Daminozide inhibits ethylene production in apple fruit by blocking the conversion of methionine to aminocyclopropane-1-carboxylic acid (ACC)

At harvest, fruit from apple trees sprayed with daminozide (+daminozide) had lower levels of aminocyclopropane-1-carboxylic acid (ACC) and produced significantly lower amounts of ethylene than untreated (–daminozide) fruit. Flesh discs from the fruit of +daminozide and –daminozide trees were fed precursors of ethylene to determine how daminozide inhibits ethylene production. ACC was metabolized to ethylene regardless of treatment. Methionine (MET), however, was only converted to ethylene by –daminozide fruit, and only after the fruit had been maintained at 4 °C for 5 months. +Daminozide fruit failed to convert MET to ethylene at harvest, as well as after cold storage. When daminozide was added to the incubation media of flesh discs it did not inhibit ethylene production or the conversion of ACC to ethylene. The addition of daminozide did, however, inhibit the metabolism of exogenous MET to ethylene. Aminooxyacetate acid (AOA) blocked both the endogenous production of ethylene and that from MET feeds. Daminozide inhibits ethylene production by preventing the conversion of MET to ACC, but it does not appear to act as a simple competitive inhibitor of ACC synthase activity.Abbreviations ACC aminocyclopropane-1-carboxylic acid - AVG aminoethoxyvinylglycine - AOA aminooxyacetic acid - CH cycloheximide - MET methionine - PUT putrescine Author for correspondence     

Significance of flower and fruit thinning on fruit quality

The effects of mechanical or chemical flower and fruit thinning on fruitquality were primarily by altering crop load. However, there were alsodirect effects of thinning agents. Fruit size was directly related tothinning intensity. In addition to crop load, age of wood, flower budquality, competition within clusters and canopy were important factorsaffecting the response to thinning. Short- and long-term thinningstudies identified two groups of quality components: Group 1characteristics include size, colour, skin performance, firmness andsugar and acid content of the fruit. Group 2 characteristics wererepresented by inorganic components, especially calcium and potassiumwhich are implicated in the susceptibility of fruit to physiologicaldisorders. While group 1 characteristics were improved by increasingthinning intensity, storability of the fruit was better at high than atlow crop loads. Therefore, a compromise between all quality requirementsmust be found for a good economic return. Establishing the trends ofthinning on the different quality parameters can help to select athinning strategy for local or regional conditions typically beingdetermined by growing and market conditions.     

\Cytokinin oxidase/dehydrogenase (CKX) activity in wild and ethylene-insensitive mutant eti5 type of Arabidopsis thaliana (L.) Heynh plants and the effect of cytokinin N1-(2-chloro-4-pyridyl)-N2-phenylurea on enzymatic activity and leaf morphology

The specific activity of cytokinin oxidase/dehydrogenase (EC 1.5.99.12) (CKX) was determined in leaves of wild type (wt) and ethylene-insensitive mutant (eti5) of Arabidopsis thaliana (L.) Heynh plants. Comparative studies showed that this mutation has lower basal CKX activity than wt. Application of 4PU-30 (N¹-(2-chloro-4-pyridyl)-N²-phenylurea) resulted in decreased CKX activity in both wt and mutant plants. The treatment increased leaf blade thickness and the volume of chlorophyll-containing cells per unit leaf area in wt but these changes were not observed in the eti5 mutant. The reduction in chlorophyll “a” and “b”, as well as in carotenoids content in the treated wt tissues resulting from altered leaf morphology was not detected in eti5 plants.     

Micropropagation of thirteen Malus cultivars and rootstocks,and effect of antibiotics on proliferation

Several factors that affect in vitro establishment, proliferation, and rooting of thirteen Malus cultivars and rootstocks were studied. Apple shoot tips (1.5±0.5 cm in length) were established using ascorbic and citric acids as antioxidants. Four proliferation media containing 1.0 mg 1^–1 BA and different concentrations of IBA and GA₃ were tested. Proliferation rates varied depending on the genotype and medium used. The highest proliferation rate was obtained for a rootstock that produced 11.6±2.5 shoots (1.5±0.8 cm in length) per tube per month. Rooting was induced with IBA for all the genotypes tested. The optimal IBA concentration was cultivar dependent (between 0.1 and 1.0 mg 1^–1 IBA), and lower concentrations were necessary to induce rooting in liquid rather than in solid medium.The effects on shoot-tip proliferation of cefotaxime, carbenicillin and kanamycin, three antibiotics commonly used for transformation studies, were also evaluated. Cefotaxime at 200 mg 1^–1 stimulated shoot growth and development, but at 500 mg 1^–1 caused abnormal shoot morphology. Carbenicillin at 500 mg 1^–1, alone or in combination with cefotaxime at 200 mg 1^–1, inhibited proliferation and caused excessive enlargement of the basal leaves, inducing callus formation and release of phenolic compounds in the medium. Kanamycin at 50 mg 1^–1 was phytotoxic and caused shoot chlorosis and necrosis. Consideration of the toxicity of these antibiotics is critical when designing transformation schemes for selection and recovery of transgenic apple plants.Abbreviations BA benzyladenine - cef cefotaxime - crb carbenicillin - GA₃ gibberellic acid - IBA indole-3-butyric acid - Kan kanamycin - ms Murashige and Skoog [19] macro- and micro-nutrients - NAA naphthalene-acetic acid     

Interaction of jasmonic acid with some plant growth regulators in the control of apple (Malus domestica) embryo germination

Embryos isolated from dormant apple seeds were treated with jasmonic acid (JA), gibberellin A₃ (GA₃), abscisic acid (ABA) and hydrogen cyanide in darkness and in light. The chemicals were present in the culture medium continuously and simultaneously or applied for 2 days and in different sequences. All treatments stimulated embryo germination except ABA, which was strongly inhibitory. Additive effects of JA with light and with GA₃ on embryo germination were observed, whereas ABA interacted synergically with JA, HCN and light. ABA and GA₃ were most effective when applied early during embryo incubation, but the late JA treatment was more stimulatory. It is concluded that JA does not act on the regulatory pathway that is initiated by light and which leads to embryo germination through gibberellin accumulation and alkaline lipase activation. ABA and HCN appear to be involved in the control of this pathway. JA and ABA may be involved in the control of alkaline lipase activity, independently of this regulatory chain.Abbreviations ABA abscisic acid - GA₃ gibberellin A₃ - JA jasmonic acid     

苹果果实糖积累特性与品质形成的关系

以'富士'和'国光'苹果为研究对象,对其果实发育过程中糖含量及其代谢关键酶活性的变化进行测定分析,以揭示糖分积累代谢特性对果实品质形成的影响.结果表明:(1)'富士'和'国光'均为己糖积累型果实, '富士'果实以积累果糖最多,果糖/葡萄糖(F/G)值为1.56,而'国光'以积累葡萄糖最多,F/G值仅为0.68;蔗糖在两品种中含量和所占比例均很低,在近成熟期'富士'高于'国光'.(2)'富士'果实蔗糖磷酸合成酶(SPS)和蔗糖合成酶(SS)活性均随果实糖的累积量增加而显著升高,酸性转化酶(AI)活性也渐趋升高,而中性转化酶(NI)活性波动不大,且其糖累积与AI和SPS活性相关性最大,而与NI相关性不大,SS的作用主要表现在发育后期;在 '国光'果实糖积累过程中SPS起主导作用,SS和NI的作用主要表现在发育前期,而AI的作用不大.(3)'富士'和'国光'果实淀粉含量变化趋势相同,在淀粉积累高峰之后,'富士'果实淀粉降解速度更快,其淀粉含量迅速下降且低于'国光',此时其相应淀粉酶活性也高于'国光'.研究发现,'富士'和'国光'果实糖积累和淀粉代谢均存在显著差异,从而直接或间接地影响着果实糖代谢过程,进而导致果实品质的显著差异.     

The source of gibberellins in the parthenocarpic development of ovaries on topped pea plants

The role and source of gibberellins (GAs) involved in the development of parthenocarpic fruits of Pisum sativum L. has been investigated. Gibberellins applied to the leaf adjacent to an emasculated ovary induced parthenocarpic fruit development on intact plants. The application of gibberellic acid (GA₃) had to be done within 1 d of anthesis to be fully effective and the response was concentration-dependent. Gibberellin A₁ and GA₃ worked equally well and GA₂₀ was less efficient. [³H]Gibberellin A₁ applied to the leaf accumulated in the ovary and the accumulation was related to the growth response. These experiments show that GA applied to the leaf in high enough concentration is translocated to the ovary. Emasculated ovaries on decapitated pea plants develop without application of growth hormones. When [³H] GA₁ was applied to the leaf adjacent to the ovary a substantial amount of radioactivity accumulated in the growing shoot of intact plants. In decapitated plants, however, this radioactivity was mainly found in the ovary. There it caused growth proportional to the accumulation of CA₁. Application of LAB 150978, an inhibitor of GA biosynthesis, to decapitated plants inhibited parthenocarpic fruit development and this inhibition was counteracted by the application of GA₃ (either to the fruit, or the leaf adjacent to the ovary, or through the lower cut end of the stem). All evidence taken together supports the view that parthenocarpic pea fruit development on topped plants depends on the import of gibberellins or their precursors, probably from the vegetative aerial parts of the plant.Abbreviations FW flesh weight - GA_n gibberellin A_n - HPLC high-performance liquid chromatography     

Photosystem 2 is more tolerant to high temperature in apple (<Emphasis Type="Italic">Malus domestica</Emphasis> Borkh.) leaves than in fruit peel

Tolerance of photosystem 2 (PS2) to high temperature in apple (Malus domestica Borkh. cv. Cortland) leaves and peel was investigated by chlorophyll a fluorescence (OJIP) transient after exposure to 25 (control), 40, 42, 44, and 46 °C in the dark for 30 min. The positive L-step was more pronounced in a peel than in leaves when exposed to 44 °C. Heat-induced K-step became less pronounced in leaves than in peel when exposed to 42 °C or higher temperature. Leaves had negative L-and K-steps relative to the peel. The decrease of oxygen-evolving complex (OEC) by heat stress was higher in the peel than in the leaves. OJIP transient from the 46 °C treated peel could not reach the maximum fluorescence (F_m). The striking thermoeffect was the big decrease in the relative variable fluorescence at 30 ms (V_I), especially in the leaves. Compared with the peel, the leaves had less decreased maximum PS2 quantum efficiency (F_v/F_m), photochemical rate constant (K_P), F_m and performance index (PI) on absorption basis (PI_abs) and less increased minimum fluorescence (F₀) and non-photochemical rate constant (K_N), but more increased reduction of end acceptors at PS1 electron acceptor side per cross section (RE₀/CS₀) and per reaction center (RE₀/RC₀), quantum yield of electron transport from Q_A ⁻ to the end acceptors (ϕ _R0) and total PI (PI_abs,total) when exposed to 44 °C. In conclusion, PS2 is more thermally labile than PS1. The reduction of PS2 activity by heat stress primarily results from an inactivation of OEC. PS2 was more tolerant to high temperature in the leaves than in the peel.     

Transformation of apple (Malus domestica Borkh.) with the stilbene synthase gene from grapevine (Vitis vinifera L.) and a PGIP gene from kiwi (Actinidia deliciosa)

The objective of the present research was to introduce genes with antifungal potential into the commercially important apple cvs. Elstar and Holsteiner Cox in order to establish resistance against fungal diseases. The gene encoding the stilbene synthase (Vst1) from Vitis vinifera L., responsible for the synthesis of the phytoalexin resveratrol in grapevine, and the gene for a polygalacturonase-inhibiting protein (PGIP) from kiwi (Actinidia deliciosa) were transferred into Holsteiner Cox and Elstar via Agrobacterium tumefaciens-mediated transformation. A total of nine transgenic Holsteiner Cox clones and one transgenic E clone carrying the stilbene-synthase gene as well as three transgenic Holsteiner Cox lines harbouring the polygalacturonase-inhibiting protein from Kiwi were identified via polymerase chain reaction and Southern blot analysis. High performance liquid chromatography analysis revealed the accumulation of a resveratrol-derivate, a glycoside, in transgenic Vst1 plants.Abbreviations BAP 6-Benzylaminopurine - E Elstar - H Holsteiner Cox - HPLC High performance liquid chromatography - IBA Indole-3-butyric acid - NAA -Naphthaleneacetic acid - TDZ Thidiazuron - YEP Yeast extract broth Communicated by H. Lörz     

Insights into secondary growth in perennial plants: its unequal spatial and temporal dynamics in the apple (Malus domestica) is driven by architectural position and fruit load

Background and Aims

Secondary growth is a main physiological sink. However, the hierarchy between the processes which compete with secondary growth is still a matter of debate, especially on fruit trees where fruit weight dramatically increases with time. It was hypothesized that tree architecture, here mediated by branch age, is likely to have a major effect on the dynamics of secondary growth within a growing season.

Methods

Three variables were monitored on 6-year-old ‘Golden Delicious’ apple trees from flowering time to harvest: primary shoot growth, fruit volume, and cross-section area of branch portions of consecutive ages. Analyses were done through an ANOVA-type analysis in a linear mixed model framework.

Key Results

Secondary growth exhibited three consecutive phases characterized by unequal relative area increment over the season. The age of the branch had the strongest effect, with the highest and lowest relative area increment for the current-year shoots and the trunk, respectively. The growth phase had a lower effect, with a shift of secondary growth through the season from leafy shoots towards older branch portions. Eventually, fruit load had an effect on secondary growth mainly after primary growth had ceased.

Conclusions

The results support the idea that relationships between production of photosynthates and allocation depend on both primary growth and branch architectural position. Fruit load mainly interacted with secondary growth later in the season, especially on old branch portions.     

Different effects of two brassinosteroids on growth,auxin and cytokinin content in tobacco callus tissue

Two synthetic brassinosteroids, 24-epibrassinolide (24-epiBR) and 2,3, 17-trihydroxy-5-androstan-6-one (THA-BR), exhibit different effects on growth of tobacco callus tissue. When added to a culture medium containing growth-limiting amounts of auxin, 24-epiBR reduced and THA-BR increased the fresh weight yield of tissue up to 53% and 207%, respectively, after 6 weeks of cultivation. The stimulatory and inhibitory effects of the two brassinosteroids on tissue growth occurred over a broad range of concentrations without a pronounced maximum corresponding to the yes or no type of response. Different effects of 24-epiBR and THA-BR on tissue growth were inversely proportional to the content of endogenous cytokinins. Maximum contents of predominant cytokinins N⁶-(²-isopentenyl)adenine (iP) and trans-zeatin (Z) in tissues supplied with 24-epiBR in growth-inhibiting concentrations were up to 3.7 fold and 3.4 fold higher, respectively, as compared to tissues grown on media containing growth-stimulating concentrations of THA-BR. Stimulation of tissue growth by THA-BR correlated with content of endogenous IAA and an inverse correlation was found between the content of endogenous IAA and cytokinins in tissues supplied with 24-epiBR. THA-BR exhibited weak cytokinin-like activity in a bioassay based on stimulation of growth of lateral buds of pea while 24-epiBR was inactive. Results indicate that the qualitatively different effects of the two brassinosteroids on growth of tobacco tissue may reflect their different influence on content of endogenous cytokinin.Abbreviations BR(s) brassinosteroid(s) - 24-epiBR 24-epibrassinolide - THA-BR 2,33, 17-trihydroxy-5-androstan-6-one - CK(s) cytokinin(s) - iP N⁶-(²-isopentenyl)adenine - [9R]iP N⁶-(²-isopentenyl)adenosine - Z trans-zeatin - [9R]Z ribosyl-trans-zeatin - ABA abscisic acid - IAA indole-3-acetic acid - NAA naphtalene-1-acetic acid - DEAE cellulose diethylaminoethyl cellulose - HPLC high performance liquid chromatography - ELISA enzyme linked immuno-sorbent assay     

Correlation of spermine levels with ovary senescence and with fruit set and development inPisum sativum L.

Separation and quantitation of polyamines from unpollinated pea (Pisum sativum L.) ovaries and young fruits induced by application of gibberellic acid to unpollinated ovaries showed, in both cases, a decrease in putrescine and spermidine levels between anthesis and 4 d later. By contrast, spermine levels increased prior to the onset of senescence of the unpollinated ovaries (3 d post anthesis) and decreased during fruit development. Low levels of putrescine, spermidine and spermine were also observed in young fruits obtained by self-pollination and by treatment of unpollinated ovaries with 2,4-dichlorophenoxyacetic acid. In-vitro culture of ovary explants in a medium containing spermine showed that a reduction of the growth of gibberellic acid-treated unpollinated ovaries was associated with a rise in the level of spermine in the fruits. The results obtained indicate that changes in spermine levels are involved in the control of ovary senescence and of fruit set and development.Abbreviations BA benzyladenine - 2,4-D 2,4-dichlorophen-oxyacetic acid - GA₃ gibberellic acid - HPLC high-performance liquid chromatography     

Synergistic activity of endochitinase and exochitinase from Trichoderma atroviride (T. harzianum) against the pathogenic fungus (Venturia inaequalis) in transgenic apple plants

Genes from the biocontrol fungus Trichoderma atroviride encoding the antifungal proteins endochitinase or exochitinase (N-acetyl--D-hexosaminidase) were inserted into Marshall McIntosh apple singly and in combination. The genes were driven by a modified CaMV35S promoter. The resulting plants were screened for resistance to Venturia inaequalis, the causal agent of apple scab, and for effects of enzyme expression on growth. Disease resistance was correlated with the level of expression of either enzyme when expressed alone but exochitinase was less effective than endochitinase. The level of expression of endochitinase was negatively correlated with plant growth while exochitinase had no consistent effect on this character. Plants expressing both enzymes simultaneously were more resistant than plants expressing either single enzyme at the same level; analyses indicated that the two enzymes acted synergistically to reduce disease. Selected lines, especially one expressing low levels of endochitinase activity and moderate levels of exochitinase activity, were highly resistant in growth chamber trials and had negligible reduction in vigor relative to control plants. We believe that this is the first report of resistance in plants induced by expression of an N-acetylhexosaminidase and is the first report of in planta synergy between an exochitinase and an endochitinase.     

Expression of MdCAS1 and MdCAS2, encoding apple beta-cyanoalanine synthase homologs, is concomitantly induced during ripening and implicates MdCASs in the possible role of the cyanide detoxification in Fuji apple (Malus domestica Borkh.) fruits

Fruit ripening involves complex biochemical and physiological changes. Ethylene is an essential hormone for the ripening of climacteric fruits. In the process of ethylene biosynthesis, cyanide (HCN), an extremely toxic compound, is produced as a co-product. Thus, most cyanide produced during fruit ripening should be detoxified rapidly by fruit cells. In higher plants, the key enzyme involved in the detoxification of HCN is β-cyanoalanine synthase (β-CAS). As little is known about the molecular function of β-CAS genes in climacteric fruits, we identified two homologous genes, MdCAS1 and MdCAS2, encoding Fuji apple β-CAS homologs. The structural features of the predicted polypeptides as well as an in vitro enzyme activity assay with bacterially expressed recombinant proteins indicated that MdCAS1 and MdCAS2 may indeed function as β-CAS isozymes in apple fruits. RNA gel-blot studies revealed that both MdCAS1 and MdCAS2 mRNAs were coordinately induced during the ripening process of apple fruits in an expression pattern comparable with that of ACC oxidase and ethylene production. The MdCAS genes were also activated effectively by exogenous ethylene treatment and mechanical wounding. Thus, it seems like that, in ripening apple fruits, expression of MdCAS1 and MdCAS2 genes is intimately correlated with a climacteric ethylene production and ACC oxidase activity. In addition, β-CAS enzyme activity was also enhanced as the fruit ripened, although this increase was not as dramatic as the mRNA induction pattern. Overall, these results suggest that MdCAS may play a role in cyanide detoxification in ripening apple fruits.     

Perfect syncarpy in apple (Malus x domestica 'Summerland McIntosh') and its implications for pollination, seed distribution and fruit production (Rosaceae: Maloideae)

BACKGROUND AND AIMS: The gynoecium of the domestic apple, Malus x domestica, has been assumed to be imperfectly syncarpic, whereby pollination of each stigmatic surface can result in fertilization within only one of the five carpels. Despite its implied effect on fruit quantity and quality, the resulting influence of flower form on seed set and distribution within the apple fruit has seldom been investigated. Instead, poor fruit quality is usually attributed to problems with pollination, such as low bee numbers and/or ineffective pollinators within apple agro-ecosystems. The objective of this study was to determine the true nature of gynoecial structure and its influence on fruit production in the apple cultivar 'Summerland McIntosh'. METHODS: A stigma-excision method was used to determine the effects of uneven pollination among the five stigmas on fruit quantity (as measured by fruit set), and quality (seed number and distribution). In addition, flowers were examined microscopically to determine pollen tube pathways. KEY RESULTS: Fruit set, seed number, seed distribution, and the microscopic examination of flower gynoecial structure reported in this study indicated that the gynoecium of the cultivar Summerland McIntosh is perfectly syncarpic and not imperfectly syncarpic as previously thought. CONCLUSIONS: Pollination levels among the five stigmas need not be uniform to obtain full seed development within Summerland McIntosh fruit; even if one stigmatic surface is adequately pollinated, a full complement of seeds is likely. The importance of perfect syncarpy in recognizing true causes of poor fruit quality in apple is discussed.     

The effects of daminozide on the levels of Indol-3yl-acetic acid and gibberellins in radish (Raphanus sativus L.) in relation to the control of storage root growth

The levels of indol-3yl-acetic acid and gibberellins were determined in shoots and storage roots of radish (Raphanus sativus L.) at various times during the vegetative growth cycle of control plants and plants in which the root to shoot ratio was modified by daminozide treatment. In control plants the onset of storage organ growth was preceded by a change in the hormone root to shoot ratio to favour the root. There was a general reduction in hormone levels in daminozide-treated plants but the pattern of their distribution in roots and shoots was very similar to that in control plants. Thus the effects of daminozide on increased storage root growth cannot be explained in terms of altered root to shoot hormone ratios.     

Impacts of bleak (Alburnus alburnus) and roach (Rutilus rutilus) on water quality,sedimentation and internal nutrient loading

Lake Vesijärvi, southern Finland, suffered sewere eutrophication by sewage effluent from the city of Lahti during the 1960's and the early 1970's. The municipal sewage loading was diverted from the lake in 1976 and the lake started to recover. However, in the 1980's blue-green algal blooms increased again and the recovery of the lake faded. Enclosure experiments demonstrated that high roach (Rutilus rutilus) biomass is one of the key factors in the fading recovery of the lake. In this study, the influence of roach and another cyprinid fish species (bleak, Alburnus alburnus) to planktonic algal productivity and biomass in Lake Vesijärvi was examined. Enclosure experiments in the field showed the impacts of planktivorous bleak on water quality; in an enclosure with a density of 1 fish m^–2 average daily algal production (1370 mg C m^–2) and chlorophyll-a concentration (50–90 µg 1^–1) were more than twice that in an enclosure without fish. Laboratory experiments showed that the availability of planktonic food affects the foraging behaviour of roach and consequently the internal nutrient loading from the sediment into the water. Roach caused the highest phosphorus loading and turbidity when there was no zooplanktonic food available in the water. The possible interactions between planktivorous and omnivorous fish species are discussed.     

Grain filling pattern and cytokinin content in the grains and roots of rice plants

Grain filling patterns and their relationships withzeatin (Z), zeatin riboside (ZR), indole-3-acetic acid(IAA) and gibberellin (GA) contents in the grains androots during grain development were examined in sixrice (Oryza sativa L.) genotypes grown in thefield and in water culture. Three grain fillingpatterns based on the filling rate of superior andinferior spikelets were observed, i.e., fastsynchronous: all spikelets started filling early andfast at the early filling stage; slowsynchronous: all spikelets filled slowly at the earlyfilling stage and reached the maximum filling ratelate; and asynchronous: superior spikeletsstarted filling and reached the maximum filling ratemuch earlier than the inferior ones. The order ofgrain filling percentage in the three types of grainfilling patterns was: fast synchronous >asynchronous > slow synchronous. Changes in Z + ZRcontents in the superior and inferior spikelets wereassociated with the grain filling patterns. Grainfilling percentage was significantly correlated withZ + ZR contents in the grains and roots at the earlyand middle grain filling stages. IAA and GA(GA¹ + GA³ + GA⁴)contents in the grains and roots were notsignificantly correlated with grain fillingpercentage. The results suggest that cytokinins in thegrains and roots during the early phase of graindevelopment play an important role in regulating grainfilling pattern and consequently influence grainfilling percentage.