Winter Buds Research Articles

Melatonin enhances the recovery of cryopreserved shoot tips of American elm (Ulmus americana L.)

Climate change and global migrations of people and goods have exposed trees to new diseases and abiotic challenges that threaten the survival of species. In vitro germplasm storage via cryopreservation is an effective tool to ensure conservation of tree species, but plant cells and tissues are exposed to multiple stresses during the cryopreservation process. The current study was designed to evaluate the potential of melatonin to improve survival through the process of cryopreservation. Shoot tips of in vitro-grown plantlets and dormant winter buds of American elm were successfully cryopreserved in liquid nitrogen (LN) at -196°C under controlled environmental conditions following melatonin treatment and cold acclimation with either vitrification or encapsulation–vitrification protocols. Explants had optimal regrowth following cryopreservation when treated with the plant vitrification solution#2 (PVS2) for 10 min. Supplementation of both preculture and regrowth media with melatonin significantly enhanced regrowth of frozen shoots compared with the untreated control (P < 0.05). Approximately 80–100% of shoot explants grew under optimized conditions using melatonin-enriched media. Shoot tips of dormant winter buds consistently produced nearly 100% regrowth with both techniques. The main steps of the optimized protocol are14-day cold-acclimated cultures exposed to preculture medium with 0.1–0.5 lM melatonin for 24 hr, application of PVS2 for 10 min, rapid cooling in LN, rapid rewarming, removal of cryoprotectants, and recovery on a medium supplemented with 0.1–0.5 lM melatonin. Our results demonstrate the usefulness of the antioxidant melatonin for long-term storage of naturally resistant elm germplasm.

Concentration of Radiocesium in the Wild Japanese Monkey (Macaca fuscata) over the First 15 Months after the Fukushima Daiichi Nuclear Disaster

Following the massive earthquake that struck eastern Japan on March 11, 2011, a nuclear reactor core meltdown occurred at the Fukushima Daiichi Nuclear Power Plant, operated by Tokyo Electric Power Company, and was followed by the release of large amounts of radioactive materials. The objective of this study was to measure the concentration of radiocesium 134Cs and 137Cs in the muscle of Japanese monkeys (Macaca fuscata) inhabiting the forest area of Fukushima City and to determine the change in concentration over time as well as the relationship with the level of soil contamination. Cesium concentrations in the muscle of monkeys captured at locations with 100,000–300,000 Bq/m2 were 6,000–25,000 Bq/kg in April 2011 and decreased over 3 months to around 1,000 Bq/kg. However, the concentration increased again to 2,000–3,000 Bq/kg in some animals during and after December 2011 before returning to 1,000 Bq/kg in April 2012, after which it remained relatively constant. This pattern of change in muscle radiocesium concentration was similar to that of the change in radiocesium concentration in atmospheric fallout. Moreover, the monkeys feed on winter buds and the cambium layer of tree bark potentially containing higher concentrations of radiocesium than that in the diet during the rest of the year. The muscle radiocesium concentration in the monkeys related significantly with the level of soil contamination at the capture locations.

Formation of long‐lasting galls by overwintered nymphs in the Japanese aphid Quadrartus yoshinomiyai (Hemiptera: Aphididae: Hormaphidinae)

AbstractThis paper examines the life history of a generation of galls created by the aphid Quadrartus yoshinomiyai (Hormaphidinae: Nipponaphidini) on its primary host plant, Distylium racemosum. First‐instar fundatrix nymphs of Q. yoshinomiyai initiated galls on stems of developing shoots in early April and incipient enclosed galls were found from later the same month. The galls lasted for up to 14 months, during which they grew to maturity, opened in early or mid‐April of the following year and dried up by the end of June. First‐instar fundatrix nymphs were found on winter buds, indicating that they hatched from eggs in autumn and overwintered as nymphs. These results suggest that Q. yoshinomiyai has a three‐year life cycle.

오디 생산용 뽕나무 품종 "대자뽕"의 특성

주로 누에사육을 위한 사료 생산용으로 이용되던 뽕나무의 용도가 다양화 되면서, 부산물로 이용되어 온 오디가 새로운 소득작목으로 정착되어 가고 있다. 이에 따라 오디 생산에 적합한 뽕 품종에 대한 수요가 늘어나고 있어, 새로운 오디용 뽕품종으로 "대자뽕"을 육성하였으며 이 품종의 육성 경위와 특성은 아래와 같다. 1. '03-'06년까지 수원, 춘천 및 진주 3개 지역에서 청일뽕을 대조품종으로 하여 지역적응시험을 거쳤으며, 직무육성신품종 심의회에서 오디 생산용으로 우수한 품종임이 인정되어 품종으로 등록하였다. 2. 대자뽕은 농업과학원 잠사양봉소재과 수원 유전자원 포장에 보존되어 있는 2배성 국상 20호 계통에 콜히친을 처리하여 만든 염색체수가 2n = 56인 4배체 품종이다. 3. 대자뽕은 노상형(Morus Lhou(Ser.) Koidz)에 속하는 암나무이며, 발아개엽기는 청일뽕과 비슷한 직립성의 중생종이다. 4. 잎의 크기는 대형으로 두꺼우며, 잎모양은 타원형이다. 5. 오디는 식재 후 3년차부터 결실하기 시작하였으며, 10a 당 연간수량은 결실 1년차에는 청일뽕보다 33% 낮았으나, 2년차에는 17% 증수되어 2년 평균 3% 증수되는 다수성 품종으로 결실성이 우수한 품종이다. 6. 오디의 단과중은 4.5 g으로 대과형이며, 완숙된 오디의 색상은 적자색으로, 오디의 당도는 청일뽕보다 높은 고품질이다. We bred new mulberry cultivar through local adaptability test, which was registered as a new cultivar for fruit production. Local adaptability test had been carried out at three places(Suwon, Chuncheon and Jinju) for four years since 2003. This is tetraploidy variety belonging to (Morus Lhou(Ser.) Koidz.) made by colchicine treatment on growing point of winter buds. Daejappong was high yielding in productivity by 3% compared to control cultivar "Chungilppong" for two years.The sugar contents of Daejappong was higher than that of control cultivar 'Chungilppong(Morus alba L.)'. It is adaptable to every where except the places where cold damage and sclerotic disease happen frequently.

오디 생산용 뽕품종 "상베리"의 특성

주로 누에사육을 위한 사료 생산용으로 이용되던 뽕나무의 용도가 다양화 되면서, 부산물로 이용되어 온 오디가 새로운 소득작목으로 정착되어 가고 있다. 이에 따라 오디 생산에 적합한 뽕 품종에 대한 수요가 늘어나고 있어, 새로운 오디용 뽕품종으로 "상베리"를 육성하였으며 이 품종의 육성 경위와 특성은 아래와 같다. 1. '05-'10년까지 6년간 4개 지역(수원, 공주, 장성 및 상주)에서 청일뽕을 대조품종으로 하여 지역적응시험을 거쳤으며, 직무 육성신품종 심의회에서 오디 생산용으로 우수한 품종임이 인정되어 품종으로 등록하였다. 2. 상베리는 농업과학원 잠사양봉소재과 수원 유전자원 포장에 보존되어 있는 2배성 카타네오 계통에 콜히친을 처리하여 만든 4배체 품종이다. 3. 상베리는 백상형(Morus alba L.)에 속하는 암나무이며, 발아개엽기는 청일뽕과 비슷한 직립성의 중생종이다. 4. 잎의 크기는 대형으로 두꺼우며, 잎모양은 타원형이다. 5. 오디는 식재 후 2년차부터 결실하기 시작하였으며, 10a당 연간수량은 식재 3년차부터 6년차까지 4년 청일뽕 대비 평균 70% 증수되는 다수성 품종으로 초기 결실성이 우수한 품종이다. 6. 오디의 단과중은 4.7g으로 대과형이며, 완숙된 오디의 색상은 적자색이다. 오디의 당도와 산도는 청일뽕 오디보다 낮다. We bred a mulberry cultivar named Sangberry, through local adaptability test, which is under registration as a new cultivar for fruit production. Local adaptability test had been carried out at four places(Suwon, Kongju, Jangseong and Sangju) for six years from 2005. This is tetraploidy variety belonging to(Morus alba L.) made by colchicine treatment on growing point of winter buds. Sangberry was high yielding cultivar in fruit productivity by 70% compared to control cultivar "Chungilppong(Morus alba L.)" for four years. Although Sangberry was a little lower in sugar content of mulberry fruits, it contains more bioactive materials like C3G, rutin and amino acids than Chungilppong. It is adaptable to every where except the places where cold damage and sclerotic disease happen frequently.

Gis-Aided Analysis of Vineyards Damaged by Winter Frosts. Case Study: Cotnari Vineyard (Romania)

Abstract Thermal regime of Cotnari vineyard, Iaşi County, Romania, was surveyed between 2011-2012 using 10 Tinytag temperature loggers TK-4014, spread in 10 different lands of the vineyard. Research is developed in the framework of the international project TERVICLIM (France), by means of was created a global network of analyses points of vineyard climate, spread in 13 countries from both hemispheres of the globe, at different latitudes. The data recorded by temperature loggers in Cotnari vineyard shows that in 2011-2012 winter, the minimum temperatures dropped to -28.6ºC, with differences of up to 7.1ºC between the coldest and the mildest lands of the vineyard. Spatial distribution of the most harmful frost (12 February 2012) shows that more than 80% from the vineyard area was exposed to temperatures lower than -24ºC, 17% to harmful temperatures between -21..- 24ºC and 3% to temperatures between -18..- 21ºC, less damaging to vines. As consequences, the damages in the vine plantations varied from less significant negative consequences for vines in the mildest location, up to massive losses of winter buds, damages of annual branches and frozen vines in the coldest land of the vineyard. Analytical data of winter buds losses spatial distribution shows that at - 28.6ºC (absolute minimum temperature for the vineyard), 0.46% (7.29 ha) of vine plantations were exposed to vines losses, 3.81% of plantations to losses of annual branches, 62.49% from vine plantations to winter buds losses exceeding 40%, difficult to compensate by pruning, 13.5% to losses of winter buds exceeding 60%, requiring reestablishing the vegetative potential of vines. The only vine plantations protected by harmful frosts were those located at over 175 m altitude, in the highest lands of the vineyards.

Bud and shoot structure may relate to the distribution area of South American Proteaceae tree species

The capacity of preformation and neoformation and the structure of winter buds are vegetative attributes that may vary between plant species and according to ontogenetic stages of the same species. The present study describes and evaluates these features for the four tree species of Proteaceae occurring in Patagonia. In particular, it analyzes the structure and development of the distal buds of the trunk and the preformed or neoformed nature of the organs involved. Two of the species, Embothrium coccineum and Lomatia hirsuta, have scaly buds, in which primordia of green leaves are covered by cataphylls. The shoots of both species may include neoformed organs, more frequently so in juvenile trees. Lomatia ferruginea and Gevuina avellana have naked buds with a low number of primordia; in juvenile and adult trees of both species trunk shoots are entirely preformed. The structure of buds and shoots suggests two different growth modalities of the axes, which would be related to ecological breadth (narrower in the species with naked buds than in those with scaly buds) and distribution area of these species. The considerable morphological differences between the two Lomatia species studied raise the question whether they ought to be included in the same genus.

Transgene stability and agronomical performance of two transgenic Basta®-tolerant lines of Populus alba L

An experimental trial was established in greenhouse with transgenic white poplars from an elite genotype (Populus alba L. cv “Villafranca”) expressing the bar gene for tolerance to Basta® herbicide. The steady-state level of bar mRNA was evaluated in different seasons, over a two-year period. Notwithstanding the presence of the 35SCaMV promoter, significant fluctuations in the amount of the bar transcript in relation to season and organ position were observed. The highest expression levels were reached in apical leaves in summer, while a consistent drop in transgene expression occurred at the onset of dormancy and in winter buds. A similar expression pattern was evidenced for the poUBI gene used as endogenous marker. The genetically modified poplars were tolerant to intense and repeated glufosinate ammonium treatments, maintaining an intact shoot sprouting and a good height increment.

Purification, Characterization, and Molecular Cloning of Lectin from Winter Buds ofLysichiton camtschatcensis(L.) Schott

A novel lectin was purified to homogeneity from winter buds of Lysichiton camtschatcensis (L.) Schott of the Araceae family. It was a tetramer composed of two non-covalently associated polypeptides with small subunits (11 kDa) and large subunits (12 kDa). Sequencing of both subunits yielded unique N-terminal sequences. A cDNA encoding the lectin was cloned. The isolated cDNA contained an open reading frame that encoded 267 amino acids. It encoded both subunits, indicating that the lectin is synthesized as a single precursor protein that is post-translationally processed into two different subunits with 45% sequence identity. Each subunit contained a mannose-binding motif known to be conserved in monocot mannose-binding lectins, but its activity was not inhibited by monosaccharides, including methyl α-mannoside. Asialofetuin and yeast invertase were potent inhibitors. Lectin activity was detected in the buds formed during the winter season but not in the expanded leaves.

Genome wide identification of chilling responsive microRNAs in Prunus persica.

BackgroundMicroRNAs (miRNAs) are small RNAs (sRNAs) approximately 21 nucleotides in length that negatively control gene expression by cleaving or inhibiting the translation of target gene transcripts. Within this context, miRNAs and siRNAs are coming to the forefront as molecular mediators of gene regulation in plant responses to annual temperature cycling and cold stress. For this reason, we chose to identify and characterize the conserved and non-conserved miRNA component of peach (Prunus persica (L.) Batsch) focusing our efforts on both the recently released whole genome sequence of peach and sRNA transcriptome sequences from two tissues representing non-dormant leaves and dormant leaf buds. Conserved and non-conserved miRNAs, and their targets were identified. These sRNA resources were used to identify cold-responsive miRNAs whose gene targets co-localize with previously described QTLs for chilling requirement (CR).ResultsAnalysis of 21 million peach sRNA reads allowed us to identify 157 and 230 conserved and non-conserved miRNA sequences. Among the non-conserved miRNAs, we identified 205 that seem to be specific to peach. Comparative genome analysis between peach and Arabidopsis showed that conserved miRNA families, with the exception of miR5021, are similar in size. Sixteen of these conserved miRNA families are deeply rooted in land plant phylogeny as they are present in mosses and/or lycophytes. Within the other conserved miRNA families, five families (miR1446, miR473, miR479, miR3629, and miR3627) were reported only in tree species (Populustrichocarpa, Citrus trifolia, and Prunus persica). Expression analysis identified several up-regulated or down-regulated miRNAs in winter buds versus young leaves. A search of the peach proteome allowed the prediction of target genes for most of the conserved miRNAs and a large fraction of non-conserved miRNAs. A fraction of predicted targets in peach have not been previously reported in other species. Several conserved and non-conserved miRNAs and miRNA-regulated genes co-localize with Quantitative Trait Loci (QTLs) for chilling requirement (CR-QTL) and bloom date (BD-QTL).ConclusionsIn this work, we identified a large set of conserved and non-conserved miRNAs and describe their evolutionary footprint in angiosperm lineages. Several of these miRNAs were induced in winter buds and co-localized with QTLs for chilling requirement and bloom date thus making their gene targets potential candidates for mediating plant responses to cold stress. Several peach homologs of genes participating in the regulation of vernalization in Arabidopsis were identified as differentially expressed miRNAs targets, potentially linking these gene activities to cold responses in peach dormant buds. The non-conserved miRNAs may regulate cellular, physiological or developmental processes specific to peach and/or other tree species.

Cloning and Characterization of WOX4 Gene from Vitis vinifera L. Involved in Stem Cell Regulation

Abstract Wuschel-related homeobox (WOX) genes play essential, specific, and sometimes redundant roles in plant embryo development, shoot and root meristem maintenance, and plant development. Though much information was quickly gained with members of the WOX gene family of Arabidopsis, monocotyledonous crops, and gymnospermous conifers, little is known about perennial woody plants. In this study, we isolated the first WOX gene family member from grape (Vitis vinifera L. cv. Cabernet Sauvignon), and named it VvWOX4 based on its characteristic domains and phylogenetic analysis. The identity of VvWOX4 was validated by MALDI-TOF MS and Western blot with polyclonal antibody against Arabidopsis thaliana Wuschel. Functional analysis showed that VvWOX4 markedly increased shoot primordia structures when overexpressed under CaMV 35S promoter in tobacco. A different expression pattern was found for VvVOX4 compared with AtWUCHEL and its expression was detected in unique organs of grapevines. Besides the expression in the vegetative shoot apical meristem (SAM) of grape shoot tips, VvWOX4 is expressed in dormant winter buds, inflorescence, young leaves, and tendril tips, but not in root tips. In young leaves, the expression of VvWOX4 is strongly upregulated by wounding, and also by plant growth regulators such as 2 mg L −1 2,4-D, 1 mg L −1 NAA and 1 mg L −1 BAP treatments, while downregulation was monitored by 1 mg L −1 IBA treatment, and there was no response to 0.5 mg L −1 GA3 treatment. Together, our results revealed the first member of grape WOX gene family and indicated different roles and regulation of VvWOX4 in the perennial woody crop grapevine.

127. Ultrastructural changes in acclimated winter buds of Malus during preparative desiccation that is needed for successful cryoconservation

127. Ultrastructural changes in acclimated winter buds of Malus during preparative desiccation that is needed for successful cryoconservation

トドマツ, エゾマツ, アカエゾマツ稚苗の冬芽の發生, 開静及伸長生長開始に就て

トドマツ, エゾマツ, アカエゾマツ稚苗の冬芽の發生, 開静及伸長生長開始に就て

Characterization and identification of leaf morphology of Populus deltoides Bartr. clones

Leaves are of fundamental importance to plants, representing their facility to generate power and are the sensing units of plants towards the environment. An attempt was made to characterize and compare the variations of leaf morphology of various Populus deltoides Bartr. clones by studying the winter buds and other leaf parameters of fully developed leaves. To achieve these objectives, forty-three exotic and indigenous clones of P. deltoides Bartr. were evaluated for different parameters. On the basis of various morphological characteristics the results reveal that each clone has a distinct color pattern of leaves. Different colors observed in these clones varied from light green through green to dark green. Two distinct lengths of the leaf apex were found, i.e., short and long; as well both acuminate and acute apex types were found. Erratic distribution of serration of leaves was also found. In this study, the morphological traits of leaves provided discriminatory grounds for separating various populations of P. deltoides Bartr. clones. Winter bud studies indicate that different clones vary considerably with regard to shape, color, shape of leaf scars and exudation.

Effect of Geographical Location, Year, and Cultivar on Survival of Malus sp. Dormant Buds Stored in Vapors of Liquid Nitrogen

Clonal woody crop germplasm collections often originate and are grown in distinct geographical locations. Because the degree of cold-hardiness is known to be a factor in the successful use of dormant bud cryopreservation for Malus, it was suggested that material from relatively warmer climates would not respond to cryopreservation as well as material from colder environments. To test this hypothesis, the effect of growing provenance on cryosurvival of dormant buds from three Malus (apple) cultivars grown in three locations (Geneva, NY; Davis, CA; and Corvallis, OR) was tested in 3 consecutive years. Dormant winter buds were harvested at the three locations, cryopreserved, and bud viability was tested by grafting. The collective 3-year mean viability for cryopreserved dormant apple buds for the three locations ranged from 63% to 81% of the buds surviving with the highest survival from the Corvallis site; however, the Geneva twigs were exposed to the lowest preharvest temperature. These results suggest that the temperature at the growing location may not hinder application of the dormant bud cryopreservation method with Malus to the extent previously speculated.

Assessing the effects of climate change on the phenology of European temperate trees

Modelling phenology is crucial to assess the impact of climate change on the length of the canopy duration and the productivity of terrestrial ecosystems. Focusing on six dominant European tree species, the aims of this study were (i) to examine the accuracy of different leaf phenology models to simulate the onset and ending of the leafy season, with particular emphasis on the putative role of chilling to release winter bud dormancy and (ii) to predict seasonal shifts for the 21st century in response to climate warming. Models testing and validation were done for each species considering 2 or 3 years of phenological observations acquired over a large elevational gradient (1500 m range, 57 populations). Flushing models were either based solely on forcing temperatures (1-phase models) or both on chilling and forcing temperatures (2-phases models). Leaf senescence models were based on both temperature and photoperiod. We show that most flushing models are able to predict accurately the observed flushing dates. The 1-phase models are as efficient as 2-phases models for most species suggesting that chilling temperatures are currently sufficient to fully release bud dormancy. However, our predictions for the 21st century highlight that chilling temperature could be insufficient for some species at low elevation. Overall, flushing is expected to advance in the next decades but this trend substantially differed between species (from 0 to 2.4 days per decade). The prediction of leaf senescence appears more challenging, as the proposed models work properly for only two out of four deciduous species, for which senescence is expected to be delayed in the future (from 1.4 to 2.3 days per decade). These trends to earlier spring leafing and later autumn senescence are likely to affect the competitive balance between species. For instance, simulations over the 21st century predict a stronger lengthening of the canopy duration for Quercus petraea than for Fagus sylvatica, suggesting that shifts in the elevational distributions of these species might occur.

Generation of expressed sequence tags, development of microsatellite and single nucleotide polymorphism markers in Primula sieboldii E. Morren (Primulaceae) for analysis of genetic diversity in natural and horticultural populations

Primula sieboldii is a self-incompatible ornamental plant that has been cultivated for more than 300 years in Japan. In order to increase the available genomic resources for this species, 5651 expressed sequence tags (ESTs) were generated from seedling and winter bud cDNA libraries. After clustering and assembling ESTs, 2960 unigene elements that contain an homologous sequence to an S locus-linked gene of P. vulgaris were identified. In total, 127 simple sequence repeats (SSRs) were found. The most frequent di- and tri-SSR motifs were GA (78.6%) and ATT (12.9%), respectively. Twenty SSR and 40 single nucleotide polymorphism (SNP) markers were developed. These markers were evaluated for polymorphisms using 24 individuals (Y) from a population in Yatsugatake and 14 individuals (PS) from across the distributional range of P. sieboldii in Japan. The average level of expected heterozygosity within Y and PS was 0.39 and 0.48, respectively for EST-SSR markers and 0.26 and 0.29, respectively for SNP markers. The level of polymorphism for EST-SSR markers was lower than that for genomic SSR markers developed in a previous study. We also surveyed the genetic diversity of 56 cultivars, which had lower allelic richness than the Y and PS populations.

The effect of low temperature on the breaking of rest for winter bud in mulberry tree

The effect of low temperature on the breaking of rest for winter bud in mulberry tree

22. Critical steps in cryopreservation of dormant winter buds collected under relatively mild winter conditions

22. Critical steps in cryopreservation of dormant winter buds collected under relatively mild winter conditions

Lipids of the meristems of the main coniferous edificators from central siberia under low-temperature adaptation: 2. Features of the fatty acid metabolism of phospholipids from winter meristems of Larix sibirica Ledeb., Picea obovata L., and Pinus sylvestris L.

The results of studies of the fatty acid composition of phospholipids from bud meristems of the main coniferous edificators from Central Siberia, Larix sibirica Ledeb., Picea obovata L., and Pinus sylvestris L., in winter (in the state of low-temperature resistance) and in spring (after the lack of low-temperature resistance) are reported. The change in the phenological state of the tree upon the transition from dormancy to vegetation has been shown to be attended by a significant transformation of the fatty acid composition of phospholipids from living tissues, which is expressed not only in the alteration of the content of individual fatty acids and the ratio of the components of one or various types, but also in the absence of some fatty acids that are present in the winter buds.