Growth of Vitis vinifera L. and Vitis aestivalis Michx. as Affected by Temperature

Abstract

Four European (Vitis vinifera L.) winegrape cvs., ‘Semillon’, ‘Pinot Noir,’ ‘Chardonnay’, and ‘Cabernet Sauvignon’, and one American (Vitis aestivalis Michx.) winegrape cv. ‘Cynthiana’, were subjected to three temperature regimes in growth chambers set at 20/15°C, 30/ 25°C, or 40/35°C, for 16/8 hr day/night to determine the influence of temperatures on vine growth and development. In general, the best temperature for shoot and root growth 28 days after temperature treatments was 20/15°C for ‘Semillon’, ‘Cabernet Sauvignon’, and ‘Cynthiana’, and 30/25°C for ‘Pinot Noir’ and ‘Chardonnay’. Although 40/35°C reduced number of leaves, shoots, tendrils, and internodes, total leaf area (LA), and total shoot biomass of all the cultivars, the reduction was more pronounced in ‘Cynthiana’ than in the European cultivars. The average reduction in number of leaves at 40/35°C for the European cultivars was 47%, compared with 92% for ‘Cynthiana’. The two types of grapes adapted differently to high temperature. Shoot growth in the European cultivars continued under high temperature, whereas growth ceased in ‘Cynthiana’. Roots of ‘Cynthiana’, however, were less susceptible to the adverse effect of high temperatures than were the shoots. This study shows that the European cultivars were relatively more tolerant to high temperature than the American cultivar and they have a potential for production of wine in the climate of south central Kansas.     

宁夏贺兰山东麓不同葡萄品种对霜霉病的抗性鉴定

为了给葡萄抗性品种的选育和葡萄霜霉病的防治提供科学依据,对16个不同酿酒葡萄品种和19个鲜食葡萄品种的抗病性进行了抗性测定。结果表明,室内回接与田间调查结果一致,16个酿酒葡萄品种可划分为4类;19个鲜食葡萄品种可划分为4类;从发病时间来看,抗性与发病时间存在一定相关性,抗性越强发病越晚。随即将测定结果采用DPS软件进行聚类分析,在酿酒葡萄抗性反应谱系图上,第1类为抗性稍强品种,为白比诺、灰比诺和赛美蓉;第2类为抗性中等品种,由白长相思和神索等8个品种组成;第3类是稍感病品种,包括赤霞珠等4个品种;第4类是感病品种,只有品丽珠1个。在鲜食葡萄谱系图上,第1类为六月紫等7个抗性稍强品种;第2类是新华1号和巨峰;第3类是稍感品种,由奥古斯特等6个品种组成;第4类为力扎马特等4个感病品种。     

The Incorporation of Growth Hormones in Seed Dressings

Summary

The consequences of using ex vitro, single-node explants from different topophysical positions in chrysanthemum (Chrysanthemum grandiflorum /Ramat./ Kitam) were determined. In particular, how explant topophysis affected the rate of propagation, which is important for the successful micropropagation of chrysanthemum. Uniform shoots of five cultivars of chrysanthemum, cultured in vitro, were each divided into three equal zones: distal, central, and proximal. Two single-node explants were isolated from each zone and cultured on MS medium without any added growth regulators. After 10 weeks of culture, 50% of the shoots that had developed from axillary buds on each single-node explant were excised and measurements were taken in order to compare those shoots that had developed from explants from the different topophysical zones. The remaining shoots were sub-cultured on rooting medium. After 4 weeks, the numbers of roots per plantlet, and the total fresh weight (FW) of roots were recorded. The cultivars fell into two groups. ‘Lady Amber’, ‘Lady Orange’, and ‘Lady Vitroflora’ explants were topophysis-dependent, while ‘Lady Bronze’ and ‘Lady Rosy’ explants were topophysis-independent. For the three topophysis-dependent cultivars, the propagation rate, growth rate, shoot length, internode length, single leaf weight, and total plantlet FW values were highest for those shoots derived from the central and proximal zones. Topophysis failed to affect the number of leaves per shoot or the number of days between the appearance of two successive leaves. The effects of topophysis on the number of roots per plantlet and on root FW were inconsistent. The unequal growth of chrysanthemum plantlets during in vitro micropropagation can be an effect of topophysis, and this phenomenon is cultivar-specific in chrysanthemum.     

Sexual compatibility within and between olive cultivars

Summary

Self- and cross-incompatibility of the olive cultivars Frantoio, Manzanillo, Kalamata, Pendolino, and Picual were investigated using a 5 × 5 diallel matrix. Pistils were collected seven days after controlled pollinations on the day of flower opening, and pollen tubes were detected by fluorescence microscopy. Diallel analysis showed significant specific combining ability, general combining ability and reciprocal effects between cultivars for pollen tube growth in the pistil. ‘Frantoio’ was cross-compatible, as either a male or female parent, with each of the other cultivars, but showed a high degree of self-incompatibility. ‘Manzanillo’, ‘Kalamata’, ‘Pendolino’, and ‘Picual’ were crossincompatible, and all except for ‘Manzanillo’, were self-incompatible. It is concluded that ‘Frantoio’ is a good general polleniser for the other cultivars investigated. Pollen tube growth decreased in discrete steps from stigma to upper style, and from upper style to lower style, with the result that only one, and rarely more, pollen tube penetrated ovules. The sex ratio of flowers, and pollen viability using fluroescein diacetate staining and in vitro germination, were examined. ‘Frantoio’, ‘Manzanillo’ and ‘Pendolino’ had more than 80% perfect flowers, while ‘Kalamata’ and ‘Picual’ had less than 30%. ‘Frantoio’ had the highest pollen viability, ‘Kalamata’ and ‘Picual’ were intermediate, and ‘Manzanillo’ and ‘Pendolino’ the lowest. Pollen staining and both in vitro and in vivo germination provided the same male fertility rankings of cultivars.     

Stilbene Levels in the Tissue and Juice of Muscadine Grapes (Vitis rotundifolia Michx.)

Abstract

Stilbenes are secondary metabolites of a class of non-flavanoid phenolics found in certain species of Vitis. This class of compounds includes resveratrol and a resveratrol glucoside called piceid that exists in trans and cis forms. This study was undertaken to determine the cis and trans forms of both resveratrol and piceid in the skin, pulp, seeds, and juice of nine cultivars of muscadine grapes and three cultivars of Vitis labruscana. Juice samples of the muscadine cultivars ‘Fry’, ‘Hunt’, ‘Magnolia’, ‘Watergate’, ‘Carlos’, ‘Noble’, and ‘Sweet Jenny’, and the V. labruscana cultivars ‘Albermarle’, ‘Miss Blanc’, ‘MidSouth’ and ‘Miss Blue’ were harvested at the full ripe stage and divided into two sub-samples. One subsample was used to extract juice and the other was divided into skins, pulp, and seeds. Sample analysis was performed using HPLC with a UV detector. Sample chromatagrams were compared with those of known standards for quantification. ‘Carlos’ and ‘Magnolia’ had the greatest total stilbene concentration in skin tissue and had bronze skin containing higher concentrations of total stilbenes than any of the black-skinned cultivars tested. Only one bronze skin cultivar, ‘Sweet Jenny’, had lower stilbenes than the dark skin cultivars. Piceid, cis piceid and resveratrol were found in the skins of all cultivars. ‘Carlos’ ‘Magnolia’, ‘Fry’ and ‘Albermarle’ muscadines contained cis resveratrol in the skins of the fruit. ‘Watergate’ was the only cultivar in which resveratrol was detected in seed tissue. ‘Sweet Jenny’ had the highest levels of cis piceid in seed tissue. ‘Albermarle’, ‘Carlos’ and ‘Sweet Jenny’ had significantly greater levels of cis picied in pulp tissue than all other cultivars.     

A 3-Year Adaptation Study of Three Distinct Grapevine Cultivars under Midwestern Field Conditions

Simultaneous responses of American (Concord), European (Pinot Noir), and American-European hybrid (Traminette) cultivars to seasonal and diurnal variation in field environmental conditions were assessed to propose suitable cultivars for potential use for selection and breeding in grapevines in dry areas. ‘Pinot Noir’ had higher gas exchange parameters, higher water potential, and chlorophyll fluorescence, while ‘Concord’ had the lowest values in all three seasons and experienced moderate stress by the end of each season. The dry conditions in 2005 caused water potential to decline sharply compared to values recorded in 2004 and 2006. ‘Pinot Noir’ and ‘Traminette’ exhibited a mild stress, while ‘Concord’ exhibited a moderate stress in 2006. Photosynthetic rate decreased slightly during the summer with a slight recovery at the beginning of September, and did not differ across the seasons among cultivars. Stomatal conductance followed the same pattern. However, in both 2004 and 2005, stomatal conductance dropped sharply late in the season because of frequently occurring high temperatures that were greater than 32 °C. Maximum morning values of Fv/Fm were within the optimal range. The study suggests the presence of marginal photoinhibition during mid-summer in Concord cultivars, because no full recovery of photochemical efficiency early in the evening was observed. The European cultivar (Pinot Noir) and the American-European hybrid (Traminette) are more adapted to field conditions in Kansas than Concord cultivars. However, high cold-mortality of Pinot Noir due to high sensitivity to cold winter temperature makes the hybrid cultivars the most suitable cultivar for Midwest regions with relatively cold winters.     

Morphology and anatomy of pollen grains from male-fertile and male-sterile cultivars of chestnut (Castanea sativa Mill.)

Summary

Pollen morphology and ultrastructure are described for four male-fertile cultivars (‘Firdola’, ‘Karamehmet’, ‘Sar?a?lama’, and ‘Hac?ömer’) and two male-sterile cultivars (‘Osmanog?lu’ and ‘Vakit Kestanesi’) of chestnut (Castanea sativa Mill.) using light microscopy and both scanning and transmission electron microscopy. Pollen grains of both male-fertile and male-sterile chestnut cultivars are tricolporate, the germinal furrow extending almost the full length of the grain axis. Pollen grains have a slightly reticulate exine. Pollen grain length varied from 13.33 – 21.30 µm, and decreased significantly in the male-sterile cultivars. Three different pollen shapes were observed among the cultivars: prolate, perprolate, and sub-prolate. The ultrastructure of the pollen grains did not differ between male-fertile and male-sterile cultivars. Intine, exine and total wall thickness (exine + intine) of pollen grains were determined as: 83.2 – 153.1 nm, 432.8 – 520.0 nm, and 516.0 – 651.6 nm, respectively; and variations were significant (P ≤ 0.05) among cultivars. The percentages of in vitro germination of pollen grains of male-fertile cultivars were between 11 – 78%, and the variations among cultivars were significant (P ≤ 0.05). The percentages of empty pollen grains observed among cultivars ranged from 3 – 32%. The correlation coefficient between the percentage of normal pollen and the germination rate was r = 0.898 (P ≤ 0.01).     

Influence of FeEDDHA on in vitro rooting and acclimatisation of red raspberry (Rubus idaeus L.) in peat and vermiculite

Summary

We investigated the dependence of in vitro rooting and acclimatisation to greenhouse conditions on the source of iron used in the shoot multiplication and rooting media using five raspberry (Rubus idaeus L.) cultivars (‘Beskid’, ‘Canby’, ‘Malling Seedling’, ‘Norna’, and ‘Veten’). Ethylenediamine di-2-hydroxy-phenyl acetate ferric (FeEDDHA) in the rooting medium led to higher chlorophyll contents, earlier and more abundant rooting (8.7 vs. 5.3 roots per shoot), 30% higher fresh and dry weights, and thus higher quality microplants than ethylenediaminetetra-acetate ferric sodium (FeEDTA). Higher quality microshoots had a beneficial effect on acclimatisation (i.e., percentage survival and length of shoots) when the microplants were planted in a peat-based substrate; however, when planted in vermiculite, the initial differences disappeared during a 4 week-long growth period in a greenhouse.     

Effect of temperature on pollen germination and pollen tube growth of four cultivars of mango (Mangifera indica L.)

Summary

The effect of a constant (10, 15, 20 or 25°C) or a diurnal maximum/minimum (15/5, 20/10, 25/15 or 30/20°C) incubation temperature on in vitro pollen germination and pollen tube growth in the pistils of two poly-embryonic (‘Kensington’ and ‘Nam Dok Mai’), and two mono-embryonic (‘Irwin’ and ‘Sensation’) mango cultivars was studied. In in vitro experiments where pollen was incubated in a liquid germination medium for 24 h in darkness, little difference was found between pollen germination of mono- and poly-embryonic cultivars. Averaged over the four cultivars, 53.9% of pollen germinated at 10°C, this increased to 76.2% when the incubation temperature was increased to 15°C, thereafter up to 25°C the percentage germination remained stable but germination decreased slightly to 68.2% at 30°C. Similarly, there was no difference in percent germination between cultivars when pollen was incubated under diurnal temperature regimes. Mean pollen germination of all four cultivars was 52.3% at 15/5°C and pollen germination increased by 10% when the temperature was raised to 30/20°C. When self-pollinated flowers were incubated for 24 h on a semi-solid agar medium at 10°C, pollen tube growth of the four cultivars was retarded and no pollen tubes reached the ovaries. As the temperature was increased from 15 to 25°C, the mean number of pollen tubes in ovaries increased from 0.04 to 0.25. At 30°C, the mean number of pollen tubes that entered ovaries decreased to 0.04. After incubation under diurnal temperature regimes, the mean number of pollen tubes in ovaries of all four cultivars at 15/5°C was 0.23 and increased to 0.42 when the temperature increased to 30/20°C. At each incubation temperature, there were significant differences in pollen tube growth between cultivars, but there were no differences between the temperature response of pollen from mono- and poly-embryonic cultivars.     

Regeneration from in-vitro leaves of ‘Conference’ and other pear cultivars (Pyrus communis L.)

Summary

Regeneration of shoots from in-vitro grown leaf tissues of the pear cv. Conference was achieved at rates of up to 40%. The highest regeneration percentage was achieved using a phytohormone combination of benzylamino purine (5 mg 1^?1) and 1-naphthalene acetic acid (1.0 mg 1^?1) on a basal medium of Murashige and Skoog, to which had been added 200 mg I^?1 of the antibiotic cefotaxime. The percentage regeneration was reduced when a higher concentration of cefotaxime (400 mg 1^?1) was used. Cefotaxime had little beneficial effect on regeneration at a lower concentration of 1-naphthalene acetic acid (0.5 mg l^?1). Regenerated shoots were easily micropropagated, rooted and transplanted to soil. To date, the plants have a true-to-type phenotypic appearance. Regeneration was also achieved with other cultivars—‘Abbé Fetel’, ‘Doyenne d’Hiver’, ‘Doyenne du Comice’ and ‘Passé Crassane’—but percentage regeneration was lower and, apart from ‘Passé Crassane’, the regenerated shoots were weak and could not be subcultured.     

Maintaining product quality in raisins throughout the introgression of powdery mildew resistance

The availability of powdery mildew-resistant raisin accessions will lower grower production costs and enhance the environment through reduced fungicide use. To achieve this objective, backcrossing has been employed with Vitis romanetii as the source of non-race-specific powdery mildew resistance. Initial crosses of powdery mildew-resistant F₁ hybrid B36-45 with seeded raisin cultivars ‘Rangspray’ and ‘Raisin de Palestine’ yielded seedless powdery mildew-resistant first-generation backcross selections C87-41 and C87-106, which were used to develop second-generation backcross populations. Principal component analyses consistently identified ‘wrinkle,’ ‘meatiness,’ ‘product attractiveness’ and ‘skin persistence’ as being the quality characteristics most important in discriminating among powdery mildew-resistant raisin accessions and commercial raisin cultivars. Raisin quality ratings were much improved across most evaluated characters in second-generation backcross families as compared with parental powdery mildew- resistant accessions C87-41 and C87-106. After two generations of backcrossing, powdery mildew-resistant raisin selections were identified with product quality characteristics similar to those of commercial raisin cultivars.     

Metabolite profiling reveals the involvement of aberrant metabolic changes in Gentiana triflora seed showing poor germination

Gentians are alpine plants and two endemic species, Gentiana triflora and G. scabra, are cultivated as ornamentals in Japan. Because most of the Japanese cultivars are F₁ hybrids, seed germination rate is an important factor for maintaining these cultivars. However, germination rates frequently differ among cultivars. Poor germination is a severe problem for gentian production, although there is almost no difference in their germination period and growth rate. In this study, we compared seed germination rates among three cultivars of G. triflora. A high rate of seed germination was observed in ‘Majel’ (MJ) and ‘Iwate’ (IW), but a low rate was recorded in ‘Iwate-Yumeminori’ (YM). We conducted a targeted metabolome analysis using these cultivars to elucidate the potential cause of inhibited seed germination. Multivariate analysis revealed that the aberrant accumulation of specific amino acids and a decrease in energy metabolites were observed in YM seed. Furthermore, protein concentrations and proteolytic activities in YM seed were lower than in MJ seed. These results imply that energy depletion was the main reason for the decreased rate of germination and that this depletion inhibited processes involved in seed germination, including de novo synthesis of the proteins necessary for germination.     

Subject Index

Prohexadione-calcium (ProCa) is a gibberellin biosynthesis inhibitor with limited persistence and low toxicity that has recently been registered as a shoot growth retardant for apples in North America (Apogee^®) and Europe (Regalis^®). We investigated the response of six different pear cultivars (‘Rosemarie’, ‘Forelle’, ‘Packham's Triumph’, ‘Flamingo’, ‘Early Bon Chretien’ and ‘Golden Russet Bosc’) to ProCa in the Western Cape, South Africa, over the period 1999–2003. ProCa treatments ranging from 50 to 250 mg l⁻¹ were applied between one and three times per season and were compared with girdling and control treatments. ProCa application reduced shoot growth in all of the cultivars. ‘Forelle’, was least responsive and showed a reduction in 2002/2003 but not 2001/2002. ‘Rosemarie’, ‘Flamingo’, ‘Packham's Triumph’ and ‘Golden Russet Bosc’ were most responsive, with shoot growth being reduced by up to 50% of the control for the latter two cultivars. The response to number of applications and ProCa concentration was not consistent across cultivars. ‘Early Bon Chretien’, for example, only responded to high ProCa concentration, whereas ‘Rosemarie’ was very responsive at the lowest ProCa concentration. ProCa application improved fruit set but decreased final fruit size in some of the cultivars, particularly in ‘Rosemarie’. It also caused a decrease in return bloom in ‘Forelle’ and ‘Packham's Triumph’. The response of different cultivars to girdling, in terms of shoot growth and fruit set was highly variable. Girdling tended to increase final fruit size and return bloom. ProCa shows potential as a shoot growth retardant for the pear cultivars investigated, but the response of reduced fruit size in some cultivars requires mitigation. Future research on combined girdling and ProCa treatments may be instructive.     

The effects of daminozide and under-soil heating on the growth of radish under winter conditions

The growth of radish cultivars ‘Saxa’, ‘Saxerre’, ‘Serra’, ‘Istar’, ‘Cherrybelle’ and ‘Robino’ was compared at 7 and 12°C under winter glasshouse conditions. ‘Serra’ alone produced marketable storage roots by 63 days at 7°C, but root weight was no greater at 12 than at 7°C; only ‘Robino’ produced significantly larger roots at the higher temperature. Under-soil heating applied to ‘Cherrybelle’ and ‘Robino’ grown at an air temperature of 7°C increased root growth, particularly in ‘Cherrybelle’. Daminozide treatment at 7°C increased total and storage root weight and root:shoot ratio of both cultivars. At 12°C, the only significant effect of daminozide treatment was reduced shoot weight in ‘Robino’.     

Changes in the competence of Quercus robur ‘Fastigiata’ to grow in vitro as affected by seedling rootstocks and differential pruning

Three physiologically distinct forms of the same genotype of Quercus robur ‘Fastigiata’, all derived from adult budwood, were identified following the growth of severely-pruned hedge plants grafted onto juvenile seedling rootstocks. Only explants from the form displaying the juvenile-like characteristic of over-winter leaf retention were capable of establishing sustainable cultures in vitro, although growth was not as rapid as those from a two year old seedling source. Pruning to release correlative inhibition of buds low on the scion framework produced vigorous shoot growth, but explants from these failed to show sustained growth in vitro. Withholding light from stockplants reduced excessive phenolic oxidation in explants taken from flushing unripe shoots, however, despite a beneficial effect on culture initiation, later growth was dependent upon the juvenile- or adult-like physiology of the original explant. A new medium was devised for this subject, which in addition to promoting growth generally, also stimulated the flushing of shoot apices. The relevance to in vitro culture of expression of juvenile habit in the stockplant is discussed.     

Cryopreservation of in vitro almond shoot tips by vitrification

Summary

Shoot tips of two almond scion cultivars, ‘Ne Plus Ultra’ and ‘Nonpareil 15-1’, and one almond/peach hybrid rootstock were successfully cryopreserved using a one-step vitrification technique. Three week old in vitro cultures were cold-hardened at 4°C on the multiplication medium (Murashige and Skoog for ‘Ne Plus Ultra’ and the hybrid rootstock; Almehdi and Parfitt for ‘Nonpareil 15-1’) for three weeks. Shoot tips, 2–2.5 mm long, were excised and precultured for 1 d at 4°C on the same basal medium, without plant growth regulators, supplemented with 0.7 M sucrose. After the preculture, the shoot tips were incubated in vitrification solution at 25°C for 45 min for the almond scion cultivars and 60 min for the hybrid rootstock, and then stored under liquid nitrogen (LN) for at least 3 d. After rapid thawing at 30°C, the shoot tips were washed with the appropriate liquid basal medium containing 1.0 M sucrose and then cultured on the same basal medium, solidified with agar, but excluding NH₄NO₃ or (NH₄)₂SO₄. Shoot regeneration was usually observed within 2–3 weeks. Survival after LN, recorded as the percentage of shoot tips that produced at least one new shoot four weeks after thawing, was 87.5, 60.0 and 72.5% for ‘Ne Plus Ultra’, ‘Nonpareil 15–1’ and the hybrid rootstock respectively. The one-step vitrification method is a promising simple technique for cryopreserving almond scion and rootstock shoot tips from in vitro cultures.     

Cross-(in)compatibility in apricot (Prunus armeniaca L.)

Summary

Eight apricot (Prunus armeniaca L.) cultivars and selections were self- and cross-pollinated in order to determine their pollen and stylar compatibility. Overall, 40 pollination combinations were examined. Pollen-tube growth in pistils pollinated in the laboratory was analysed using fluorescence microscopy. Three inter-incompatiblity groups of cultivars were found, of which two had not been described previously, while an existing group was expanded with one additional cultivar. The first group consisted of three Hungarian cultivars (‘Ligeti Orias’, ‘Cegledi Orias’, and ‘Szegedi Mammut’) and a Moldavian cultivar (‘Kostjuzhenskyi’). The second group consisted of two American cultivars (‘Stark Early Orange’ and ‘Nugget’). The third group consisted of two Serbian selections (‘Novi Sad Early’ and ‘Frushka Gora Early’). In the incompatible cultivar combinations, pollen-tube growth stopped in the style with the formation of the characteristic swelling. In the compatible combinations, the pollen tubes reached the ovary in the majority of the pistils examined.     

The physiology of hyacinth bulbs. XV. The effect of gibberellic acid and silver nitrate on dormancy release and growth

The effect of gibberellic acid (GA₃), supplied to dormant hyacinth bulbs of cultivars ‘Lady Derby’ and ‘L'Innocence’ by vacuum infiltration, on growth and flowering was investigated. Results showed that GA₃ in all applied concentrations (50, 500, 1000 and 5000 mg/l) accelerated growth and flowering in both cultivars, after chilling for 42 days in a garden frame in natural conditions or chilled dry in cold storage at 5°C. Bulb infiltration with 10 mg/l AgNO₃ resulted in the acceleration of flowering only in ‘L'Innocence’, but stimulated the growth of the inflorescence stalk and leaves in both cultivars regardless of the mode of chilling.The infiltration method was confirmed to be very promising.     

Selection-pressure effects of medium pH during regeneration on successive performances of leaf-derived Tomuri’ and ‘Hayward’ kiwifruit (Actinidia deliciosa) somaclones cultured on proliferation culture media with variable pH

Summary

The aims of the work were in vitro selecting for tolerance to high culture pH (CpH) in kiwifruit and evaluating the effects of selection pressure exerted by pH variations during regeneration. Leaf-derived ‘Tomuri’ and ‘Hayward’ kiwifruit (Actinidia deliciosa) somaclones, which had been regenerated at pH 5.7, 7 and 7.5, were compared to each other and to controls (i.e. cultures derived from ‘Tomuri’ and ‘Hayward’ mother plants through standard micropropagation techniques) for shoot growth, proliferation and appearance when cultured at up to pH 8. The proliferation rate (PR), the number of nodes (NN) produced by each shoot and axillary shoot length (ASL) decreased in all ‘Tomuri’ and ‘Hayward’ somaclones and in controls as culture pH increased. At CpH 8, ‘Tomuri’ somaclones regenerated at pH 7.0 generally tended to have higher average PR and NN than the other somaclones and control; their ASL also exceeded that of the other somaclones and was just slightly higher than in control. Regardless of CpH, ‘Hayward’ somaclone 7.5-1 tended to have higher PR, NN and ASL than other female somaclones. When shoot growth and appearance (tissue hyperhydricity, leaf yellowing or browning) were considered together through a multivariate data analysis, most ‘Tomuri’ and ‘Hayward’ somaclones which appeared less sensitive than controls to high CpH had been regenerated at pH 7 and 7.5. These regeneration and culture methods, especially the selection pressure exerted by the high pH during regeneration, thus seem to be effective in obtaining ‘Tomuri’ and ‘Hayward’ variants tolerant to high culture pH.     

Association of different groups of phytoplasma in flower malformation,phyllody, foliar yellowing,and little leaf disease of rose (Rosa sp.)

An extensive survey on rose cultivars in New Delhi showed phytoplasma-suspected symptoms such as leaf chlorosis, phyllody, virescence, and little leaf in 35 rose cultivars. Vegetative propagation through stem and tissue culture was performed. The cuttings/budded plants raised from infected plants showed the symptoms of phytoplasma disease. Phytoplasma was detected in 13 symptomatic plants using nested polymerase chain reaction (PCR) with phytoplasma-specific primer pairs P1/P7 and R16F2n/R16R2. Nested PCR amplicons from the 13 most severely infected rose cultivars (Arunima, Sindoor, Taboo, Sunsation, Raktgandha, Jawahar, Surkhab, Pusa arun, Marco Polo, Perfume de French Comete, Green Rose, Snowfire, and Gladiator) were cloned and sequenced. BLASTn analysis of 16S rRNA gene sequences, virtual RFLP and phylogenetic analysis revealed that the phytoplasma sequences amplified from rose belonged to ‘Candidatus Phytoplasma asteris’ subgroup 16SrI-B and ‘Ca. P. australasia’ subgroup 16Sr II-D. Our results suggest the inclusion of phytosanitary measures in planting materials for management of these phytoplasma diseases.