Epidemiology of stamen blight of raspberry

Stamen blight, caused by Hapalosphaeria deformans Syd., is a disease of a number of Rubus species. It is widespread on commercial raspberries in Angus and east Perthshire. Evidence indicates that spread is from diseased flowers to the axillary buds of young canes during the summer and entry into these buds is complete by autumn, after which it is unlikely that the pathogen can be reached even by an eradicant fungicide. Attempts to infect young canes artificially, however, have failed. Rain splash appears to be of considerable importance in dispersal, but some spread by wind and pollinating insects could occur. Possible control measures are discussed and the importance of planting canes from spawn beds, where no flowering canes are kept, is emphasized.     

Resistance to Botrytis cinerea in canes of Rubus idaeus and some related species

Mycelial inoculation of canes with Botrytis cinerea proved a useful method for assessing Rubus material for resistance. When canes were inoculated in summer resistant genotypes developed relatively small lesions which produced few small sclerotia in the following spring. The size of lesions in autumn generally provided the best discrimination between genotypes. Very strong resistance was found in several species, of which Rubus pileatus and R. occidentalis are the most useful for breeding; hybrids of raspberry with these species or with R. crataegifolius also had strong resistance. The resistance shown by a derivative of red raspberry cv. Chief was less strong, as was the resistance conferred by gene H, which determines cane pubescence.     

Heritability of adventitious rooting of grapevine dormant canes

One of the most important viticultural characteristics of a grapevine rootstock is the ability to form roots on dormant lignified canes (rootstrike). North American species of Vitis are the primary source of germplasm for grapevine rootstocks and vary widely in their rate of rootstrike. Breeders have hybridized grape species in order to introgress traits to produce commercial rootstocks. A combination of 26 parents consisting of improved and wild accessions of Vitis spp. was used to generate 27 families. The percentage of rootstrike of dormant canes was observed over several years for 552 individuals. A logistic generalized linear mixed model (GLMM) method was used to estimate the narrow sense heritability (h ²) of rootstrike. Heritability was found to be moderate (h ²?=?0.307?±?0.050). The model also estimated breeding values of all parents and progeny. A GLMM method can be used to estimate breeding values of germplasm to identify individuals with commercially acceptable rates of rootstrike with a defined probability of transmitting this trait to progeny. This is useful for the introgression of traits into potentially new commercial rootstocks. The pattern of normal distribution of rooting indicates that it is possible to identify individuals with good rootstrike from Vitis species that are generally considered to have low rootstrike. Selection of individuals with a higher breeding value will increase the efficiency of rootstock breeding.     

<Emphasis Type="Italic">In vitro</Emphasis> organogenesis of <Emphasis Type="Italic">Pelecyphora aselliformis</Emphasis> erhenberg (cactaceae)

Summary In vitro propagation of Pelecyphora aselliformis, a Mexican cactus which is considered rare and is highly valued in the commercial market, was initiated using seeds as explants. The longitudinal explants from seedlings germinated in vitro were cultivated on Murashige and Skoog medium containing 8.8 μM benzyladenine (BA) or 4.6 μM kinetin at pH 7.0. After 120 d, each explant gave rise to five shoots and this number of shoots increased 20–25% after subculture. The hyperhydricity was similar in both media, but callus formation was lower on the medium with BA. The shoot development, in terms of epicotyl length, and fresh and dry weight after 6 wk, was also recorded. The epicotyl length was similar on shoot-forming media but the quality of shoots was better on media containing BA. In about 1 yr, 500–600 well-defined shoots were obtained. The rooting of shoots was very slow and a vigorous radical system was observed after 1 yr of culture.     

Induction,growth and direct rooting of adventitious shoots of Begonia x hiemalis

The efficiency of commercial micropropagation programs for Begonia x hiemalis depends on the production of large adventitious shoots for easy handling and on effective rooting and acclimatization procedures. Maximum induction of adventitious buds on petiole segments occurred in response to NAA (0.1 mg, l^-1) and BA (0.5 mg l^-1), but continued shoot growth was limited. With a lower concentration of BA (0.1 mg l^-1) fewer shoots were produced but shoot growth was enhanced. With a combined agar/liquid culture program the low BA (0.1 mg l^-1) medium produced 50 percent more shoots larger than 1 cm than did the high BA (0.5 mg l^-1) medium. In vitro rooted explants developed weak root systems and acclimatization losses occurred during adaptation to greenhouse conditions. Adventitious shoots treated with commercial rooting powder and placed directly in mist frames produced much stronger root systems and could be adapted to greenhouse conditions without loss. The elimination of the in vitro rooting stage also simplifies the micropropagation program.Contribution No. 743     

Agrobacterium tumefaciens-mediated transformation of severalRubus genotypes and recovery of transformed plants

Experiments in shoot regeneration and virulentAgrobacterium tumefaciens-mediated transformation were used to develop a protocol forRubus transformation. This protocol was then used to produce transformedRubus plants fromin vitro internodes inoculated with anAgrobacterium tumefaciens encoding neomycin phosphotransferase on its disarmed T-DNA. Two transformed plants were selected from 800 inoculations on a medium containing 10 µg ml^–1 kanamycin. Results indicated that this level of kanamycin successfully selected against non-transformed cells but did not reduce the number of transformed, kanamycin-resistant, shoots formed. Enzyme assays and Southern blot analysis verified the presence of the -glucuronidase gene in the plant genome.     

Effects of short light-dark regimes on in vitro shoot rooting of some fruit tree rootstocks

Experiments were carried out to evaluate the effects of 4/2 light-dark cycles (4 h of light followed by 2 h of dark) on the rooting responses of shoots cultivated in vitro of the fruit tree rootstocks GF 677 (peach × almond hybrid), Mr.S. 2/5 (Prunus cerasifera), MM 106 (apple Nothern Spy × Paradise M1) and BA 29 (Cydonia oblonga). Under this light regime rooting percentage of GF 677, Mr.S. 2/5 and MM 106 shoots reached 100 % as in the control treatment (16/8), while in BA 29 shoots, short light-dark cycles increased rooting response by about 65 %. Moreover, the shoots of all rootstocks submitted to the 4/2 cycle showed an appreciable increase in root number and length, and an earlier root emergence of about 4 – 5 d compared to the 16/8 cycle. Finally, rooting percentage of BA 29 shoots submitted to the 4/2 light regime and treated with 0.2 mg dm⁻³ indolebutyric acid (IBA), was equal to that reported with 0.4 mg dm⁻³ IBA under the 16/8 regime, indicating that the former light regime also amplified the rhizogenic effect of auxin.     

Evidence for an interaction effect during <Emphasis Type="Italic">in vitro</Emphasis> rooting of oil palm (<Emphasis Type="Italic">Elaeis guineensis</Emphasis> Jacq.) somatic embryo-derived plantlets

In vitro rooting of oil palm shoots derived from somatic embryos was achieved through a single-phase protocol in which three shoots are cultured in the same culture tube on an α-naphtaleacetic acid-enriched culture medium. Rooting performance was dependent on both the genetic origin and initial size of the shoot explants. All shoots from a given tube showed a tendency to give roots of the same type, independent of the original size of the explant. Whatever the clonal line, longer-size shoots (L-type: >9 cm) showed higher rooting rates than medium-size (M-type: 7–9 cm) and short-size ones (S-type: 5–7 cm). When groups of three shoots from the same clonal line were rooted together in the same culture tube, the combination of plant size within the group impacted overall quality of rooting. Within triplets of shoots containing more than one short individual, the probability of obtaining adequate rooting was low. Similarly, when more than one long shoot was included in the triplet rooting, quality was also poor. By avoiding such combinations, the rate of well-rooted plantlets increased by 25%, with a maximum of 66% when triplets of S/M/L combination were used. Smaller shoots, which usually showed poor rooting performance, were therefore found to benefit from the presence of their neighbors. This interaction between the sizes of individuals in a given tube was found to be associated with a within-tube correlation effect, a phenomenon previously described as “event coupling,” which was estimated using a distorted binomial-type distribution of probabilities. The resulting calculation of a coupling factor (average r = 0.60) explains the behavior of shoots within the same culture tube and their average rooting performance. Modeling of the interactions that occurred during in vitro rooting is described here and is recommended for improvement of this critical step in micropropagation.     

裸果木再生体系建立与不定芽发生研究

为探究裸果木再生体系建立的影响因素,确定其不定芽发生的起源,该研究以裸果木健壮植株的茎段为外植体,采用6 BA和IBA不同浓度组合,筛选愈伤增殖及不定芽再生的最佳浓度组合,确定生根诱导的关键影响因素,建立再生体系,并对其不定芽分化进程进行解剖结构分析,以确认其起源。结果表明：(1)裸果木茎段的最佳愈伤增殖培养基为MS+1 mg·L^-1 IBA+1 mg·L^-1 6 BA+30 g·L^-1蔗糖+7 g·L^-1琼脂,主体间效应分析表明IBA为关键影响因素;愈伤大小随IBA浓度增加呈现先升高后下降的趋势。(2)最佳不定芽诱导培养基为MS+0.5 mg·L^-1 6 BA+30 g·L^-1蔗糖+7 g·L^-1琼脂,诱导不定芽数量为4.9个/块,生芽率达92.3%。(3)生根诱导中,SH基本培养基和蔗糖浓度为关键因素,最佳生根培养基为SH+0~10 g·L^-1蔗糖+7 g·L^-1琼脂,生根率达91.3%。(4)解剖结构观察发现,不定芽起源于愈伤表层的分生细胞,为外起源。该研究通过器官发生途径建立了裸果木的再生体系,确定了不定芽为外起源,为裸果木这一珍稀濒危的林木种质资源保护及可持续利用奠定了研究基础,并为其未来的发展利用提供了有效途径。     

Radial growth responses to gap creation in large,old Sequoiadendron giganteum

Questions: Do large, old Sequoiadondron giganteum trees respond to the creation of adjacent canopy gaps? Do other co‐occurring tree species and younger S. giganteum adjacent to gaps also respond? What are the likely factors affecting growth responses? Location: Mixed‐conifer forests of the southern Sierra Nevada, California, USA. Methods: We measured the growth response of large, old S. giganteum trees (mean DBH=164 cm; ages estimated >1000 yr) to gap creation by coring trees and comparing growth after gap creation to growth before gap creation. We also measured young Abies concolor, Pinus lambertiana, and young S. giganteum. Gap‐adjacent trees were compared with non‐adjacent reference trees. Tree rings were analysed for carbon isotope discrimination and for longer‐term growth trend correlations with climate. Results: Following gap creation gap‐adjacent old S. giganteum grew more than reference trees. Abies concolor trees also exhibited a growth response to gap creation. No response was detected for young S. giganteum or P. lambertiana, although detection power was lower for these groups. There was no difference in carbon isotope discrimination response to gap creation between gap‐adjacent and reference trees for old S. giganteum and radial growth was positively correlated with winter precipitation, but not growing season temperature. Conclusion: It is unclear what caused the growth release in old S. giganteum trees, although liberation of below‐ground resources following removal of competing vegetation appears to be a significant contributor. Sequoiadondron giganteum, the third‐longest lived and the largest of all species, remains sensitive to local environmental changes even after canopy emergence. Management activities that reduce vegetation surrounding individual specimen trees can be expected to result in increased vigor of even these very old and large trees.     

Effects of Didymella applanata and Botrytis cinerea on axillary buds, lateral shoots and yield of red raspberry

The viability of axillary buds and the growth and potential yield of lateral shoots at nodes of red raspberry (Rubus idaeus) infected naturally by Didymella applanata or Botrytis cinerea were measured on excised nodes, decapitated nursery canes or on canes from fruiting plantations. In comparison with lesion-free nodes, buds at infected nodes were smaller and fewer of them were capable of growth when excised and ‘forced’, although the difference in growth between infected and uninfected nodes decreased during late winter. After February, those buds at infected nodes which were capable of forced growth did so as early and with a similar growth rate as those at lesion-free nodes. In April, 70% of buds at infected nodes were capable of growth compared with 94% of those at lesion-free nodes. When nursery canes of cv. Mailing Delight were decapitated above infected nodes the emergence of lateral shoots from the terminal infected node did not differ significantly from that at lesion-free nodes. On a range of farm sites in Scotland the emergence of shoots at infected nodes in the cropping region of canes was significantly poorer than from uninfected nodes but substantially better at infected nodes of cv. Glen Clova than at those of cvs Mailing Jewel and Mailing Orion. It is suggested that cv. Glen Clova is relatively tolerant of spur blight and cane botrytis. The length and potential yield of laterals which developed at infected nodes in the cropping region of canes in these three cultivars did not differ significantly from those at lesion-free nodes. In all tests there was no significant difference in growth at nodes infected by D. applanata and B. cinerea which may indicate a common mechanism for suppression of buds.     

Sanitizing long-term micropropagated grapes from covert and endophytic bacteria and preliminary field testing of plants after 8 years <Emphasis Type="Italic">in vitro</Emphasis>

Summary Micropropagated grape (Vitis vinifera L.) cv. Arka Neelamani cultures showed a decline in root and shoot growth performance after 6–7 yr of continuous in vitro culture. Indexing the culture medium using nutrient agar or 523 bacteriological medium (Viss et al., 1991) revealed covert bacteria in 75–100% cultures. Testing the tissue from different parts of in vitro plantlets on nutrient agar showed bacteria comprising of six or more morphotypes in 100% of root and collar tissue samples but less frequently in stem segments. The shoot tips had the lowest incidence of bacterial association. The whole shoots treated with NaOCl (4% chlorine) or HgCl₂ (0.1%) showed endophytic bacterial survival. Culturing the HgCl₂-treated (5 min) shoot tips on antibiotic overlaid medium (1 ml of 50 mg l⁻¹ gentamycin and/or cefazolin) in culture tubes (150×25 mm) for 1 mo. facilitated the cleansing of cultures with 75% recovery of contaminant-free shoots as monitored through indexing for the next 2 yr. Repeated indexing of medium and tissue from various plant parts during the first two to four subculture cycles following antibiotic treatment was instrumental in reliably identifying clean cultures and preventing bacterial re-emergence. Antibiotic incorporation in the medium was detrimental to grape microcuttings. Bacteria-freed cultures showed 80–100% rooting and a high number of plantlets that could be acclimatized. The plants put in the field after 8 yr of active micropropagation showed some juvenile characteristics initially, which disappeared in 6–8 mo., and the pruned shoots showed flowering and bunch development within 1 yr of field planting. This indicated the feasibility of keeping grape plants in vitro for long periods if covert microbes were eliminated.     

In vitro propagation of Centaurea spachii from inflorescence stems

Procedures for micropropagation of Centaureaspachii (Compositae), an endangered rosulate plantendemic from the mediterranean Spain area, have beendeveloped using inflorescence nodal segments asexplants for in vitro establishment. Only 15%of explants remained contaminated using this materialto start the in vitro axenic cultures. Higherproliferation of shoots and multiplication coefficientwas obtained on Murashige and Skoog (MS) mineralmedium supplemented with 1.0 mg l^{minus 1}6-benzyladenine. However, shoot elongation decreasedwith the addition of this cytokinin.Rooting of shoots with only one auxin was very lowafter 6 weeks on the majority of rooting media tested.The best rooting result (60%) was obtained on MSmedium with a combination of 2 mg l^{minus 1}indole-3-acetic acid plus 2 mgl^{minus 1} indole-3-butyric acid. Moreover, in thisculture medium 50% of shoots rooted during the thirdweek of culture. High survival, over 80%, wasobtained when the plantlets were transferred togreenhouse conditions. The endangered Centaureaspachii can be successfully micropropagatedbeginning with a single inflorescence stem and withoutsignificant damage to the mother plant.     

Stock-plant etiolation causes drifts in total soluble sugars and anthraquinones, and promotes adventitious root formation in teak (Tectona grandis L. f.) coppice shoots

The effects of stock-plant etiolation on coppice-shoot growth, drifts in total soluble sugars and anthraquinones (AQs; C₁₄H₈O₂), and rooting potentiality of shoot cuttings were examined in Tectona grandis L. f. (clone FG1). When seedlings were one-year-old, they were coppiced and maintained in the dark for etiolation, with a parallel set kept under natural light in an open environment. Coppice shoots were made into single-node leafy cuttings (SNCs), which were cultured under intermittent mist for rooting. These SNCs were treated with different concentrations of NAA (0, 2000 and 3000 mg l⁻¹). Etiolation significantly increased the coppice-shoot length, internode length, number of coppice shoots, number of leaves, number of nodes and total soluble sugars. The HPTLC analysis showed qualitative and quantitative differences in AQs in coppice shoots obtained from etiolated and non-etiolated stock plants. The study showed that AQs could be used as a marker for maturity and juvenility in teak. Stock-plant etiolation caused a significant increase in percent rooting and sprouting, shoot length, number of shoots and number of leaves per SNC, but a decrease in callusing at the base of the SNC. NAA at 2000 and 3000 mg l⁻¹ had inhibitory effects on rooting and sprouting of SNCs. The result showed that stock-plant etiolation fostered rooting by rejuvenating the coppice shoots.     

Copper uptake and translocation in chicory ( Cichorium intybus L. cv. Grasslands Puna) and tomato (Lycopersicon esculentum Mill. cv. Rondy) plants grown in NFT system. I. Copper uptake and distribution in plants

The uptake and distribution of copper was examined in chicory (Cichorium intybus L. cv. Grasslands Puna) and tomato (Lycopersicon esculentumMill. cv. Rondy) plants grown in a Nutrient Film Technique System (NFT) with addition of 0.05, 5, 10 and 20 mg Cu L^-1. Biomass production of shoots and roots of both chicory and tomato was strongly depressed by Cu concentrations higher than 5 mg Cu L^-1 in the rooting media. Although Cu concentrations in both shoots and roots of both species increased with increasing Cu concentrations in the rooting media, the increase in roots was very much greater than that in shoots, in which the range of concentrations was small. A large proportion of total Cu uptake was retained by roots except when plants were grown in solution Cu concentrations of 0.05 mg Cu L^-1. Copper retention by roots limited Cu translocation to xylem and shoots. Copper adsorption by the root appears to buffer against increases of Cu in the rooting media. A cupric-sensitive electrode used in conjunction with total Cu analysis by graphite furnace atomic absorption spectrophotometry (GFAAS) indicated that more than 99.6% of total Cu in xylem sap was in a complexed form. Large differences between measured and predicted Cu accumulation by shoots of tomato (0.134–0.243 mg Cu plant^-1, 0.660–4.274 mg Cu plant^-1, respectively) and chicory (0.095–0.203 mg Cu plant^-1, 0.626–1.620 mg Cu plant^-1, respectively) suggest that some xylem transported Cu is recirculated to roots via the phloem. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genetic stability of cryopreserved shoot tips of <Emphasis Type="Italic">Rubus</Emphasis> germplasm

Questions often arise concerning the genetic stability of plant materials stored in liquid nitrogen for long time periods. This study examined the genetic stability of cryopreserved shoot tips of Rubus germplasm that were stored in liquid nitrogen for more than 12 yr, then rewarmed and regrown. We analyzed the genetic stability of Rubus grabowskii, two blackberry cultivars (“Hillemeyer” and ‘Silvan’), and one raspberry cultivar (“Mandarin”) as in vitro shoots and as field-grown plants. No morphological differences were observed in greenhouse-grown cryopreserved plants when compared to the control mother plants. In the field, cryopreserved plants appeared similar but were more vigorous than mother plants, with larger leaves, fruit, and seeds. Single sequence repeats (SSR) and amplified fragment length polymorphism (AFLP) analyses were performed on shoots immediately after recovery from cryopreservation and on shoots subcultured for 7 mo before analysis. Ten SSR primers developed from “Marion” and “Meeker” microsatellite-enriched libraries amplified one to 15 alleles per locus, with an average of seven alleles and a total of 70 alleles in the four genotypes tested. No SSR polymorphisms were observed between cryopreserved shoots and the corresponding mother plants regardless of subculture. Although no polymorphisms were detected in shoots analyzed immediately after recovery from cryopreservation, AFLP polymorphisms were detected in three of the four Rubus genotypes after they were subcultured for 7 mo. Field-grown plants from the polymorphic shoot tips of R. grabowskii and ‘Silvan’ displayed the same AFLP fingerprints as their corresponding mother plants. Only long-cultured in vitro shoot tips displayed polymorphisms in vitro, and they were no longer detected when the plants were grown ex vitro. The transitory nature of these polymorphisms should be carefully considered when monitoring for genetic stability.     

The effect of cane management techniques on raspberry yellow rust (Phragmidium rubi-idaei)

The effects of various cane removal treatments on the epidemiology of Phragmidium rubi-idaei were studied in four experiments in infected raspberry plantations in Wales and Scotland. In 1981, removal of 15 cm high young canes either by cutting or by spraying dinoseb-in-oil reduced infection of a second flush of canes (replacement canes) compared to that of the first flush of canes on untreated plots of cv. Mailing Delight in Wales and of cv. Glen Clova in Scotland. In 1982, removal of 20 cm high young canes reduced infection of cv. Mailing Delight in Wales but in Scotland only removal of 60 cm high canes by cutting reduced infection of replacement canes. Removal of all fruiting canes and old cane stubs reduced rust infection of young canes, but removal of successive flushes of replacement canes did not reduce infection of lateral shoots on the fruiting canes. Dinoseb-in-oil applied to the bases of fruiting canes before emergence of young canes had no effect on infection of young canes in Wales or Scotland. The production of basidiospores was inhibited in vitro by dinoseb-in-oil at concentrations over 1.0 μg a.i./ml.     

Effect of genotype on rooting of hypocotyls and in vitro-produced shoots of Pinus radiata

Significant genotypic variation at both the family and clone-within-family levels was seen for hypocotyl rooting and the rooting of adventitious shoots produced in vitro for Pinus radiata D. Don. High rooting frequencies for hypocotyls were obtained in the absence of exogenous auxin; auxin greatly stimulated the rooting of adventitious shoots. No correlation was seen between the rooting of hypocotyls and shoots; families whose hypocotyls rooted at high frequencies did not necessarily produce shoots that rooted at high frequency. No correlation was seen between adventitious shoot production and subsequent rooting at either the family or clone level. The lack of a negative correlation indicated that selecting families or clones for high levels of shoot production will not automatically select for low rooting ability, obviating a possible bottleneck for commercial propagation of Pinus radiata. Significant family by replication interaction suggests that rooting protocols could be optimized through manipulations of the rooting environment for each family.     

继代培养次数对杜梨生根能力和叶形态及其激素水平的影响

为了探索杜梨组培复幼变化规律,对10年生杜梨进行连续继代培养,统计不同继代次数杜梨丛生芽繁殖系数和生根率,观察记录叶片形态变化并测定内源激素含量。结果表明:(1)通过连续继代培养,杜梨丛生芽生根率由0提升到66.70%,繁殖系数由第1代的2.13提升到第10代的4.20。(2)叶片在继代第3次时出现裂刻且随后裂刻程度逐代加深;在继代过程中,丛生芽叶面积和叶脉数显著降低,叶周长和叶形指数呈先下降后上升的变化趋势。(3)丛生芽叶片内源IAA含量在继代第6次时达到46.39 ng·g-1,且显著高于初代丛生芽;随着继代次数的增加,叶片内源ZR呈先上升后下降的变化趋势,内源GA3含量没有发生显著性变化,而内源ABA含量逐渐降低;叶片IAA/ABA和IAA/ZR的值随着继代次数的增加而增加。(4)丛生芽叶片ABA含量和IAA/ZR与其生根率分别呈显著负相关和显著正相关关系,叶片裂刻数和IAA/ABA与生根率均呈现极显著正相关关系,而叶脉数与生根率则呈现极显著负相关关系。研究认为,连续继代培养可显著提高杜梨丛生芽的生根能力,并且与丛生芽叶形和激素含量及其比值有密切的关系,该研究结果为难生根植物无性繁殖以及树木复幼提供了重要技术借鉴。     

Factors affecting the efficiency of micropropagation from lateral buds and shoot tips of <Emphasis Type="Italic">Rubus</Emphasis>

Factors affecting micropropagation efficiency of 32 selections of Rubus, including the pre-treatment and initiation culture stages, were investigated. Chilling at 4°C for 6 weeks as a pre-treatment significantly promoted in vitro initiation culture; up to 65% of initiation cultures post chilling were successful. The cytokinin 6-benzyladenine (BA) was the most effective of the three tested for promotion of multiple shoot development in culture, with an average of three to seven shoots or plantlets developed from each single node. Alternating the concentration of BA between 4.44 and 13.31 μM from sub-culture to sub-culture improved recalcitrant Rubus micropropagation, and reduced the problems associated with long-term culture in the presence of high concentrations of BA. Reduction of the strength of macro- and micro-elements in the basal medium from half to one-third, with addition of 0.49 μM indole-3-butyric acid and 0.05% activated charcoal, and placing the cultures under reduced light intensity (17 μmol m⁻² s⁻¹), was found to alleviate chlorosis and improve micropropagation with high quality Rubus plantlets. Response to in vitro culture differed greatly and consequently different methods of micropropagation were required by different genotypes. From these selections, three types of micropropagation including micro-cutting, micro-shoots and multi-shoots were observed, and their efficiency was characterized.