Molecular and physiological identification of new S-alleles associated with self-(in)compatibility in local Spanish almond cultivars

The S-allele characterisation of ‘Alzina’ and ‘Garondès’, two local almond cultivars from the island of Majorca, by a multidimensional approach has allowed the confirmation of the presence of the S_f-allele and the identification of a new allele not previously described in almond, S₃₆. When these cultivars were phenotypically evaluated, both showed a self-incompatible phenotype and were cross-incompatible, as assessed by artificial self- and cross-pollinations and fruit sets after field pollinations, confirming that their S_f-allele is in its active form, S_fa. Thus a new CGI group in almond is proposed and named XXVIII. These results confirm the wide diversity of S-alleles in almond both at genotypic and phenotypic levels, as well as their similarity with the S-alleles from other close Prunus species. This similarity suggests the possibility of allele introgression between species or allele identity by descent from a common ancestor.     

Physiological Breakdown in Stored Monarch Plums

Summary

Several pollination studies carried out on different, self-incompatible almond cultivars and seedlings have shown the presence of variable levels of fruit set following self-pollination that could be attributed to partial self-incompatibility (PSI). PSI is an intermediate reproductive behaviour, described in some angiosperm species, which indicates that self-incompatibility is a quantitative and plastic trait. The present study was performed to substantiate the occurrence of PSI in almond by studying ten cultivars that are traditionally considered to be self-incompatible (four of which had previously shown fruit set after self-pollination). These cultivars were analysed by microscopic observations of the progression of pollen tubes through the pistil following controlled self-pollination, determinations of fruit set after bagging flower buds, and molecular identification of the parentage of the fruits obtained using consensus PCR-primers for Prunus S-RNase alleles. The results showed that, in nearly all cases, the pollen tubes did not enter the ovary and, from a total of 5,349 bagged flower buds, only 17 fruits were obtained. In all cases, PCR analysis of the plants obtained after germination of these seeds showed a band corresponding to an S-RNase allele not present in the maternal progenitor, clearly indicating that they could only have arisen from cross-pollination.Therefore, the low fruit set values observed in some of the cultivars studied were not due to a breakdown in the self-incompatibility response that confers PSI, but to very low rates of contamination with foreign pollen. These results corroborate the self-incompatibility phenotype of the cultivars studied here, and highlight the importance of ascertaining the identity of the parentage in any progeny obtained after bagging.     

The almond Sf haplotype shows a double expression despite its comprehensive genetic identity

Since self-compatibility has become the primary objective of most almond breeding programmes, search for new self-compatibility sources has acquired a great importance in almond research. The local Spanish cultivar ‘Vivot’, identified as showing the genotype S₂₃S_f, thus presumably self-compatible, was found to be unexpectedly self-incompatible in spite of the presence of the S_f allele, as also observed in other almond cultivars. However, not only the coding sequences of both the S_f-RNase and the SFB_f of ‘Vivot’ and ‘Blanquerna’, a confirmed self-compatible cultivar, were identical, but also the 5′ regulatory sequence of the S_f-RNase of both. Thus, the reason for the different expression of the S_f is independent of the complete genetic identity found in the whole chromosome region bordering the S-locus in the almond cultivars sharing the S_f allele.     

Year and enclosure effects on fruit set in an autogamous almond

Fruit set was studied over 2 years in ‘Guara’, an autogamous almond (Prunus amygdalus Batsch) cultivar, following four different pollination treatments: open pollination, open pollination of a reduced number of flowers, self-pollination, and cross-pollination with a cross-compatible pollen. The treatments were applied to trees growing in the open air, to a tree inside an insect-proof cage, and to bagged branches in the open air. Fruit sets inside the cage were in general lower than at the open air, thus pointing to the possibility that a reduction of the light intensity inside the cage may reduce the level of fruit set. However, artificial pollinations resulted in higher sets inside the cage than in the open air in one of the years. Fruit sets on bagged branches after artificial pollinations were always lower than for the open air and inside the cage, showing that besides a general enclosure effect, as observed inside the cage, a bag effect is responsible for the reduction of sets, possibly due to the branch manipulation and flower damage by the bag. The reduction of the number of flowers did not always result in an increase in set of the remaining flowers. There were no consistent differences between the two types of artificial pollination, confirming ‘Guara’ self-compatibility. Year effects were observed, but not affecting all the treatments in the same direction nor at the same level, thus stressing the need for multiyear evaluation of autogamy.     

Self-fertilization in homozygous and heterozygous self-compatible almonds

In homozygous self-compatible genotypes 100% of the pollen grains are potentially able to grow through their own pistil, and thus the rate of self-fertilization could be higher than in heterozygous self-compatible genotypes. To evaluate the advantages of growing homozygous self-compatible almonds, pollen tube growth along the pistil at different times following self-pollination, and fruit set were studied in four homozygous and four heterozygous self-compatible seedlings. The results showed important differences between homozygous and heterozygous individuals for the percentages of pollen tubes in the third section of the style at 24 and 48 h, the pollen tube growth rate being higher in the homozygous. Twenty-four hours following self-pollination only the homozygous individuals showed pollen tubes in the ovary. However, at 72 and 96 h those values were similar for both genotypes, suggesting that space and availability of nutrients become the main limiting factors, overcoming the genetic interactions between pollen and pistil. In general, fruit set was similar in homozygous and heterozygous individuals. Interestingly, one of the homozygous individuals showed problems of fruit development, which might be explained by its inbred origin.     

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.     

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.     

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。     

Pollen tube growth in styles of apple and almond flowers after spraying with pesticides

Summary

The timing of fungicide and antibiotic applications in fruit crops often overlaps flowering and pollination. Numerous studies report detrimental effects of chemical applications on pollination, fruit set and yield. However, field results can be quite variable even when similar compounds are tested on the same crop. Problematic is that field variables unrelated to the effects of pesticides on reproductive activities can mask results. In the current study, we specifically investigated the effects of selected pesticide sprays applied to stigmas, on subsequent pollen germination and tube growth. Using two model systems, apple and almond, sprays were applied to detached emasculated flowers under constant laboratory conditions using an electronically controlled robotic apparatus which simulated a field sprayer. flowers were pollinated at specific times after spraying, and pollen tube numbers and growth assessed using fluorescence microscopy. The pesticides evaluated were for apple: captan, myclobutanil and streptomycin; and for almond: azoxystrobin, myclobutanil, iprodione and cyprodinil. In apple, captan significantly reduced pollen germination, by 20% compared with water controls when pollination was conducted 18 h after treatment. The number of tubes that reached the basal part of the style within 20 h was not affected. In contrast, myclobutanil and streptomycin had no significant effect on pollen tube growth. With almond, none of the sprays significantly affected pollen germination or tube growth.     

The response of different almond genotypes to moderate and severe water stress in order to screen for drought tolerance

In order to screen almond genotypes for drought tolerance, three different irrigation levels including moderate and severe stress (Ψs = −1.2 and −1.8 MPa respectively) and a control treatment (Ψs = −0.33 MPa) were applied for five weeks to six different cultivated almond seedlings. A factorial experiment was conducted with a RCBD which included 3 irrigations factors, 6 genotype factors and 3 replications. Seeds were prepared from controlled pollination of the bagged trees (after emasculation and flower isolation using isolator packets in the previous year). Genotypes included: homozygote sweet (Butte), heterozygote sweet (SH12, SH18, SH21 and White) and homozygote Bitter (Bitter Genotype). Leaf and root morphological and physiological traits including; midday relative water content, midday leaf (xylem) water potential, shoot dry weight and growth, total leaf area, leaf size, total leaf dry weight, specific leaf area, leaf greenness (SPAD), stomatal size and density, root and leaf nitrogen content and chlorophyll fluorescence were measured throughout the study. Results showed the six genotypes had different reactions to water stress but all genotypes showed an ability to tolerate the moderate and severe stresses and they showed different degrees of response time to drought stress. Almond seedling leaves could tolerate Ψ_w between −3 and −4 MPa in short periods. Water availability did not significantly affect stomatal density and size of young almond plants. The analysis of leaf anatomical traits and water relations showed the different strategies for almond genotypes under water stress conditions. Although almond seedlings even in severe stress kept their leaves, they showed a reduction in size to compensate for the stress effects. All genotypes managed to recover from moderate stress so Ψ_w = −1.2 could be tolerated well by almond seedlings but Ψ_w = −1.8 limited young plant growth. Leaf greenness, leaf size, shoot growth, shoot DW, TLDW and stomatal density were not good markers for drought resistance in almond seedlings. Root DW/LA, lower stomatal size and lower SLA might be related to drought resistance in cultivated almonds. Butte had the least resistance and White showed better performance during water stress while other genotypes were intermediate. Bitter seedlings showed no superiority in comparison with other genotypes under water stress conditions except for better germination and greater root DW which might make them suitable as rootstocks under irrigation conditions.     

Fruit set after self-pollination at different floral stages and its relation to pollen-tube growth and stylar S-RNase content in Japanese apricot (Prunus mume Sieb. et Zucc.)

Summary

To investigate fruit set after self-pollination at different floral stages, and its relationship to the S-RNase content of the style in Japanese apricot, which exhibits gametophytic self-incompatibility (GSI), the cultivar ‘Xiyeqing’ was selfand cross-pollinated at –6, –4, –2, 0 and +2 d after anthesis (DAA). At –6 and –4 DAA, the percentage fruit set was low, suggesting that the propensity to seed set was weak during this period. At –2 DAA, the percentage fruit set, after self-pollination, reached a maximum of 5.97%. Thus, the effective time of self-pollination to obtain inbred seed was –2 DAA. Soluble protein extracts of styles at each floral stage were subjected to iso-electric focussing. The results showed that S-RNase began to be synthesized from –6 DAA, reached its maximum expression 2 – 3 d before anthesis, and was degraded slowly after anthesis. The number of pollen-tubes arriving at the stylar base, as detected by fluorescence microscopy, varied at –2, 0, and +2 DAA. The number of pollen-tubes arriving at the stylar base, and the percentage fruit set after self-pollination, exhibited a positive correlation (r = 0.8615; P = 0.1), while the S-RNase content per style showed a negative relationship to fruit set percentage (r = –0.9640; P = 0.01), and to the number of pollen tubes arriving at the stylar base (r = –0.9380; P = 0.01) during this period. This implied that the S-RNase content in the style was mainly responsible for the low percentage fruit set and the low number of pollen-tubes arriving at the stylar base after self-pollination.     

Comparison of the use of morphological,protein and DNA markers in the genetic characterization of Iranian wild Prunus species

In this study, the genetic diversity of four Iranian wild Prunus species including Prunus eleagnifolia, Prunus hauskenchtii, Prunus scoparia and Prunus lycioides were investigated using morphological, protein and DNA markers. DNA markers included nuclear and chloroplast SSRs and self-incompatibility (S) allele amplification. At the morphological level, leaf width showed significant differences between the four wild Prunus species. Concerning fruit and kernel characters, their values are relatively similar indicating the high degree of homoplasy described in Prunus. Nuclear SSR markers have been the most abundant markers with a higher polymorphism in comparison with morphological, protein and chloroplast SSR markers. Results also indicated the high variability present in the S locus. On the other hand, the correlation between the clustering based on DNA markers and protein were in general low. Dendogram performed using nuclear and chloroplast SSR indicated a more diffuse clustering between the wild almond species probably due to the natural introgression of genes observed in these wild almond species. Data from the analysis of the total protein seems to be more accurate to establish taxonomy relationships in these very close wild species.     

Determination of self-incompatible genotypes in 21 cultivated sweet cherry cultivars in Greece and implications for orchard cultivation

Summary

Sweet cherry is self-incompatible due to having a gametophytic self-incompatibility system. S alleles in the style and pollen determine possible crossing relationships. Knowledge of the S allele constitution of cultivars is important for sweet cherry growers and breeders. Recently, molecular methods have been developed to distinguish between S alleles in sweet cherry.The S allele genotypes of 21 sweet cherry cultivars widely grown in Greece, including 19 not previously genotyped, were determined based on their S-RNase gene sequences using PCR analysis. Eight different S alleles in ten combinations were distinguished and two new S-genotypes (S₁S₁₃ and S₄S₃₀) were documented. Four alleles, S₁, S₃, S₄, and S₉ were widespread and together were responsible for 85% of the S-haplotypes. Therefore many of the cultivar combinations were semi-compatible. In Greece, semi-compatibility was shown to correlate with low yields. However, the cultivar ‘Hybrid Tragana Edessis x Unknown’ (S₃S₁₃) and the cultivar ‘Kapsiotika’ (S₂S₅) carry rare S-haplotypes and are therefore fully cross-compatible with most of the cultivars analysed.     

Observations on the Effect of Potash Supply on the Tension of the Tracheal Contents in Fruit Trees and Bushes

Summary

The effects of pollen source on fruit set and fruit quality characteristics (xenia effects) were examined in three commercial cultivars of mandarin, Imperial, Ellenor, and Murcott. Six trees of each cultivar received six pollination treatments: three cross-pollen sources, self-pollination, bagging and bagging with emasculation. Pollen tube growth, percentage fruit set, fruit weight, seediness, sugar and acid content were assessed in all treatments. Significant xenia effects, including effects on sugar content, were found in all cultivars. `Imperial' was found to be self-incompatible as self-pollen tubes were inhibited in the upper style. This resulted in a lower fruit set in self-pollinated fruits (P<0.01), a very low fruit weight (33–55 g, compared with 92–103 g, P<0.01), and fruits containing few or no seeds. In addition, `Ellenor' and `Murcott' pollen significantly increased sugar content of fruit by between 0.9–1.6% compared with self-pollinated and unpollinated treatments (P<0.05). Widespread problems of variable production and poor fruit quality in `Imperial' may be alleviated by interplanting with appropriate pollen sources such as `Ellenor' and `Murcott'. Pollen source significantly affected fruit set, seed number and sugar content but not fruit weight of cv. Ellenor. In particular, `Murcott' pollen produced a significantly higher fruit set, relatively low seed number, and the highest mean sugar content (13.2%), significantly higher than `Imperial' pollen, bagged, and unpollinated treatments (>12.5%, P<0.01). Fruit production, seediness and sugar content of `Ellenor' mandarin may be improved by interplanting with `Murcott'. Cross-pollination significantly increased seed number of `Murcott' (15–21 seeds per fruit, compared with 13–17 seeds per fruit, P<0.01). `Murcott' could be planted in pure blocks since self-pollinated `Murcott' fruit had slightly fewer seeds than crosses, without any significant loss of size or quality. These results demonstrate the need for careful consideration of interplanting of cultivars and management of pollinators such as the honeybee, Apis mellifera L. to maximize fruit production and quality in mandarins.     

Development of a male sterile eggplant by utilizing the cytoplasm of Solanum virginianum and a biparental transmission of chloroplast DNA in backcrossing

To develop a male sterile line of eggplant (Solanum melongena L.) utilizing the cytoplasm of S. virginianum, an interspecific F₁ hybrid between S. virginianum and S. melongena ‘Senryo Nigou’ was continuously backcrossed to S. melongena ‘Uttara’ using ‘Uttara’ as a recurrent pollen parent and four backcross generations, BC₁, BC₂, BC₃ and BC₄ were produced. All the plants in backcross generations were anther indehiscent although the F₁ hybrid, S. virginianum and S. melongena were dehiscent. Pollen stainability with acetic carmine and in vitro germination ability of pollen in all the backcross progenies were quite lower than those of the parental species. Fruit set percentage, number of seeds per fruit and seed germination rate were high in all the backcross progenies. The present results indicate that anther indehiscent type of functional male sterile line of eggplant could be developed by utilizing the cytoplasm of S. virginianum. PCR-RFLP analysis of chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) were performed to identify the organelle inheritance. The F₁ hybrid and all the backcross progenies displayed maternal inheritance of mtDNA. The cpDNA of the examined single BC₁ plant exhibited the recombinant cpDNA pattern of the parental F₁ hybrid and ‘Uttara’, indicating occurrence of the biparental inheritance of cpDNA. As all the BC₂, BC₃ and BC₄ progenies showed the same recombinant cpDNA patterns of the BC₁ plant, the recombinant cpDNA might be stable and harmonize with the nuclear genome of S. melongena. The present male sterile line can contribute to expand the male sterility source of eggplant.     

Agronomic,cytogenetic and molecular studies on hermaphroditism and self-compatibility in the Greek kiwifruit (Actinidia deliciosa) cultivar ‘Tsechelidis’

The development of hermaphrodite cultivars of kiwifruit (Actinidia deliciosa) to overcome dioecism is a priority for all breeding programmes, worldwide. If successful, the number of pollinators in the orchard could be reduced, allowing more space for reproductive vines. Hermaphroditism and the expression of self-compatibility were studied in A. deliciosa, using the new kiwifruit cultivar, ‘Tsechelidis’, as a model. This plant material was chosen for investigation as its female flowers produced viable pollen. The aim of the present study was to investigate the expression and nature of self-compatibility in ‘Tsechelidis’ and to evaluate its potential for breeding purposes. The expression of self-compatibility in ‘Tsechelidis’, between years, was investigated by controlled self-pollination of female flowers in the field and by microscopic studies on pollen viability in the laboratory. The commercial cultivar ‘Hayward’ and the pollinator ‘Matua’ were used as controls. Controlled self-pollination in the field confirmed that ‘Tsechelidis’ exhibited a degree of hermaphroditism and self-compatibility. Its flowers produced viable pollen and could set fruit of an acceptable size and quality after self-pollination, compared to open-pollinated fruit. The average fruit set percentage for ‘Tsechelidis’ was 34%, and ranked from 20% to 60% per vine, while the corresponding rate for ‘Hayward’ was only 4%. Microscopic studies on ‘Tsechelidis’ pollen revealed variations in pollen viability between different vines, which were even greater between different flowers on the same vine, ranging, in some cases, from 0% (absolutely sterile) to 70% (significantly fertile) per flower. Female control ‘Hayward’ vines were found to be practically sterile (0%), while male control ‘Matua’ vines were almost 100% fertile. The nature of self-compatibility in ‘Tsechelidis’ was investigated further by molecular analysis using the sex-linked sequence characterised amplified region (SCAR) markers, SmX, SmY, and SmY1, and by single nucleotide polymorphism (SNP) analysis. Hermaphroditism in ‘Tsechelidis’ was related to sex expression, as observed by SNP analysis, but complete genome mapping would be required to fully understand the mechanisms involved and to exploit this new genetic material for breeding purposes.     

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.     

Influence of self-pollination in fruit quality of autogamous almonds

Due to the several advantages expected from growing self-compatible almond cultivars, breeding programmes have recently introduced self-compatibility as an important trait. However, doubts have arisen regarding the capability of the newly obtained self-compatible almonds to produce good yields in monovarietal plantations, and the commercial quality of the fruits from self-pollination. For this reason, in the last years several studies have been conducted to evaluate fruit set and fruit traits following self- and cross-pollination of self-compatible almonds. Their results showed inconsistencies for fruit set and for some of the fruit traits studied. Thus, while some studies showed no differences between both pollination types for any of the fruit traits, others clearly showed important differences with the detriment of self-pollination for commercially important fruit traits. The fact that some of these studies evaluated a low number of individuals and/or fruit traits, implies the need of a more complete study. In this work, 10 fruit traits were studied in 26 self-compatible almonds following self-pollination by bagging and open pollination treatments. In general, and despite a few individuals showed differences between both pollination types for two of the traits, the results showed no influence of self-pollination in fruit quality.     

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.     

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.