar and shoot mass for pollarded black locust.

Anti-transpirant activity in xylem sap from flooded tomato (Lycopersicon esculentum Mill.) plants is not due to pH-mediated redistributions of root- or shoot-sourced ABA

In flooded soils, the rapid effects of decreasing oxygen availability on root metabolic activity are likely to generate many potential chemical signals that may impact on stomatal apertures. Detached leaf transpiration tests showed that filtered xylem sap, collected at realistic flow rates from plants flooded for 2 h and 4 h, contained one or more factors that reduced stomatal apertures. The closure could not be attributed to increased root output of the glucose ester of abscisic acid (ABA-GE), since concentrations and deliveries of ABA conjugates were unaffected by soil flooding. Although xylem sap collected from the shoot base of detopped flooded plants became more alkaline within 2 h of flooding, this rapid pH change of 0.5 units did not alter partitioning of root-sourced ABA sufficiently to prompt a transient increase in xylem ABA delivery. More shoot-sourced ABA was detected in the xylem when excised petiole sections were perfused with pH 7 buffer, compared with pH 6 buffer. Sap collected from the fifth oldest leaf of "intact" well-drained plants and plants flooded for 3 h was more alkaline, by approximately 0.4 pH units, than sap collected from the shoot base. Accordingly, xylem [ABA] was increased 2-fold in sap collected from the fifth oldest petiole compared with the shoot base of flooded plants. However, water loss from transpiring, detached leaves was not reduced when the pH of the feeding solution containing 3-h-flooded [ABA] was increased from 6.7 to 7.1 Thus, the extent of the pH-mediated, shoot-sourced ABA redistribution was not sufficient to raise xylem [ABA] to physiologically active levels. Using a detached epidermis bioassay, significant non-ABA anti-transpirant activity was also detected in xylem sap collected at intervals during the first 24 h of soil flooding.