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1.
We characterized SCARECROW-LIKE genes induced by auxin in rooting-competent cuttings of two distantly related forest species (Pinus radiata D. Don and Castanea sativa Mill.) before the activation of cell division that results in adventitious root formation. The predicted protein sequences contain domains characteristic of the GRAS protein family and show a strong similarity to the SCARECROW-LIKE proteins, indicating conserved functions of these proteins. Quantitative RT-PCR analysis showed that these genes are expressed at relatively high levels in roots. Induction of increased mRNA levels in rooting-competent cuttings of both species in response to exogenous auxin was observed within the first 24 h of the root induction process, a time when cell reorganization takes place, but before the resumption of cell division and the appearance of adventitious root primordia. These results suggest that SCARECROW-LIKE genes play a role during the earliest stages of adventitious root formation.  相似文献   

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This paper reports on the effect of various micropropagation factors of Gmelina arborea Roxb. through multiple shoot induction. Factors like the source and age of explants, plant growth regulators (PGRs), media composition, and carbon source affected multiple shoot-ing in the present study. Among all the explants used, only shoot tips derived from one, two, and three week old seedlings could form multiple shoots. Besides, the formation of multiple shoots depended on the con-centration and combination of PGRs. Among all the PGRs, BAP (6-benzylaminopurine) alone gave the highest regeneration efficiency. Simi-larly, IBA (Indole-3-butyric acid) was the most efficient PGR in inducing root formation in the microshoots. Media composition and carbon source also affected the regeneration efficiency. MS (Murashige and Skoog medium) proved to be the best media for regeneration followed by B5, SH (Schenk and Hilderbrandt medium) and WPM (Woody plant medium) in that order. Similarly, among sugars, only sucrose and glucose sup-ported induction of microshoots. Based on this study we recommend the use of glucose in place of sucrose in MS medium for maximum regenera-tion efficiency.  相似文献   

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【目的】为了解生长素诱导杜梨不定根发生效应,明确不定根诱导过程与氧化酶活性间的关系,从而为杜梨等难生根植物的生根研究提供理论指导和技术借鉴。【方法】以连续继代培养的杜梨不定芽为材料,调查了不同种类(NAA、IBA和IAA)和浓度(0.1、0.5、1.0、2.0 mg·L-1)的生长素对不定芽生根的影响情况,分别在生根诱导0、4、8、12、16 d后观察不定根发育的内外组织形态,测定并分析相关酶活性的变化情况。【结果】外源生长素均能促进不定芽不定根的发生,在相同种类的生长素处理下,浓度分别为0.1和2.0 mg·L-1处理的各项生根指标均显著低于浓度分别为0.5和1.0 mg·L-1处理的。以NAA诱导产生的愈伤组织较大且松软易脱落,不利于其后期的移栽成活;以IBA处理的生根率显著高于NAA和IAA处理的,其中以浓度为0.5 mg·L-1的IBA处理的各项生根指标均最优。对其组织形态的观察结果表明,杜梨不定根发生类型属于诱生根原基,经IBA处理4 d后,其茎基部表皮颜色变红,内部根原基发生于韧皮部和维管束形成层交界处,诱导8 d后茎段外部出现了愈伤组织且组织内部根原基已形成,诱导12 d后不定根突破表皮向外生长。在杜梨不定芽生根过程中,SOD和PPO活性在诱导的0~4 d内均快速上升,在诱导的4~12 d内均下降,而在诱导的12~16 d内均呈现缓慢上升的变化趋势;IAAO活性总体呈现先下降后上升的趋势,且每个时期的活性均显著高于对照组;POD活性呈现出上升→下降→上升→下降的变化趋势,2个峰值分别出现在诱导4 和12 d之时。【结论】0.5 mg·L-1的IBA可有效促进连续继代培养的杜梨不定芽的生根,其不定根的形成过程与氧化酶有密切的关系。  相似文献   

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Stabilized shoot cultures initiated from crown material of six adult Quercus robur L. trees and from basal epicormic shoots of a Quercus rubra L. tree showed good in vitro rooting capacity. An initial five-day dark period generally improved the rooting response but was detrimental to plantlet quality. There were clonal differences in rooting capacity. The concentration and exposure time of the indolebutyric acid (IBA) treatment were critical for root induction. In both species, best rooting efficiency was achieved by culture in medium containing 25 mg l(-1) IBA for 24 h and subsequent transfer to an auxin-free medium containing 1% activated charcoal. For all clones tested, the charcoal benefited both shoot quality and root system development, the latter being enhanced by the formation of many lateral roots. Total root system area and length, measured with a digital image analyzer, were significantly greater in medium containing charcoal than in medium lacking charcoal. Because darkening the basal part of the shoots with aluminum foil during the rooting phase only caused a small increase in rooting, we conclude that the large effect of charcoal on rooting was the result of adsorption of inhibitory compounds from the medium or the explant or both, rather than of basal darkening. Other factors affecting the rooting response of Q. robur were: (a) the position on the tree of the material from which cultures were initiated (the topophysical effect); and (b) shoot quality. Recycling the same horizontally placed explant on multiplication medium allowed three successive crops of shoots to be obtained, and rootability was typically maintained from crop to crop.  相似文献   

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SELBY  C.; SEABY  D.A. 《Forestry》1982,55(2):125-135
Primary lateral root production on Pinus contorta seedlingswas increased 5–10 fold by immersion in auxin solutions.Watering auxin solutions onto P. contorta seedlings growingin soil or compost induced the development of large numbersof primary lateral roots in the root collar region. The numberof primary lateral roots which developed depended mainly uponthe auxin used and its application rate, seedling age and toa lesser extent on the soil characteristics. Instability oftransplanted pines is partly due to their lack of primary lateralroots and their inability to regenerate new ones after plantingout; this instability may be overcome by using auxin treatedseedlings. Potential practical applications of this techniqueare discussed.  相似文献   

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霍山石斛组培丛生芽诱导增殖及生根技术   总被引:1,自引:0,他引:1  
以霍山石斛无根试管苗为试材,研究了丛生芽诱导增殖和生根技术.丛生芽诱导增殖通过4因素3水平正交试验,筛选出的最适培养基为1/2MS 0.2 mg/LKT 0.2 mg/L NAA 0.3 mg/L 6-BA.4个因子对霍山石斛丛生芽诱导的影响顺序依次为:KT>基本培养基>6-BA>NAA,其中,KT对霍山石斛丛生芽的诱导效果极为显著.添加不同浓度的生长素及香蕉泥进行生根诱导,其生根效果排序为:香蕉泥>IBA>NAA,最佳的生根培养基是MS基本培养基添加20%香蕉泥,生根率可达94.3%.  相似文献   

10.
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.  相似文献   

11.
Leader shoots from juvenile succulent Norway spruce seedlings were used as cuttings 11 weeks after sowing. The seedlings were grown for 6 additional weeks and then apically dominant and lateral shoots were also used as cuttings. These three cutting categories, leaders, dominants and laterals, rooted 99.4, 99.0 and 97.5 % respectively and performed differently with respect to rooting development, number of roots per cutting and response to IBA and PIBA treatments (synthetic auxins). A large individual variation in number of roots per cutting, which auxin treatments did not reduce, was present within each cutting categoty. Laterals from the three different families included in the study showed a different variation than did leaders and dominants. Generally, the juvenile succulent cuttings were easy to root. Seedlings grown at a rather high light intensity (62 W/m2) had the potential to give good rooting even without hormonal treatments.  相似文献   

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为建立三叶青毛状根的诱导和液体培养体系,采用发根农杆菌株系C58C1和ATCC15834进行毛状根的诱导,通过无菌三叶青叶片预培养、浸染、共培养、筛选、形态和PCR鉴定等过程获得三叶青毛状根根系,并对三叶青毛状根和两年生块根总黄酮进行比较分析。试验结果表明,发根农杆菌C58C1能从三叶青叶片诱导出毛状根,三叶青毛状根在不添加生长素的1/2MS液体培养基中生长状态良好,其总黄酮含量约是2年生三叶青块根的1/5。  相似文献   

13.
Auxin polar transport genes PIN(PINFORMED)determine the concentration gradient of auxin in plants.To understand the relationship between the development of different tissues in Betula pendula‘Dalecartica’,BpPIN gene expression and indole-3-acetic acid(IAA)content were analyzed using qRT-PCR,ELISA,and GUS staining.Gene expression of BpPIN genes and IAA levels in the leaves,buds,stems,xylem,and roots of B.pendula‘Dalecartica’and B.pendula as a control were measured.BpPIN1,BpPIN5 and BpPIN6 were upregulated during development in both species,suggesting a dominant role in the development of B.pendula‘Dalecartica’leaves.Moreover,BpPIN1 gene expression was positively associated with IAA levels during leaf,vein and petiole development in B.pendula‘Dalecartica’only.The correlation coefficient of the first three leaves was 0.69(P=0.04),while that of the first three petioles was 0.85(P=0.001).In addition,GUS staining of the pro-DR5::GUS transgenic line of cultivar was correlated with the results of BpPIN1 expression.Overall,these findings suggest that BpPIN1 is associated with the formation of lobed leaves in B.pendula‘Dalecartica’.  相似文献   

14.
An in vitro regeneration system was developed using organogenic callus derived from in vitro grown cotyledonary explants of Gleditsia caspica Desf., an important leguminous tree. Murashige and Skoog (MS) basal medium augmented with 0.2 g L?1 myo-inositol and various concentrations of either 2,4-dichlorophenoxyacetic acid (2,4-D), naphthaleneacetic acid, or indole-3-butyric acid (IBA) alone as well as combined with cytokinins was used for callus induction. The highest frequency of organogenic yellowish-white and nodular callus (93 %) was obtained from explants grown on medium supplemented with 13.5 μM 2,4-D and 4.4 μM benzyladenine (BA). The yellowish-white and nodular callus when transferred to MS medium supplemented with BA (2.2–17.7 μM) or kinetin (KT; 2.3–18.8 μM) solely or in combination with 2.3 μM 2,4-D produced several microshoots after 5 weeks culture. The calli cultured on MS medium with 4.4 μM BA singly showed superior growth response and produced both maximum shoot regeneration (94 %) and the highest mean number (4.3) of microshoots per callus. Transfer of regenerated microshoots onto modified MS basal medium fortified with 5.8 μM gibberellic acid and 4.4 μM BA resulted in the maximum number of internodes per shoot and the highest shoot elongation after a period of 6 weeks. Optimum rooting of 90 %, an average 6.1 roots per shoot, and a mean root length of 3.6 cm was observed when half-strength MS medium was supplemented with 9.8 μM IBA and 0.92 μM KT. The regenerated healthy plants with well-developed shoots and roots showed a survival rate of 77 % after acclimatization and transplanting to garden soil for a 10-week hardening period under ex vitro conditions.  相似文献   

15.
Adventitious rooting is essential for vegetative propagation of woody species. We studied the effects of auxin and light on the development of adventitious roots in cuttings obtained from seedlings of Eucalyptus saligna Smith and E. globulus Labill in an attempt to characterize the adventitious rooting process and identify factors controlling rhizogenesis. Root development was scored as rooting percentage, root density (roots per rooted cutting), mean rooting time and root length. In both species, rooting time was reduced in the presence of auxin. Cuttings from 2-month-old E. saligna seedlings were responsive to lower auxin concentrations than comparable cuttings from E. globulus seedlings. Cuttings from 3-month-old E. saligna seedlings rooted promptly and rooting was not significantly affected by light conditions. In contrast, rooting of cuttings from 3-month-old E. globulus seedlings exhibited recalcitrant behavior and no roots were formed if illuminated during the root formation phase. Effective root regeneration of E. globulus cuttings was obtained by a 4-day exposure to 10 mg l(-1) IBA and culture in darkness during the root formation step. Loss of rooting capacity with seedling age was more pronounced in E. globulus than in E. saligna. The possibility of switching adventitious rooting off and on by manipulating light regime and exogenous auxin supply in E. globulus, and the constitutive nature of rooting in E. saligna may provide useful models for examining the rooting process at the biochemical and molecular levels in Eucalyptus.  相似文献   

16.
We determined the effects of removal of leaves, stem axillary buds, or the entire shoot on root suckering (adventitious shoot formation by roots) and basal stem sprouts in 3- and 4-year-old potted seedlings of aspen (Populus tremuloides Michx.). The greatest number of root suckers (67.9 +/- 8.5 per plant) emerged after excision of the entire shoot. Defoliated and debudded stems were the major source of inhibitory agents for root suckering, although axillary buds and developing new leaves also exerted a significant inhibitory effect. Removal of mature leaves had only a minor effect on root suckering. Removal of a continuous band of bark (girdling) at the base of the stem consistently stimulated growth of adventitious shoots from the stem below the girdle and occasionally promoted root suckering. Exogenous application of indole-3-acetic acid to excised stumps inhibited root suckering and basal stem sprouting. Naphthylphthalamic acid (NPA), an auxin polar transport inhibitor, had no effect on root suckering or stem sprouting when it was applied to the bark of the basal stem. However, NPA significantly increased root suckering when it was applied to the exposed surface of xylem after girdling. These results suggest that polar transport of auxin in the xylem parenchyma is an important inhibitor of root suckering. On decapitated stems, vacuum extraction of xylem sap from the root system lowered the frequency of root suckering compared with decapitation alone, indicating that substance(s) originating in the root system also play a significant role in controlling root suckering.  相似文献   

17.

Adventitious rooting of Scots pine (Pinus sylvestris L.) hypocotyl cuttings was promoted by binucleate Rhizoctonia (BnR) and to a lesser extent by ectomycorrhizal fungi. Four BnR isolates (251, 266, 268 and 269) differentiated root meristems and significantly induced adventitious rooting in young derooted seedlings. Rooting rates were significantly higher in BnR treatments than in either indol-3-butyric acid (IBA) pretreatment (200 µm) or co-cultivation with the ectomycorrhizal fungi Suillus bovinus or Laccaria bicolor. In pre-IBA-treated cuttings, adventitious root numbers were higher in the treatments with isolates 251 and particularly 268, while more similar in other BnR treatments. In the 251, roots emerged from distal positions along the hypocotyl and not from the cut base as in the other three BnR isolate treatments. Loss of turgor and hypocotyl wilt was commonly detected up to the point of root initiation. Lateral roots and dichotomous short roots subsequently appeared at high frequency from primary adventitious roots. Mechanisms involved in root meristem differentiation, e.g. auxin production, wound response and oligosaccharide signals, are discussed with respect to host-fungal signalling mechanisms. These beneficial BnR and ectomycorrhizal fungi could provide promising new tools in the development of efficient clonal propagation methodology for this highly recalcitrant Pinus species.  相似文献   

18.
In-vitro-cultured shoots of clones initiated from shoots of the basal parts (BS) and the crown (C) of two mature Quercus robur L. trees were subjected to rooting experiments to relate rooting with shoot topophysical origin. The BS shoots exhibited morphologically juvenile characteristics and rooted more easily after indole-3-butyric acid (IBA) treatment than C shoots. When naphthylphthalamic acid (NPA) was applied to BS shoots, rooting capacity decreased and root emergence was delayed at least 2 days compared with shoots treated with IBA only. During the first days of the rooting process, endogenous indole-3-acetic acid (IAA) concentration was higher in C shoots than in BS shoots, regardless of whether the shoots were treated with NPA. Mitotic figures were observed in cells from the basal part of both BS and C shoots 24 h after IBA treatment. After 4 days of IBA treatment, the occurrence of histological events differed between BS shoots and C shoots. Cells of BS shoots became meristematic, giving rise to meristemoids and root primordia, whereas no differentiation of root meristemoids occurred in cells of C shoots. Thus, although adult oak material (C shoots) is capable of responding to the initial stimulus of auxin during the adventitious rooting process, the endogenous IAA concentration is not the factor limiting rooting in adult material.  相似文献   

19.
[目的]分生组织对器官发生和形态建成起到至关重要的作用,其活性受多种转录因子的调控。KNOTTEDlike homeobox(KNOX)基因家族由2个亚类即I类和II类KNOX组成,在模式植物拟南芥中I类KNOX成员主要在茎的顶端分生组织区域表达,对维持顶端分生组织分化及侧生器官的形态建成过程起关键作用。木本植物生长发育过程主要包含以顶端分生组织(Shoot apical meristem,SAM)为中心的初生生长与以形成层为中心的次生生长过程,本研究通过分析杨树I类KNOX基因在SAM、根顶端分生组织(Root apical meristem,RAM)的形成过程及形成层相关区域的表达,探讨I类KNOX基因在杨树分生组织形成与分化过程中的功能。[方法]以拟南芥I类KNOX基因STM的蛋白序列在毛果杨基因组中进行同源比对分析,获取杨树I类KNOX成员的核酸和氨基酸序列。根据核酸及氨基酸序列信息构建系统发育树,并绘制基因结构与蛋白质结构图谱。利用84K杨(Populus alba×P.glandulosa)不定芽与不定根诱导实验体系模拟顶端分生组织发育过程,通过实时定量PCR(RT-PCR)技术,对杨树I类KNOX成员在不定芽与不定根形成过程及形成层相关区域中的表达进行分析。[结果]本研究通过同源比对分析鉴定出10个杨树I类KNOX成员,根据进化关系与基因结构差异将其分为三组:组1、组2、组3,其中,组1为拟南芥KNAT2和KNAT6同源基因,组2为STM和BP同源基因,组3为杨树所特有的I类KNOX基因。杨树I类KNOX基因在不定芽与不定根发生过程中表达量均发生较大的变化,在不定芽发生过程中,组1基因在芽原基分化产生不定芽的关键期上调表达,而组2与组3基因多在分生组织分化产生芽原基阶段高表达;在不定根发生过程中,组1成员多在根原基分化产生不定根的过渡期上调表达,组2与组3基因则多在发育后期不定根形态建成阶段高表达。杨树I类KNOX成员在形成层相关区域均表达,组1中KNAT2/6与组2中STM、BP同源基因的表达量明显高于其他成员。[结论]以上结果说明,杨树I类KNOX除了与拟南芥同源的2个组外,还进化产生新的组别,分别参与分生组织形成和分化过程中不同阶段的调控,且Pt KNAT2/6b、ARK1与ARK2在形成层区域高表达,提示以上3个基因可在形成层功能维持以及木质部分化中发挥主要作用。  相似文献   

20.
奇异核桃组织培养嫩枝试管外生根苗移植3个月后进行地上及地下部分的生长情况调查.结果表明,根系修剪对植株地上部分的生长及根系的再生量没有显著影响,但是修剪处理明显减少了曲根的产生,尤其是重度修剪的处理根本没有曲根现象发生,半量修剪的处理有少量曲根发生,而不剪则出现很多曲根植株.  相似文献   

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