The effects of temperature on in vitro pollen germination and pollen tube growth of Pistacia spp.

The pollen germination and pollen tube growth among nine Pistacia genotypes was quantified in order to identify differences in the tolerance of pollen to temperature variations. The effect of temperature on in vitro pollen germination and pollen tube growth was investigated in Pistacia vera (Uygur, Atli, Kaska, Sengel, Kavak), P. atlantica, P. khinjuk, P. terebinthus and P. palaestina. When pollen was incubated in a germination medium for 24 h in darkness, distinct differences were observed in pollen germination and pollen tube growth at different temperatures.     

Pollen biology of ornamental ginger (Hedychium spp. J. Koenig)

An improved in vitro pollen germination assay was developed to assess the viability of stored Hedychium pollen. The effect of polyethylene glycol (PEG) (10, 15, and 20%, w/v) on pollen germination and tube growth was evaluated for Hedychium longicornutum and two commercial Hedychium cultivars, ‘Orange Brush’ and ‘Filigree’. Overall, the inclusion of PEG 4000 in the medium improved both pollen germination and tube growth for the three different genotypes tested and the results varied depending on genotype. In vitro germination was used to assess the viability of Hedychium pollen stored up to two months. Pollen nucleus status was determined for four Hedychium cultivars, ‘Orange Brush’, ‘Anne Bishop’, ‘Filigree’, and ‘Daniel Weeks’. Pollens of ‘Orange Brush’, ‘Anne Bishop’, and ‘Daniel Weeks’ were found to be binucleate but ‘Filigree’ was shown to possess both binucleate and trinucleate pollens. High pollen:ovule ratio values were obtained in several Hedychium taxa. The results obtained on the nuclear pollen status and pollen:ovule ratios will further our understanding of the pollination biology and help clarify the taxonomy and phylogeny of Hedychium species.     

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.     

Screening Capsicum species of different origins for high temperature tolerance by in vitro pollen germination and pollen tube length

Successful fruit set depends on several reproductive processes including pollen germination and tube growth processes. An experiment was conducted to determine the effects of temperature on pollen germination characteristics and to identify species/genotypic differences in Capsicum using the cumulative temperature response index (CTRI) concept. Pollen was collected from plants of seven genotypes from five Capsicum species, adapted to various parts of the world and grown outdoors in large pots. The pollen was subjected to in vitro temperatures ranging from 15 to 50 °C at 5 °C intervals. Pollen germination and tube lengths were recorded for all species after 24 h of incubation at the respective treatments. Species/genotypes differed significantly for in vitro pollen germination percentage and pollen tube length with mean values of 78% and 734 μm, respectively. The mean cardinal temperatures (T_min, T_opt, and T_max) averaged over genotypes, were 15.2, 30.7, and 41.8 °C for pollen germination and 12.2, 31.2, and 40.4 °C for pollen tube growth. The CTRI of each species/genotype calculated as the sum of eight relative individual stress response values, such as maximum pollen germination, maximum pollen tube length; T_min, T_opt, and T_max temperatures of pollen germination, and pollen tube lengths, identified species tolerance to high temperatures. Capsicum annum cv. Mex Serrano from Mexico was identified as tolerant, C. chacoense cv. 1312 and C. spp. cv. Cobanero from Argentina and Guatemala, respectively as intermediate and C. frutescens cv. Early Spring Giant from China, C. annum cv. Long Green from South Korea, C. spp. cv. NM89C130 and C. pubescens cv. 90002 from Guatemala as sensitive to high temperatures. The tolerant species/genotypes can be used in breeding programs to develop new genotypes that can withstand high temperature conditions both in the present climate and particularly in a future warmer climate.     

Influence of pistil extract and temperature on in vitro pollen germination and pollen tube growth of olive cultivars

In vitro tests for pollen germination and pollen tube growth of olive cultivars were carried out with the addition of pistil extracts. Low pistil extract concentrations (.0.3–0.6 mg ml^-1 f.w.) enhanced pollen germination and pollen tube growth for the same or different cultivars. Different responses to temperature were observed when pollen and pistil extract belonged to the same or to different cultivars. The results suggest that this in vitro test could be used to assess compatibility groups between olive cultivars.     

In vitro asymbiotic seed germination and seedling growth of Ansellia africana Lindl.

The effects of sodium hypochlorite treatment and growth medium on in vitro seed germination and seedling growth of the leopard orchid (Ansellia africana Lindl.) were investigated. Forty minutes treatment of seeds with 1.75% sodium hypochlorite stimulated germination (70.6%) and seedling development when measured at Stage 5 (emergence of first leaf) on P668 medium after 8 weeks of culture. The growth medium played a significant role in determining the germination response of A. africana seeds. Dark pretreatment of seed cultures significantly enhanced the germination percentage and the growth of rhizoids on the protocorms. Leaf growth in terms of length was substantially enhanced on P668 medium. It was significantly reduced in modified Knudson C medium after 16 weeks of culture. Seedlings developed on P668 medium showed a significantly better growth performance in relation to leaf length, leaf number, root length, fresh and dry weights per plant in vermiculite after 12 weeks of ex vitro growth in a mist house with 90% relative humidity. Further studies developing a suitable method for in vitro symbiotic germination could assist in reintroduction of this threatened orchid species into the wild.     

杏不同品种花粉量及花粉萌发特性的研究

以17个鲜食杏品种、4个仁用杏品种和4个杏李杂交品种为试材,对单个花药花粉量、花粉萌发率、花粉管长度进行测定,并观察统计了这25个杏品种的完全花比例,为杏主栽品种选择适宜授粉树。结果表明:不同杏品种间单个花药花粉量、花粉萌发率、花粉管长度和完全花比率差异显著;不同果实类型(鲜食、仁用和杏李杂交)间花粉量和完全花比例差异显著,花粉萌发率和花粉管长度则没有果实类型间的显著差异。花粉量与花粉萌发率和完全花比率之间无显著相关性,而与花粉管长度显著相关(r=0.417,P0.05);花粉萌发率与花粉管长度极显著正相关(r=0.691,P0.01),与完全花比率负相关但不显著。聚类分析发现,花粉量较多且花粉管生长状况较好的杏品种有"串枝红"、"沙金红"、"金太阳"和"胡安娜",适宜作为授粉树;花粉粒较少和花粉管生长状况较弱的有"恐龙蛋"、"味帝"和"苏联4号",而它们的完全花比例都很高,需要配置授粉树。     

精胺对梨花粉原位萌发及花粉管生长的影响

为进一步研究活体条件下精胺对梨花粉萌发及花粉管生长的影响,以幸水、新雪、长-二十世纪为材料,采用荧光显微镜观测了不同浓度精胺处理的花粉萌发及花粉管生长情况。试验结果表明:较低浓度的精胺能促进花粉萌发,而超过一定浓度时则表现抑制作用,适宜于花粉萌发的精胺浓度是0-0.025mmol/L。精胺对花粉管生长的促进作用因品种而异,0-0.05mmol/L,精胺促进幸水、新雪花粉管生长的作用主要表现在花柱中上部;而到达基部的花粉管仍较少,与对照间差异不显著。在0-0.025mmol/L的适宜浓度下,精胺可促进长-二十世纪花粉管生长并到达花柱基部。不同浓度的精胺对幸水、新雪坐果影响不大;而适宜浓度的精胺可促进长-二十世纪坐果。     

Improvement of caper (Capparis spinosa L.) propagation using in vitro culture and gamma irradiation

Some of the factors influencing the propagation of caper (Capparis spinosa L.) plants in vitro and germination of the seed were studied. The number of adventitious shoots emerging from caper stems cultured in vitro increased from 2.2 shoots per explant when the growth medium contained 2 mg/L of gibberellic acid (GA3) to 5.5 when the growth medium contained 2 mg/L zeatin riboside (ZR) and 1 mg/L naphthalene acetic acid (NAA). The best medium for callus formation from leaf and stem parts contained the growth regulators 1 mg/L 6-benzylaminopurine (BAP) and 0.1 mg/L NAA and the best medium for plant regeneration contained 1 mg/L kinetin and 0.1 mg/L indole-3-acetic acid (IAA). The effect of gamma irradiation on the growth of caper shoots in vitro was also studied. A 10 Gy dose of gamma irradiation stimulated growth of shoots up to 200% and increased shoot rooting percentage from 75 to 100%.     

Effects of cadmium and copper on pollen germination and fruit set in pea (Pisum sativum L.)

In this work, pea (Pisum sativum) plants exposed to increasing cadmium and copper concentrations were tested for heavy metals accumulation in flowers and for ‘in vivo’ pollen germination. Based on the Cd and Cu accumulation amounts in the flowers, an evaluation of the same metals effects on ‘in vitro’ pollen germination was achieved. Moreover, the effects of both metals on fruits number and weight and on seed set and yield at individual plant level were examined. While cadmium concentrations did not affect ‘in vivo’ pollen germination, only higher copper concentrations rendered a significant reduction. This is in contrast with the clear negative effect on pollen germination in vitro and might be explained by the different dynamic and bioavailability of both metals. A clear effect of Cd and Cu was observed on two important yield components ie, fruit weight and seed set. Although results obtained herein cannot give a clear cut relationship between the effect of Ca and Cu on reproductive development and its consequences on yields, they represent emerging results on the potential consequences of metals contamination on reproductive development in plants.     

Factors affecting the quality and in vitro germination capacity of strawberry pollen

Summary

Poor pollen quality and germination capacity curtails early yield in strawberry. The aim of this study was to establish a reliable method for in vitro assessment of strawberry pollen germination ability and to investigate further the effects of photoperiod and gibberellin on pollen germination and quality. In the first part of the study, pollen from seven strawberry cultivars (Chandler, Selva, Tudla, Camarosa, Eris, Pajaro and Irvine) was collected and its germination capacity and incidence of deformed pollen grains assessed in vitro using the hanging-drop technique. Highest germination rates, in ‘Selva’, were observed in a nutrient medium of 10% sucrose. Addition of calcium nitrate to the medium decreased the germination percentages of all cultivars. There was no significant difference, on average, between the germination rate at 20° and 25°C. Genetic factors affected the incidence of deformed pollen grains significantly, with ‘Pajaro’ showing the highest percentage (76%). In the second part, groups of young strawberry plants, cultivar Seascape, grown either under natural early spring conditions or under long-day or short-day conditions were sprayed once with GA₃ at 0, 50, or 200 mg l^–1. Pollen germination and deformation and stamen length were assessed three months later. In plants of the first group, GA₃ at 50 mg l^–1 increased pollen germination and decreased the incidence of deformed pollen grains, while GA₃ at 200 mg l^–1 decreased pollen germination without affecting the formation of deformed pollen grains. Plants of the second group showed a higher rate of pollen germination under long than under short days. GA₃ at 200 mg l^–1 decreased pollen germination under either short- or long-day conditions compared with the controls but doubled the percentage of deformed pollen only under short days. Stamens in control plants grew four times as long under long- than short-day conditions. GA₃ did not affect stamen length under long days but significantly enhanced their growth under short days.     

GA_3、2,4-D、B和蔗糖对梨花粉生活力及花粉生长速度的影响(英文)

以5个亚洲梨品种为试材,通过离体培养研究了花穗中花的着生部位,不同浓度的GA3、2,4-D和蔗糖对花粉生活力、花粉管生长速度的影响。结果表明,不同品种的花粉生活力差异明显,黄花花粉萌发快;新世纪花粉生活力强,花粉管生长迅速。花的着生位置及花器官的发育程度对花粉生活力有明显影响,边花的生活力比中心花强。不同浓度的B、2,4-D、GA3对花粉萌发及花粉管生长均有不同程度的影响,高浓度的GA3、2,4-D抑制花粉管的生长,但浓度较高的蔗糖能促进花粉粒的萌发和生长,且两者呈正相关(R2=0.983)。     

台湾青枣不同品种花粉萌发和生活力测定

以5个台湾青枣品种盛花期花粉为试材，应用正交试验测定不同品种花粉萌发率及适宜的萌发条件，并用4种不同方法测定花粉生活力。结果表明：（1）不同品种的花粉萌发率存在显著差异，其中大世界的花粉萌发率最高。（2）硼酸浓度、培养温度等因素对花粉萌发率有极显著影响。最适培养基为含琼脂1％、蔗糖15％、硼酸0．02％的组合，适宜温度为30—32℃。（3）花粉生活力的测定中，离体萌发法是最佳选择，I—KI染色法、醋酸洋红染色法和TTC染色法不适宜用作台湾青枣花粉生活力的测定。     

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.     

Rapid regeneration of plants of Dendrobium lituiflorum Lindl. (Orchidaceae) by using banana extract

Effects of banana extract (BE) and 6-benzylaminopurine (BAP) were evaluated on asymbiotic seed germination and an early differentiation of protocorms and plant regeneration of Dendrobium lituiflorum Lindl. High percentage germination was achieved by culturing seeds on modified Knudson C medium supplemented with 10% (v/v) BE. Rapid regeneration was observed within 60 days of culture on 10% (v/v) BE supplemented KC medium where maximum percentage propagules showed development of leaves and root formation. Propagules on BAP supplemented KC medium showed no further development beyond one leaf stage. In another experiment, culture of shoots on 12.5% (v/v) BE supplemented KC medium led to multiplication, shoot elongation as well as vigorous rooting. Shoots cultured on 10 μM BAP supplemented MS medium showed maximum multiplication but these were stunted. Plants with well expanded deep green leaves and elongated roots from BE media were first hardened in vitro followed by ex vitro hardening on cocopeat:perlite (9:1) in the greenhouse conditions and exhibited 90% survival. The study emphasizes the role of BE as a natural additive at different stages of development from seed germination to plant regeneration.     

Micrografting of almond (Prunus dulcis Mill.) cultivars “Ferragnes” and “Ferraduel”

The success of various in vitro micrografting techniques, establishment of the rootstock, size of the microscion, and the effects of culture medium on the grafted seedling development for almond cultivars “Ferragnes” and “Ferraduel” were studied. In vitro germinated wild almond seedlings developed from seeds were used as rootstocks. Shoot culture initiation was successfully achieved from the above almond cultivars by culturing mature shoot tips from forced nodal buds, about 3–5 mm, on 0.7 mg/L BA and 0.01 mg/L NAA containing a MS medium. The regenerated adventitious shoots from in vitro cultures were maintained and proliferated by sub-culturing on a fresh medium every three to 4 weeks. Regenerated shoot tips, which were micrografted onto in vitro seedlings, resulted in the restoration of shoot proliferation. The results indicated that the most successful method for the grafting of tested almond cultivars was slit micrografting. High levels of micrograft take were achieved with all ranges of scions (4–15 mm) obtained from the regenerated shoot tips. Slow growth and lack of axillary shoot development on the micrografts were noticeable when the micrografts were cultured on hormone-free germination medium. In vitro micrografted plantlets were successfully acclimatized and no problems were encountered with the establishment of micrografted plants in vivo. The developed technique has demonstrated a high potential for application in the micropropagation of almond cvs. “Ferragnes” and “Ferraduel” and thereby, represents a feasible method for the renewal of almond orchards in Turkey and elsewhere in the world.     

不同蔗糖浓度对辣椒花粉萌发的影响

分别用3%、5%、7%、10%、13%的蔗糖液体培养基,对红秀2002、红火二金条、冷江1号、宁椒5号和天胜朝天椒5个辣椒品种的花粉萌发率和花粉管生长情况进行了测定。结果表明,3%和5%的蔗糖浓度最适合花粉萌发和花粉管生长,冷江1号辣椒花粉萌发率最高,红秀2002花粉管生长最好。     

Effects of post-harvest foliar boron and calcium applications on subsequent season's pollen germination and pollen tube growth of pear (Pyrus pyrifolia)

The primary objective of this study was to evaluate the effects of foliar boron and calcium application after harvest on the quantity and activity of pollen in the ‘Housui’ and ‘Wonwhang’ pears on a subsequent year. Pollen grains of the ‘Housui’ pear were cultured on germination medium, to which had been added boric acid (0, 25, 75, 100, 200, 300, 400, and 500 mg L⁻¹) and calcium nitrate (0, 10, 25, 50, 100, 150, 200, 250, 300, 400, and 500 mg L⁻¹). Boric acid, which was added to the germination media, exerted a significant stimulatory effect on both pollen germination and pollen tube growth, although pollen tube growth was inhibited at higher concentrations than 300 mg L⁻¹. Calcium nitrate addition stimulated pollen germination, except at concentration of 500 mg L⁻¹. However, pollen tube growth was significantly inhibited with increasing concentrations of calcium nitrate. In the orchard experiment, boron and calcium were sprayed at concentrations of 0, 100, 200, 500 or 1000 mg L⁻¹ onto leaves after harvest, respectively. Boron and calcium content in the tissues as well as pollen production and growth were determined after these treatments. The foliar application of boron mainly resulted in an increase of boron concentration in buds. It also induced an increase in the weight of the anther and pollen in the following year. On the other hand, the foliar application of calcium resulted in an increase of calcium concentration mainly in the leaves, but pollen weight was decreased at high concentration treatment in the following year. The germination rate and tube growth of collected pollen were highest in the trees which had received boron treatment at a concentration of 200 mg L⁻¹. In contrast, the germination rate and tube growth of collected pollen were decreased by calcium application at concentrations of 500 and 1000 mg L⁻¹ without significant increase at lower concentrations. Consequently, the accumulation of boron in the developed buds of pear trees subjected to post-harvest foliar boron application generated positive effects on both the quantity and quality of pollen in the following year.     

不同发育阶段甜樱桃花授粉后花粉萌发和花粉管生长的差异

选用自交不亲和性程度不同的2个甜樱桃品种莫利和拉宾斯为试材,分别在开花前、开花当天和开花后进行莫利自花、拉宾斯自花和莫利×拉宾斯异花授粉,并于授粉后12、24、48、72、96h切取花柱,用FAA固定,荧光染色后压片观察。结果表明,3个组合的花粉萌发率都以开花当天授粉的最佳,拉宾斯自交和莫利×拉宾斯的花粉管生长状况也以开花当天授粉的最佳,但莫利自交的花粉管生长状况却以花前第4天最佳,花后第4天次之,而当天授粉的最差。因此认为,甜樱桃自交不亲和程度与其雌蕊的发育状态密切相关,蕾期授粉和延迟授粉都能够在一定程度上克服甜樱桃的自交不亲和性,但蕾期授粉的作用更加明显。     

Conservation of Vanilla species, in vitro

Vanilla (Vanilla planifolia) is a crop of great commercial importance as the source of natural vanillin, a major component of flavor industry. The primary gene pool of V. planifolia is narrow and is evidently threatened due to destruction of its natural habitats making the secondary gene pool important as a source of desirable traits especially for resistance to diseases. Many species of vanilla are considered rare and endangered hence an urgent need to conserve them, arises. Effective procedures for micropropagation and in vitro conservation by slow growth in selected species of vanilla, are described. Synthetic seed technology was standardized by encapsulating 3–5 mm in vitro regenerated shoot buds and protocorms in 4% sodium alginate, which could be stored up to 10 months with 80% germination in sterile water at 22 ± 2 °C. In vitro conservation technology of Vanilla was standardized and shoot cultures could be maintained for more than 1 year without subculture, on slow growth medium, i.e. Murashige and Skoog medium supplemented with 15 g l⁻¹ each of sucrose and mannitol in sealed culture vessels at 22 ± 2 °C. These cultures were maintained in vitro for more than 7 years with yearly subculture. The conserved material could be retrieved and multiplied normally in MS medium with 1.0 mg l⁻¹ BA and 0.5 mgl ⁻¹ IBA. The in vitro conserved plants showed good growth and developed into normal plants. This synseed and in vitro conservation system can be utilized for conservation and exchange of vanilla genetic resources.