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.     

Comparison of S-RNase,RNase T1, T2, and A effects on growth inhibition and RNA degradation of in vitro-cultured pear pollen

To obtain the basic information on fruit set regulation, effects of several RNases including S-RNase on pollen tube growth and RNA degradation in the tube were studied in the pear. Purified S-RNase from the Japanese pear ‘Kosui’ (S₄S₅) predominantly inhibited the growth of ‘Kosui’ pollen tubes (self) in vitro at 0.28 unit μL⁻¹, but it inhibited ‘Chojuro’ (S₂S₃) pollen (cross) only slightly. The same unit of RNase T₁ (EC 3.1.27.3) clearly inhibited the pollen tube growth, but the action was significantly weaker than that of the S-RNase against the self-pollen. Inhibitory effect of RNase T₂ (EC 3.1.27.1) and RNase A (EC 3.1.27.5) was only slight. The proteins other than the S-RNase extracted from pear style did not have any inhibitory action, though they possessed RNase activity 3.8 times higher than S-RNase. Thus, RNases tested here could not substitute for the S-RNase in specific inhibition against the self-pollen tube growth. Total RNA degradation by each RNase occurred in the pollen tubes as following order; S-RNase (self) ≥T₁ > T₂ ≥ A > S-RNase (cross). Degradation degree of 28S and 18S rRNA was as follows; S-RNase (self) > A > T₁ > T₂ > S-RNase (cross). The degradation of 5.8S and 5S rRNA was; S-RNase (self) > S-RNase (cross) > A > T₂ > T_1. The degree of rRNA degradation was, thus, not always in parallel with the degree of pollen growth inhibition. The S-RNase may degrade not only rRNA but also mRNA essential for pollen tube growth, and may be specifically adapted to inhibit the growth of self-pollen tubes. Therefore, controlling S-RNase amount in the style will produce self-thinning cultivars efficiently, which are unnecessary not only for hand-pollination but fruit-thinning practices in the pear. Practically, cultivar with weak self-incompatibility and small amount of S-RNase, such as ‘Okusankichi’, may be an expecting candidate for breeding self-thinning cultivars.     

梨不同花龄自花与异花授粉的花柱内花粉管生长及坐果率的比较

对自交不亲和强度不同的梨品种丰水和菊水的不同花龄花进行自花授粉和丰水×菊水异花授粉后,用荧光显微镜观察花粉萌发及生长特性。结果表明,开花当天及以后的自花和异花授粉花粉在柱头上均有较高的萌发率;自花和异花花粉管在花柱内的生长速度及停止生长的位置不同,丰水自花授粉后花粉管生长的短且慢,菊水自花花粉管生长的长且快,而异花授粉的花粉管生长最快。自花授粉后,不同花龄对花粉管生长影响表现为:开花4d前的蕾期授粉以及开花4d后的延迟授粉,花粉管生长长度显著的长。开花6d前的各种授粉组合,花粉管生长均较差。这与田间授粉坐果率的高低基本一致,即整体表现为各个花龄的授粉都以异花授粉坐果率最高,其次为菊水自花授粉,丰水自花授粉坐果率最低。另外,自花授粉中,蕾期授粉和延迟授粉(开花2d后除外)坐果率都有提高,而异花授粉中蕾期授粉和延迟授粉的坐果率却有所下降。     

离体梨花柱S2RNa se对自体花粉管超微结构变化的影响

 采用透射电子显微镜研究了离体条件下‘今村秋’、‘丰水’梨花柱S2RNase对‘今村秋’花粉管生长过程中花粉管超微结构的影响。‘丰水’花柱S2RNase处理的‘今村秋’花粉管(异体, 亲和)和对照花粉管结构在生长过程中表现正常, 即花粉管顶端生长区域充满细胞质和细胞器, 结构完整, 花粉管壁上没有胼胝质层分布。而‘今村秋’花柱S-RNase处理的‘今村秋’花粉管(自体, 不亲和) 超微结构随时间的延长出现衰退现象。处理3 h时, 自体花粉管内充满细胞质和细胞器, 结构完整, 但在花粉管的极顶端分泌小泡融合, 并且此区域的细胞壁比后部的薄; 处理8 h后, 细胞器出现一定程度的衰退现象,线粒体膨大, 嵴减少或消失, 内质网膨大并包围在液泡和其它细胞器周围, 液泡融合并侵蚀周围的细胞质和细胞器; 到24 h时, 自体花粉管内线粒体、内质网等细胞器消失, 只有靠近花粉管前端有少量细胞质存在, 细胞壁增厚, 并且有一层厚厚的胼胝质层紧靠细胞壁。结果表明, 花柱S2RNase能引起自体花粉管的衰退, 从而抑制自体花粉管生长。     

离体梨花柱S-RNase对自体花粉管超微结构变化的影响

采用透射电子显微镜研究了离体条件下‘今村秋’、‘丰水’梨花柱S-RNase对‘今村秋’花粉管生长过程中花粉管超微结构的影响。‘丰水’花柱S-RNase处理的‘今村秋’花粉管(异体,亲和)和对照花粉管结构在生长过程中表现正常,即花粉管顶端生长区域充满细胞质和细胞器,结构完整,花粉管壁上没有胼胝质层分布。而‘今村秋’花柱S-RNase处理的‘今村秋’花粉管(自体,不亲和)超微结构随时间的延长出现衰退现象。处理3h时,自体花粉管内充满细胞质和细胞器,结构完整,但在花粉管的极顶端分泌小泡融合,并且此区域的细胞壁比后部的薄;处理8h后,细胞器出现一定程度的衰退现象,线粒体膨大,嵴减少或消失,内质网膨大并包围在液泡和其它细胞器周围,液泡融合并侵蚀周围的细胞质和细胞器;到24h时,自体花粉管内线粒体、内质网等细胞器消失,只有靠近花粉管前端有少量细胞质存在,细胞壁增厚,并且有一层厚厚的胼胝质层紧靠细胞壁。结果表明,花柱S-RNase能引起自体花粉管的衰退,从而抑制自体花粉管生长。     

梨43个品种花粉生活力及4种测定方法的比较

用花粉离体培养法测定比较了43个梨品种花粉离体萌发率和花粉管生长长度;从中选择花粉相对生活力低、中、高3组6个品种(今村秋和雪花梨、金二十世纪和丰水、青松和满天红)的花粉,采用I-KI染色法、蓝墨水染色法、MTT染色法、过氧化物酶染色法4种花粉生活力测定方法,以花粉离体萌发法为对照。结果表明,花粉培养24h后,43个梨品种的花粉萌发率平均为66.15%,其中花粉萌发率≥60%的品种约占84%,满天红萌发率最高为82.79%;花粉管长度平均为1.77mm,花粉管长度≥1mm的品种约占89%,其中奥萨二十世纪最长为3.25mm,说明多数梨品种花粉生活力较强。与对照相比,4种花粉生活力测定方法中I-KI染色法的测定值极显著偏低,MTT(噻唑蓝)染色法的测定值显著偏高,二者不适于梨花粉生活力的测定;过氧化物酶染色法测定值与对照差异显著,也不适宜用来测定梨花粉生活力;蓝墨水染色法测定值与对照没有显著性差异,可用于梨花粉生活力的快速测定。     

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.     

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.     

Biocontrol of postharvest blue mould rot of pear by Cryptococcus laurentii

Summary

Cryptococcus laurentii was evaluated for its activity in reducing postharvest blue mould decay of pear caused by Penicillium expansum. Washed cell suspensions of yeast controlled blue mould better than yeast in culture broth. Treatment of wounds with autoclaved cultures or cell-free culture filtrate did not prevent decay. There was a negative correlation between concentrations of the yeast cells and infectivity of the pathogens: at concentrations of yeast at or above 1 × 10⁹ colony forming units (CFU) ml^–1 and pathogen spores suspension of 5 × 10⁴ spores ml^–1, blue mould was completely inhibited after 7 d of incubation at 25°C, and was almost inhibited after 30 d incubation at 2°C followed by 7 d at 20°C. The interval between inoculation with the pathogen and application of the antagonist significantly influenced the efficacy of biocontrol. Efficacy was maintained when C. laurentii was applied simultaneously or prior to inoculation with P. expansum but reduced if applied after inoculation with P. expansum. Rapid colonization of the yeast in wounds was observed during the first 3 d at 20°C, and then the populations stabilized for the remaining storage period. On pear wounds kept at 4°C, the increase in population density of C. laurentii was lower than at 20°C, but rose over 15 d after application of the antagonist to approx. 10⁷ CFU wound^–1.     

Li~+、肌醇与Ca~(2+)对苹果、梨、山楂花粉萌发和花粉管生长的影响

观测了苹果（金冠）、梨（鸭梨）、山植（面值）的花粉萌发和花粉管生长情况,结果表明,Li2SO4的浓度越高、处理时间越早,花粉萌发和花粉管生长的抑制程度越大,Ca2+和肌醇可不同程度地逆转上述效应。因此认为,细胞内一定量的肌醇和Ca2+是果树花粉萌发和花粉管生长所必需的,并对维持正常的肌醇磷脂代谢是至关重要的。     

Fructose inhibits pear pollen germination on agar medium without loss of viability

Pollen of Japanese pear (Pyrus pyrifolia Nakai) germinated well on agar medium containing 10% sucrose or glucose, but not on agar containing fructose. The inhibitory effect of fructose was dose-dependent. Sucrose enhanced pollen tube growth much more effectively than glucose. Addition of 5% fructose to 5% or 10% sucrose or glucose media suppressed germination completely. Ungerminated pollen, however, showed similar respiration rate and stainability against acetocarmine dye as germinated pollen. When pollen was transferred onto fructose medium after culturing it on glucose or sucrose medium for 1–2 h, germination was completely impeded. Reversely, pollen transferred to sucrose or glucose medium from fructose medium germinated at almost the same ratio as pollen on sucrose or glucose medium without transfer. Thus, pollen inhibition by fructose is reversible. Compared with uncultured pollen, cultured pollen contained less than half amount of total sugars, even if failed in germination on fructose medium. Germinated pollen on sucrose and glucose media contained sucrose and glucose, but ungerminated pollen on fructose medium contained only trace levels of these sugars, suggests that pollen on fructose medium predominantly uses sucrose and glucose as respiration substrates and cannot maintain the constant levels of these sugars. However, as pollen germination occurred on agar medium without any sugar, fructose may impede a physiological factor that triggers germination, and once the trigger is impeded, many physiological pathways including sugar biosynthesis may be blocked.     

Germination and growth of sponge gourd (Luffa cylindrica) pollen tubes and FTIR analysis of the pollen tube wall

Emergence of multiple pollen tubes from single pollen grains occurred both in vitro and in vivo in sponge gourd (Luffa cylindrica (L.) Roem). The frequency with which pollen grains produced multiple pollen tubes in vivo (7.2%) was lower than that under in vitro conditions (14.9%). In pollen grains germinated in vitro, the total length of the multiple pollen tubes was greater than that of single pollen tubes, but individual tubes among the multiple tubes did not reach the same length as single tubes. Moreover, the growth of the single pollen tubes continued for a longer period in vitro than that of the multiple tubes. Fluorescence microscopy showed that callose was present throughout the pollen tube wall except in the apical part of growing pollen tubes, and nuclei moved into the longest of the multiple tubes. Results of Fourier transform infrared microspectroscopy indicated that abnormal cell wall components (peaks at 800–1000 cm⁻¹) were more frequent in multiple pollen tubes lacking nuclei, and the pectin content (1733 cm⁻¹) in multiple pollen tubes was much lower than that in single pollen tubes. These findings suggested that there were significant differences in pollen tube growth and wall composition between single and multiple pollen tubes, and that multiple pollen tubes had much less opportunity than single pollen tubes to reach the embryo sac and achieve double fertilization.     

Effect of pistil age and pollen parent on pollen tube growth and fruit production of pistachio

Female pistachio (Pistacia vera) trees were hand pollinated with pollen from three different male genotypes 0,1,2,3 and 4 d after anthesis. Results were measured by mature fruit set and by fluorescence microscopy of pollen germination and tube growth in the pistil. Maximum pollen tube growth and fruit set of split nuts was achieved following pollination within two days of anthesis. Three day old pistils supported pollen tube growth, but fruit set was low. Four day old pistils supported little pollen tube growth or fruit set. Significant differences were also apparent between pollen parents, with low pollen tube growth and fruit set following interspecific pollination with P. atlantica pollen. All pollen tubes were observed to penetrate chalazogamously, and parthenocarpic production of blank fruits occurred in both unpollinated and pollinated treatments. The results indicate that for optimum fruit production of pistachio, pollen transfer must be achieved within 2 d of anthesis and pollination by P. atlantica should be avoided.     

Fenoxycarb,a selective insecticide for inclusion in integrated pest management systems for pear in the UK

The effects of fenoxycarb, an insect growth regulator, on predatory anthocorids, were tested in comparison with the broad-spectrum insecticide deltamethrin, in pear orchard trials. The results confirmed the effectiveness of fenoxycarb against pear sucker, Cacop- sylla pyricola and revealed no damaging effects on larval or adult anthocorids (mainly Anthocoris nemoralis). Deltamethrin reduced numbers of both C. pyricola and anthocorids, allowing a resurgence of C. pyricola later in the season. Fenoxycarb is shown to be suitable for use in integrated pest management programmes in pear.     

Analyses of pollen-tube growth and biological action of S-RNase in the style of self-compatible Japanese pear

The Japanese pear ‘Osa-Nijisseiki’ (S₂S₄^SM) (SM = stylar-part mutant) is a self-compatible bud mutant that originated from self-incompatible ‘Nijisseiki’ (S₂S₄). The S₄-allele of the pear is deleted and it shows unilateral incompatibility to cultivar with an S₂S₄ genotype. However, when pollen-tube growth was compared between cross-compatible [‘Osa-Nijisseiki’ × ‘Okusankichi’ (S₅S₇)], unilateral-compatible [‘Osa-Nijisseiki’ × ‘Kikusui’ (S₂S₄)], self-compatible (‘Osa-Nijisseiki’ × ‘Osa-Nijisseiki’), and unilateral-incompatible pollination (‘Kikusui’ × ‘Osa-Nijisseiki’), pollen-tube growth clearly showed the following order: cross-compatible > unilateral-compatible > self-compatible > unilateral-incompatible. This indicates that the ‘Osa-Nijisseiki’ style produces specific inhibitor(s) not only to S₂- and S₄-pollen but also “self-pollen”, because the phenotype of S₄^SM-pollen is the same as S₄-pollen. Stylar protein analysis demonstrated that ‘Osa-Nijisseiki’ produces S₂-RNase (RNase associated with S₂-allele) together with a small amount of S₄-RNase. The purified S₄-RNase possessed almost the same inhibitory action on the growth of S₄-pollen-tubes in vitro at 1 μg μl⁻¹ as that from original ‘Nijisseiki’. These results suggest that the depressed growth of unilateral-compatible and self-pollen-tubes in ‘Osa-Nijisseiki’ is due to this biologically active S₄-RNase. Growth of self-pollen-tubes may also be depressed by inhibitor(s) specific to “self-pollen” unrelated to S-alleles.     

Growth and compositional changes during the development of prickly pear fruit

Growth and compositional changes in prickly pear (Opuntia spp.) fruit during the development of four species (O. ficus-indica, O. inermis, O. hyptiacantha and O. lind- heimeri) were studied. The weight and size of fruits belonging to O. ficus-indica, O. hyptiacantha and O. lindheimeri generally followed a single sigmoid growth pattern while fruits belonging to O. inermis followed a double sigmoid growth pattern. In all species, soluble solids and ascorbic acid contents increased with a concomitant decrease in titrat- able acidity as fruits matured. O. hyptiacantha consistently had the highest soluble solids and ascorbic acid contents and the lowest titratable acidity while O. lindeimeri consistently had the lowest soluble solids and ascorbic acid contents and highest titratable acidity throughout ontogeny. Although significant differences in soluble solids and ascorbic acid contents were observed, all four prickly pear species exhibited similar trends in compositional changes with growth. Attributes that contributed to fruit flavour such as sugar to acid ratio positively correlated with attributes that contributed to eating quality such as sugar and ascorbic acid content. The attainment of the characteristic fruit colours is highly species-specific and dependent on the number of days after fruit set.     

Water stress induces in pear leaves the rise of betaine level that is associated with drought tolerance in pear

Summary

Mild water stress signi®cantly induced the accumulation of betaine in the leaves of young pear plants (Pyrus bretschneideri Rehd. `Suly'). The betaine induced by water stress in mature leaves was maintained for about two weeks of re-watering after water stress, and gradually declined two weeks after the relief of water stress, dropping to the normal level one month later. Exogenously foliar-applied betaine slightly improved the shoot growth and the ability of the leaves to resist dehydration, alleviated wilting of leaves under water stress, and promoted recovery of leaves from wilting after re-watering. These ®ndings indicate that the accumulated betaine induced by water stress was associated with drought tolerance in pear.