The Physiology of Rooting Populus Cuttings

Activity of endogenous cytokinin of cuttings was investigated in relation to the process of rotting. The levels of cytokinin in hardwood cuttings of easy-to-root Populus × euramericana (Dode) Guinier and difficult-to-root Populus tremula L. were monitored at various stages of root formation. In cuttings of P. tremula was found a higher cytokinin activity than in P. × euramericana and this condition persisted throughout the rooting period. There was a general initial decrease of cytokinin activity in both upper and lower halves of cuttings of both species. The declining level was reversed in the easy-to-root species when roots had formed and to a lesser extent in the difficult-to-root species when leaves expanded. In both cases this increase was relatively higher in the lower halves than in the upper halves.     

Causes of Centrifugal Root Promotion

Role of water in centrifugal root promotion, effect of centrifugation on ethylene concentration of cuttings, and effect of ethylene on root formation were investigated using Salix jragilis softwood cuttings. When cuttings were centrifuged with water, more roots formed with increasing water depth. Soaking of cuttings upright in water for 24 h stimulated root formation and produced more roots as the depth of water was increased, with submerged cuttings producing the highest number of roots. Soaking of cuttings upright in hot water for 1 h also stimulated root formation with the best root formation occurring at 40°C. Submerging in, or centrifuging with water increased ethylene concentration in cuttings. Ethylene gas and Ethrel treatments stimulated root formation. It is suggested that submerging in water increases the ethylene concentration in Salix fragilis softwood cuttings which in turn stimulates root formation of the cuttings. Centrifuging cuttings with water increased water content of the cuttings. It is suggested that this increase in water content plays a role similar to the submersion of cuttings as described above.     

Compared responses of poplar cuttings and in vitro raised shoots to short-term chilling treatments

 In woody plants, chilling stress occurs during the early spring growth and can have important economic consequences. The aim of this study was to compare the reliability of two different experimental systems, 3-month-old softwood cuttings and in vitro-grown shoots, to study chilling effects in a poplar clone (Populus tremula×P. tremuloides cv. Muhs1). Different parameters were recorded: lignin content, sucrose concentration and protein and fresh weight variation, during a 2-week treatment at 10  °C. Two families of polypeptides of high molecular weight (110 and 116 kDa) were shown to accumulate in response to chilling in both cuttings and microshoots. For some of the parameters studied, i.e. appearance of some groups of polypeptides and reduction of fresh weight gain, both in vitro and ex vitro systems were suitable and produced similar results. In contrast, for some other observations, i.e. on sucrose concentration and lignin content, the systems led to different conclusions. While sucrose and lignin contents were shown to increase in cuttings submitted to chilling, no variation in lignin and only a small temporary peak of sucrose could be observed in microshoots kept under chilling. These parameters seem not to be suitable for studying the response of poplar in the in vitro system in such a short-term study. Received: 18 November 1999 / Revision received: 3 March 2000 / Accepted: 7 March 2000     

Light effects on root formation in aspen and willow cuttings

The effect of light on rooting of leafy cuttings of aspen (Populus tremula × tremuloides) and a willow hybrid (Salix caprea × viminalis) was investigated under controlled conditions in water culture. Two levels of irradiance were used, 40 and 8 W m^?2. The lower level gave the best rooting of aspen cuttings, both when applied to the stock plants before the cuttings were taken and when given to the cuttings during the rooting period. Irradiation of the cutting base during the rooting period inhibited rooting almost completely in aspen and decreased the number of roots formed in the Salix hybrid. Net photosynthesis in the cuttings of Salix decreased considerably after excision and increased again after formation of roots. Indirect evidence indicated that photosynthesis was even more affected in aspen cuttings. The possible roles of carbohydrates and inhibitors in the light effects are discussed.     

Conditions for rooting of leafy cuttings of Alnus incana

The rooting of softwood cuttings of Alnus incana (L.) Moench in nutrient solution was studied under controlled conditions. Cuttings consisting of one internode with the leaf and axillary bud attached rooted easily and more rapidly than shoot tip cuttings. Light was necessary for rooting but good rooting was obtained in photon flux densities of both 40 and 190 μmol m^-2s^-1. Root number and root length was reduced when light reached the base of the cuttings. Treatment with indolebutyric acid (10^-6–10^-4M) increased the number of roots but 10^-4M delayed rooting and decreased the root length. Debudded internode cuttings rooted as well as intact cuttings, and detached leaves also contained sufficient substances for rooting.     

Production, turnover and mycorrhizal colonization of root systems of three Populus species grown under elevated CO2 (POPFACE)

A fast growing high density Populus plantation located in central Italy was exposed to elevated carbon dioxide for a period of three years. An elevated CO₂ treatment (550 ppm), of 200 ppm over ambient (350 ppm) was provided using a FACE technique. Standing root biomass, fine root turnover and mycorrhizal colonization of the following Populus species was examined: Populus alba L., Populus nigra L., Populus x euramericana Dode (Guinier). Elevated CO₂ increased belowground allocation of biomass in all three species examined, standing root biomass increased by 47–76% as a result of FACE treatment. Similarly, fine root biomass present in the soil increased by 35–84%. The FACE treatment resulted in 55% faster fine root turnover in P. alba and a 27% increase in turnover of roots of P. nigra and P. x euramericana. P. alba and P. nigra invested more root biomass into deeper soil horizon under elevated CO₂. Response of the mycorrhizal community to elevated CO₂ was more varied, the rate of infection increased only in P. alba for both ectomycorrhizal (EM) and arbuscular mycorrhizas (AM). The roots of P. nigra showed greater infection only by AM and the colonization of the root system of P. x euramericana was not affected by FACE treatment. The results suggest that elevated atmospheric CO₂ conditions induce greater belowground biomass investment, which could lead to accumulation of assimilated C in the soil profile. This may have implications for C sequestration and must be taken into account when considering long‐term C storage in the soil.     

Detection of quantitative trait loci influencing growth trajectories of adventitious roots in Populus using functional mapping

The capacity to root from cuttings is a key factor for the mass deployment of superior genotypes in clonal forestry. We studied the genetic basis of rooting capacity by mapping quantitative trait loci (QTLs) that control growth rate and form of root traits in a full-sib family of 93 hybrids derived from an interspecific cross between two Populus species, P. deltoides and P. euramericana. The hybrid family was typed for different marker systems (including SSRs, AFLPs, RAPDs, ISSRs, and SNPs), leading to the construction of two linkage maps based on the female P. deltoides (D map) and male P. euramericana (E map) with a pseudotestcross mapping strategy. The two maps were scanned by functional mapping to detect QTLs that control early growth trajectories of two rooting traits, maximal single-root length and the total number of roots per cutting, measured at five time points in water culture. Of the six QTLs detected for these two growth traits, only one is segregating in P. deltoides with poor rooting capacity, while the other five are segregating in P. euramericana showing good rooting capacity. Tests with functional mapping suggest different developmental patterns of the genetic effects of these root QTLs in time course. Five QTLs were detected to change their effects on root growth trajectories with time, whereas one detected to affect root growth consistently in time course. Knowledge about the genetic and developmental control mechanisms of root QTLs will have important implications for the genetic improvement of vegetative propagation traits in Populus.     

Proteins responding to drought and high-temperature stress in <Emphasis Type="Italic">Populus </Emphasis>× <Emphasis Type="Italic">euramericana</Emphasis> cv. ‘74/76’

Proteomic analysis provides a powerful method of studying plant responses to stress at the protein level. In order to study stress-responsive molecular mechanisms for Populus × euramericana cv. ‘74/76’, one of the most important forest plantation tree species in subtropical and temperate regions, we analyzed the response of 2-year-old cuttings of P. × euramericana cv. ‘74/76’ to drought and high temperature using two-dimensional gel electrophoresis. More than 1,000 reproducible leaf proteins were detected in the controls and treatments, and 26 proteins were found to change notably in abundance. We identified 13 proteins affected by drought stress and 11 proteins affected by high temperature. These proteins are mainly involved in photosynthesis such as ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit and putative photosystem I reaction center subunit II precursor, and detoxification (manganese superoxide dismutase and methionine sulfoxide reductase A). Furthermore, the level of the photosynthesis proteins affected greatly by the imposed stress conditions was consistent with the observed noticeable decrease in net photosynthesis rate. These studies provides a fundamental data for future research on responses to drought and high temperature, two major factors limiting the growth of forest trees during summer under recent climatic warming.     

Interspecific and intraspecific variation in tree seedling survival: effects of allocation to roots versus carbohydrate reserves

We examined interspecific and intraspecific variation in tree seedling survival as a function of allocation to carbohydrate reserves and structural root biomass. We predicted that allocation to carbohydrate reserves would vary as a function of the phenology of shoot growth, because of a hypothesized tradeoff between aboveground growth and carbohydrate storage. Intraspecific variation in levels of carbohydrate reserves was induced through experimental defoliation of naturally occurring, 2-year-old seedlings of four northeastern tree species –Acer rubrum, A. saccharum, Quercus rubra, and Prunus serotina– with shoot growth strategies that ranged from highly determinate to indeterminate. Allocation to root structural biomass varied among species and as a function of light, but did not respond to the defoliation treatments. Allocation to carbohydrate reserves varied among species, and the two species with the most determinate shoot growth patterns had the highest total mass of carbohydrate reserves, but not the highest concentrations. Both the total mass and concentrations of carbohydrate reserves were significantly reduced by defoliation. Seedling survival during the year following the defoliation treatments did not vary among species, but did vary dramatically in response to defoliation. In general, there was an approximately linear relationship between carbohydrate reserves and subsequent survival, but no clear relationship between allocation to root structural biomass and subsequent survival. Because of the disproportionate amounts of reserves stored in roots, we would have erroneously concluded that allocation to roots was significantly and positively related to seedling survival if we had failed to distinguish between reserves and structural biomass in roots. Received: 14 December 1999 / Accepted: 2 June 1999     

Morphactin and Adventitious Root Formation in Pea Cuttings

Stock plants of pea (Pisum sativum L. cv. Alaska) were grown at different controlled levels of irradiance (4, 16 or 38 W m^?2) for 11 days from sowing. Morphactin (CFM, methyl-2-chloro-9-hydroxy-fluorene-9-carboxylate) was applied to the apex of the stock plants 3 days before cuttings were excised. The cuttings were rooted at 16 W m^?2. High levels of morphactin (>5 × 10^?3 mg l^?1) inhibited root formation in the cuttings. Low concentrations of CFM (5 × 10^?5 mg l^?1) promoted the formation of adventitious roots in cuttings from plants grown at all three levels of irradiance, with the most pronounced effect in cuttings from 4 W m^?2. Measurements of ethylene evolution by CFM-treated plants 3 days after application, revealed a stimulatory effect on ethylene production by high CFM concentrations.     

Responses of nonstructural carbohydrates to shoot removal and soil moisture treatments in <Emphasis Type="Italic">Salix nigra</Emphasis>

Aboveground disturbances are common in dynamic riparian environments, and Salix nigra is well adapted with a vigorous resprouting response. Soil moisture stresses are also common, and S. nigra is flood tolerant and drought sensitive. The objective of this study was to quantify nonstructural carbohydrate (NSC) reserves in S. nigra following shoot removal and soil moisture treatments. NSC reserves provide energy for regeneration of shoot tissue until new functional leaves are developed. Three soil moisture treatments: well-watered (W), periodic flooding (F) and drought (D); and three shoot removal treatments: no shoots removed (R0), partial shoot removal (R1), and complete shoot removal (R2) were applied. Plants were harvested when new shoot development was observed (day 13). Statistical significance in the 3 × 3-factorial design was determined in two-factor ANOVA at P < 0.05. Both roots and cuttings were important reservoirs for NSC during resprouting response, with decreases in root (31%) and cutting (14%) biomass in R2 compared to R0. Rapid recovery of photosynthetic surface area (from 15 to 37% of R0) was found in R1. A clear pattern of starch mobilization was found in roots in R0, R1 and R2, with lowest root starch concentration in W, F higher than W, and D higher than F. Shoot starch concentration was lower in F and D compared to W in R0, however, in R1 shoot starch was reduced in W compared to F and D, possibly indicating reduced rates of translocation during soil moisture stress. Evidence of osmotic adjustment was found in roots and shoots with higher total ethanol-soluble carbohydrates (TESC) during soil moisture stress in F and D treatments. Total plant NSC pool was greater in F and D treatments compared to W, and progressively reduced from R0 to R1 to R2. Results indicated negative effects of drought, and to a lesser extent periodic flooding on resprouting response in S. nigra, with implications for reduced survival when exposed to combined stresses of aboveground disturbance and soil moisture.     

Classification of rhizosphere components using visible–near infrared spectral images

To establish new techniques for automatic classification of rhizosphere components, we investigated the utility of visible (VIS) and near–infrared (NIR) spectral images of the rhizosphere under two soil moisture conditions (mean volumetric water content: 0.39 and 0.16 cm³ cm^?3). Spectral reflectance images of the belowground parts of hybrid poplar cuttings (Populus deltoides × P. euramericana, I45/51) grown in a rhizobox were recorded at 120 spectral bands ranging from 480 to 972 nm. We examined which wavelengths were suitable and the number of spectral bands needed to accurately classify live roots of four age classes, dead roots, leaf mold, and soil. VIS reflectance (<700 nm) of live roots first increased and then decreased with age, whereas NIR reflectance (≥700 nm) was stable in mature roots. The reflectance of dead roots was lower than that of mature roots in both the VIS and NIR spectral regions. VIS reflectance did not differ among dead roots, leaf mold, and soil, but the NIR reflectance was clearly lower in soil than in the other materials. The reflectance of leaf mold and soil increased mainly in the NIR spectral region with reducing soil moisture, but this increase did not affect the order of reflectance intensity among the rhizosphere components in general. Although the most suitable spectral bands statistically selected for classifying rhizosphere components differed somewhat between moist and dry conditions, the spectral bands 580–679 nm (VIS) and 848–894 nm (NIR) provided high reliability under both conditions. Classification accuracy was higher when using two to five VIS–NIR images (overall accuracy ≥87.8%) than three VIS images (red, green, and blue; accuracy <67.1%). The high accuracy with VIS–NIR was mainly due to successful separation of leaf mold and soil. Irrespective of soil moisture condition, the overall accuracy tended to be stable at 92–94% with use of four VIS–NIR images. The spectral bands effective in wet soil conditions could also be used for classification in dry conditions, with overall accuracies >86.9%. These results suggest that automatic image analysis using VIS–NIR images at four spectral bands, including red and NIR, allows for accurate classification of the growth stage or live/dead status of roots and distinguishes between leaf mold and soil.     

山杨杂种无性系的SSR分子标记遗传多样性

采用5对SSR引物对52个山杨杂种无性系进行了遗传多样性检测,结果表明在研究的5个位点上SSR标记多态位点百分率为100%,平均等位基因数为4.4个,有效等位基因数最多的位点是PTR7,最少的位点为PTR12;欧美山杨杂种的遗传多样性最丰富,相比之下,中美山杨杂种遗传变异最低;聚类分析表明,在一定的遗传距离基础上,欧美山杨杂种和欧洲山杨首先聚为一类,然后又与中美山杨杂种聚类,最后是中国山杨。研究表明来自芬欧美山杨杂种具有较高的遗传多样性,这对我国山杨遗传资源的扩大,以及未来山杨杂交育种,杂种优势的利用都是重要的。     

Evidence for the signaling role of methyl jasmonate, methyl salicylate and benzothiazole between poplar (Populus simonii × P. pyramidalis ‘Opera 8277’) cuttings

Interplant communication has been widely demonstrated in plants, especially in herbaceous plants. In this study, mechanical damage was shown to affect the levels of pyrochatechol, chlorogenic acid, gallic acid and p-hydroxyl benzoic acid in poplar (Populus simonii × P. pyramidalis ‘Opera 8277’) cuttings, indicating the activation of defense response. In neighboring intact cuttings, the levels of those phenolics also varied when compared to the control, suggesting the interplant communication between poplar cuttings. Three volatiles, methyl jasmonate, methyl salicylate and benzothiazole, were detected in volatiles emitted from mechanically damaged poplar cuttings. All of them can induce changes in the levels of four phenolics. Therefore, they could act as airborne signals between P. simonii × P. pyramidalis ‘Opera 8277’ cuttings. The different change patterns of phenolic contents induced by different volatiles imply that the defense response activated in neighboring plants may be regulated by multiple signaling pathways. The results also suggest that the entire defense response of plants involves a variety of airborne signals in wound-induced volatiles.     

Rooting and vitality of poinsettia cuttings was increased by arbuscular mycorrhiza in the donor plants

In this paper, we provide evidence that the rooting performance of cuttings can be improved by the arbuscular mycorrhizal (AM) symbiosis of donor plants. Poinsettia stock plants were inoculated with the Glomus intraradices isolate H510 and grown in three different cultivation systems (two organic and one conventional). AM colonization was not related to P availability in the substrate. Decay of the excised cuttings in response to unfavorable postharvest storage conditions was significantly reduced by AM colonization of the stock plants. In most cases, AM significantly promoted the formation of adventitious roots in the stored cuttings. The strongest effect of AM was found when donor plants were grown in a modified organic substrate; then AM-conditioned cuttings showed higher leaf sugar levels and a changed kinetic of carbohydrates during storage. Analyses of N, P, and K in cuttings did not indicate a nutritional effect. The results support the idea that an altered carbohydrate metabolism and plant hormones can contribute to improved rooting performance of cuttings excised from mycorrhizal donor plants.     

Photosynthesis, water use efficiency and stable carbon isotope composition are associated with anatomical properties of leaf and xylem in six poplar species

Although fast‐growing Populus species consume a large amount of water for biomass production, there are considerable variations in water use efficiency (WUE) across different poplar species. To compare differences in growth, WUE and anatomical properties of leaf and xylem and to examine the relationship between photosynthesis/WUE and anatomical properties of leaf and xylem, cuttings of six poplar species were grown in a botanical garden. The growth performance, photosynthesis, intrinsic WUE (WUE_i), stable carbon isotope composition (δ¹³C) and anatomical properties of leaf and xylem were analysed in these poplar plants. Significant differences were found in growth, photosynthesis, WUE_i and anatomical properties among the examined species. Populus cathayana was the clone with the fastest growth and the lowest WUE_i/δ¹³C, whereas P. × euramericana had a considerable growth increment and the highest WUE_i/δ¹³C. Among the analysed poplar species, the highest total stomatal density in P. cathayana was correlated with its highest stomatal conductance (g_s) and lowest WUE_i/δ¹³C. Moreover, significant correlations were observed between WUE_i and abaxial stomatal density and stem vessel lumen area. These data suggest that photosynthesis, WUE_i and δ¹³C are associated with leaf and xylem anatomy and there are tradeoffs between growth and WUE_i. It is anticipated that some poplar species, e.g. P. × euramericana, are better candidates for water‐limited regions and others, e.g. P. cathayana, may be better for water‐abundant areas.     

THE ANNUAL CARBOHYDRATE CYCLE OF ALPINE PLANTS AS RELATED TO GROWTH

Mooney, H. A., and W. D. Billings. (Duke U., Durham, N. C.) The annual carbohydrate cycle of alpine plants as related to growth. Amer. Jour. Bot. 47(7): 594–598. Illus. 1960.—Analyses were made of the carbohydrate content of roots, rhizomes, and shoots of certain plants in the alpine tundra region of the Medicine Bow Mountains, Wyoming, from the beginning of one growing season until the start of the following one. Principal species investigated were Saxifraga rhomboidea, Polygonum bistortoides, and Geum turbinatum. Results were correlated with phenological events as observed in the field. Growth of these plants was found to be very rapid, commencing in some instances under a cover of old snow. The underground organs contained relatively large amounts of carbohydrate reserves. A great part of this stored carbohydrate was utilized in growth prior to snowmelt and during the grand period of shoot growth immediately following snowmelt. In Polygonum, 50% of the rhizome reserves was used in a 1-wk. period in early growth. Except for this short period of rapid depletion in rhizomes and roots during early growth, high carbohydrate levels were maintained both in the shoot and in the underground organs during most of the growing season. Generally, the lowest carbohydrate reserve level in both root and shoot occurred before flowering, a relatively high level was maintained in the shoot from flowering until after fruiting, while peak storage in underground parts was reached at the start of fall dormancy. The carbohydrate cycle in these alpine plants is quite similar to that in certain arctic plants.     

木质素含量对四倍体刺槐嫩枝插穗扦插生根的影响

以四倍体刺槐1年生嫩枝插条为试验材料,分析插穗木质素含量与其横截面剪切强度、相关酶活性和激素关系,并探究不同木质素含量的插穗扦插生根性状的效应,为四倍体刺槐扦插选择合适插穗提供理论参考。结果表明:(1)插穗的木质素含量与其横截面剪切强度呈极显著正相关关系(相关系数为0.99),根据剪切强度可以间接地估算其木质素含量。(2)插穗的木质素含量与其POD和PPO活性呈极显著正相关关系(POD相关系数为0.98,PPO相关系数为0.92),也与激素ABA含量呈极显著的正相关关系(相关系数为0.97),而同IAA、IBA呈不显著的相关性,根据POD和PPO活性以及激素ABA含量对生根的影响可以推测木质素含量对生根有一定的影响。(3)不同木质素含量的插穗生根性状差异显著,木质素含量为19.47%时插穗生根能力最强,其插穗生根率为60.39%,平均每株生根量为9.70个,根长为4.85cm;木质素含量为10.60%时插穗生根能力最差。(4)生根性状最佳时的木质素含量为19.47%,其对应的剪切强度范围为40～50kg。     

The Utilization of Carbohydrate and Nitrogen Reserves by Taxus During Its Spring Growth Period

The spring growth and the utilization of carbohydrate and nitrogen reserves in this growth was studied in Taxus media cv. Hicksii plants 0, 2, 4 and 6 weeks after the plants started growing in the spring. The effect of nitrogen applied the previous season on the storage and utilization of the carbohydrate and nitrogen reserves during spring growth was determined. The plants were separated into buds (all new growth), stems, needles (those produced the previous season) and roots and analyzed for changes in total nitrogen, basic and non-basic amino acids, total available carbohydrate, sugars, hemicelluloses, organic acids and chlorophyll. The bulk of the soluble nitrogen reserves were stored as arginine in the stems and old needles. With the onset of spring growth, arginine nitrogen was converted to other amino acids which accumulated in the new growth (buds). The roots, stems and needles of plants grown under high nitrogen levels always contained more total nitrogen than those grown under low nitrogen levels. The bulk of the carbohydrate reserves were stored as hemicelluloses. The plants grown under high nitrogen levels utilized the bulk of the carbohydrate reserves from the roots and smaller amounts from the stems and old needles, while plants grown under low nitrogen levels used only the reserves in the roots. In the low nitrogen plants, carbohydrates accumulated in the needles and stems. Both the carbohydrate and nitrogen reserves were important in the dry weight increase due to spring growth. However, the nitrogen reserves were the limiting factor and the high nitrogen plants grew twice as much, produced more chlorophyll, and utilized more nitrogen and carbohydrate reserve in spring growth than low nitrogen plants. The additional chlorophyll allowed the production of more carbohydrates and these additional carbohydrates were used in increased growth rates, while in the low nitrogen plants the carbohydrate produced was less and accumulated within the plant.     

Effect of Wild-Type and Mutant Plant Growth-Promoting Rhizobacteria on the Rooting of Mung Bean Cuttings

Mung bean cuttings were dipped in solutions of wild type and mutant forms of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2 and then incubated for several days until roots formed. The bacteria P. putida GR12-2 and P. putida GR12-2/aux1 mutant do not produce detectable levels of the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, whereas P. putida GR12-2/acd36 is an ACC deaminase minus mutant. All bacteria produce the phytohormone indole-3-acetic acid (IAA), and P. putida GR12-2/aux1 overproduces it. Treatment of cuttings with the above-mentioned bacteria affected the rates of ethylene production in the cuttings in a way that can be explained by the combined effects of the activity of ACC deaminase localized in the bacteria and bacterial produced IAA. P. putida GR12-2 and P. putida GR12-2/acd36-treated cuttings had a significantly higher number of roots compared with cuttings rooted in water. In addition, the wild type influenced the development of longer roots. P. putida GR12-2/aux1 stimulated the highest rates of ethylene production but did not influence the number of roots. These results are consistent with the notion that ethylene is involved in the initiation and elongation of adventitious roots in mung bean cuttings. Received October 21, 1998; accepted January 3, 1999