Exopolygalacturonase protein accumulates late in peach fruit ripening

Two exo-acting polygalacturonase enzymes (exoPG, EC 3.2.1.67) increase in activity as peach ( Prunu persica L. Batsch cvs Coronet and Flavorcrest) fruits ripen. By examining populations of fruit, we show that the increase in activity occurs late in ripening when the fruit are very soft (below 2 kgf). The more abundant form of the enzyme, exoPG 2, was extensively purified and analyzed for its amino acid content and N-terminal amino acid sequence. ExoPG 2 is a polypeptide of M, 66 000 and has a substantial excess of basic over acidic amino acids. Polyclonal antisera to exoPG 2 were raised in mice. The antisera inhibited the enzyme activity and recognized a M_r 66 000 polypeptide in Western blots. Western blot analyses of extracts of fruit ranked for softness revealed a M_r 66 000 polypeptide only in the softest fruit (less than 2.5 kgf). We conclude that the increased in exopolygalacturonase activity that occurs in very soft fruit is due to an increase in the amount of enzyme protein.     

A set of EST‐SSRs isolated from peach fruit transcriptome and their transportability across Prunus species

Twenty‐one expressed sequence tag–simple sequence repeat (EST–SSR) markers were developed in peach from a mesocarp cDNA library. Eighteen of them gave successful amplification in 22 peach genotypes and produced one to three alleles each with an average of 1.8 alleles per locus. The average value of expected and observed heterozygosities was 0.24 and 0.20, respectively. All the primers gave successful amplification in other six Prunus species (almond, apricot, sweet cherry, Japanese plum, European plum and Prunus ferganensis).     

Quantifying sink and source limitations on dry matter partitioning to fruit growth in peach trees

We describe an approach for determining the degree of sink and source limitations on peach ( Prunus persica L. Batsch) fruit growth during several growth periods. Source limitations on fruit growth may be due to either a shortfall in assimilate supply within the tree (supply limitation) or to a deficiency in the capacity of the translocation system to deliver assimilates in sufficient quantity to support the maximum fruit growth rate (transport/competition limitation). To ascertain the potential maximum rate of fruit growth, fruit thinning treatments were used. One month after bloom, the number of fruits per tree was adjusted to between 50 and 700 on an early and a late maturing peach cultivar (cvs Spring Lady and Cal Red, respectively). Rates of potential sink demand, potential source supply and actual fruit growth were estimated from sequential harvests of all fruits on 42 trees on two (Spring Lady) and three (Cal Red) dates. These values were used to estimate the proportion of potential growth achieved, and the supply and transport/competition limitations on fruit growth. The results indicated that source limitations were significant on trees with moderate to high fruit numbers. These source limitations were due to supply limitations during all harvest intervals and to transport/competition limitations during the early harvest intervals. Sink limitations occurred to the greatest extent during the mid-period of fruit growth on the later maturing cultivar.     

Two forms of exopolygalacturonase increase as peach fruits ripen

Abstract. Freestone peach cultivars are distinguished from clingstone cultivars by a more extensive softening of the mesocarp tissue, and by the separation of mesocarp and endocarp during ripening. Cultivars of both types have been reported to develop exopolygalacturonase activity during ripening, but the enzyme has not been characterized in any detail. During development of freestone peaches ( Prunus persica L. var Coronet), two exopolygalacturonase enzymes were detected 42, 65 and 85 d after full bloom and in ripe fruit. During ripening one enzyme (exoPG 1) increased 36-fold and the other (exoPG 2) 90-fold but exoPG 2 accounted for a 73% of the total activity in ripe fruit. ExoPG 1 was purified 24-fold and exoPG 2 540-fold. ExoPG 2 is a slightly acidic glycoprotein. ExoPG 1 and exoPG 2 differ slightly in their pH optima and in their responses to calcium: each produces monogalacturonic acid as a reaction product. Similar enzymes were found in Flavorerest, a semi-freestone peach.     

Light environment, growth and morphogenesis in a peach tree canopy

Morphogenic and growth processes were studied in relation to photosynthetically active radiation (PAR), and red, far red and blue spectral bands monitored at three different heights of a peach canopy during the vegetative season. The PAR intercepted by the bottom of the tree was significantly lower than that at the top, and blue fluence rate changed with height and season in a manner similar to PAR. Phytochrome photoequilibria indicated spatial and temporal differences in the three layers of the canopy: significantly lower values were detected at the bottom in correspondence with the maximum leaf area index. In this layer, a stimulation of internode elongation and a decrease of flower density were detected. Higher shoot growth rates and about double number of lateral shoots were found at the top of the canopy, where a greater number of sun leaves was present. Possible explanations in terms of different growth strategies induced by shade, depletion of blue, and low phytochrome levels at the bottom of the canopy are given.     

Effect of abscisic acid (ABA) on sugar accumulation in the flesh tissue of peach fruit at the start of the maturation stage

Experiments were conducted on¹⁴C-sorbitol, fructose, and glucose uptakeinto flesh discs, and sorbitol efflux from thediscs, with and without ABA application toexamine the effect of abscisic acid (ABA) onsugar accumulation in peach fruit flesh at thestart of the maturation stage in relation tomembrane transport. Total uptake of¹⁴C-sorbitol, fructose, and glucose intoflesh discs was effectively promoted by ABA ata concentration of 10^–5 M. PCMBS(p-chloromercuribenzensulfonicacid)-sensitive uptake, which was considered ascarrier-mediated uptake, of sorbitol into thediscs was clearly stimulated by ABA at10^–5 M, compared with glucose andfructose uptake. Sorbitol efflux from the discsacross the tonoplast was restricted by ABA at10^–5 M. ABA application todeveloping fruit increased sugar accumulationin the fruit. Estimated ABA concentration inthis fruit was approximately 10^–5 M. These results indicate that sugar accumulationin peach fruit flesh is stimulated by ABA at aconcentration of 10^–5 M both invitro and in vivo. ABA stimulatesuptake of sugars, especially sorbitol, into theflesh by enhancing carrier-mediated transportpossibly across both tonoplast and plasmamembrane.     

遮光对桃幼树形态及一些生理指标的影响

对不同遮光条件下,桃（Prunus persica L.）不同品种（‘朝晖’、‘早露蟠桃’和‘南方早红’）1年生幼树的形态和生理反应进行了研究。结果表明,在中度遮光条件下,品种‘朝晖’和‘南方早红’的叶面积增大;在重度遮光条件下,3个供试品种的新梢直径、新梢长度、叶面积、比叶鲜重和比叶干重均减小,且不同品种的变化幅度不同。以干物质增加量为耐弱光能力的判定指标,可以看出品种‘朝晖’较耐弱光,‘南方早红’耐弱光能力差。遮光能引起3个品种可溶性糖含量的下降。叶绿素a／b值的变化可用于判定桃品种的耐弱光能力。     

Sucrose phosphate synthase and other sucrose metabolizing enzymes in fruits of various species

Recent reports have suggested that sucrose phosphate synthase (EC 2.4.1.14), a key enzyme in sucrose biosynthesis in photosynthetic “source” tissues, may also be important in some sucrose accumulating “sink” tissues. These experiments were conducted to determine if sucrose phosphate synthase is involved in sucrose accumulation in fruits of several species. Peach (Prunus persica NCT 516) and strawberry (Fragaria x ananassa cv. Chandler) fruits were harvested directly from the plant at various stages of fruit development. Kiwi (Actinidia chinensis), papaya (Carica papaya), pineapple (Ananas comosus) and mango (Mangifera indica) were sampled in postharvest storage over a period of several days. Carbohydrate concentrations and activities of sucrose phosphate synthase, sucrose synthase (EC 2.4.1.13), and acid and neutral invertases (EC 3.2.1.26) were measured. All fruits contained significant activities of sucrose phosphate synthase. Moreover, in fruits from all species except pineapple and papaya, there was an increase in sucrose phosphate synthase activity associated with the accumulation of sucrose in situ. The increase in sucrose concentration in peaches was also associated with an increase in sucrose synthase activity and, in strawberries, with increased activity of both sucrose synthase and neutral invertase. The hexose pools in all fruits were comprised of equimolar concentrations of fructose and glucose, except in the mango. In mango, the fructose to glucose ratio increased from 2 to 41 during ripening as sucrose concentration more than doubled. The results of this study indicate that activities of the sucrose metabolizing enzymes, including sucrose phosphate synthase, within the fruit itself, are important in determining the soluble sugar content of fruits of many species. This appears to be true for fruits which sweeten from a starch reserve and in fruits from sorbitol translocating species, raffinose saccharide translocating species, and sucrose translocating species.     

Purification of an anionic isoperoxidase from peach seeds and its immunological comparison with other anionic isoperoxidases

A soluble anionic isoperoxidase (EC 1,11,1,7) was purified from peach ( Prunus persica L. Batsch cv. Merry) seeds. Purification was achieved by DEAE-Sephacel, Sephacryl S-300 and CM-cellulose chromatography. The purified isoperoxidase de-carboxylated indole-3-acetic acid (S_0.5 0.13 m M , Hill coefficient 1.7). Molecular mass, determined by gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis, was ca 60 kDa. Polyclonal antibodies were raised in rabbit against this isoperoxidase. Using immunoprecipitation this isoenzyme was found to be immunologically different from other soluble anionic isoperoxidases isolated from peach seeds.     

Partial deglycosylation of an anionic isoperoxidase from peach seeds – effect on enzyme activity, stability and antigenicity

An anionic isoperoxidase (EC 1.11.1.7) purified from peach seeds ( Prunus persica L. Batsch cv. Merry) was partially deglycosylated by glycopeptidase F (EC 3.2.2.18) treatment. A 40% deglycosylation resulted in an activity loss of 50% when assayed with o -dianisidine. 60% with guaiacol and 78% with 2,2'-azino-bis(3-ethyl)benzethiozoline-6-sulfonic acid (ABTS) as substrate. The indole-3-acetic acid oxidase activity loss was close to 55%. The partially deglycosylated isoperoxidase also showed a higher K_m value for H₂O₂ and higher values for Arrhenius activation energy and enthalpy of activation. There was a decrease in enzyme stability at 4°C after deglycosylation. Native and partially deglycosylated isoperoxidase reacted equally well in an enzyme-linked immunosorbent assay (ELISA) with rabbit polyclonal antibodies raised against the native enzyme. The carbohydrate moiety of this peach seed isoperoxidase appears to be important for enzyme activity and stability.     

Deciphering the metabolic pathways influencing heat and cold responses during post‐harvest physiology of peach fruit

Peaches are highly perishable and deteriorate quickly at ambient temperature. Cold storage is commonly used to prevent fruit decay; however, it affects fruit quality causing physiological disorders collectively termed ‘chilling injury’ (CI). To prevent or ameliorate CI, heat treatment is often applied prior to cold storage. In the present work, metabolic profiling was performed to determine the metabolic dynamics associated with the induction of acquired CI tolerance in response to heat shock. ‘Dixiland’ peach fruits exposed to 39 °C, cold stored, or after a combined treatment of heat and cold, were compared with fruits ripening at 20 °C. Dramatic changes in the levels of compatible solutes such as galactinol and raffinose were observed, while amino acid precursors of the phenylpropanoid pathway were also modified due to the stress treatments, as was the polyamine putrescine. The observed responses towards temperature stress in peaches are composed of both common and specific response mechanisms to heat and cold, but also of more general adaptive responses that confer strategic advantages in adverse conditions such as biotic stresses. The identification of such key metabolites, which prime the fruit to cope with different stress situations, will likely greatly accelerate the design and the improvement of plant breeding programs.     

桃果实成熟前后细胞壁成分和降解酶活性的变化及其与果实硬度的关系

比较桃品种‘双久红’和‘川中岛白桃’果实成熟前后20 d内果肉硬度、细胞壁成分和细胞壁降解酶活性变化的结果表明,桃果实成熟5 d后,‘双久红’桃果实的硬度、纤维素含量和原果胶含量均极显著高于‘川中岛白桃’:从成熟前15 d开始,‘双久红’的水溶性果胶含量、多聚半乳糖醛酸酶活性和纤维素酶活性均极显著低于‘川中岛白桃’;整个成熟期间,‘双久红’的果胶甲酯酶活性明显低于‘川中岛白桃’。相关分析表明,果实硬度与原果胶、纤维素含量呈极显著正相关,而与可溶性果胶含量、多聚半乳糖醛酸酶活性和纤维素酶活性呈极显著负相关。     

中国武夷山脉地区野生毛桃资源收集与初步评价

为明确中国武夷山脉地区野生毛桃(Prunus persica(L.) Batsch)资源分布现状,对浙江、江西、福建武夷山脉地区野生毛桃资源进行了系统的考察、收集和评价。此次考察共收集野生毛桃资源244份,来自于武夷山脉地区24个采集点。物候期评价发现这些资源多态性丰富。相关性分析显示,叶芽萌动期、花期与采集点海拔、纬度呈极显著正相关。果实评价显示果实性状多态性丰富。通过2年抗涝性评价,初步筛选出53株较抗涝野生毛桃资源后代单株。净光合速率、气孔导度、蒸腾速率与淹水时间相关性分析显示淹水时间与净光合速率、气孔导度、蒸腾速率均呈极显著负相关,而净光合速率、气孔导度、蒸腾速率三者之间呈极显著正相关关系。     

Sucrose metabolism and fruit growth in parthenocarpic vs seeded blueberry (Vaccinium ashei) fruits

Although fruit set and development are induced by applications of gibberellins, final fruit weight of gibberellin-induced parthenocarpic fruit is often less than that of pollinated fruit. We examined changes in the activities of sucrose-metabolizing enzymes and sugar accumulation in developing fruits of cultivated blueberry (Vaccinium ashei Reade) and their correlation with fruit growth upon pollination or exogenous applications of gibberellic acid (GA₃). The objective was to determine if differences in fruit growth could be attributed to differences in enzyme activities and subsequent sugar accumulation in fruits. The fruit development period of GA₃-treated fruits was 15 days longer than that of pollinated fruits. At maturity, GA₃-treated fruit accumulated an average of 180 mg dry weight while pollinated fruit accumulated 390 mg dry weight. Dry weight accumulation in nonpollinated fruits was negligible and these fruits abscised by 45 days after bloom (DAB). The total carbon (C) cost (dry weight C + respiratory C) for fruit development was 109 and 244 mg C fruit^-1 for GA₃-treated and pollinated fruits, respectively. Hexose concentration increased to 100 mg (g fresh weight)^-1 at ripening in both GA₃-treated and pollinated fruits. Nonpollinated fruits reached a maximum hexose concentration at 45 DAB. Sucrose phosphate synthase (EC 2.4.1.14) and sucrose synthase (EC 2.4.1.13) activities reached a maximum of ≤5.0 μmol (g fresh weight)^-1 h^-1 in both GA₃-treated and pollinated fruits. Soluble acid invertase (EC 3.2.1.26) activity increased to about 60 μmol (g fresh weight)^-1 h^-1 in both GA₃-treated and pollinated fruits at ripening, while in nonpollinated fruits, a maximum soluble acid invertase activity of 0.12 μmol (g fresh weight)^-1 h^-1 was measured at 24 DAB. Insoluble acid invertase activity declined during the early stages of fruit growth and remained relatively low throughout fruit development. Neutral invertase activity was low throughout development, increasing to 5 μmol (g fresh weight)^-1 h^-1 at ripening in GA₃-treated and pollinated fruits. Our studies demonstrate that blueberry fruit development does not appear to be limited by sucrose metabolizing enzyme activity and/or the ability to accumulate sugars in either GA₃-treated or pollinated fruits.     

Molecular cloning, identification, and chromosomal localization of two MADS box genes in peach （Prunus persica）

MADS box proteins play an important role in floral development. To find genes involved in the floral transition of Prunus species, cDNAs for two MADS box genes, PpMADS1 and PpMADS10, were cloned using degenerate primers and 5'- and 3'- RACE based on the sequence database of P. persica and P. dulcis. The full length of PpMADS1 eDNA is 1, 071bp containing an open reading frame (ORF) of 717bp and coding for a polypeptide of 238 amino acid residues. The full length of PpMADS10 cDNA is 937bp containing an ORF of 633bp and coding for a polypeptide of 210 amino acid residues. Sequence comparison revealed that PpMADS1 and PpMADS10 were highly homologous to genes AP1 and PI in Arabidopsis, respectively. Phylogenetic analysis indicated that PpMADS1 belongs to the euAP1 clade of class A, and PpMADS10 is a member of GLO/PI clade of class B. RT-PCR analysis showed that PpMADS1 was expressed in sepal, petal, carpel, and fruit, which was slightly different from the expression pattern of AP1; PpMADS10 was expressed in petal and stamen, which shared the same expression pattern as PI. Using selective mapping strategy, PpMADS1 was assigned onto the Bin 1:50 on the G1 linkage group between the markers MCO44 and TSA2, and PpMADS10 onto the Bin 1:73 on the same linkage group between the markers Lap-1 and FGA8. Our results provided the basis for further dissection of the two MADS box gene function.