The influence of ethylene on in vitro rooting of GF 677 (Prunus persica × Prunus amygdalus) hybrid peach rootstock

Summary Treatments differing from each other for the type of tube closure (i.e., cotton plug for free gas exchange, airtight rubber cap, and rubber cap with ethysorb) and/or rooting culture medium (i.e., enriched or not by 25 to 100 μM acetylsalicylic acid) were compared for their effects on gaseous composition of the culture atmosphere and microcutting rooting of the GF 677 (Prunus persica × Prunus amygdalus) hybrid. Rubber capping, which leads to rapid ethylene accumulation inside tubes, strongly reduced rooting time and in some cases enhanced final rooting percentage over that of cotton plugs. Ethysorb almost completely absorbed ethylene produced by shoots, which showed lower rooting percentages within 9 d than microcuttings cultured in the absence of ethysorb. In contrast, no significant difference in rooting was found between the two treatments after 14 d. Carbon dioxide concentration was similar in all treatments within 5 to 9 d and seemed to be ineffective for rooting. The influence of acetylsalicylic acid on rooting was unclear. Root number and length were not significantly influenced by the treatments. These results demonstrate that the use of airtight closures, leading to rapid ethylene accumulation, can reduce time of rooting expression for GF 677 microcuttings. However, free gas exchange towards the end of the rooting period (from Day 9 to Day 14) is advisable to prevent leaf yellowing. No significant difference in plantlet survival and growth after transfer ex vitro was found among treatments.     

Enhanced seed dispersal of Prunus africana in fragmented and disturbed forests?

Forest destruction and disturbance can have long-term consequences for species diversity and ecosystem processes such as seed dispersal. Understanding these consequences is a crucial component of conserving vulnerable ecosystems. In the heavily fragmented and disturbed Kakamega Forest, western Kenya, we studied seed dispersal of Prunus africana (Rosaceae). In the main forest, five forest fragments, and differently disturbed sites, we quantified the overall frugivore community as an indicator for species diversity. Furthermore, we determined the frugivores on 28 fruiting P. africana trees, estimated seed dispersal, crop size and the general fruit availability of surrounding trees. During the overall frugivore census we recorded 49 frugivorous species; 36 of them were observed visiting P. africana trees and feeding on their fruits. Although overall frugivore species richness was 1.1 times lower in fragments than in main forest sites and 1.02 times higher in highly disturbed than in less disturbed sites, P. africana experienced 1.1 times higher numbers of frugivores in fragments than in main forest sites and 1.5 times higher numbers of frugivores in highly disturbed than in less disturbed sites. Correspondingly, seed dispersal was 1.5 times higher in fragments than in main forest sites and 1.5 times higher in more disturbed than less disturbed sites. Fruit availability of surrounding trees and crop size influenced the number of visitors to some degree. Thus, the number of dispersed seeds seemed to be slightly higher in fragmented and highly disturbed sites. This indicates that loss of single species does not necessarily lead to a decrease of ecosystem services. However, loss of diversity could be a problem in the long term, as a multitude of species might act as buffer against future environmental change.     

Evaluation of the Hydroxynitrile Lyase Activity in Cell Cultures of Capulin （Prunus serotina）

Enzymatic preparations obtained from young plants and cell cultures of capulin were screened for hydroxynitrile lyase activity. The threeweek old plants, grown under sterile conditions, were used to establish a solid cell culture. Crude preparations obtained from this plant material were evaluated for the transformation of benzaldehyde to the corresponding cyanohydrin (mandelonitrile). The results show that the crude material from roots, stalks, and leaves of young plants and calli of roots, stalks, internodes and petioles biocatalyzed the addition of hydrogen cyanide (HCN) to benzaldehyde with a modest to excellent enantioselectivity.     

Fungal biomass production in response to elevated atmospheric CO2 in a Glomus mosseae–Prunus cerasifera model system

Biomass and length of intraradical and extraradical mycorrhizal mycelium under ambient (aCO₂) and elevated (eCO₂ ) atmospheric CO₂ was investigated using a non-destructive in vivo experimental model system. Time-course experiments allowed measurements of intact extraradical mycelium spreading from mycorrhizal roots of Prunus cerasifera micropropagated plants inoculated with the arbuscular mycorrhizal fungus Glomus mosseae, in controlled environmental chambers. The length of extraradical mycelium was significantly increased at the highest CO₂ concentration, ranging from 10.7 to 20.3 m at aCO₂ and eCO₂, respectively. The biochemical determination of mycelial glucosamine content allowed the evaluation of intraradical and extraradical fungal biomass, which were 2 and 3 times larger at eCO₂ than at aCO₂. Present data show that Glomus mosseae responds to increases of CO₂ concentrations producing larger mycorrhizal networks which may potentially represent carbon sink agents in soil ecosystems.     

Does Prunus serotina act as an aggressive invader in areas with a low propagule pressure?

Since most studies on Prunus serotina in Western Europe focused on heavily invaded areas, we wondered whether P. serotina also acts as an aggressive invader in areas with a low propagule pressure. Based on long-term data for the Liedekerke forest reserve, we found that connectivity to seed sources and light availability were the major drivers of P. serotina presence: long-distance dispersal events and ‘windows of opportunity’ seem to direct P. serotina colonization. In the studied forest, P. serotina could not be considered an aggressive invader since its spread slowed down rather quickly and did not hamper the establishment of native tree species. Furthermore, understory P. serotina showed low growth and seed production, while the high Rubus cover hampered germination and establishment. Nonetheless, calamities opening up the canopy layer in the few areas with high P. serotina sapling density might alter the course of the invasion process.     

Detection and Characterization of UDP-Glucose:Flavonoid O-Glucosyltransferases in the Leaves of Prunus ? yedoensis Matsum

A novel O-glucosyltransferase (I4'GT) which catalyzes the transferof D-glucose from UDP-D-glucose to position 4' of prunetin (4',5-dihydroxyl-7-methoxyisoflavone)was isolated from the leaves of Prunus ? yedoensis Matsum. andpurified 66-fold by precipitation with ammonium sulfate andchromatography on DEAE-cellulose. UDP-glucose:flavonol 3-O-glucosyltransferase(F3GT) was also isolated and purified 50-fold in the same manner.The molecular weights of both I4'GT and F3GT were estimatedby elution from a column of Sephadex G-100 to be about 51,000Da. The pH optima for I4'GT and F3GT activities were 8.0 and7.5, respectively. The specificities of I4'GT and F3GT for thesugar donor were quite strict, and only UDP-glucose could serveas glucosyl donor, both ADP-D-glucose and GDP-D-glucose beingineffective. The apparent K_m values for UDP-glucose and prunetinwere 10.0µM and 1.20µM, respectively, for I4'GT.The K_m values for UDP-glucose and quercetin were 9.8 µMand 1.21 µM, respectively, for F3GT. The activities ofboth I4'GT and F3GT were stimulated by 1 mM Mg*⁺ and stronglyinhibited by 1 mM Cu²⁺, 1 mM Zn²⁺ and various reagents thatreact with sulfhydryl groups. (Received May 16, 1990; Accepted September 3, 1990)     

Simple Sequence Repeat Analysis of Genetic Diversity in Primary Core Collection of Peach （Prunus persica）

In this study, the genetic diversity of 51 cultivars in the primary core collection of peach （Prunus persica （L.） Batsch） was evaluated by using simple sequence repeats （SSRs）. The phylogenetic relationships and the evolutionary history among different cultivars were determined on the basis of SSR data. Twenty-two polymorphic SSR primer pairs were selected, and a total of 111 alleles were identified in the 51 cultivars, with an average of 5 alleles per locus. According to traditional Chinese classification of peach cultivars, the 51 cultivars in the peach primary core collection belong to six variety groups. The SSR analysis revealed that the levels of the genetic diversity within each variety group were ranked as Sweet peach 〉 Crisp peach 〉 Flat peach 〉 Nectarine 〉 Honey Peach 〉 Yellow fleshed peach. The genetic diversity among the Chinese cultivars was higher than that among the introduced cultivars. Cluster analysis by the unweighted pair group method with arithmetic averaging （UPGMA） placed the 51 cultivars into five linkage clusters. Cultivar members from the same variety group were distributed in different UPGMA clusters and some members from different variety groups were placed under the same cluster. Different variety groups could not be differentiated in accordance with SSR markers. The SSR analysis revealed rich genetic diversity in the peach primary core collection, representative of genetic resources of peach.     

QTLs for powdery mildew resistance in peach ×Prunus davidiana crosses: consistency across generations and environments

Powdery mildew, caused by Sphaerotheca pannosa var. persicae is one of the most important diseases in European peach orchards. Quantitative trait loci controlling powdery mildew resistance were detected using three related F1, F2 and BC2 populations derived from the cross between the resistant parent P. davidiana clone P1908 and the susceptible peach cultivar Summergrand. Powdery mildew resistance of each population was evaluated under natural exposure, in several locations and over several years. Thirteen QTLs were detected. For nine of them, the favourable allele came from the resistant parent. Five QTLs were consistently detected across the three populations. The F1 hybrid used to produce F2 and BC2 populations had not inherited the favourable allele from P1908 for QTL detected on LG3 and LG8 in F1 population. QTLs were not detected in the corresponding regions in F2 and BC2 populations. In two other genomic areas, significant substitution effects between P1908 alleles were evidenced in the F1 population, but the favourable allele came from Summergrand in the F2 and BC2 populations. Analysis of phenotypic data suggested an important qualitative change in the distribution of powdery mildew resistance after 1996, confirmed by QTL analysis. Indeed, a dramatic decrease of the effect of the major QTL previously detected on LG6 was observed after 1996, while the QTL on LG8 was increasingly involved in the control of powdery mildew resistance. Consequences for peach breeding strategies to improve powdery mildew resistance are discussed.     

Physiological and transcriptional responses in the iron–sulphur cluster assembly pathway under abiotic stress in peach (Prunus persica L.) seedlings

As one of the most indispensable element in mineral nutrition of plants, iron (Fe) is closely related to fruits quality and yield. However, molecular mechanisms towards Fe metabolism in fruit trees is largely unclear. In higher plants, iron–sulphur (Fe–S) cluster assembly occurs in chloroplasts, mitochondria and cytosol involving dozens of genes. In this study, we identified 44 putative Fe–S cluster assembly genes in peach (Prunus persica cv. ‘Xiahui6’), and analyzed Fe–S cluster assembly gene expression profiles in response to abiotic stresses. Peach seedlings were more sensitive to iron deficiency, drought and salinity stress, evidenced in reduced photosynthetic performance and altered activity of nitrite reductase, succinate dehydrogenase and aconitase. In addition, Fe–S cluster assembly genes are differentially regulated by abiotic stresses. Iron depletion and drought stress are likely to affect Fe–S cluster assembly genes in leaves. Excess iron toxicity mainly induces Fe–S cluster assembly gene expression in roots, whereas salinity stress massively inhibits Fe–S cluster assembly gene expression in roots. Interestingly, we found that un-functional scaffolds are more prone to disappear during the long-term evolution in perennial woody plants. Our findings directly provide molecular basis for Fe metabolism in peach, and favorably reveal potential candidate genes for further functional determination.     

Effects of exogenous l-arginine on in vitro rooting, chlorophyll, carbohydrate, and proline concentrations in the sweet cherry rootstock M × M 14 (Prunus avium L. × Prunus mahaleb L.)

The effects of indole-3-butyric acid (IBA) alone and in combination with l-arginine on the morphogenic and biochemical responses in shoot tip explants of the cherry rootstock M × M 14 (Prunus avium × Prunus mahaleb) were examined. The maximum root number per rooted explant (16), root fresh (FW) and dry (DW) weights, as well as the rooting percentage (100 %) were recorded when 2 mg l^?1 IBA (alone) were applied. Including the lowest IBA concentration (0.5 mg l^?1) with the lowest and highest l-arginine concentrations (0.5 and 2 mg l^?1, respectively) resulted in the greatest root length. The maximum leaf chlorophyll concentration and shoot length of the initial explant were recorded when 0.5 mg l^?1 IBA plus 2 mg l^?1 l-arginine were applied. In addition, l-arginine in combination with IBA (1 and 2 mg l^?1) was found to suppress shoot FW and DW. On the other hand, l-arginine enhanced the promoting effect of IBA on both root length and leaf chlorophyll concentration. The carbohydrate and proline concentrations in leaves were not significantly altered with the application of IBA alone or in combination with l-arginine. On the other hand, the carbohydrate and proline concentrations in roots were decreased with the application of 1 and 2 mg l^?1 IBA with l-arginine, resulting in the suppression of the promoting effects of IBA. It is clear from the findings that l-arginine has a direct effect on the in vitro rooting of M × M 14 explants, is involved in the function of the photosythetic apparatus, influences leaf chlorophyll content, participates in carbohydrate biosynthesis and metabolism, and is involved in proline accumulation both in leaves and roots.     

Induction of Haploid Morphogenic Calluses from in vitro Cultured Anthers of Prunus Armeniaca cv. ‘Harcot’

Apricot (Prunus armeniaca) ‘Harcot’ anthers, were cultured in vitro for the production of haploid plants. The best androgenic response was achieved with Nitsch and Nitsch (1969) medium, supplemented with 4.52 μM 2,4-D, 4.52 μM zeatin, 2.85 μM IAA and 40 g l⁻¹ sucrose. Cultures were maintained in the dark for 8 days, at 28°C, followed by transfer to a 16-h photoperiod, with 35 μm m⁻² s⁻¹ light intensity and 24/22°C day/night temperature. The androgenic response was correlated with the floral bud size, its phenologic stage and the level of microspore evolution. Anthers containing microspores at the tetrad/uninucleate stage were the most appropriate. The ploidy level of the calluses was evaluated by flow cytometry revealing that they range from haploid to octaploid. Mixoploid calluses have also been identified. Histological studies showed that the haploid calluses have their origin in the microspores. Nodular structures consisting of cells with dense cytoplasm and differentiated xylem elements were observed and were surrounded by an autofluorescent layer, probably due to cutin deposition.     

Fe-EDDHA Promotes Rooting of Rootstock GF-677 (Prunus amygdalus × P. persica) Explants in vitro

The effect of organic (Fe-EDTA and Fe-EDDHA) and inorganic (FeCl₃) iron substances on rooting of the rootstock GF-677 (Prunus amygdalus × Prunus persica) in vitro was studied. Full rooting (100 %) was observed in explants nourished with Fe-EDDHA, while less rooting was found in the absence of iron or in the presence of FeCl₃. On the contrary, no root formation was observed in explants nourished with Fe-EDTA, which showed extremely lower chlorophyll and high iron contents at the end of the experiment. This revised version was published online in July 2006 with corrections to the Cover Date.     

A highly conserved α-tubulin sequence from Prunus amygdalus

The sequence of an -tubulin from Prunus amygdalus has been obtained by cDNA cloning. When this sequence is compared to that of the Tub1 gene from maize it shows a very high degree of similarity, much higher than any of the -tubulin sequences reported so far from plants. The expression of this gene is high in the stages of seed development where a high divisional activity is present. It is preferentially expressed in the radicular tissues as it is gene Tub1 in maize. Southern analysis indicates that this gene may from a subfamily of -tubulin genes having similar sequence and tissue specificity and existing at least in maize and in Prunus.     

Influences of Cross Pollination on Pollen Tube Growth and Fruit Set in Zuili Plums （Prunus salicina）

Zuili plum （Prunus salicina L.） trees usually set fruit poorly, although they produce high quality fruit. To elucidate the causes of the poor fruit set, pollen tube growth into pistils and fruit set percentage were investigated after cross-, self- and open-pollination. Ovule development in Zuili pistils was also investigated. Pollen tube penetration into the ovules via the obturator and micropyle was best when Zuili pistils were pollinated by cv. Black Amber （P. domestica） pollen grains, although cross-pollinations with Hongxinli and Miili （P. salicina） pollen were more effective than self- and open-pollination. The fruit set percentage was also highest in pistils pollinated with Black Amber pollen grains. Morphological observation of Zuili pistils revealed that the trees produce ＂double pistils＂, developing two ovaries from a basal pistil, at a rate as high as 28%. In such abnormal pistils, most ovules were lacking an embryo sac or were entirely degenerated. The percentage of normally developed ovules was 24.3% and 8.9% in normal and double pistils, respectively. From these results, we conclude that the main causes of poor fruit set of Zuili plums are a lack of effective cross-pollination and the production of high percentages of double pistils in which normally developed ovules are scarcely formed.     

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.     

Cross-linked enzyme aggregates of β-glucosidase from Prunus domestica seeds

Cross-linked enzyme aggregates (CLEAs) of β-glucosidase were prepared and characterized. Under the optimum conditions, the activity recovery of CLEAs reached 84?%. The reduction by NaBH(4) resulted in slightly lower activities of CLEAs, while their thermostability was enhanced. CLEAs were more thermally stable than free enzyme (half lives, 973 vs. 518?min at 50?°C), while less stable than seed meal (half life, 1,090?min). In 90?% (v/v) t-butanol, the half lives of CLEAs and free enzyme were 53 and 6.7?h, respectively. Besides, the catalytic efficiency (V (max)/K (m)) of CLEAs was comparable to free enzyme (0.42 vs. 0.47?min(-1) mg(-1)). This carrier-free immobilized enzyme had a network structure with multiple layers. The productivity of salidroside using CLEAs reached 150?g/l?g catalyst, while being 6.3?g/l?g with seed meal.     

Phylogeny and Classification of Prunus sensu lato （Rosaceae）

The classification of the economically important genus Prunus L. sensu lato （s.L） is controversial due to the high levels of convergent or the parallel evolution of morphological characters. In the present study, phylogenetic analyses of fifteen main segregates of Prunus s.I. represented by eighty-four species were conducted with maximum parsimony and Bayesian approaches using twelve chloroplast regions （atpB- rbcL, matK, ndhF, psbA-trnH, rbcL, rpL 16, rpoC1, rps16, trnS-G, trnL, trnL-F and ycfl） and three nuclear genes （ITS, s6pdh and Sbel） to explore their infrageneric used to develop a new, phylogeny-based classification relationships. The results of these analyses were of Prunus s.I. Our phylogenetic reconstructions resolved three main clades of Prunus s.I. with strong supports. We adopted a broad-sensed genus, Prunus, and recognised three subgenera corresponding to the three main clades： subgenus Padus, subgenus Cerasus and subgenus Prunus. Seven sections of subgenus Prunus were recognised. The dwarf cherries, which were previously assigned to subgenus Cerasus, were included in this subgenus Prunus. One new section name, Prunus L. subgenus Prunus section Persicae （T. T. yu ＆ L. T. Lu） S. L. Zhou and one new species name, Prunus tianshanica （Pojarkov） S. Shi, were proposed.     

甜樱桃(Prunus avium L.)品种S基因型鉴定

根据蔷薇科S-RNase基因(S基因)高度保守区C2和RC4区设计一对特异引物PruC2和PruC4R,对甜樱桃品种的基因组DNA进行S基因特异PCR扩增。克隆S基因的扩增片段,核酸序列在GenBank上搜索,确定了4种S基因的核酸序列和大小。结果表明,在琼脂糖凝胶上位置相同的扩增带其核酸序列相同,是同一种S基因。4种S基因扩增片段的大小分别是：S1为677bp,S3为762bp,S4为945bp,S6为456bp。参试的自交不亲和品种的S基因型分别是：红灯、红艳、早红宝石和先锋相同,为S1S3;抉择、红丰和那翁相同,为S3S4;大紫为S1S6;长把红为S1S4;养老为S2S6;自交亲和品种外引7号和斯太拉为S3S4。     

Phylogeny and spatio‐temporal diversification of Prunus subgenus Laurocerasus section Mesopygeum (Rosaceae) in the Malesian region

The goals of this study were to reconstruct the phylogeny of Prunus subgenus Laurocerasus section Mesopygeum and to provide a preliminary assessment of its spatio‐temporal diversification in the Malesian region. We inferred the phylogeny using nuclear ITS and ETS and plastid psbA‐trnH, rps16, rpl16, and trnC‐petN sequences. Our analyses support the monophyly of sect. Mesopygeum. Within sect. Mesopygeum, we identified four main subclades: (i) Prunus lancilimba from continental Asia; (ii) Prunus ruthii from Malay Peninsula; (iii) a subclade comprising species from areas centered on the Sunda shelf and also a few species from continental Asia and Wallacea; and (iv) a subclade composed of species from areas of the Sahul shelf, with a small number of taxa also from areas of the Sunda shelf, continental Asia, and the Philippines. We estimated that sect. Mesopygeum originated in continental Asia at c. 44.71 Mya (95% HPD: 31.66–46.90). Nine dispersals between major geographic areas were inferred. From continental Asia, three and two dispersals were inferred to the Sunda shelf and the Sahul shelf in the mid‐Oligocene, respectively. Two dispersals were inferred from the Sahul shelf region to the Sunda shelf in late Oligecene and early Miocene, respectively. There were also two dispersals inferred from the Sunda shelf region, one to the Philippines and one to Wallacea, in the middle and late Miocene, respectively. The diversification in sect. Mesopygeum was likely driven by active geologic events and orogenies in the Neogene in the Malesian region.