Role of Ethylene in Avocado Fruit Development and Ripening: I. FRUIT DROP

A survey of avocado (Persea americana Mill.) fruit drop andethylene production was carried out during fruit development.Natural drop of avocado fruitlets started soon after set (May)and continued at a gradually decreasing rate until September,except for a temporarily increasing rate in late July. Fruitletsweighing up to 0?2 g dropped at a rate of over 30 percent perweek. With larger fruits, the rate was under 1 percent per week.Fruit drop ceased after September, when fruit growth declinedand the seed coat began to shrivel. A positive correlation was found between the rate of fruitletand fruit drop and ethylene production. Fruitlets with defectiveseeds produced ethylene at a very high rate of 7–10 timesmore than apparently normal fruits. The high incidence of defectiveseeds might be the cause of the very high levels of ethyleneproduction by young avocado fruitlets. The seed was found to be the main site of ethylene productionin fruitlets. Abscisic acid (ABA) levels in young abscissing fruits were 7times as high as those in non-abscissing fruits.     

Light Microscope Study of Pollen Tube Growth, Fertilization and Early Embryo and Endosperm Development in the Avocado Varieties Fuerte and Hass

Pollen tube growth, fertilization and early embryo and endospermdevelopment were studied using light microscopy in the avocadovarieties Fuerte and Hass. The ovule was penetrated by a pollen tube by 24 h after pollination.On reaching the ovary, the pollen tube grew along the surfaceof the inner ovary wall. It then grew around the funicle, throughthe micropyle in the inner integument and between the papillatecells at the apex of the nucellus. It entered the embryo sacvia a synergid. Sperm nuclei were present in the embryo sacat 48 h after pollination and fusion of the polar and spermnuclei took place before fusion of the egg and sperm. The endospermnucleus was the first to divide and cell wall formation occurredfollowing division. The first division of the zygote occurredat 5 or 6 days after pollination. In the variety Fuerte less than 20 per cent of the 1- and 2-day-oldembryo sacs had been penetrated by a pollen tube although tubeswere often observed in the integument or nucellus. In the varietyHass over 60 per cent of the embryo sacs were penetrated. Inwas concluded that low yields of the variety Fuerte may be partlyattributable to the failure of the pollen tube to penetratethe embryo sac. Persea americana Mill, avocado, pollen tube, fertilization, embryo, endosperm     

Combined Effects of Auxin Transport Inhibitors and Cytokinin: Alterations of Organ Development in Tobacco

We have examined the effects of the auxin transport inhibitors1-naphthylphthalamic acid (NPA) and 2,3,5-triiodobenzoic acid(TIBA) on leaf morphogenesis of transgenic Nicotiana tabacum(cv. Xanthi) plants expressing the Agrobacterium tumefacienscytokinin biosynthetic gene, ipt. We have observed the formationof saucer-shaped leaf-like organs at the shoot apex and at lateralbuds. The formation of apical saucer-shaped leaf-like organscan be duplicated by the application of exogenous NPA and cytokininto wild-type tobacco seedlings. We have also observed adventitiousleaf-like organs with altered petiole and blade morphology inthe transgenic plants treated with auxin transport inhibitors.These results suggest that the combination of diminished auxintransport and elevated cytokinin can lead to alterations inleaf development in tobacco. ⁴Present address: Genesis Research and Development Corporation,P.O. Box 50, Auckland, New Zealand     

Ratio of Dietary n-6/n-3 Polyunsaturated Fatty Acids Independently Related to Muscle Mass Decline in Hemodialysis Patients

Background

n-3 polyunsaturated fatty acids (PUFAs) might be useful nutritional strategy for treating patients with sarcopenia. We evaluated the effect of the intake of dietary n-3 PUFAs on the skeletal muscle mass (SMM), appendicular skeletal muscle mass (ASM), and its determinants in patients receiving standard hemodialysis (HD) treatment for the management of end stage renal disease.

Methods

In this cross-sectional study, data of 111 HD patients were analyzed. Anthropometric and bioelectrical impedance measurements used to estimate the muscle mass were performed the day of dialysis immediately after the dialysis session. Routine laboratory and 3-day dietary data were also collected. The cutoff value of adequate intake (AI) for both n-3 PUFAs and alpha-linolenic acid (ALA) was 1.6 g/day and 1.1 g/day for men and women, respectively.

Results

The mean age, mean dietary n-3 PUFAs intake, ALA intake, ratio of n-6/n-3 PUFAs intake, SMM, and ASM of patients were 61.4 ± 10.4 years, 2.0 ± 1.3 g/day, 1.5 ± 1.0 g/day, 9.5 ± 6.7 g/day, 23.9 ± 5.5 kg, and 17.5 ± 4.5 kg, respectively. A higher SMM and ASM significantly observed in patients who achieved an AI of n-3 PUFAs. Similar trends appeared to be observed among those patients who achieved the AI of ALA, but the difference was not significantly, except for ASM (P = 0.047). No relevant differences in demographics, laboratory and nutritional parameters were observed, regardless of whether the patients achieved an AI of n-3 PUFAs. Multivariate analysis showed that the BMI and equilibrated Kt/V were independent determinants of the muscle mass. Moreover, the ratio of n-6/n-3 PUFAs was an independent risk determinant of reduced ASM in HD patients.

Conclusion

Patients with an AI of n-3 PUFAs had better total-body SMM and ASM. A higher dietary ratio of n-6/n-3 PUFAs seemed to be associated with a reduced muscle mass in HD patients.     

Role of Ethylene in Avocado Fruit Development and Ripening: II. ETHYLENE PRODUCTION AND RESPIRATION BY HARVESTED FRUITS

Avocado (Persea americana Mill.) fruits were harvested at successivedevelopment stages during a period of 10 months. Ethylene productionand respiration were determined during the post-harvest period. Detached immature fruits were found to have a preclimactericincrease in ethylene production and respiration without anysigns of ripening. In fruits larger than 20 g a second phaseof climacteric ethylene production and respiration, associatedwith ripening, ensued. The preclimacteric ethylene was produced mainly by the seedcoat. It is suggested that the high ethylene production potentialof the seed coat may serve as a means for inducing abscissionin young fruits.     

The Effects of Fusicoccin,Abscisic Acid,and Benzyladenine on the Transport and Partitioning of Substances as Related to Cytokinin Sink Activity

We tested the possible cytokinin effect on the functioning of the active transport system involved in the assimilate loading into the phloem as a cause for the cytokinin sink and retention effect. This effect is manifested in the deceleration of substance export from and the stimulation of substance import to the sites of local phytohormone application to the mature detached leaf from untreated leaf areas. To affect the membrane mechanisms of the substance transport, we used leaf treatment with the phytotoxin fusicoccin, an enhancer of plasmalemmal H⁺-ATPase and a potential stimulator of assimilates export, and with the phytohormone ABA affecting transport, metabolism, and plant growth. However, fusicoccin did not enhance ¹⁴C-sucrose export from the leaf blade and did not interfere with the cytokinin-induced export deceleration. ABA reduced substantially ¹⁴C export from the leaf but eliminated the cytokinin effect on this process. Similar results were obtained for broad bean (Vicia faba L.) leaves with apoplastic phloem loading, involving H⁺-ATPase activity, and pumpkin (Cucurbita pepo L.) leaves with symplastic phloem loading, that is, occurring without sucrose transmembrane translocation and without H⁺-ATPase involvement. The conclusion is that the cytokinin-induced development of sink zones in source leaves is not related to the membrane mechanisms of the substance transport in the mesophyll–phloem system. The data obtained support the idea that the cause for the cytokinin sink and retention effect is the enhancement of elongation growth and total activation of metabolism in the mesophyll cells of the detached leaf.     

Symplastic Transport of Carboxyfluorescein in Staminal Hairs of Setcreasea purpurea Is Diffusive and Includes Loss to the Vacuole

The kinetics of symplastic transport in staminal hairs of Setcreasea purpurea was studied. The tip cell of a staminal hair was microinjected with carboxyfluorescein (CF) and the symplastic transport of this CF was videotaped and the digital data analyzed to produce kinetic curves. Using a finite difference equation for diffusion between cells and for loss of dye into the vacuole, kinetic curves were calculated and fitted to the observed data. These curves were matched with data from actual microinjection experiments by adjusting K (the coefficient of intercellular junction diffusion) and L (the coefficient of intracellular loss) until a minimum in the least squares difference between the curves was obtained. (a) Symplastic transport of CF was governed by diffusion through intercellular pores (plasmodesmata) and intracellular loss. Diffusion within the cell cytoplasm was never limiting. (b) Each cell and its plasmodesmata must be considered as its own diffusion system. Therefore, a diffusion coefficient cannot be calculated for an entire chain of cells. (c) The movement through plasmodesmata in either direction was the same since the data are fit by a diffusion equation. (d) Diffusion through the intercellular pores was estimated to be slower than diffusion through similar pores filled with water.     

Succinoxidase Activity of Avocado Fruit Mitochondria in Relation to Temperature and Chilling Injury throughout the Climacteric Cycle

Mitochondria were isolated from `Fuerte' avocado fruit (Persea americana Mill.) at four different stages of the respiratory climacteric. Preclimacteric fruit had the highest rate of succinate oxidation and the postclimacteric mitochondria the lowest. Subsequently, successive additions of ADP increased the respiratory control ratio.     

Kinetics of Endogenous Cytokinin, IAA and ABA Levels in Relation to the Growth and Morphology of Tobacco Crown Gall Tissue

The kinetics of endogenous cytokinin, IAA and ABA levels duringthe growth cycle of a wild-type tobacco crown gall (W₃₈-B₆S₃)were compared with that of a shoot-inducing (Shi^-) mutant. Inboth tumor types, high IAA and cytokinin (essentially ribosyl-trans-zeatinand its corresponding glucoside) levels were built up by theend of the linear growth phase and maintained during the greaterpart of the exponential growth period. The stationary phasewas preceded by a very drastic decrease in the endogenous levelof both hormones. Quantitatively, the wild-type tumour showed a higher IAA leveland a reduced cytokinin level compared with the Shi^- mutant.No significantly different endogenous ABA pattern was observed.The reduced cytokinin level might correspond to the ratio oftransformed/untransformed cells in the wild-type tumour whereasthe reduced IAA level in the Shi^- mutant may be correlated withthe deletion of gene 2 in the T-DNA of the pGV 2206 Ti plasmid. The elevated cytokinin/IAA ratio induced shooting mainly ofthe untransformed cells in the Shi^- mutant tissue whereas inthe wild-type, the shoot suppression was compatible with thereduced cytokinin/IAA ratio. ⁴Senior Research Associate Nationaal Fonds WetenschappelijkOnderzoek (N.F.W.O.). ⁵Research Associate N.F.W.O. ⁶Recipient of an Instituut voor Wetenschappelijk Onderzoek inNijverheid en Landbouw grant. (Received February 23, 1984; Accepted June 19, 1984)     

Pattern and Rate of Cognitive Decline in Cerebral Small Vessel Disease: A Prospective Study

Objectives

Cognitive impairment, predominantly affecting processing speed and executive function, is an important consequence of cerebral small vessel disease (SVD). To date, few longitudinal studies of cognition in SVD have been conducted. We determined the pattern and rate of cognitive decline in SVD and used the results to determine sample size calculations for clinical trials of interventions reducing cognitive decline.

Methods

121 patients with MRI confirmed lacunar stroke and leukoaraiosis were enrolled into the prospective St George’s Cognition And Neuroimaging in Stroke (SCANS) study. Patients attended one baseline and three annual cognitive assessments providing 36 month follow-up data. Neuropsychological assessment comprised a battery of tests assessing working memory, long-term (episodic) memory, processing speed and executive function. We calculated annualized change in cognition for the 98 patients who completed at least two time-points.

Results

Task performance was heterogeneous, but significant cognitive decline was found for the executive function index (p<0.007). Working memory and processing speed decreased numerically, but not significantly. The executive function composite score would require the smallest samples sizes for a treatment trial with an aim of halting decline, but this would still require over 2,000 patients per arm to detect a 30% difference with power of 0.8 over a three year follow-up.

Conclusions

The pattern of cognitive decline seen in SVD over three years is consistent with the pattern of impairments at baseline. Rates of decline were slow and sample sizes would need to be large for clinical trials aimed at halting decline beyond initial diagnosis using cognitive scores as an outcome measure. This emphasizes the importance of more sensitive surrogate markers in this disease.     

橄榄果实发育过程中细胞壁物质和相关酶活性变化

为了解橄榄(Canarium album)果实质地差异形成的原因,以鲜食型橄榄‘清榄1号’和加工型橄榄‘长营’为材料,对果实发育过程中细胞壁物质含量和相关酶活性进行了测定。结果表明,随着橄榄果实的成熟,‘清榄1号’较‘长营’维持较高的果胶甲酯酶(PME)活性,促进了果胶的水解,离子型果胶(ISP)含量较高而共价型果胶(CSP)含量较低。2个橄榄品种纤维素含量均较高,‘清榄1号’果实的半纤维素含量低于‘长营’。‘清榄1号’木质素含量低于‘长营’,较高的苯丙氨酸解氨酶(PAL)和过氧化物酶(POD)活性促进了木质素含量的增加。因此,ISP、CSP、半纤维素和木质素含量的不同可能是2个橄榄品种果实质地差异形成的原因。     

综述: 糖基化对斑马鱼发育及再生作用的研究进展

对有机体发育和再生的研究一直是生命科学领域探索热点之一,斑马鱼由于具有发育速度快、胚胎透明便于观察以及其尾鳍、心脏、神经系统等组织器官可以再生等众多优点,已经成为应用最为广泛的模式生物之一．在斑马鱼发育及再生过程中,细胞发生增殖、分化、形态建成等代谢活动,而对其中调控机制的研究尚未完全明确,对其深入探究的意义不言而喻,可以为很多疾病的认识治疗、组织工程的发展奠定理论基础．研究已经表明,在此过程中,有很多蛋白质参与发育及再生的调节,而蛋白质糖基化是重要的蛋白质翻译后修饰,糖基化的变化会影响细胞识别、黏附、信号传导等,从而导致炎症、肿瘤等恶性疾病的发生．近年来,糖组学的发展为发育生物学和再生医学提供了新的思路和研究成果,本文就糖基化变化在斑马鱼的发育过程中发挥的作用做一综述,并对今后它对再生过程的作用进行展望．     

Regulation of Cyclic Photophosphorylation during Ferredoxin-Mediated Electron Transport : Effect of DCMU and the NADPH/NADP Ratio

Addition of ferredoxin to isolated thylakoid membranes reconstitutes electron transport from water to NADP and to O₂ (the Mehler reaction). This electron flow is coupled to ATP synthesis, and both cyclic and noncyclic electron transport drive photophosphorylation. Under conditions where the NADPH/NADP⁺ ratio is varied, the amount of ATP synthesis due to cyclic activity is also varied, as is the amount of cyclic activity which is sensitive to antimycin A. Partial inhibition of photosystem II activity with DCMU (which affects reduction of electron carriers of the interphotosystem chain) also affects the level of cyclic activity. The results of these experiments indicate that two modes of cyclic electron transfer activity, which differ in their antimycin A sensitivity, can operate in the thylakoid membrane. Regulation of these activities can occur at the level of ferredoxin and is governed by the NADPH/NADP ratio.     

Water Transport Properties of the Grape Pedicel during Fruit Development: Insights into Xylem Anatomy and Function Using Microtomography

Xylem flow of water into fruits declines during fruit development, and the literature indicates a corresponding increase in hydraulic resistance in the pedicel. However, it is unknown how pedicel hydraulics change developmentally in relation to xylem anatomy and function. In this study on grape (Vitis vinifera), we determined pedicel hydraulic conductivity (k_h) from pressure-flow relationships using hydrostatic and osmotic forces and investigated xylem anatomy and function using fluorescent light microscopy and x-ray computed microtomography. Hydrostatic k_h (xylem pathway) was consistently 4 orders of magnitude greater than osmotic k_h (intracellular pathway), but both declined before veraison by approximately 40% and substantially over fruit development. Hydrostatic k_h declined most gradually for low (less than 0.08 MPa) pressures and for water inflow and outflow conditions. Specific k_h (per xylem area) decreased in a similar fashion to k_h despite substantial increases in xylem area. X-ray computed microtomography images provided direct evidence that losses in pedicel k_h were associated with blockages in vessel elements, whereas air embolisms were negligible. However, vessel elements were interconnected and some remained continuous postveraison, suggesting that across the grape pedicel, a xylem pathway of reduced k_h remains functional late into berry ripening.In grape (Vitis vinifera), fruit growth by water accumulation follows a double sigmoid pattern and is influenced by the diurnal and developmental changes in water flow between fruit and the parent plant (Matthews and Shackel, 2005). Until the onset of fruit ripening (i.e. veraison), water enters the fruit predominantly via the xylem and thereafter mainly through the phloem (Greenspan et al., 1994, 1996). Choat et al. (2009) showed that the hydraulic conductance (i.e. 1/resistance) of the grape berry and pedicel declines substantially at later ripening stages predominantly due to a decline in pedicel conductance. Significant developmental changes in pedicel hydraulic properties were also reported for tomato (Solanum lycopersicum) and were found to be associated with xylem anatomical changes (Lee 1989; Van Ieperen et al., 2003; Rancić et al., 2008, 2010). Due to its position along the vascular transport pathway between fruit and the parent plant, the pedicel can play an important role in affecting fruit growth, as in kiwi (Actinidia deliciosa; Mazzeo et al., 2013). However, for grape, it needs to be elucidated how pedicel hydraulic properties change developmentally in relation to xylem anatomy and function.The location and nature of the loss in hydraulic conductance between the parent plant and the fruit is unclear and may differ among fruits. For tomato, Malone and Andrews (2001) showed that most of the loss of hydraulic conductance occurs in the fruit per se, but Van Ieperen et al. (2003) reported important and decreasing hydraulic conductance in the pedicel abscission zone over fruit development. For Citrus spp., Garcia-Luis et al. (2002) reported that xylem vessels in the pedicel remain largely functional late into fruit ripening. For grape, although vessel breakage in the berry was thought to lead to xylem dysfunction (Coombe and McCarthy 2000), several studies and methods have shown that xylem vessels in the fruit remain functional (Rogiers et al., 2001; Bondada et al., 2005; Chatelet et al., 2008a, 2008b). In line with these findings, data by Keller et al. (2006) suggest that the pedicel xylem also remains at least partially functional in ripening grape berries and can conduct water to and from the parent plant. Nevertheless, a reduction in the ability to transport water during ripening has been reported for grape (Tyerman et al., 2004; Choat et al., 2009) and other fleshy fruits, such as apple (Malus domestica; Lang and Ryan, 1994) and kiwi (Mazzeo et al., 2013), and it still remains unclear what causes this loss in xylem hydraulic conductance. For the grape pedicel, Choat et al. (2009) detected higher concentrations of xylem solutes postveraison and proposed that this is related to the deposition of gels into the xylem vessel lumen. However, direct evidence for the presence of xylem blockage and/or embolism formation in the grape pedicel is missing.This study of the grape ‘Cabernet Sauvignon’ pedicel was conducted with the goal to obtain a comprehensive understanding of how changes in hydraulic properties relate to changes in xylem structure and function over fruit development. Over the course of fruit development from 20 to 90 d after anthesis (DAA), water transport properties of pedicels were investigated under osmotic and hydrostatic driving forces using a modified pressure-probe system. This was combined with analyses of spatial and temporal changes in pedicel xylem anatomy and function using fluorescent light microscopy and x-ray computed microtomography (microCT; Brodersen et al., 2010, 2013; Rancić et al., 2010).     

A rice orthologue of the ABA receptor, OsPYL/RCAR5, is a positive regulator of the ABA signal transduction pathway in seed germination and early seedling growth

Abscisic acid (ABA) is a phytohormone that positively regulates seed dormancy and stress tolerance. PYL/RCARs were identified an intracellular ABA receptors regulating ABA-dependent gene expression in Arabidopsis thaliana. However, their function in monocot species has not been characterized yet. Herein, it is demonstrated that PYL/RCAR orthologues in Oryza sativa function as a positive regulator of the ABA signal transduction pathway. Transgenic rice plants expressing OsPYL/RCAR5, a PYL/RCAR orthologue of rice, were found to be hypersensitive to ABA during seed germination and early seedling growth. A rice ABA signalling unit composed of OsPYL/RCAR5, OsPP2C30, SAPK2, and OREB1 for ABA-dependent gene regulation was further identified, via interaction assays and a transient gene expression assay. Thus, a core signalling unit for ABA-responsive gene expression modulating seed germination and early seedling growth in rice has been unravelled. This study provides substantial contributions toward understanding the ABA signal transduction pathway in rice.     

The Rate of Decline in Small Fibre Function Assessed Using Axon Reflex-Mediated Neurogenic Vasodilatation and the Importance of Age Related Centile Values to Improve the Detection of Clinical Neuropathy

Background

The LDIflare technique (LDIflare) is a simple non-invasive test of small fibre function in dorsal foot skin involving skin heating and measuring the size of the resulting axon reflex-mediated vasodilator (flare) response using a laser Doppler imager (LDI). This study establishes age-related normative reference ranges for the test and determines the rate of decline in small fibre function per decade. Additionally, the potential value of using age related centiles rather than Receiver Operator Curves (ROC) was explored by comparison of the sensitivity and specificity of each analytic technique in identifying clinical neuropathy.

Methods

LDIflare areas were assessed in 94 healthy controls and 66 individuals with diabetes with (DN+, n = 31) and without clinical neuropathy (DN-, n = 35); neuropathy defined as a Neuropathy Disability Score ≥3. The age specific 5th centile values were used as the ‘cut-offs’ for the diagnosis of neuropathy from which sensitivity and specificity were calculated.

Results

There was a significant age dependant decrease in LDIflare size (r = −0.42, p<0.0001) with no significant gender differences. The LDIflare size reduced 0.56 cm² per decade which gives a percentage reduction of approximately 5.5% per decade. Using the normative 5th centiles as the cut-offs, the technique had a sensitivity of 77%, specificity of 90%, positive predictive value of 82% and negative predictive value of 87%.The ROC analysis gave a threshold of <3.66 cm² for the cut-off, resulting in a sensitivity of 75%, specificity of 85%, positive predictive value of 74% and negative predictive value of 86%.

Conclusions

There is an age dependent decrease in small fibre function in the foot of 5.5% per decade. Both analytic techniques demonstrate good sensitivity and specificity for detecting clinical neuropathy but the technique based on age centiles offers better diagnostic accuracy and is therefore proposed as the method of choice.     

皱皮木瓜果实发育后期品质变化及其成熟阶段的划分初探

以湖北长阳产皱皮木瓜为材料,测定果实发育后期果实鲜质量、果长、果径、果色、果实硬度以及果肉干物质量、可溶性糖含量、总酸含量和总黄酮含量等品质指标的动态变化,划分不同成熟阶段,为判断果实适宜采收期、实现优质生产提供理论参考。结果表明:(1)皱皮木瓜果实发育后期果实鲜质量、果长、果径、果肉干物质量和可溶性糖含量均呈现上升趋势;果色由绿色、黄绿色渐变为淡黄色到黄色;果实硬度、果肉总酸和总黄酮含量呈先上升后下降趋势。(2)各品质指标快速变化的时间区域存在差异,果实鲜质量在花后105~150d增加较快,果色在150d后逐渐变黄,果实硬度在花后135~165d快速下降,果肉总酸、总黄酮含量则在花后105~120d快速增加至峰值。(3)根据主成分分析结果和各品质指标的变化特点,可初步将皱皮木瓜果实发育后期划分为未成熟(花后105d之前)、早期成熟(花后120~150d)和成熟(花后165~180d)3个阶段。研究表明,随着果实成熟度的提高,皱皮木瓜果实鲜质量、果色、果肉干物质量、可溶性糖含量等指标不断升高,果实硬度逐渐下降,其食用加工品质不断提升,而在早期成熟阶段(花后120~150d)果实的药用品质则相对较高。     

黄瓜茎端ABA/GA_4比值与花芽性别分化的关系(英文)

在同一环境条件下,单雌性株茎端ABA含量显著高于雌雄同株的,而GA_4水平却正好相反。 较高的ABA/GA_4比值有利于黄瓜雌性分化,反之则有利于雄性分化。促进黄瓜雄性分化的生长物质GA_3和AgNO_3会引起黄瓜茎端ABA/GA_4比值的下降,而促进其雌性分化的生长物质乙烯利(Ethrel)则会引起ABA/GA_4比值的上升。