Mechanism of seedlessness in Iranian seedless barberry (Berberis vulgaris L. var. asperma)

Species of barberry (Berberis vulgaris L. var. asperma) is cultivated in arid and semi arid areas of Iran (South Khorasan province). It is widely used as a food additive. Fruits of this species are seedless, while wild type barberries produce seeds in the same area. In this study, we investigated the mechanism of seedlessness in seedless barberry by pollen viability test, field pollination experiments and microscopic observation of pollen tube growth in pistil and ovule development. For comparison, we also examined ovule development in wild type barberry (B. crataegina DC). In seedless barberry pollen germination was about 54%. Seedless barberry produced 20% seeded fruits when pollinated with pollen of wild type barberry. There was a sharp decrease in fruit set in emasculated unpollinated flowers of seedless barberry. In seedless barberry, a large number of pollen grains (about 370) were observed on stigma of each flower at 12 h after balloon stage (ABS). Most of them germinated and penetrated intracellular area of stigma surface, but no pollen tube reached ovary. In seedless barberry, many ovules did not have any embryo sac or had a very small incomplete embryo sac. In addition, unfused polar nuclei were clearly recognized in some cases at 14 days after full bloom (AFB). However, in wild type, double fertilization was accompanied by disappearance of polar nuclei. In seeded barberry, the cellularized endosperm became apparent at seven days AFB. At 21 days AFB, all ovules of seedless barberry were degenerated, while at the same time in wild type, one or two ovules of each flower were normal and were developing into complete seeds. Results showed that self-incompatibility has a main role in seedlessness of seedless barberry. However, the high frequency of abnormal ovules and single fertilization can be considered as two other reasons of seedlessness. Due to our results, fruits of seedless barberry were set by stimulative parthenocarpy.     

A pollen extender medium technology for seedless watermelon production by pollination with soft X-ray irradiated pollen

The inactivation of pollen by soft X-ray irradiation and subsequent artificial pollination are time-intensive practices used in the production of seedless watermelons (Citrullus lanatus L.). Watermelon generally has a lot of staminate flowers; however, they only have a small amount of pollen. Watermelon pollen cannot be used in pollination under the present situation. Therefore, the objective of this study was to determine which is the most effective pollen extender medium for cultivation of watermelon with soft X-ray irradiated pollen. In this experiment, ‘Agar,’ ‘Marriage powder,’ and ‘Sekishoshi’ (Lycopodium dyed red with safflower pigment) were used as extender media with soft X-ray irradiated pollen at equal or twice the weight of the pollen. When ‘Sekishoshi’ was used as an extender medium, fruit set was very low. A lot of deformed fruit was produced when agar was used with the pollen. On the other hand, when ‘Marriage powder’ was used in equal proportions with the pollen, fruit set was about 70% and, moreover, Brix was high. Thus, the mixture of ‘Marriage powder’ with an equal amount of pollen was the best for use in actual cultivation. We conclude, then, that soft X-ray irradiated pollen in an extender medium can be effectively adapted for producing seedless watermelons.     

南丰蜜橘无核成因的研究

 观察和分析南丰蜜橘[Citrus reticulata Blanco var. kinokuni（Tanaka）H. H. Hu]的花粉育性、成熟胚囊结构以及胚胎发育情况,探讨其产生无核现象的原因。结果表明：南丰蜜橘的花粉具有一定的生活力和萌发率,属于中育性花粉;南丰蜜橘胚囊发育正常;授粉4周后大部分合子胚发育停滞并且出现败育迹象;饱满种子在授粉8周后胚发育到心形胚阶段时,部分种子的胚乳开始瓦解,到授粉16周后退化形成瘪籽和痕迹,另一部分发育成正常种子和充满胚乳的饱满种子;到成熟期时,小果系、早熟系南丰蜜橘种子全部退化,大果系南丰蜜橘种子大部退化,形成无核果实。胚胎早期败育是南丰蜜橘无核的主要原因。     

Ovules that failed to form seeds in zinnia (Zinnia violacea Cav.)

The structure of ovules that failed to form seeds after compatible pollination in double-flowered zinnias (Zinnia violacea Cav.) was investigated by observing histological sections, and comparing the normal and abnormal development of the seeds. In an embryo sac of zinnia at anthesis, a large nucleus of fused polar nuclei was clearly recognized. Success in double fertilization was determined by the disappearance of this nucleus. In normally developing seeds, the cellular endosperm became vague at 5–6 days after pollination (DAP) and the developing embryos occupied the entire portion of the seeds up to 10 DAP. A number of ovules failed in fertilization for the lack of an embryo sac. In most of the aborted seeds, the embryo sacs degenerated, whereas the aborted embryos were still alive in a small number of aborted seeds. A small percentage of the aborted seeds exhibited an aborted embryo and an unfertilized fused nucleus of two polar nuclei or ‘single fertilization’. The three major problems suggested for the failure in seed formation in pollinated florets of zinnia included failure in fertilization, ovules lacking embryo sacs, and abortion of developing seeds.     

GA_3诱导枇杷无核的组织解剖学观察

为了阐明GA3处理枇杷导致无核的原因,以枇杷品种大五星为材料,采用石蜡切片法和花粉发芽试验等方法分别对花前经GA3处理枇杷花穗后的枇杷胚胎发育过程进行了研究。结果表明,1)枇杷的胚珠为倒生胚珠,双珠被,核型胚乳;2)花前用GA3处理枇杷导致无核的原因主要在于胚乳败育或者珠心退化导致胚退化,由于胚乳败育使胚发育所需要的营养不足,引起合子胚分裂异常,最终胚解体退化。但珠被在初期能够发育,所以形成败育种子(秕子)。     

Water spray as a potential thinning agent for date palm flowers (Phoenix dactylifera L.) c.v. ‘Lulu’

Flower or fruit thinning is a critical cultural practice in the date palm production chain that affects fruit development, quality and yield and regulates tree yearly bearing. Development of a save and economic thinning agent for date palm is critically required especially under harsh conditions. During 2004 and 2005 seasons, water spray for 3 min was applied at different times following mechanical pollination on ‘Lulu’ date palm cultivar growing under Al-Ain oasis conditions. The results showed that water spray following pollination generally decreased fruit set percentage to different extent depending of the time of application. In this respect, the most effective treatment was water spray after 4 h following pollination. This treatment significantly decreased fruit set percentage (48%) compared to the control (79%), as the mean of both seasons. However, the other treatments including water spray 1 h before pollination also decreased fruit set percentage but to a lesser extent. Fruit quality characteristics especially fruit and flesh weight, length and diameter were significantly increased by water spray treatments during the 2004 season, in contrast to the 2005 season. Thinning with water significantly decreased both bunch weight and total yield per tree at the Tamr stage especially when applied after 4 h following pollination. To the best of our knowledge, this is a pioneer study investigating the role of water as a potential thinning agent for date palm flowers. The use of water spray as a save and economic thinner needs, however, more research work to justify the time following pollination and duration of application which may vary upon cultivar, method of pollination and district.     

Contributing to the knowledge of the fertilisation process in four apricot cultivars

Causes of poor crops in apricot were investigated by studying the reproductive process after pollination. The evolution of the ovules and the growth of the pollen tubes were determined. Also the fertilisation of the ovules and the fruit set obtained, when pollination was delayed, were examined in two groups of cultivars with different fruiting behaviour. ‘Beliana’ and ‘Palstein’, two cultivars that consistently set good yields, presented ovules that mature earlier and in larger number than those of ‘Guillermo’ and ‘Bergeron’, characterised for year-to-year low fruit production. Pollen tubes also grew faster in the first group of cultivars. Differences found between both groups of cultivars, seem to be related to the ability of the ovules to develop and mature after anthesis, and around 50% of them only had a mother cell without an embryo sac, suggesting that development of those ovules was stopped at an early stage. This fact critically influenced fertilisation and, therefore, fruit set. There seem to be a good correlation between percentages of mature ovules a few days after anthesis, the percentages of fertilised ovules and final percentages of fruit set.     

辣椒受精过程及其经历时间的研究

 运用常规石蜡制片技术与光学显微镜技术研究了辣椒（Capsicum annuum L.）受精过程及其经历的时间。结果表明：辣椒为自花授粉植物;成熟雄配子体为二细胞型花粉,生殖细胞在花粉管内分裂产生二个精子;花粉管破坏一个助细胞并释放精子,二精子同时移向卵细胞与次生核,精卵融合速度较慢,精子和次生核融合较快;辣椒为细胞型胚乳。自花授粉后2–4 h在柱头上大量花粉萌发;授粉后8–12 h左右花粉管长过花柱进入子房腔;授粉后12–20 h左右花粉管在助细胞里释放精子;授粉后14–30 h左右雌雄性细胞融合;授粉后18–40 h初生胚乳核分裂,初生胚乳核休眠期约为4–6 h;授粉后4 d左右合子分裂,合子休眠期为3 d左右。     

Spray pollination as a labor-saving pollination system in Japanese pear (Pyrus pyrifolia (Burm.f.) Nakai): Development of the suspension medium

Effects of suspension media used for spray pollination on pollen grain viability were investigated in Japanese pear (Pyrus pyrifolia (Burm.f.) Nakai ‘Kosui’). The suspension media tested in this study consisted of pectin methylesterase (PME) and polygalacturonase (PG) combined with either 0.1% agar or 0.04% xanthangum (XG). We also evaluated the influences of spray pollination on fruit set and fruit quality. Pollen grain viability, as measured by germination and pollen tube growth rates, was maintained in media supplemented with 0.1 mg L⁻¹ PME or 0.1 mg L⁻¹ PG. The level of fruit set after spray pollination using media containing PME or PG in combination with either 0.1% agar or 0.04% XG was almost the same as the level after artificial pollination by hand (hand pollination). The media containing XG combined with either PME or PG seemed to show better results for pollen grain viability and fruit set, although the results were variable from year to year. With regard to fruit size, shape and other parameters for fruit quality, spray pollination and hand pollination gave comparable results, irrespective of the medium composition. The time required for spray pollination was less than half of that required for hand pollination, and the amount of pollen grain required for spray pollination was one-third or less than the amount required for hand pollination. Thus, spray pollination may be a time- and labor-saving pollination system for the cultivation of Japanese pear.     

Pollen germination as affected by pollen age in cherimoya

Cherimoya (Annona cherimola (Mill.)) is a subtropical fruit tree, which is cultivated in a good range of subtropical regions. In most of these areas the crop relies on hand pollination. However, following this practice, erratic fruit set is often produced, which could be related to problems in pollen handling. Indeed, very little is known of the time that the pollen remains viable and on which is the best stage to collect the anthers or pollen from the flower. The aim of this work is to evaluate pollen germinability prior and after anther dehiscence and also how the age of pollen affects pollen vigor, understood as speed of germination. Pollen samples at different times following anther dehiscence were germinated in vitro and in vivo. Pollen up to 90 min following dehiscence performed as well as freshly dehisced pollen. However, the pollen taken 120 min following dehiscence, showed a clear reduction in vigor and germinated much slower in vivo. To overcome this short pollen germinability, pollen was taken from anthers 30 and 5 h prior to natural anther dehiscence and compared with pollen taken at anther dehiscence and 20 h later. However, a reduction in germination rate was obtained in pollen taken prior to anther dehiscence. The narrow stage at which pollen can be collected together to its ephemeral germinability explains erratic results obtained following hand pollination in this crop and these results provide the clues for an adequate pollen handling.     

Effect of distance from early flowering pollinizers ‘Maypole’ and ‘Dolgo’ on ‘Fiji’ fruit set

An apple orchard consisting of a single cultivar under the condition of natural pollination must have suitable pollinizers such as Crab apples to ensure stable fruit production. We selected ‘Maypole’ and ‘Dolgo’ as pollinizers for the cultivar ‘Fuji’, and investigated the rate of fruit and seeds in ‘Fuji’ fruits produced by pollen of the pollinizers. We developed a method for tracing pollen flow based on the leaf color of progeny and S-RNase allele of ‘Maypole’, and on Simple Sequence Repeat (SSR) analyses of ‘Maypole’ and ‘Dolgo’. These were powerful tools for determining the distance insects (mainly Osmia cornifrons) carry pollen from the pollinizers to ‘Fuji’. Although the fruit set of ‘Fuji’ apples was not reduced with increasing distance between ‘Fuji’ apple trees and pollinizers (probably due to pollen flow from other commercial cultivars planted outside the area), the rate of ‘Fuji’ apple fruit produced by the pollen of the pollinizers decreased with increasing distance. The rate of fruit produced by the pollinizers was 84% and 77% when ‘Fuji’ was 2.5 m from ‘Maypole’ and ‘Dolgo’, respectively, and 71% and 64% when ‘Fuji’ was 5 m from ‘Maypole’ and ‘Dolgo’, respectively, but was reduced to 47% and 39% when ‘Fuji’ was 10 m from ‘Maypole’ and ‘Dolgo’, respectively. However, the spacing could cause reduced fruit size, and require extra fruit thinning for producing large fruits. It could also cause reduction of the yield for increasing the planting area of pollinizers. We recommend that pollinizers should be planted not more than 10 m from ‘Fuji’.     

Changes in endogenous gibberellins during fruit growth in guava,with a general discussion

Levels of endogenous gibberellins were determined in seeded and seedless fruits of guava (Psidium guajava L.) in relation to fruit development. The gibberellin content in the seeded fruit was at all times higher than that in the seedless fruit. The peak concentrations of gibberellins in both kinds of fruits occurred at 15 days after pollination and thereafter declined rapidly. This suggests that these substances are involved in the early phase of fruit growth.     

Effect of pollination intensity,frequency and pollen source on fig (Ficus carica L.) productivity and fruit quality

Yield and fruit quality in fig (Ficus carica L.) are highly dependent on cultural practices especially caprification (pollination). However, this technique remains not well controlled in Tunisia. This study was conducted during two successive years, 2009 and 2010, to investigate different pollination conditions, i.e. number of caprifigs and repetitions of caprification on fruiting of two cultivars Bouhouli (San Pedro type) and Zidi (Smyrna type). In addition, the efficiency of four pollen sources was evaluated to identify the most effective pollinator for female cultivar Zidi. The following parameters were recorded: fruit number per shoot, fruit set, productivity, fruit characteristics and vegetative growth.     

Rain effect on pollen–stigma adhesion and fertilization in almond

The fact that rainy weather negatively affects production in almond is well-known by growers. However, as far as we know, no previous study has been carried out in this species to determine if the rain is able to wash off pollen grains from the stigma surface and consequently affect fertilization efficiency. To answer this question, flowers of the self-compatible almond cultivar ‘Antoñeta’ were emasculated on branches of trees in the field, pollinated by hand with pollen of the cultivar ‘Marcona’, and sprayed with water to simulate the rain effect at 4, 8, 24 and 48 h after pollination. A control for which no treatment was applied was also included in the assay. Seven days after spraying the flowers, the number of germinated pollen grains on the stigma was recorded in 10 pistils for each treatment by fluorescent microscopic observation. In addition, for each treatment the initial and final fruit set were determined at 30 and 60 days after pollination, respectively. The following year additional assays were performed in the field and in the laboratory in order to complete the study with earlier treatment times. Thus, an assay of rain simulation in the field at 0 and 2 h, and other consisting on immersing a branch with flowers and shaking it in a water container in the laboratory at 0, 30, 60 and 120 min after pollination were performed. One sample more was immersed, shaken, then pollinated and again immersed and shaken. The results from field assays showed that adhesion of pollen grains to the stigma was very quick and strong, and the treatments did not significantly affect pollen–stigma adhesion and subsequent fruit set. On the other hand, laboratory results showed that all washing treatments decreased the number of germinated pollen grains on the stigma, mainly when the flowers were immersed before pollination. In conclusion, the results showed that the simulated rain was not able to completely wash off pollen grains from the stigma surface, although it seems to affect adhesion in forthcoming pollinations.     

English walnut fruit growth and development

Samples of the ‘Ashley’, ‘Hartley’ and ‘Franquette’ cultivars of English walnut (Juglans regia L.) were collected at weekly intervals from bloom to harvest, and development of the fruit, embryo and surrounding tissue was followed. The first division of the zygote occurred approximately one week after pollination, by which time the endosperm, nucellus and integument had shown considerable growth. The endosperm first formed cell walls at the 8-celled embryo stage and was completely cellular by the time the embryo contained approximately 64 cells. Rapid growth of the cotyledons started as the shell began to harden at Week 7. Measurement of fruit growth indicated that fresh weight gain in ‘Ashley’ and ‘Hartley’ fruits followed a double sigmoid pattern, characterized by a 3-week period of slowed growth that began approximately 7 weeks after bloom. Rate of increase in total length and diameter also slowed greatly at Week 7. Fresh weight gain in ‘Franquette’ appeared to follow a single sigmoid curve, with no prolonged period of slowed growth corresponding to that in the other cultivars. Kernel growth also described a double sigmoid curve, with its initial, rapid phase nearly coincident with the beginning of Stage III of the whole-fruit, fresh-weight curve at Week 10.     

大白菜受精过程及其各阶段持续时间的研究

 对3个大白菜(Brassica campestris ssp. pekinensis) 品种的受精过程及其各阶段持续的时间进行了研究。其结果如下: 1. 授粉后2～3 h, 花粉在柱头上萌发。2. 授粉后4～8 h, 花粉管在花柱中生长。3.授粉后8～14 h, 花粉管在子房中生长并通过珠孔进入胚珠。4. 授粉后16 h, 花粉管进入胚囊, 破坏1个助细胞, 并将内含物释放入该助细胞内。5. 授粉后16～18 h, 精核向卵细胞移动并进入卵细胞。授粉后18 h,精核附在卵核的核膜上。6. 授粉后20 h, 精核进入卵细胞核, 精核染色质逐渐松解; 授粉后24 h, 出现雄性核仁。受精卵核内有一明显大的雌性核仁和1个较小的雄性核仁, 此时形成合子。精核染色质在卵核内分散延续的时间长达4 h左右。7. 授粉后32～34 h, 合子进行第1次分裂。大白菜合子静止期8～10 h左右。8.受精前二极核合并成次生核或不合并, 位于卵细胞的合点端。9. 授粉后16～18 h精核向极核或次生核移动。授粉后18 h, 精核附在极核或次生核的核膜上。10. 授粉后20 h, 精核进入1个极核或次生核, 二者进行融合。精核与极核或次生核的融合过程同精核与卵核的融合过程相同, 只是精核与极核或次生核融合速度较快。精核染色质在极核或次生核里分散的时间大约为2 h。11. 授粉后22 h, 形成初生胚乳核, 在分裂前常常看到初生胚乳核内的雌雄性核仁仍未合并。12. 授粉后24 h, 初生胚乳核进行第1次有丝分裂。     

Quality fruit improvement in sweet pepper culture by bumblebee pollination

The percentage of ovules that set seeds per fruit has been used to evaluate the bumblebee activity of a two campaigns greenhouse trial. The experiment was carried out with two commercial cultivars of sweet pepper, ‘bell’ type, called Bardenas and Vergasa. The averages of the percentages of seed set per fruit of the two campaigns were 49.8% (Bardenas) and 40.7% (Vergasa) on the pollination treatment, whereas the averages of the control (self-pollination) treatment were 27.5% and 25.7%, respectively. Percentages of seed set per fruit was strongly related with the number of bee visits, but weakly with duration of visits.     

Breeding of disease-resistant seedless grapes using Chinese wild Vitis spp.: I. In vitro embryo rescue and plant development

Seedless grape cultivars (Vitis vinifera L.) are widely grown in Europe, America and Asia. Fungal diseases are a great threat of them. Several wild Chinese Vitis species showed high resistance to many fungal diseases. Therefore, an investigation was conducted to assess the potential to incorporate these species in a breeding project for development of the disease-resistant seedless cultivars. Hybridization was conducted using V. vinifera as female parents and the wild Chinese Vitis spp. as male parents. In-ovulo embryo rescue was used to develop hybrid plants from the seedless females. An efficient protocol is reported here for the in vitro embryo rescue and plant development from the cross Emerald Seedless × Beichun. Ovules were excised from immature fruits 7 weeks after pollination (WAP) and cultured in the double-phase ER medium supplemented with 6.0% sucrose and 0.3% activated charcoal (AC). Following 8–12 weeks of culture, embryos were removed from the ovules and transferred onto WPM supplemented with 1.0 μM 6-benzyladenine (6-BA), 2.0% sucrose and 0.2% AC and solidified with 0.6% agar. After 8 weeks of culture, the embryos germinated and subsequently grew into whole plantlets. With the optimized parameters developed in the present study, about 34.0%, 91.2% and 77.4% of embryo formation, embryo germination and plant development were obtained, respectively. When this protocol was applied to 11 other cross combinations, genotype was found to significantly influence embryo formation, embryo germination and plant development, with different frequencies of hybrid plants successfully obtained in all crosses.     

用于无核葡萄选育的胚挽救技术研究

通过4a的连续试验,观察了无核葡萄果粒及胚珠发育变化,对胚挽救的多个影响因素包括品种、接种时间、培养基和培养方式进行了研究。结果表明,红脸无核、底来特、黎明无核作母本的胚珠萌发率较高;各品种适宜的接种时间为无核白花后26d,红脸无核花后35d,爱莫无核×火焰无核花后48d,底来特×红宝石无核花后56d;用Nitsch作基本培养基优于B5;添加BA0.2mg/L获得的胚珠萌发率最高;常温黑暗、固体培养有利于胚珠的发育。已定植结果的无核葡萄杂交胚挽救后代的无核比率平均为58.6%。将胚挽救技术用于无核葡萄育种是十分有效的。