Morphological and physiological markers of juvenility and maturity in shoot cultures of oak (Quercus robur and Q. petraea)

Evaluation of potential techniques for rejuvenating oak shoots requires robust quantitative markers for juvenile and mature plants. To identify suitable in vitro markers, shoot cultures of Quercus species of juvenile, adolescent and mature origin were screened for a range of morphological and physiological markers of juvenility and maturity. Criteria examined included angle of the shoot to the horizontal, stem length, stem diameter (tip, mid, base), leaf number, scale leaf number and shoot number. Image analysis was also carried out to determine leaf area, size, perimeter and breadth and length of leaves. Mature Q. robur L. clones had a larger mid-stem and tip diameter than juvenile clones, whereas mature Q. petraea ex Liebl. clones were characterized by plagiotropic growth and larger mid-stem and tip diameters compared with juvenile clones. Based on discriminant analysis of the data, we propose the following formulae for discrimination of juvenile and mature shoots, where a negative value for the discriminant score (D) indicates juvenility. For Q. petraea: D = -1.308 - 0.0351 SA + 2.206 TD + 1.435 MSD, where SA is stem angle ( degrees ), TD is tip diameter (mm) and MSD is mid-stem diameter (mm). For Q. robur: D = -3.546 + 2.418 TD + 2.202 MSD. Quercus robur clones derived from stump sprouts and designated as juvenile had a negative D value suggesting a juvenile status for these clones. Clones sourced from a hedged, grafted Q. robur tree of mature origin had a positive D value indicating a mature status. Clones initiated from a 20-25-year-old Q. petraea tree displayed morphology in vitro consistent with a mature status and had a positive D value; however, these clones displayed other traits such as vigor suggesting that vestiges of juvenility remain. Multiplication rate and leaf size and shape were variable among clones and did not provide suitable markers for juvenility or maturity for these Quercus species.     

Influence of temperature on the dynamics of ATP,ADP and non-adenylic triphosphate nucleotides in vegetative and floral peach buds during dormancy

The nucleotides test of endodormancy, which is based on the capacity of tissues to synthesize ATP and non-adenylic triphosphate nucleotides (NTP), cannot be used for floral buds, and it is of questionable use for vegetative buds. In an attempt to find an alternative test, we examined whether the dormancy state of vegetative and floral buds of trees exposed to different temperature conditions during the rest period is directly related to their ATP, ADP and NTP concentrations and ATP/ADP ratio. Once the buds had entered endo- or paradormancy, the nucleotide concentrations and the ATP/ADP ratio were low in the vegetative primordia and very low in the floral primordia. Only after the action of chilling, when the buds were considered to have completed the endodormancy and paradormancy phases, did the nucleotide concentrations increase, accompanied by a steep rise in ATP/ADP ratio. We conclude that the ATP/ADP ratio could be used to characterize the bud dormancy state by comparison with critical values of 1.5 for vegetative primordia and 1.0 for floral primordia.     

不同强度修剪对幼龄期油茶无性系生长及结实的影响

2004年冬在江西中部新余市设置两片油茶幼林修剪试验林。结果表明,在生长势良好林分中,修剪处理对次年春梢的抽生长度、次年新梢数量以及次年产量均具有显著或极显著影响,但对全年新梢长度及粗度,以及次年的花芽数影响不显著。修剪虽然对无性系间的新梢长度和粗度影响不大,但对无性系间的新梢数量、花芽数量及产量却有显著影响。修剪对亚林4号和亚林56号无性系有较好的促进作用,而对亚林27号无性系有较明显的抑制作用。其新梢数量分别比对照增加87.20%和6.47%,而亚林27号无性系则比对照低49.33%;次年结果数则分别比对照增加153.72%和5.49%,而亚林27号无性系则比对照低44.39%。在生长势一般的林分中,强度修剪明显抑制了幼林的生长。因此,油茶幼林修剪需要考虑无性系和长势因素。     

Shoot culture dynamics of six Populus clones

Shoot tips of five genotypically diverse Populus clones, P. alba x P. grandidentata 'Crandon,' P. nigra 'Betulifolia' x P. trichocarpa, P. nigra x P. laurifolia 'Strathglass,' P. maximowiczii x P. trichocarpa 'Androscoggin' and P. deltoides x P. nigra 'Eugenei,' were collected from hardwood cuttings, sterilized,and established in vitro. Stable shoot cultures were obtained from all clones except P. deltoides x P. nigra 'Eugenei'. The four poplar clones that formed stable shoot cultures together with a previously established P. tremula 'Erecta' clone were placed as two-node explants on either Murashige and Skoog medium or Woody Plant Medium containing benzyladenine to determine the rate of shoot multiplication, shoot growth and other responses of the clones. All five poplar clones showed rapid shoot multiplication when cultured in the presence of 0.4-1.0 microM benzyladenine on Murashige and Skoog medium, although P. tremula 'Erecta' produced a greater number of healthy shoots when grown on Woody Plant Medium. Individual shoot growth of all clones was more vigorous when the medium contained 0-0.1 microM benzyladenine, and 100% of such shoots rooted ex vitro.     

J 16: An apex protein associated with juvenility of Sequoiadendron giganteum

Shoot apex proteins from 11 seedlings, two juvenile clones and three mature grafted clones of Sequoiadendron giganteum (Lindl.) Buchh. were examined throughout the year by SDS-polyacrylamide gel electrophoresis. A membrane-associated protein known as J 16, with an approximate molecular weight of 16 kD, was always detected in juvenile tissue, but was absent in tissue of the mature clones. The J 16 protein was also present in extracts of a mature clone that had been morphologically rejuvenated by micrografting. Protein extracts from the rejuvenated clone, were separated by electrophoresis and assayed for J 16 using an immunoblotting procedure with anti-J 16 serum. This test revealed two bands, J 16 and another protein with a lower molecular weight. The small molecular weight protein was not detected in juvenile clones.     

Polyphenols as potential markers to differentiate juvenile and mature chestnut shoot cultures

The phenolic contents of eight in-vitro-cultivated chestnut clones (Castanea sativa Mill. and C. sativa x C. crenata Siebold & Zucc. hybrids) were analyzed both qualitatively and quantitatively. The aim of the work was to identify potential phenolic markers of: (i) juvenile or mature state; (ii) topophysical origin; and (iii) rooting capacity. A condensed tannin was detected in mature material but not in juvenile material, indicating that it could be used as a qualitative marker. Other qualitative phenolic differences were found between basal shoots and crown shoots of some clones, but it was not possible to discriminate among these materials in a general way. Canonical discriminant analysis was used for the study of quantitative markers. Differentiation between mature and juvenile material, and between materials differing in in vitro rooting capacity was possible according to the results of the analysis. Nevertheless, no significant quantitative differences were found between the phenolic content of material of basal shoot origin and that of crown shoot origin, indicating that the greater juvenility of material of basal origin compared with that of crown origin was not reflected in differences in phenolic content.     

Fall frost resistance in willows used for biomass production. I. Characterization of seasonal and genetic variation

Fast-growing willow clones (six clones of Salix viminalis L. and one clone each of S. viminalis x S. schwerenii E. Wolf and S. dasyclados Wimm.) were compared with respect to growth rhythm and frost hardening in the fall. Frost resistance of stem tissues was assessed by controlled freezing followed by analysis of chlorophyll fluorescence and scoring of visible cambial discoloration. The fluorescence method proved superior to scoring based on visible cambial discoloration because it was more rapid and less subjective, but needed calibration against cambial damage. Frost hardening in mature parts of stems did not start until growth cessation was initiated in the shoot apices, irrespective of whether growth cessation occurred early or late in the fall. Frost resistance varied because of clonal variations in: (1) pre-hardening frost resistance; (2) timing of growth cessation and hence start of frost hardening; and (3) rate of frost hardening. Compared with coastal and southern clones, continental and northern clones started hardening earlier, and a continental clone proceeded through hardening more rapidly at a given temperature. A cross between a continental and coastal clone was intermediate in timing. The pre-hardening frost resistance, however, was unrelated to both growth and frost hardening characteristics.     

Carbohydrate metabolism of vegetative and reproductive sinks in the late-maturing peach cultivar 'Encore'

Activities of NAD(+)-dependent sorbitol dehydrogenase (SDH), sorbitol oxidase (SOX), sucrose synthase (SS), acid invertase (AI), and neutral invertase (NI) in 'Encore' peach (Prunus persica L.) fruits and developing shoot tips were assayed during the growing season to determine whether carbohydrate metabolizing enzymes could serve as indicators of sink strength. In fruit flesh, SS activity was detected during Stage I of growth, when cells were actively dividing, and SDH activity was detected during Stage III, when cells were actively enlarging. Acid invertase activity was detected during Stage I and showed a closer correlation with relative increase in fruit weight during the growing season than SS activity. During seed filling and pit hardening (Stage II), when relative fruit growth rate was slowest, activities of carbohydrate metabolizing enzymes in fruit flesh were not detectable. No SOX activity was detected during Stages I and II. The highest sucrose content occurred near the end of fruit development when the activities of sucrose metabolizing enzymes were low. In developing shoot tips, the sorbitol:sucrose ratio was 2:1 (w/w) and SDH activity was low at the beginning and end of the season when vegetative growth was slowest. The sorbitol:sucrose ratio changed to 1:1 (w/w) along with an increase in SDH activity in shoot tips during the mid-growing season. In 'Nemaguard' peach, SDH exhibited higher activity in root tips than in other organs. Among the sorbitol- and sucrose-metabolizing enzyme activities, only SDH activity was positively correlated with shoot growth in 'Nemaguard' plants.     

Influence of nitrogen,photoperiod, cutting type,and clone on root and shoot development of rooted stem cuttings of sweetgum

This study was designed to test the effects of nitrogen fertilization, photoperiod, cutting type, and clone on root and shoot development of stem cuttings of sweetgum (Liquidambar styraciflua). Differences in the growth of roots and shoots were observed as a result of weekly applications of varying levels of nitrogen (N) fertilization (0, 25, 50, 100, or 200 mg N/liter). As the concentration of N increased, new-shoot dry weight increased, but root dry weight decreased at rates greater than 50 mg N/liter. The percentage of rooted cuttings surviving also decreased as N concentrations increased past 50 mg N/liter. A night-interruption light treatment did not significantly affect survival percentages or the amount of root and shoot growth. Across all treatments, only 46% of all cuttings produced new-shoot growth within a 15-week period following rooting. Cutting type (terminal or sub-terminal) affected rooted cutting development. A higher percentage of terminal cuttings survived and were deemed plantable. In contrast, sub-terminal cuttings produced more shoot and root growth. Differences among clones were observed for all traits measured.Manipulating N fertilization, in conjunction with using clones that propagate well, has the potential of producing rooted cuttings of a size adequate for plantation establishment. However, higher percentages of cuttings that produce new shoot growth shortly after rooting must be achieved.     

Changes in host chitinase isoforms in relation to wounding and colonization by Heterobasidion annosum: early and strong defense response in 33-year-old resistant Norway spruce clone

We studied the defense reactions of 33-year-old susceptible and resistant clones of Norway spruce (Picea abies (L.) Karst.) to the major root-rot fungus Heterobasidion annosum (Fr.) Bref. and determined if tissue cultures can be used as a model system for studying defense responses of mature trees at the molecular level. Quantitative PCR analysis of genomic DNA obtained from samples taken at different times along the lesion length in living bark indicated that the fungus was present in higher amounts and extended further into the host tissue in the susceptible clone than in the resistant clone. In protein extracts from the same lesion samples, there were differences in temporal and spatial changes in host chitinase isoform profiles between the resistant and susceptible clones. Host chitinase isoforms with pI values approximately 4.8, 4.4 and 3.7 increased more during the first 7 days after wounding and inoculation and extended further along the lesion length in the resistant clone than in the susceptible clone. These results suggest that the time from wounding and infection to induction of defense-related expression is shorter in the resistant clone indicating a more efficient host defense response than in the susceptible clone. Tissue cultures from the same clones were not resistant to H. annosum and showed no difference in the timing of the increase in chitinase isoforms in response to the pathogen. However, tissue cultures from both clones showed an increase in chitinase isoforms within 6 to 24 h past inoculation, indicating that increased chitinase expression in response to the pathogen is part of a general defense response common to both mature clones and tissue cultures.     

Regeneration and multiplication of Albizia procera Benth. through organogenesis

Vegetative propagation techniques are recognized as indispensable tools for mass multiplication of important multipurpose trees adopted in different agroforestry systems. Albizia procera, one among important species, is difficult to propagate commercially either by stem / root cuttings or layering. A study was undertaken to develop procedure for its in vitro regeneration through organogenesis. Explants collected from 15±2 yr-old mature plus trees and from 15 days old juvenile seedlings were regenerated with exogenous application of different hormones. Epicotyl and hypocotyl explants excised from juvenile seedlings showed higher callusing than axillary bud and shoot tip explants derived from mature trees. Benzylaminopurine (BA) at 3 μg/l was most effective, which induced hundred percent callusing in epicotyl and hypocotyl explants in 1/2 Murashige and Skoog (MS) medium. Callus originated from axillary buds and apical shoot tips of mature trees failed to form organs, however callus derived from epicotyl and hypocotyl explants proliferated and formed de novo shoots and leaflets. A concentration of 3 μg/l of BA was found effective for shoot proliferation. Shoots grew vigorously in 2 μg/l gibberellic acid (GA₃) treatment and rooted in 1/2 MS medium supplemented with indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). Rooting was most successful on medium supplemented with 6 μg/l IBA alone on which 93.3% of the shoots formed roots. Sand or vermiculite supplemented with 4 ml of yoshida solution proved as best hardening media, which recorded 70-80% survival of plantlets. One year old tissue culture raised plants had comparatively more height, collar diameter, biomass, and root shoot ratio than plants raised from cuttings and seeds of the same age. The procedures enumerated provide a basis for the development of in vitro techniques for rapid multiplication of A. procera. This revised version was published online in June 2006 with corrections to the Cover Date.     

Comparison of photosynthetic induction and transient limitations during the induction phase in young and mature leaves from three poplar clones

We tested the hypothesis that leaf age affects photosynthetic induction, because conductance to CO2 diffusion usually decreases with increasing leaf age. Photosynthetic inductions, primarily determined by the light modulation of Rubisco activity and stomatal opening, were investigated in both young and mature leaves, as defined by leaf plastochron index (LPI), from three poplar clones: Populus alba L., P. nigra L. and P. x euramericana (Dode) Guinier. In all clones, maximum assimilation rates (A max), maximum stomatal conductance (G Smax) and dark respiration rates (RD) were higher in young leaves (LPI = 3-5) than in mature leaves (LPI = 10-14), and A max decreased from P. alba via P. x euramericana to P. nigra. The clones with high photosynthetic capacity had low induction states 60 s after leaf illumination (IS60; indicating a slow initial induction phase), and required less time to reach 90% photosynthetic induction (T90). In contrast, the clone with the lowest photosynthetic capacity (P. nigra) exhibited high IS60 (high initial induction state) but a long induction time (high T90). A comparison of mature leaves with young leaves revealed significantly (P < 0.01) lower IS60 values in mature leaves of P. nigra only, and significantly higher T90 values in mature leaves of P. alba only. In all clones, young leaves exhibited a lower percentage of maximum transient stomatal limitation during photosynthetic induction (4-9%) compared with mature leaves (16-30%). Transient biochemical limitation, assessed on the basis of the time constants of Rubisco activation (tau), was significantly higher in mature leaves than in young leaves of P. alba; whereas there were no significant differences in tau between young and mature leaves of the other poplar clones. Thus, our hypothesis that leaf age affects photosynthetic induction was confirmed at the level of transient stomatal limitation, which was significantly higher in mature leaves than in young leaves in all clones. For the induction parameters IS60, T90 and tau, photosynthetic induction was more clone-specific and was dependent on leaf age only in some cases, an observation that may apply to other tree species.     

Evidence for increased sensitivity to nutrient and water stress in a fast-growing hybrid willow compared with a natural willow clone

The hypothesis that fast-growing breeds of willow (Salix spp.) are more sensitive to nutrient and water stress and less efficient in nutrient- and water-use than slower-growing natural willow clones was tested. Cuttings of a natural clone of S. viminalis L. collected in Sweden (L78183) and a hybrid clone of S. schwerinii E. Wolf. x S. viminalis L. ("Tora") were grown outdoors in pots under various experimental conditions in a full-factorial design. The experimental conditions included three fertilization, two irrigation and two temperature regimes. Classical growth analysis techniques, based on an initial and a final harvest, were used as a screening method, together with calculation of intrinsic water-use efficiency (foliar carbon isotope ratio; delta13C). In addition, nitrogen-use efficiency was calculated as the product of nitrogen productivity and mean residence time of nitrogen on an annual basis. There were significant differences in plant structural parameters (leaf area ratio, specific leaf area) and water-use efficiency between the clones. Furthermore, several clone x treatment interaction effects on various growth parameters indicated that the clones adapted to specific environments in different ways. "Tora" plants produced up to 25% more shoot biomass than plants of the natural clone in response to high rates of fertilization and irrigation, whereas clone ranking was reversed in most other treatments. The results support the hypothesis that fast-growing hybrids are more sensitive to nutrient and water stress than slower-growing natural clones. The hypothesis that natural clones have higher resource-use efficiency than fast-growing hybrids was supported with respect to water, but not nitrogen.     

A comparison of pre-planting treatments on hardwood cuttings of four hybrid poplar clones

Rooting and early growth of four hybrid poplar clones (Populus spp.) planted in a greenhouse were examined after applying 40 pre-rooting treatment combinations to dormant cuttings. Treatments included 2 cutting lengths (5 and 10 cm), 5 soaking times (0, 2, 4, 8, and 12 days), and 4 dips (chitosan, rooting hormone powder, liquid rooting hormone added to the soaking water, and none). Significant differences in both rooting percentages and growth were shown between clones after 7 weeks of growth. Ten cm cuttings had 29% greater rooting success, 28% more above-ground growth, and 12% lower root/shoot ratios than 5 cm cuttings. Cuttings planted without soaking had the lowest rooting success, at less than 45% on average. Commercial rooting hormones decreased the number of rooted cuttings but increased root/shoot ratios. For optimal rooting, we recommend using 10 cm cuttings, soaked for 2 days in water (4 days for the Jackii10 clone) without any additional dipping/hormone substance.     

赤桉人工种子制备过程中微芽繁殖体转株的研究

实现包裹后的微芽繁殖体的生根和萌发生长即转株是以微芽为繁殖体制备人工种子中较为重要的环节之一。本试验系统地研究了生长调节剂种类、浓度以及微芽发育状态与赤桉（Ｅｕｃａｌｙｐｔｕｓｃａｍａｌｄｕｌｅｎｓｉｓ）微芽转株的关系，研究了包裹基质中直接添加诱导生根的生长调节剂及微芽生根预诱导对包裹后微芽转株的影响。试验结果表明：ＮＡＡ较ＩＢＡ和ＩＡＡ有较好的诱导微芽生根的效果；选取幼嫩及活跃生长的微芽是获得较高生根率的重要环节；包裹基质中直接添加诱导生根的生长调节剂未能解决包裹后微芽转株的问题；微芽经生根预诱导再附带生根介质包裹对微芽转株有良好的促进作用。微芽包裹后二周生根率可达８１３％，三周后转株率可达８３３％。     

湿地松、加勒比松及湿加松大树嫁接促花效果分析

通过大树嫁接促进接穗提早开花是缩短林木育种周期的重要手段。研究以湿地松（Pinus elliottii）精选种子园 3 个无性系为中间砧木,湿地松、加勒比松（P. caribaea）3 个变种和他们的杂交子 代湿加松（P. elliottii×P. caribaea）共 10 个无性系大树的新梢为接穗,研究砧木无性系、砧木无性系分 株、接穗树种、接穗无性系及其交互作用对接穗成活率、开花性别及数量的影响。结果显示：嫁接 258 条接穗,共成活 240 条,砧木 AE19 对 5 个树种的接穗都表现出最好的亲和力;嫁接 10 个月后,成活接 穗中有 129 条接穗开花,64.3% 的穗条产生了雌球花,17.8% 产生了雄花序,17.8% 接穗产生了雌雄花; 砧木无性系、接穗无性系均对雌球花量、雄花序量有显著的影响,一些砧木无性系对某一性别花量促进 有倾向性;砧木无性系分株、接穗树种对雌球花量有显著的影响。     

Water stress, photosynthesis and early growth patterns of cuttings of three Populus clones

Photosynthetic attributes, leaf area and early root growth patterns were studied in three Populus clones to identify traits associated with superior growth potential on sites where water could be a limiting factor. It was found that early root growth and superior leaf area production were more closely related to growth potential than were photosynthetic capacity or carboxylation efficiency. A hybrid clone of Populus nigra var. charkowiensis (syn. P. nigra var. plantierensis) x P. nigra cv. 'Incrassata' (NE308) had more leaf area production and greater root system development in both wet and dry soil than did a P. trichocarpa clone (T6) and a P. balsamifera clone (B3). Despite greater above- and below-ground productivity, plants of clone NE308 had significantly lower photosynthetic capacity and carboxylation efficiency and a slightly higher CO(2) compensation point than plants of clones T6 and B3. Rapid early leaf and root growth appear to be key attributes associated with productivity in these clones regardless of soil water availability.     

摘除花果对岑溪软枝油茶春梢生长的影响

对岑溪软枝油茶采穗圃进行控花控果对比试验,结果表明：摘除花果能促进采穗圃春梢早发芽,春梢的梢枝数、小枝长和小枝基径分别比对照增加7．03％～31．30％、32．63％～64．20％和31．12％～36．47％;摘除花果对不同品系油茶春梢的影响也不同,摘除花果对无性系3有较好的促进作用,而对无性系1和无性系2的效果不如无性系3;对油茶进行控花控果在阴坡的效果好于阳坡。     

桉树无性系顶梢伸长节律及其在良种选择中的潜在作用

分析63个桉树无性系种植后第2年的顶梢伸长过程,并用一个Sigmoid-Logistic混合生长方程对每一无性系分别予以拟合。所有拟合结果均极为理想,拟合度(R2)高达0.97以上,拟合显著性水平P<0.0001,4个方程参数在拟合中作用突出,大多数情况下P<0.0001。顶梢伸长节律和林木生长均呈现出无性系间的变异;所有无性系均明显地呈现出早晚缓慢生长期和中间速生期;大部分无性系,特别是快速生长者,顶梢在气温较低的冬天仍然继续生长。秋冬季顶梢伸长是否停止或快慢可能说明桉树无性系林木对低温危害的敏感程度。根据顶梢伸长特征,可以将桉树无性系分成不同类型,选择适合不同需求的无性系。建议将种植区分成3类区域,并采用相应的无性系选育策略。