Effects of IBA on the rooting of branch cuttings of Chinese jujube (<Emphasis Type="Italic">Zizyphus jujuba</Emphasis> Mill.) and changes to nutrients and endogenous hormones

‘Zhongqiusucui’ jujube secondary shoots were treated with 3-indolebutyric acid (IBA) at three concentrations, 500, 1000 and 1500 mg/L. Results show that IBA could significantly enhance rooting and root characteristics of cuttings and were best with IBA at 1500 mg/L. In the rooting process, the formation of adventitious roots was related to the consumption and accumulation of nutrients (soluble sugars and proteins) and the changes in endogenous hormones in phloem, leaf tips and leaf bases. The rooting of cuttings had a positive correlation with the consumption of soluble sugars during the period of callus formation and with the accumulation of soluble sugars during adventitious root formation and growth. Rooting was positively related to the breakdown of soluble proteins in the phloem when the callus formed, and had a positive correlation with its accumulation during adventitious root formation and growth. Leaf tips and leaf bases showed a reverse trend in changes of soluble protein. However, together with the phloem, leaf tips and leaf bases regulated and controlled the formation and development of adventitious roots. The main activities of soluble proteins exist in the leaf tips as this was the main source of soluble proteins. The relation between rooting and IAA (indole-3-acetic acid) content in phloem was positive and thus a high concentration of IAA could benefit the induction and formation of adventitious roots. However, rooting was negatively related with ABA (abscisic acid) and GA (gibberellic acid) and a high concentration of both could inhibit the induction and formation of adventitious roots. Rooting had a positive correlation with phloem IAA/ABA ratios, and higher ratios could improve rooting. Low concentrations of ZR (zeatin riboside) triggered the induction of adventitious roots, while higher concentrations promoted root growth. Endogenous hormones in leaf tips and bases had an impact on rooting. The activities of endogenous hormones mainly existed in leaf tips because they play a major role in the production and consumption of IAA and its ABA content increased during rooting. The ZR in leaf tips influenced the rooting of cuttings, especially in the callus formation and rooting stage. Leaf tips were the main source of GA.     

兴安圆柏嫩枝插穗生根相关内源激素特征分析

为了提高兴安圆柏扦插生根率,探讨兴安圆柏的扦插生根机理,以改进的HPLC法测定了兴安圆柏嫩枝插穗生根过程中4种内源激素的含量变化。结果表明,外源激素处理促进了插穗内IAA合成,显著提高了插穗内IAA含量、IAA/ABA值,ABA含量变化不明显;低浓度的ZT促进生根,插穗生根过程中ZT含量较低,NAA处理使ZT含量比对照显著降低;外施激素降低了生根初期插穗内GA含量,促进了愈伤组织和根原基形成,提高了不定根形成期插穗内GA含量,有利于不定根生长。     

Metabolic changes during adventitious root primordium development in <Emphasis Type="Italic">Tectona grandis</Emphasis> Linn. f. (teak) cuttings as affected by age of donor plants and auxin (IBA and NAA) treatment

Aging of the donor tree decreased adventitious root formation in shoot cuttings of Tectona grandis Linn. f. (teak). Exogenous application of auxins, i.e., α-naphthalene acetic acid (NAA) and indole-3-butyric acid (IBA) has a significant positive effect on the percentage of rooting. The maximum percent rooting was obtained with 4,000 ppm IBA as compared to other treatment. Significant increase in root number was recorded in shoot cuttings treated with 4,000 ppm NAA. The overall rooting response was better in the treatment with IBA rather than with NAA. Further periodic samples (0, 10, 20, and 30 days) were taken to assess the total soluble sugar, starch, protein, and peroxidase (PER) activity in the rooting zone of shoot cuttings of teak during adventitious root formation. Application NAA and IBA to shoot cuttings resulted in an increase in the level of total soluble sugar, starch, protein, and PER-activity in the rooting zone. The stored carbohydrates were utilized during adventitious root formation. Hence, total soluble sugar and starch contents of cuttings, irrespective of age of donor plants, decreased with the passage of time in cuttings planted for rooting. Significant fluctuations were observed in the protein content of cuttings during the time of root induction. There was an increase in the protein content with the passage of time from the day of planting up to its 20th day, followed by a sharp decline in the protein content of cuttings at the 30th day of planting, irrespective of the age of donor plants or the treatment of cuttings with auxins. Irrespective of donor plant age, PER-activity in the cuttings increased from the day of their planting for rooting up to the 20th day, and then declined at its 30th day of planting. It was interesting to note that PER-activity remained higher at all stages in the cuttings of 2-month-old seedlings which rooted profusely as compared to the cuttings of 15- and 30-year-old donor plants those rooted poorly. This study suggested that the exogenously applied NAA and IBA at different concentration seems to activate sugar metabolism for release of energy, protein and PER-activity which are necessary for cellular division and differentiation during adventitious root primordium initiation or development in the rooting zone of shoot cuttings.     

Influence of growth hormones on adventitious root formation in semi-hardwood cuttings of <Emphasis Type="Italic">Celasturs paniculatus</Emphasis> Willd.: a contribution for rapid multiplication and conservation management

Celastrus paniculatus Willd. (Celastraceae) is an important medicinal tree valued for its seeds. Indiscriminate collection of this plant from the habitat zones has posed a serious threat to its existence in the wild, results in globally a threatened plant species. Ex situ conservation is a prioritized agenda of research for redlisted plant species. Semi-hardwood stem cuttings collected from 10 to 15 year old trees were treated with different concentrations (0, 0.5, 1.0, 2.0, 3.0, 4.0 and 5.0 g l⁻¹) of growth hormones; indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and 1-naphthalene acetic acid (NAA) for positive effect on root induction. After 90 days of treatment, results were analyzed for percentage of rooting, root number and average root length per rooted cuttings. The highest rooting (57%) response with highest number of roots (~77.2) was obtained in indole-3-acetic acid at 3.0 g l⁻¹. Overall, high significant (P ≤ 0.05) rooting response was noticed in IAA treatment comparative to hormones IBA and NAA. Rooted cuttings exhibited 100% survival in the experimental field. This study reveals the propagation of semi-hardwood cuttings derived from partially matured branches is possible and it provides a basis for conservation of this threatened plant species by reducing pressure on its natural populations.     

Effect of branch position and auxin treatment on clonal propagation of <Emphasis Type="Italic">Tectona grandis</Emphasis> Linn. f.

The main objective of our clonal propagation program is the production of high quality timber and faster tree growth. Teak (Tectona grandis Linn. f.) is an important plantation species in the tropics but the demand of teak timber is increasing globally. Teak planting stock is still produced from seeds of unselected elite genotypes which may lead to poor stand establishment. The number of seeds per fruit and their germination ability are also variable and limited by several factors. Therefore an alternative method i.e., clonal propagation is required for mass multiplication of elite trees. It is an established fact that the clonal propagation or adventitious root formation in cuttings is markedly affected by several external and internal factors. Considering these, an experiment was conducted on leafy soft wood shoot cuttings of 3-year-old hedged teak stock plants grown in a hedge garden to study the effects of branch position and auxin treatment on adventitious root formation. A factorial completely randomized design was used for experimentation. Coppice shoots were collected from basal, middle and upper positions of the hedge garden; and mono-nodal leafy cuttings were prepared. These cuttings were treated with different concentrations of IBA and NAA; and were cultured in a mist chamber for rooting where the relative humidity was maintained at 85 ± 2% with maximum and minimum day-night temperature at 32 ± 1°C and 26 ± 1°C respectively. After 45 days the cuttings were removed from the rooting medium and observations were made. It was observed that the rooting parameters viz., percent rooting and percent sprouting, mean number of leaves, mean number of shoots and length, and mean number of roots and length per cutting were significantly influenced by the branch position and auxin treatment. Treatment with 4000 ppm IBA increased percent rooting and percent sprouting whereas NAA suppressed it. Treatment with 4000 ppm IBA also increased the mean number of leaves and shoots, and the length, and the mean number of roots per cutting. The highest mean number of roots and length were observed in cuttings taken from the branch emerging at the upper position followed by branches of middle and lower position. Cuttings of branches at a middle position showed the highest values of percent rooting, percent sprouting, mean number of leaves and shoots and mean shoot length per cutting. Furthermore, the interactive effect of branch position and auxin treatment on rooting and sprouting response was also found to be significant. Overall, the findings of this investigation suggest that middle branch position in hedge plants and 4000 ppm IBA can be made to induce maximum rooting and to produce high quality planting stock material for clonal forestry program of teak.     

Effects of mother tree ages,different rooting mediums,light conditions and auxin treatments on rooting behaviour of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> branch cuttings

Dalbergia sissoo Roxb. is one of the promising multipurpose tree species of South Asia. Most of the plantations of D. sissoo from seeds are facing severe threats due to the die-back disease, which ultimately causes death of this potential tree-species within a few months. Vegetative propagation could avoid the die-back disease. Thirty mother trees of different age-groups of D. sissoo were selected for evaluating the rooting behaviour of branch cuttings from D. sissoo as influenced by auxins (IAA or IBA at 100, 200, 500 mg·L⁻¹), ages of mother trees (10, 4 and 2 years old) and different environment conditions, i.e., different mediums (soil and sand) or light conditions (in shade and open condition). The results show that application of IAA and IBA induced more numbers of cuttings (collected from 10-year-old mother trees) to root compared to control. Branch cuttings of D. sissoo collected from 10-year-old mother trees and planted in soil bed in open conditions had 100.0% of cuttings to root in IAA (100 mg·L⁻¹) and IBA (200 mg·L⁻¹) treatments. Both rooting medium (Soil and sand) influenced significantly (p<0.05) on rooting response of branch cuttings. Soil medium was found to achieve maximum no. of branch cuttings to root, compared to sand medium.     

金叶银杏硬枝扦插繁殖生根过程及其生根机制研究

【目的】研究不同生长调节剂处理对金叶银杏硬枝扦插生根率和成活率的影响,探讨金叶银杏的生根机制,为金叶银杏苗木扦插快繁技术体系的建立提供技术支持和理论指导。【方法】通过NAA和IBA处理金叶银杏硬枝插穗,检测它们对插穗的生根状态、生根指标及IBA对扦插过程中插穗皮部可溶性糖、抗氧化酶、激素含量的影响,探究金叶银杏硬枝扦插繁殖技术和生根机理。【结果】1)金叶银杏插穗愈伤组织诱导期和形成期、不定根发生期和形成期分别在插后的20~26、26~42、42~53、53~79 d。2)800 mg·L^-1 IBA和400 mg·L^-1 NAA+400 mg·L^-1IBA(混合)处理的插穗愈伤组织和不定根出现期及生根率与根数都较其他相应处理的效果好,其中生根率分别达64.4%和51.1%;不定根数分别达8.6条和9.3条。3)插穗生根过程中(500 mg·L^-1 IBA处理)基部韧皮部的SOD、PPO活性在插后40、60 d达到高峰,均比对照提前20 d达到峰值;插穗的POD活性出现上升-下降-升高的趋势,插穗可溶性糖也出现升高-下降-升高的趋势,但时间点不一致;而淀粉的含量出现下降-上升的趋势。4)生根期间插穗基部韧皮部的IAA含量出现了2个峰值和一个谷值;而ZR含量在第40天达到峰值后下降;GA含量先下降后上升;ABA含量在插后的前期高后期低,且值都低于同期对照。IAA/ABA比值在插后的第40天(愈伤组织形成期)达到谷底,后又上升(不定根形成期)。IAA/GA在插后的40~60 d内快速上升,80 d后又开始大幅下降;IAA/ZR值在扦插愈伤组织形成期(40 d)呈下降趋势,到60 d时(不定根发生期)又急剧上升到峰值。【结论】高活性的SOD和POD有利于插穗初期愈伤组织的形成(40 d),高活性的POD与PPO有利于不定根的形成(60 d)。较高的IAA含量是保证愈伤组织形成(20~40 d)的必要条件,不定根形成期(60 d)需要较高的IAA/ZR、IAA/ABA、IAA/GA比值。本研究为金叶银杏扦插繁殖技术和生根机理的研究及金叶银杏苗木的快速繁殖生产提供技术支持。     

Vegetative propagation of <Emphasis Type="Italic">Litsea monopetala</Emphasis>, a wild tropical medicinal plant: Effects of indole-3-butyric acid (IBA) on stem cuttings

In this study we investigated the rooting ability and the growth performance of juvenile single-node leafy stem cuttings of Litsea monopetala (Roxb) Pers. collected from two mature mother trees preserved in the hill forest of Chittagong district, Bangladesh. The rooting ability of cuttings was studied under 0%, 0.1%, 0.2% and 0.4% indole-3-butyric acid (IBA) treatments. Significantly better rooting response (p ≤ 0.05) was observed with 0.1% IBA compared to control (0% IBA). The mean number of roots and the length of the longest root of cuttings in different treatments showed no significant difference (p≤0.05). After transfer into polythene bags from non-mist propagator, rooted cuttings treated with 0%, 0.1% and 0.2% IBA demonstrated the highest (100 ± 0.00%) survival capacity. The mean number of shoots developed in cuttings in the polythene bags in first three weeks varied significantly (p≤0.05) among the treatments. Effects of three fertilizer treatments, viz. T0 (no fertilizer), T1 (10g Urea, 20g TSP, 10g MOP dissolved in 1 L water) and T2 (10g Urea, 20g TSP, 10g MOP dissolved in 2 L water) on initial growth ofstecklings were also measured over a 90-days period. The increment of leaf area of stecklings was significantly higher (p≤0.05) under T0 compared with that under T1 and T2 while the increment of stem length, collar diameter and root biomass varied insignificantly among different fertilizer treatments. The results suggest that rooting juvenile single-node leafy stem cuttings could be an effective mean of regenerating L. monopetala. The application of 0.1% IBA concentration is recommended for rooting of juvenile leafy stem cuttings and application of fertilizer appeared unnecessary for the subsequent growth of stecklings in polythene bags.     

Root biomass and distribution of a <Emphasis Type="Italic">Picea–Abies</Emphasis> stand and a <Emphasis Type="Italic">Larix–Betula</Emphasis> stand in pumiceous Entisols in Japan

Root biomass and root distribution were studied in Entisols derived from the thick deposition of volcanic pumice on Hokkaido Island, Japan, to examine the effect of soil conditions on tree root development. The soil had a thin (<10 cm) A horizon and thick coarse pumiceous gravel layers with low levels of available nutrients and water. Two stands were studied: a Picea glehnii–Abies sachalinensis stand (PA stand) and a Larix kaempferi–Betula platyphylla var. japonica stand (LB stand). The allometric relationships between diameter at breast height (DBH) and aboveground and belowground biomass of these species were obtained to estimate stand biomass. The belowground biomass was small: 30.6 Mg ha₋₁ for the PA stand and 24.3 Mg ha₋₁ for the LB stand. The trunk/root ratios of study stands were 4.8 for the PA stand and 4.3 for the LB stand, which were higher than those from previous studies in boreal and temperate forests. All species developed shallow root systems, and fine roots were spread densely in the shallow A horizon, suggesting that physical obstruction by the pumiceous layers and their low levels of available water and nutrients restricted downward root elongation. The high trunk/root ratios of the trees may also have resulted from the limited available rooting space in the study sites.     

Stress responses of <Emphasis Type="Italic">Salix gracilistyla</Emphasis> and <Emphasis Type="Italic">Salix subfragilis</Emphasis> cuttings to repeated flooding and drought

To compare the responses to repeated flooding and drought of Salix gracilistyla, which grows on coarse gravel substrates, and Salix subfragilis, which grows on fine silt or clay substrates, we measured pre-dawn leaf water potential (Ψw _pd), osmotic adjustment (Ψw _tlp, Ψo _sat), and biomass production of cuttings under greenhouse conditions. The experimental design involved a control and four treatments that crossed 1 or 3-week flooding (F) with 1 or 2-week droughts (D). Ψw _pd was reduced after 2 weeks of drought when preceded by 1 week of flooding. Neither species increased osmotic adjustment in response to increased duration of drought between repeated 3-week flooding. Moreover, a decrease in the ratio of leaf biomass to total biomass or an increase in the ratio of root biomass to total biomass with longer drought repetitions was not observed for either species. The root ratio of S. gracilistyla was more strongly inhibited by flooding than that of S. subfragilis. The shoot-to-root ratio of S. subfragilis was higher than that of S. gracilistyla in all F combinations. The hypertrophied lenticel ratio of S. gracilistyla after 1 week of flooding was nearly the same as that after 3 weeks of flooding, whereas values for S. subfragilis after 1 week of flooding were lower than those after 3 weeks of flooding. The low allocation to roots and the generation of hypertrophied lenticels by S. gracilistyla in response to flooding, as compared with S. subfragilis, seem to be related to the different habitat substrate conditions of the two species.     

无患子硬枝扦插进程插穗内源激素和多酚类物质含量变化

为了解无患子硬枝扦插生根机制,以1年生无患子硬枝作为扦插材料,在扦插第0—80天调查愈伤组织发育和根系形成情况,测定插穗的腋芽(嫩叶)以及基部2cm长度的韧皮部内源激素和多酚类物质的含量。结果表明,无患子硬枝扦插第10天开始出现愈伤组织,第50天愈伤率达到最高值83.33%;第40天开始出现生根插穗,生根率为6.67%,随后不定根数量迅速增加,第70天生根率达到86.67%,此时平均有5.33条。之后生根数量不变,根系仍然在生长,且扦插后根系效果指数持续增高。插穗韧皮部内源激素含量变化较复杂。整体来看,高含量赤霉素(GA3)抑制愈伤组织和不定根形成,且在根原基发生期和不定根形成关键期达到峰值,在不定根速生期持续下降;高含量生长素(IAA)促进不定根的形成,且在扦插后第20天,即根原基发生期,达到峰值(92.7μg/g);玉米素核苷(ZR)对无患子硬枝扦插过程的生理作用较复杂,低含量的ZR有利于根原基的发生和不定根速生,但高含量的ZR促进不定根形成。韧皮部ZR/IAA在扦插后第20—40天呈下降趋势,第40—60天快速上升,促进不定根生长;GA3/IAA整体呈现抛物线形下降趋势,特别是在根原基发生期和不定根速生期,下降速率更快,以促进不定根的生长。无患子硬枝插穗生根进程中,多数多酚类物质对愈伤组织形成、根原基发生及不定根形成有极显著抑制作用,没食子酸抑制效果稍弱;插穗内多种激素和多酚类物质的含量均发生了变化,且对插穗生根产生重大影响。总之,插穗根原基发生和不定根形成等关键时期,IAA含量升高,GA3、ZR以及多酚类物质含量降低,植物体内多种内源物质此消彼长,达到动态平衡,共同促进插穗生根。     

Clonal propagation of <Emphasis Type="Italic">Dalbergia sissoo</Emphasis> Roxb. and associated metabolic changes during adventitious root primordium development

A study was conducted on the twelve clones of shisham (Dalbergia sissoo Roxb.). These clones were obtained from India and Nepal. Single-node leafy cuttings were prepared from the vegetative multiplication garden to examine the clonal variation, effect of IBA treatment on rooting response and associated metabolic changes during adventitious root formation. A remarkable and significant variation was observed due to treatment of 2,000 ppm IBA in the rooting parameters. Clonal variations were also significant for root and shoot growth while length of root was insignificant. Among the twelve clones studied; C3 (Tulsipur, Gonda, Uttar Pradesh, India) and C4 (Laxmipur, Gonda, Uttar Pradesh, India) clone cuttings have given the highest rooting response. Interaction (clone × IBA) was significant only for production of number of roots per cutting. Periodic sampling for clone C3 was performed at 0, 7, 14, 21, 28 and 35 days to examine the contents of total soluble sugars, starch, protein and peroxidase (PER) activity in the rooting zone of cuttings (∼0.5 cm) during adventitious root primordium development. A significant increase in all the metabolic activities was noted due to IBA. Total soluble sugars and starch contents of cuttings decreased with the passage of time. Protein content and PER-activity started to increase in the early stage and reached the highest level on day 21, followed by a decline at the 35th day of sampling. These trends were common for both IBA treated and untreated cuttings. Protein content and PER-activity remained higher in the rooting zone of IBA treated cuttings. Overall these findings suggested that exogenous application of IBA may have activated carbohydrate metabolism for release of energy, while protein and PER-activity were necessary for cell division and differentiation during adventitious root primordium initiation and development in the rooting zone of cuttings.     

Mini-cuttings: an effective technique for the propagation of <Emphasis Type="Italic">Pinus pinaster</Emphasis> Ait.

Pinus pinaster Ait. is one of the main forest tree species planted in Spain, Portugal and France. Due to its high economic relevance, there is considerable interest in developing techniques for vegetative breeding aimed at mass propagation. In this study we present a mini-propagation protocol in order to define an efficient method to propagate families or clones of P. pinaster. We carried out three experiments using mini-cuttings of 3–5 cm in length with the aim of evaluating the effects of temperature (4°C vs. 25°C), plant growth regulator (IBA) and shoot age on rooting ability. Percentage of rooted cuttings and morphological root variables were recorded. The percentage of rooted cuttings per treatment ranged from 68 to 97%. Treatment with IBA significantly influenced the rooting process at 25°C but not at 4°C. The number of apexes, length, area and volume of roots were all positively affected by temperature treatment. Shoot age also had a positive effect on rooting capacity of cuttings, with the cuttings from the youngest shoots (70 days after pruning) having higher rooting percentages, ranging from 84.7 to 98.3%. The use of juvenile material, good environmental conditions and IBA all benefited the rooting of clonal material, resulting in high rooting capacity. This study presents an innovative propagation protocol for P. pinaster that can be used as a tool in breeding programs.     

Clonal propagation of guava （Psidium guajava Linn.） by stem cutting from mature stockplants

Introduction Guava (Psidium guajava Linn.), belonging to the Family Myrtaceae, is originated in the tropical South America (Hayes 1970; Pathak and Ojha 1993) and grows wild in Bangladesh, India, Thailand, Brazil, Florida, West Indies, California and also …     

细柄蕈树嫩枝扦插生根过程中内源激素动态特性研究

以3年生细柄蕈树实生苗嫩枝为试验材料,运用酶联免疫吸附测定法(ELISA),研究其在扦插生根过程中内源激素含量的变化,旨在揭示细柄蕈树扦插生根的机理。结果表明,与CK相比,以质量浓度300 mg/L的NAA处理插穗,能有效缩短其生根周期;扦插生根过程中,NAA处理与CK的几种内源激素(IAA、ZT和ABA)含量随时间变化的趋势相同,均呈正"V"形变化;NAA处理可调节内源激素,促进不定根的形成和发育;不定根形成期间,NAA处理与CK的IAA/ZT、IAA/GA3和IAA/ABA谷值的出现时间(20 d和30 d)与愈伤组织形成期重合,内源激素的这种节奏性变化反映了生根与生理之间的协同性。     

杂种落叶松连续繁殖与插穗生根关系的生理研究

观察了日×长杂种落叶松连续繁殖插穗不定根发育进程,研究了连续繁殖对插穗生根力衰退的阻滞效应,并分析了一轮采穗圃与原株采穗圃插穗不定根发育期内源激素含量的动态变化。结果表明:扦插后13 31 d是愈伤组织形成和不定根原始体分化发育的关键期,此时一轮分生株愈伤组织形成和不定根发育均优于原株。连续繁殖对插穗生根性状有显著作用,尤其是生根率很低的原株,经过一轮繁殖后,生根性状有极显著提高。连续繁殖影响插穗自身激素含量,尤其是IAA,经过一轮繁殖后,含量明显高于原株。从不定根发育过程中激素的动态变化看,一轮分生株插穗(IAA+GA3+ZR)/ABA比值在不定根发育前期明显高于原株,与生根率的变化一致,可用来衡量不同繁殖次数插穗生根性状的优劣。     

垂丝海棠插穗的内源激素水平及其与扦插生根的关系

垂丝海棠扦插繁殖的试验结果表明,嫩枝扦插效果好于硬枝扦插,并且母树年龄是影响其扦插生根的重要因子。母树年龄为2年生时,生根率达77%以上。本文在此试验的基础上,对嫩枝和硬枝插穗中内源植物激素水平进行了测定。结果发现嫩枝插穗中IAA、CTKs的含量要高于硬枝插穗中IAA、CTKs的含量,并且随母树年龄的增加而降低;相反,嫩枝插穗中ABA含量要低于硬枝插穗中ABA的含量,并且随着母树年龄的增加而增加。实验结果表明:IAA、ABA的含量和扦插生根有密切关系。IAA促进生根,ABA抑制生根。IAA/ABA比值可以用来衡量垂丝海棠的生根能力。     

The role of antioxidant system in freezing acclimation-induced freezing resistance of Populus suaveolens cuttings

We investigated the changes in the contents of H2O2, malonaldehyde (MDA) and endogenous antioxidants, the activities of protective enzymes and some critical enzymes involved in the ascorbate-glutathione (ASA-GSH) cycle as well as freezing resistance (expressed as LT50) and correlations mentioned above, in detail using Populus suaveolens cuttings. The purpose was to explore the physiological mechanism of the enhancement of freezing resistance induced by freezing acclimation at –20°C, and to elucidate the physiological mechanisms by which trees adapt to freezing. The results showed that freezing acclimation enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), monodehydroascorbate reductase (MDAR), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and glutathione reductase (GR). And it increased the contents of reduced ascorbate (ASA), reduced glutathione (GSH), dehydroascorbate (DHA) and oxidized glutathione (GSSG). However, H2O2 and MDA contents and LT50 of cuttings were decreased. LT50 in cuttings was found to be closely correlated to the levels of SOD, POD, CAT, APX, DHAR, MDAR, GR, H2O2, MDA, ASA, GSH, DHA and GSSG during freezing acclimation. This suggested that the enhancement of freezing resistance of cuttings induced by freezing acclimation may relate to the distinct increase for the levels of SOD, POD, CAT, APX, DHAR, MDAR, GR, ASA, GSH, DHA, and GSSG. In addition, the observed levels of APX, DHAR, MDAR, GR, ASA, DHA, GSH and GSSG were higher than those of SOD, POD and CAT during freezing acclimation. It indicated that a higher capacity of the ASA-GSH cycle is required for H2O2 detoxification, and growth and development of cuttings. Based on the obtained results, it can be concluded that the ASA-GSH cycle plays an important role in enhancement of freezing resistance of P. suaveolens cuttings during freezing acclimation.     

The impact of shade on morphology,growth and biomass allocation in <Emphasis Type="Italic">Picea sitchensis</Emphasis>, <Emphasis Type="Italic">Larix</Emphasis> × <Emphasis Type="Italic">eurolepis</Emphasis> and <Emphasis Type="Italic">Thuja plicata</Emphasis>

To determine the effect of shade on morphology, growth and biomass allocation in Picea sitchensis, Larix × eurolepis and Thuja plicata, seedlings were grown in the open or under shadehouses providing 25%, 50% and 75% reductions of full-light for two growing seasons. For most of the characteristics assessed there was no significant interaction between species and shade indicating that the morphological responses to changing shade treatments were not species-dependent. After two growing seasons the mean height increment for the three species was significantly greater in 25% (76.1 cm) and 50% shade (74.9 cm) than in the open (69.5 cm). Root collar diameter increment, shoot, root and total biomass declined significantly with increasing shade while the opposite was true for the height:diameter ratio. In both western red cedar and hybrid larch the shoot:root ratio was significantly greater in the shade while in Sitka spruce this characteristic was not influenced by shade. While all species had significantly greater specific shoot areas in 75% shade than in 0% shade, this trend was particularly pronounced in hybrid larch. In hybrid larch and western red cedar, the normalised specific projected shoot area increased significantly with increasing shade. The opposite trend was observed for Sitka spruce. We conclude that in the main the species studied demonstrated similar shade acclimation responses despite their reported differences in shade tolerance.     

Characterization of expressed sequence tags obtained by SSH during root formation in Larix cuttings

SSH was used to analyze gene transactivation during root formation of Larix cuttings. Two subtractive cDNA libraries