利用微嫁接技术促进麻疯树再生不定芽的生长

该研究以麻疯树种子实生苗的小芽和芽条为接穗,以带有胚根的实生苗下胚轴为砧木进行无菌微嫁接,试图建立新的有效微嫁接方法,解决农杆菌介导的麻疯树遗传转化体系中再生的转化不定芽难以顺利发育成完整植株的问题。结果表明:(1)抗生素对嫁接苗的生长有显著的抑制作用。(2)进行微嫁接所用砧木的苗龄以5d为宜。(3)进行微嫁接时适宜采用的砧木类型为带有部分胚根的下胚轴。(4)嫁接苗在0.3mg/L IBA+2mg/L谷氨酰胺+1/2MS培养基上的生长效果最好。(5)嫁接苗的移栽成活率最高可达76.40%。(6)以小芽或芽条为接穗的嫁接苗均可正常生长。该研究建立的麻疯树微嫁接体系,为解决麻疯树转化不定芽或芽条生长发育困难的问题提供了一条新的途径。     

Effect of Abscisic Acid and Other Plant Hormones on Growth of Apical and Lateral Buds of Seedlings

The effect of abscisic acid (AbA) on the growth of lateral and apical buds was studied in seedlings of Pisum sativum and some other species. The hormone was applied in three different ways: 1) directly to the lateral bud on the second node of decapitated pea seedlings as 5 μI droplets in an ethanolic solution; 2) to the cut surface of decapitated seedlings: 3) to the apical bud of intact plants. AbA directly applied in amounts of 5 to 0.1 μg to the lateral bud of the second node of decapitated seedlings had a strong inhibitory effect on the bud. Application to the cut surface of seedlings decapitated about 5 mm above the second node resulted in slight inhibition of the lateral bud on the second node and in growth promotion of the bud on the first node. When AbA at 10 to 0.1 μg was applied to the apical bud of intact seedlings, the growth of this bud was inhibited but the lateral buds grew out. It is concluded that the release of the lateral buds from apícal dominance is the result of the inhibitory effect of AbA on growth of the apical bud and of low transport of AbA. This conclusion is supported by application of GA₃ and IAA, individually and each combined with AbA.     

The Promotion of Indole-3-acetic Acid Oxidation in Pea Buds by Gibberellic Acid and Treatment

Terminal buds of dark-grown pea (Pisum sativum) seedlings have an indole-3-acetic acid oxidase which does not require Mn(2+) and 2,4-dichlorophenol as cofactors. Oxidase activity is at least 50 times higher in buds of tall peas than in dwarf seedlings. Administration of gibberellic acid to dwarf peas stimulates both growth and indoleacetic acid oxidase activity to the same levels as in tall seedlings. By contrast, indoleacetic acid oxidation assayed in the presence of Mn(2+) and 2,4-dichlorophenol proceeds at similar rates regardless of gibberellin application. Treatment of tall peas with the growth retardant AMO-1618 reduces growth and oxidase activity. Such treated seedlings are indistinguishably dwarf. The enzyme does not appear to be polyphenol oxidase, nor do the results suggest that reduced activity in dwarf buds is due to higher levels of a dialyzable inhibitor. The peroxidative nature of the oxidase is probable.     

Execution of the auxin replacement apical dominance experiment in temperate woody species

The classic Thimann-Skoog or auxin replacement apical dominance test of exogenous auxin repression of lateral bud outgrowth was successfully executed in both seedlings and older trees of white ash, green ash, and red oak under the following conditions: (1) decapitation of a twig apex and auxin replacement were carried out during spring flush, (2) the decapitation was in the previous season's overwintered wood, and (3) the point of decapitation was below the upper large irrepressible lateral buds but above the lower repressible lateral buds. Although it has been suggested that neither auxin, the terminal bud, nor apical dominance have control over the outgrowth of the irrepressible buds during spring flush, there is evidence in the present study that indicates that such control over the repressible buds exists. In seedlings, second-order branching, which resulted from decapitation of elongating current shoots, was also inhibited by exogenous auxin in the three species. Hence, the auxin replacement experiments did work on year-old proleptic buds (of branches of older trees) that would have entered the bud bank and also on current buds of seedlings. Cytokinin treatments were ineffectual in promoting bud growth.     

Bud dormancy breaking affects respiration and energy balance of bilberry shoots in the initial stage of growth

Respiration and heat production in the shoots of bilberry (Vaccinium myrtillus L.) were studied at the beginning of growth after breaking bud dormancy by means of transfer of the shoots to indoor conditions (November–April) and upon natural sprouting in spring (May). The buds released from dormancy at the beginning of winter sprouted slower and showed lower respiratory activity than the buds that started growing in May. In May, cytochrome respiratory pathway in sprouting buds was 1.3 times more active than energetically ineffective alternative pathway, whereas activity of cytochrome pathway in December was 1.4 times lower as compared with the alternative. In November–December, the rate of heat evolution by the buds was 3–5 times lower than in April–May. In case of early breaking of bud dormancy, the share of respiration energy dissipated as heat was 30% on average. In the buds whose growth was induced later, the value of this parameter was twice as much. The ratio between heat evolution and respiration depended on temperature. High temperature more intensely activated heat evolution than respiration, which caused a decrease in the level of metabolic energy available for growth. In the temperature range of 5–15°C characteristic of the beginning of vegetation, the share of respiration energy dissipated as heat was 2–3 times lower than at 20–30°C, which reflects a great adaptability of V. myrtillus to climatic conditions of the region. Our data suggest that progression through a full cycle of winter dormancy is physiologically important for shoot growth. Early dormancy release brought about changes in respiration and energy balance of the shoots in the initial stage of extra-bud growth.     

Predicting spring frost sensitivity by bud development and temperature sum in Norway spruce seedlings

Nurseries would benefit greatly if frost hardiness (FH) of seedlings could be predicted by some environmental variable or by bud development in spring. We investigated the FH of 1-year-old Norway spruce (Picea abies (L.) Karst.) seedlings of local origin. The seedlings were stored frozen until incubated in the growth chamber at six different temperature sums (TSs) (0, 55, 88, 142, 185 and 240 d.d., >5°C) from mid-February to mid-March. FH of the buds, stems and previous year needles was assessed on three occasions. When the TS was 88 d.d. or less, buds exhibited only microscopic signs of development, even when seedlings tolerated temperatures below −10°C. As TS increased, primordial needles and primordial stems of buds grew while FH weakened, especially in previous year needles. When the TS was at least 142 d.d., all plant parts were frost hardy to approximately −6°C. Monitoring TS and bud development can help predict FH of Norway spruce seedlings in spring. However, more studies with seedlings of different ages and from multiple locations are necessary to appreciate the generality of our results.     

光照条件对蒙古栎幼苗生长及形态特征的影响

对生长于不同光照环境(林内和林外)下的蒙古栎幼苗的形态特征进行了比较.结果表明,光照条件对蒙古栎幼苗的生物量及其分配具有明显影响,林外幼苗单株生物量为林外的4.48倍,其中以根系生物量的差别为最大,林外为林内的6.23倍;林外与林内幼苗根冠比分别为2.70和1.11.在不同的光照条件下,蒙古栎幼苗的叶片形态表现出明显不同,林外叶片比叶面积明显低于林内,分别为139.55cm2/g 和284.94cm2/g,林内约为林外的2.04倍;一次生长叶面积林外明显低于林内,总叶面积则高于林内.二次生长使蒙古栎幼苗的叶面积有明显增加,但是经过二次生长后,幼苗的单株生物量没有明显增加,说明二次生长叶片对于蒙古栎幼苗生物量的积累没有明显贡献;光照对于蒙古栎主干及根系的形态具有明显影响,林外幼苗高(包括二次生长)明显低于林内幼苗,而地径则相反,林外幼苗根系的长度和直径则明显高于林内幼苗;林外幼苗顶芽体积明显大于林内幼苗.同时,林外幼苗的二次生长导致顶芽体积明显下降,从而对翌年的高生长量和未来干型产生影响.认为,蒙古栎幼苗通过形态上的一系列可塑性变化来适应不同的光环境以获取最大的碳收益.     

The antagonism of (2-Chlorethyl) trimethylammonium chloride and indole 3-acetic acid in epicotyl and axillary buds growth in the pea seedlings

The 0·03–0·06% indole-3-acetic acid (IAA) in lanoline paste smeared on the cut surface of decapitated epicotyl of the pea seedling inhibited the growth of the axillary buds of the cotyledons. 0·03–0·06% solution of (2-Chlorethyl) trimethylammonium chloride (CCC) considerably weakened the inhibitory effect of IAA when applied simultaneously to the roots. In a similar way CCC diminishes even the growth inhibition of intact pea epicotyls caused by 0·06% IAA lanoline paste. 0·03% solution of CCC administered to the roots of the decapitated pea seedlings significantly increased growth of the axillary buds. This effect may play a role in the demonstrated antagonism between the low concentration of CCC and IAA.     

Flower bud differentiation in Salix pentandra as affected by photoperiod, temperature and growth regulators

Flower bud initiation in seedlings and vegetatively propagated plants of Salix pentandra from different locations has been studied under controlled conditions. In mature plants flower bud formation was induced by 2-chloroethyltrimethylammoniumchloride (CCC) and by short day treatment. The effect of CCC was antagonized by GA₃. The critical photoperiod for flower bud formation was about 18 h for a southern clone (from 49°48'N), but cuttings of a northern ecotype (from 69°39'N) formed flower buds even at 24 h photoperiod. Generally, flower bud formation occurred simultaneously with apical growth cessation. However, apical growth cessation was not a prerequisite for floral initiation and flower buds were also found in elongating plants. Seedlings up to 60 days old did not form flower buds in growth chamber studies. The length of the juvenile phase has not been studied in detail, but cuttings taken from seedlings approximately 20 cm high and 60 days old were able to develop flower buds when treated with CCC. A gradual transition from the juvenile to the mature phase was obtained by repeated pruning of seedlings grown at 18°C and 24 h photoperiod.     

Carotenoid composition and down regulation of photosystem II in three conifer species during the winter

Seedlings and coppice shoots of Betula pubescens Ehrh. were grown under controlled conditions designed to simulate the annual growth cycle, and a water stress was introduced during the short day (SD). Alleviation of hud dormancy after increasing periods at chilling temperatures was tested under long day (LD) conditions. Abscisic acid (ABA) was analysed in leaf and bud samples by gas chromatography-mass spectrometry using [²H₄]ABA as the internal standard. Elongation growth of coppice shoots was faster than that of seedlings under both LD and SD conditions, while the final growth cessation occurred in a similar manner and was not affected by water stress, which significantly reduced growth rate in both plant types. Bud dormancy gradually decreased with increasing length of chilling, starting from the basal parts of the plant axis. Water stress did not retard hudhurst. but rather improved it in the chilled coppice shoots and in the non-chilled and partially chilled seedlings. Water content of buds was higher in coppice shoots than in seedlings, but after exposure to SD. it gradually decreased to 45% in both plant types and was not affected by water stress or chilling. The ABA level in both leaves and buds increased during SD treatment and was" enhanced by water stress. No clear differences in bud ABA level were found between the seedlings and coppice shoots under SD conditions, although coppice shoots had less ABA during the preceding LD conditions. There was, in general, no clear effect of chilling on bud ABA level. Budbursl in chilled, single-node cuttings was inhibited by external ABA treatment, which raised the internal ABA levels 10 to 150 times above normal. The observed correlation between ABA level and water content in buds during induction of dormancy under SD and water stress conditions indicates a possible role for ABA in the regulation of dormancy.     

Increases in cytokinin levels in Scots pine in relation to chilling and bud burst

Levels of isopentenyladenosine and zeatin riboside were monitored in buds and needles of Scots pine ( Pinus sylvestris L.) seedlings grown under controlled climatic conditions and in field-grown trees. Extracts were purified by immunoaffinity chromatography and high-performance liquid chromatography. Cytokinin levels were quantified with an enzyme-linked immunosorbent assay. The cytokinin content was low in buds and needles of dormant seedlings but increased during dormancy release, reaching peak values in buds just before resumption of shoot growth. Samples collected in the field also showed a marked increase in the levels of cytokinins just prior to bud burst. Further, the biological activity of applied cytokinins in activating the dormant buds of Scots pine is shown. The results indicate a probable role of cytokinins in seedlings during dormancy release.     

Efficient protocol for <Emphasis Type="Italic">in vitro</Emphasis> mass micropropagation of slash pine

An efficient protocol was developed for large-scale micropropagation of slash pine (Pinus elliottii Engelm. var. elliottii). For that, explants consisting of shoot apices 1.5–2.0 cm in length from 4 wk-old seedlings were cultured on two different basal media (Murashige & Skoog (MS) and Westvaco WV5 (WV5)). All media were supplemented with 6-benzylaminopurine (BAP), to stimulate the formation of axillary buds. Best bud induction was achieved, after 4 wk, on Westvaco WV5 medium (with 10-μM BAP) with rates close to 100%, and an average number of ~?7 new buds formed per explant. Elongation took place for 6 wk on Westvaco WV5 medium containing activated charcoal (0.2% (w/v)) and without growth regulators. Rooting took place on half-strength Westvaco WV5 medium containing 9.8-μM indole-3-butyric acid (IBA). After 6 wk, root primordia were visible in ~?43% of shoots. The acclimatization protocol was also optimized by controlling relative humidity, light/photoperiod, temperature, and nutrition, which led to an acclimatization success of ~?89%. Flow cytometry analysis of DNA-ploidy did not show any variation between micropropagated plants and seedlings. With the protocol here described, it is possible to obtain a high number of genetically uniform plants per explant, 1 yr after in vitro germination of slash pine seeds.     

Direct Organogenesis in Hypocotyl Cultures of Tamarindus Indica

Direct differentiation of shoot buds from hypocotyl segments of 12-d-old seedlings of Tamarindus indica was obtained on Murashige and Skoog (MS) medium with or without growth regulators. The highest regeneration (66 %) and the maximum number of shoots (3 - 4) per explant were obtained from the explants on MS medium containing 6-benzylaminopurine (5 × 10-6 M). A maximum roots per shoot were produced on medium containing 3-indole butyric acid (5 × 10-6 M). The resulting plantlets were hardened and transferred to soil in pots where 75 % of them survived and resumed growth. Histological examination of explants suggests that the shoots were of de novo origin which would make this system suitable for transformation experiments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Vegetative propagation of western hemlock (Tsuga heterophylla) through tissue culture

Western hemlock cultures derived from cotyledon explants of2–5 week-old seedlings were established on a chemicallydefined medium containing both cytokinin and auxin. Adventitiousbud formation required a relatively high concentration of cytokininwith respect to that of auxin. The growth of buds was stimulatedby a culture medium devoid of all plant growth regulators. Plantswere successfully established in soil by rooting shoots producedfrom somatic cells directly in rooting medium "Mica-Peat." (Received July 7, 1976; )     

特大‘新红宝’西瓜的快速繁殖研究

利用新红宝西瓜种子、茎段、茎尖及腋芽为外植体,接种子改良的ＭＳ培养基上。种子经常规消毒灭菌后,７—１５ｄ后可获得９５％以上的健壮无菌苗．苗高３ｃｍ─５ｃｍ时即可用于有关实验。接种６ｄ后,茎段两端切口处膨大,２８—３５ｄ后出现愈伤组织,出愈率达到６８％。接种５ｄ后,茎尖明显长大。在本实验的激素组合中．３５ｄ后,茎尖周围均出现了不同数量的腋芽。最好的组合为改良的ＭＳ＋２．５ｍｇ／ＬＢＡ＋ＩＡＡ１ｍｇ／Ｌ,可使一个茎尖在３５—４２ｄ后扩增为１８─２１个芽并逐渐长成无根苗。４７ｄ后,转入改良ＭＳ＋ＮＡＡ０．５ｍｇ／Ｌ培养基中．生根率达到９４％以上。２８ｄ后,将苗移到田间,第一批移苗５００株,成活４３８株,成活率达到８７．６％。     

Transport of Benzyladenine and Gibberellin A(1) from Roots in Relation to the Dominance between the Axillary Buds of Pea (Pisum sativum L.) Cotyledons

In etiolated, 5-day-old pea (Pisum sativum L.) seedlings a significantly more intensive growth of buds situated in the axil of the excised cotyledons was observed as early as 4 hours after decapitation and excision of one cotyledon of each pair. If [8-¹⁴C]benzyladenine ([¹⁴C]BA) was applied to roots of intact plants 10 hours prior to such decapitation and excision, significantly higher both total and specific ¹⁴C activities were observed in buds situated on the side of the excised cotyledons as early as 4 hours after decapitation and excision. Although the removal of a substantial part of the root system carried out simultaneously with decapitation and excision of one cotyledon resulted in a decrease in total ¹⁴C activity of buds, nevertheless a higher accumulation of ¹⁴C activity was maintained in buds situated on the side of excised cotyledon. If [¹⁴C]BA was applied to roots of seedlings after they were decapitated and deprived of one cotyledon, both total and specific ¹⁴C activities of buds situated on the side of excised cotyledons were significantly higher as early as the end of uptake of [¹⁴C]BA by roots, i.e. after 10 hours. On the other hand, [1,2-³H]gibberellin A₁ applied to roots of intact and/or decapitated and one-cotyledon-deprived seedlings in the same way as [¹⁴C]BA did not appear in the buds until very much later and only in negligible amounts (i.e.³H activity). This indicates that the release of buds from apical dominance represents an active and selective process which can result from the ability of buds to utilize and/or synthesize only certain growth substances within a certain time interval.     

Micropropagation of Capparis decidua through In Vitro Shoot Proliferation on Nodal Explants of Mature Tree and Seedling Explants

In vitro clonal propagation of Capparis decidua was achieved using nodal explants from mature trees, and cotyledonary node, cotyledon and hypocotyl explants taken from the seedlings. Explants cultured on MS medium supplemented with BAP showed differentiation of multiple shoots and shoot buds in 4–5 weeks in the primary cultures. The medium with BAP (5 mg/l) was the best for shoot bud proliferation from the nodal as well as seedling explant. Shoot multiplication was best on cotyledonary node. In the nodal explants shoot multiplication was best on medium supplemented with 5 mg/l BAP and after second subculturing further multiplication of shoot buds was highest on the medium containing 3 mg/l BAP. Shoots were separated from mother cultures in each subculturing for rooting. Rooting was best achieved using 1 mg/l IBA in the medium. Rooted plantlets were transferred td earthen pots with garden soil and peat moss mixture.     

Shoot Apical Meristems of Picea sitchensis Seedlings Accelerate in Growth Following Bud-set

First-year seedlings of Picea sitchensis were induced to setbuds by transferring them from long days (17 h) to short days(10 h at 20 °C, 12 h night at 12 °C). After 20 shortdays, the seedlings stopped growing in height, the shoot apicaldomes began enlarging and the transverse growth rates of theshoot apical dome tissues increased. For about 25 days thereafterprimordia were produced faster within the developing buds thanat the apices of seedlings which had been kept in long daysto prevent bud-set. Accelerated primordia production after bud-setenables the largest possible buds to be formed during the frost-freeperiod before winter. Picea sitchensis, apical meristem, bud-set     

Effect of (2-Chloroethyl)phosphonic Acid on Flower Induction in Juvenile Mango (Mangifera indica) Seedlings

Spray applications of 1000 mg/l (2-chloroethyl)phosphonic acid (CEPA) five times, at weekly intervals, during November and early December induced heavy flowering in ringed and nonringed juvenile mango seedlings. The treated plants produced flower buds which opened by the end of December, while the control and ringed seedlings produced only a new flush of vegetative growth. The results show that CEPA could be profitably used for early evaluation of hybrid seedlings in mango breeding programme.     

Xylem changes in the correlative inhibition of lateral bud growth in flax (Linum usitatissimum L.)

Xylem or tracheary changes at the base of the cotyledonary buds of flax seedlings (Linum usitatissimum L.), released from inhibition by decapitation of the main apex were studied. The differentiation of xylem strands and/or tracheary elements was correlated with the growth in length of the lateral buds, especially 48–72 hr after the removal of the main apex. The xylem strands, connected to the hypocotylary stele or not, and the tracheary elements increased with age within and outside the strands of both non-decapitated and decapitated seedlings. In the latter, the differentiation of these structures, however, occurred much earlier and in greater abundance in the same regions. The early growth in length of lateral buds, 1 or 2 hr after decapitation, was correlated with the early development of tracheary perforations in the xylem strands. The xylary strands with perforated elements are known to be more efficient than those without them. Therefore, it is suggested that the inhibition of lateral-bud growth was due, in fact, to a lack of appropriate tracheary perforations in the bud xylem strands that were connected with the hypocotylary stele of flax seedlings.