Temporal and spatial variation of pollen yield in natural populations of Pinus roxburghii

Male components of the reproduction process in Pinus roxburghii were investigated for their variation in time and space as well as pollination,viz.flowering phenology,pollen production variability and pollen dispersal,for five successive years at two different locations (at the lower and higher elevations).The study reveals that elevation and the hour of the day are the main determinants of anthesis and microsporangium dehiscence,because both are related to temperature and humidity.The receptivity of ovulate cone strobili or female strobili occurred earlier at the lower elevation than at the higher elevation,with a longer receptivity period in the latter case.The phenology of male and female cones varied significantly between years and elevations.The determinations of pollen yield considered various sources of variability,i.e.,the number of pollen strobili per branch,strobili per tree,microsporangia per tree and pollen grains per tree.Each of these parameters revealed significant year-to-year and elevation effects.Year-to-year variation in the production of pollen cone and pollen grains with mass production after a three-year period revealed a three-year cycle of masting in pollen production in P.roxburghii.The pollen dispersal decreased quickly with distance from the pollen source.As a consequence,a distance of 600 m was proposed as a minimum to prevent contamination by pollen in the management of seed orchards.The results ultimately suggest that the ample production of pollen grains per tree along with flowering synchrony and long dispersal of pollen grains results in homogeneity in large populations.     

马尾松二代种子园无性系开花物候观测

[目的]在用材林高世代种子园建园材料选择时,最初重点考虑的是其速生性,而对优树的开花物候关注较少。在高世代种子园营建时,无性系的配置常根据其种源在大范围内考虑其开花物候的遗传因素,但即使是小范围内相同种源的无性系,其开花物候也不可能绝对同步。因此,了解种子园无性系的开花物候,可为种子园的集约经营、产量预测、种子遗传品质调控等提供依据。[方法]于2012—2013年,在桂阳县苗圃马尾松二代种子园Ⅰ大区、Ⅴ大区分别随机选取不同类型的无性系10个和25个,对其开花物候进行观测。[结果]表明:不同无性系的雌、雄球花花期起止日期和持续时间均有差异,但无性系间雌、雄球花花期值差异均未达显著水平,35个无性系总体雌、雄球花花期具有一定的同步性;35个无性系雌、雄球花的初花期基本同步,雌、雄球花的盛花期和末花期同步。株间花期差异相对于无性间要大,但雄球花花期较早的植株,其雌球花花期也较早,同一株建园亲本的雌、雄球花花期有一定的同步性。除雌球花的末花期与雄球花的初花期呈微弱负相关外,雌、雄球花的初花期、盛花期、末花期间均呈极显著或显著正相关。各花期的相关性呈现初花期和盛花期的相关盛花期和末花期的相关初花期和末花期的相关,雌球花初花期与雄球花初花期的相关系数最大。坡位对母树的花期有极显著影响,生长在不同坡位的植株,其雌、雄球花花期值差异明显,其大小均为上坡中坡下坡。坡位对花期影响最大的是雌球花的初花期,其次是雄球花的初花期、雌球花的盛花期、雄球花的盛花期,而对雌、雄球花的末花期影响最小。偏北产地无性系的花期相对较早,不同的无性系类型,其雄球花的盛花期和雌球花的初花期的差异显著;雄球花的初花期、末花期以及雌球花的盛花期和末花期差异均不显著。不同的年份,雌、雄球花花期的早晚差异很大,2012年最早初花期较2013年最早初花期早25 d。[结论]桂阳马尾松二代种子园35个无性系总体雌、雄球花花期具有较好的同步性;从开花物候方面考虑,该种子园建园材料选择较合理,对种子园种子产量和遗传品质没有影响。     

华山松种子园无性系开花习性研究

云南楚雄华山松种子园的无性系,其雌球花着生在树冠外围枝条,而雄球花主要着生在树冠的内膛枝条。在水平方向上,雌球花多着生在树冠南部,雄球花的分布比较均衡;在垂直方向上,雄球花基本分布在树冠的中下部,雌球花则分布在树冠的中上部。大多数无性系的雄球花散粉期比雌花可授期早2~4 d;不同无性系间的雌雄花期具有一定的同步性。各无性系间雌、雄球花数量存在极显著差异,且有明显的偏雌或偏雄特性。雄球花量与无性系生长性状间存在显著正相关,雌球花量与树体生长相关性不显著。     

油松无性系开花物候特点的研究

在河南卢氏油松种子园对46个无性系结实初期和结实盛期开花物候作了观测，结果表明，一般雌球花可授期比授期比雄球花散粉期来得早，持续时间长，散粉期含在可授期内，散粉期和可授期同步性较好；在不同年份开花始期前后相差2-3d，开花始期早晚与当年≥10℃的活动积温有关，由于受花期温度和降雨等因素的影响，不同年份花期长短可相差10d左右，但不存在花期不遇，不同无性系在开花物候上存在很大差异，但在不同年份各无性系在始花时间上序次则相对稳定，无性系内分株间开花物候仅有1-2d的差异，位于树冠北向的雌雄球花各日期在开花频率上与其它3个方位有较大差异。始花期推后1d左右，此外，还研究了开花物候对同步指数的影响，以及不同物候类型授粉效率的差异。     

马尾松二代无性系种子园的花期同步性

【目的】研究马尾松二代种子园无性系的开花物候特征、花期同步性、结实状况及气候影响因子,评价无性系花期同步性规律,指导种子园经营生产及提高种子遗传品质与产量。【方法】连续2年对浙江省淳安县姥山林场马尾松二代无性系矮化种子园的22个无性系进行花期、花量和结实性状调查,分析其开花物候与花期同步指数。【结果】各无性系雌雄球花整体花期持续时间为8～17天, 2014年始花期相对于2013年滞后约6天,且持续时间缩短约6天。整体水平上,年度内雌球花及雄球花的各花期时长差异较小;无性系的始花期和盛花期的物候特征年度间存在差异, 2014年无性系内雌雄球花开花的同步性更强。无性系的开花物候受气候因素影响较大,这使得无性系间开花同步程度降低,花期同步指数变异增加, 2013年和2014年花期同步指数变化范围分别为0.041～0.556和0.284～0.802,无性系的平均变异系数分别为32.3%和16.8%。各无性系分别作为父、母本时,年度内无性系花期整体同步性水平差异较小,表现出无性系作为父本时的散粉期整体的同步性要高于作为母本时的可授粉期的同步性。年度间无性系作为父、母本及自交的花期同步指数均有极显著差异, 2014年各无性系的花期同步指数在整体水平均高于2013年,但自交指数在年度间无显著差异。相关分析表明,雌球花越多的无性系的球果数越多;而雌球花花期持续时间越长的无性系座果率越高;不同年度的雌球花盛花期和花期的持续时间均与千粒质量指标呈显著或极显著正相关。【结论】浙江姥山林场马尾松二代无性系种子园雌、雄球花的花期具有较好的同步性,无性系间没有出现花期不遇现象。无性系交配组合的雌球花可授粉期和雄球花的散粉期影响了种子园无性系的种实品质及座果率。连续2年的观测表明无性系间花期整体同步性水平存在差异,且这种差异与二代种子园无性系的亲本遗传背景有关。各无性系分别作为父、母本时,表现出无性系作为父本时的散粉期整体的同步性要高于作为母本时的可授粉期的同步性,无性系作为父本和母本时花期同步指数呈负相关,且种子园内存在1/3无性系具有高于异交平均水平的自交可能性,是马尾松二代种子园管理和无性系再选择应综合考虑的因素。     

Mating System and Pollen Contamination in a Pinus brutia Seed Orchard

Mating system parameters and pollen contamination were estimated in an 11-year-old Pinus brutia Ten. clonal seed orchard, located in Antalya, Turkey, with the aid of isozymes. Isozyme analysis was performed on both maternal (megagametophyte) and embryo tissues of seeds collected from the seed orchard and the surrounding natural stand. Fourteen loci encoding nine enzyme systems were assayed. Based on a multilocus estimator, the proportion of viable seeds originating from outcrossing (tˆ_m) in the seed orchard was found to be 0.947; the remainder (0.053) was due to selfing. We estimated that 85.7% of the orchard seed resulted from pollen contamination by trees in surrounding stands. This may be related to the young age and subsequent low pollen production of the orchard. Only 9% [i.e., 1.0−0.857 (contamination)−0.053 (selfing) = 0.090] of the orchard offspring was due to cross-pollination among clones within the orchard. Due to the high level of pollen contamination, it is estimated that genetic gain in this seed crop is less than 57% of that expected if all offspring resulted from random mating among orchard clones (i.e., no contamination).     

Lateral female strobili production in a Japanese red pine (Pinus densiflora Sieb. et Zucc.) clone by exogenous cytokinin application

Application ofN ⁶-benzylaminopurine (BAP) and gibberellin (GA₃) to a grafted Japanese red pine (Pinus densiflora Sieb. et Zucc.) clone promoted flowering. BAP (250 mg/l) was sprayed five times to the top of branches in July or September 1992. GA₃ (100 or 500 mg/l) was applied five times to the whole plants by spray treatment during the above period, or GA₃ solution of the same concentration was injected once into a small incision made in the main stem of clone trees at the beginning of July or the middle of September 1992. In May 1993, no apical female strobili were observed at the top of the new shoots in any treatment. However, lateral female and bisexual strobili were produced in the lower part of new shoots by BAP application in September, regardless of GA₃ application. BAP applied in September without GA₃ application was sufficient to produce lateral female and bisexual strobili. In such treatment, production of female and bisexual strobili was 47 and 25, and they occurred in 16 out of 68 new shoots, 11 out of 15 treated branches and 5 out of 5 treated trees. The average number of female and bisexual strobili per new shoot was 1.06. BAP treatment either in July or in September significantly decreased (p<0.05) the production of male strobili. These results indicate that BAP application induces the development of lateral female strobili in Japanese red pine when applied during the period of floral differentiation. A part of this paper was presented at the 105th annual meeting of the Japanese Forestry Society.     

Supplemental mass pollination in a seed orchard of Pinus Sylvestris L. investigated by isozyme analyses

Isozyme‐analysis techniques have been applied to test the efficiency of supplemental mass pollination in a seed orchard of Pinus Sylvestris. Mass pollination was done on one single occasion before the general pollen dispersal. The results of isozyme analyses of seed after mass pollination demonstrate that it is possible to introduce desirable genes into the seed orchard crop. Mass pollination without isolation of female strobili gave an average contribution of 4% to the total fertilizations and after isolation of female strobili 26.5%. The main conclusion is that supplemental mass pollination offers very interesting possibilities for breeding, particularly if some kind of isolation of the female flowers is made. It can increase the seed quality as well as improve the breeding value of the seed orchard. More attention should be paid to increasing the efficiency of the controlled pollination. It is suggested that in the future, more effort is invested in improving the technique of mass pollination.     

思茅松无性系种子园雌雄球花分布特性及人工辅助授粉的研究

在云南省林业科学院普文试验林场思茅松种子园开展了 16个无性系的雌雄球花空间分布特性及人工辅助授粉的研究 ,结果表明 :各无性系间雌雄球花数量及雌雄比均差异极显著 ,存在偏雌偏雄的变异 ;雌雄球花数量的空间分布存在一定的规律性 ,在垂直方向上 ,79%以上的雌球花分布于树冠的中上部 ,80 %的雄花分布在树冠的中下部 ;在水平方向 ,东、西、南、北 4个方位球花数量的分布基本一致。种子园的落花现象比较严重 ,且不同无性系的落花率不同 ;人工辅助授粉有助于雌球花的保存和座果率的提高     

Reduction of pollen contamination in Scots pine seed orchard crop by tent isolation

This study examined the differences in mating system between indoor versus outdoor Scots pine (Pinus sylvestris L.) seed orchard using DNA markers. By analyzing the paternities in seed crops from isolation tents, we found a significant reduction in contamination rate, from 23% in the outdoor open blocks to 0% inside the tents. Increase in selfing rate and a decrease in genetic diversity in the crops were also observed inside the tents. These undesirable effects can be mitigated to some extent by the supplementation of extra pollen genotypes, e.g. selfing rate decreased from 14.4% to 6.6% and the effective number of fathers increased from 6 to 11.4 in the tents without and with supplemental mass pollination, respectively. Our study showed that tent isolation may become an effective rescuer for seed orchards where pollen contamination is severe.     

Female fertility variation in mature Pinus sylvestris clonal seed orchards

Abstract

Female fertility is the basis for the output of seeds from clonal orchards and its variation is of major interest for the economics and biology of seed orchards, especially for the efficiency and diversity of seed orchard crops. Assessments of female fertility variation in 10 mature (>15 years old) seed orchards of Scots pine (Pinus sylvestris L.) were evaluated and compared. Depending on the individual orchard, fertility variation for each clone was assessed in slightly different ways, e.g. number of strobili, cones, seeds or litre of cones per ramet. Assessments in five of the orchards were made over consecutive years. The main result was that the clonal variation in mean female fertility per surviving ramet was lower than expected from the literature; the Kang–Lindgren sibling coefficient (Ψ) within individual years averaged 1.35. The variation between ramets within clones and years was of similar magnitude as that between clones. Clone by year interactions were noticeable, but were slightly lower than the variation between as well as that within clones in individual years. There was considerable variation in the variance components between years. The limited variation in female fertility indicated that it should not be a selection criterion when selecting clones for a seed orchard. Furthermore, it will not result in large differences between clones in seed set or large reductions in gene diversity in productive Scots pine seed orchards.     

Temporal and spatial variation of flowering among Pinus nigra Arn. clones under changing climatic conditions

Until recently, the most important factors affecting the economics and genetics of the seed crop from seed orchards were considered to be the timing and duration of flowering, variation in fertility and the total number of clones used to establish the seed orchards. Change in climatic conditions however is an emerging factor that could prove crucial regarding the timing of flowering and synchronization among clones and thus, the quality and quantity of seed production. The temporal and spatial variation in flowering phenology and the duration of flowering were studied in consecutive years in a Pinus nigra Arn. seed orchard. Sixty plus trees representing the distribution of the species in Northern Greece were used to establish the seed orchard, and nineteen ramets per clone were planted in a honeycomb experimental design in order to avoid kinship. Temporal variation among clones, as well as spatial variation among ramets within clones growing at different sites of the orchard were recorded, for initiation and duration of male and female flowering. The majority of clones were synchronized in dates of flowering during the year with weather conditions close to the long-term climatic conditions, except for a limited number of clones that were precocious or late flowering. The pronounced variability in climatic conditions over the 2 years strongly affected the flowering and synchronization among clones, resulting in almost complete asynchrony during the xerothermic year, which was characterized by a prolonged mean monthly temperature increase of 2.3 °C and a water deficit of 53% in a 7-month-period (November to May). These results suggest that one of the effects of a warmer and drier climate may be the lack of flowering synchronization, as pollen shedding might be completed before female conelets reach the phase of receptivity. The restriction of male parentage to a limited number of clones severely violates the panmixia assumption and could result in fertilization failure. The projected climate change for the Mediterranean region could potentially prove detrimental for fertility and flowering synchronization of forest trees, having consequences on the quantity and genetic diversity of the seed crop in seed orchards, and the natural regeneration of forest trees in forest ecosystems due to the reduced percentage of sound seed.     

Reproduction of postfire Pinus halepensis Mill. stands six years after silvicultural treatments

In Spain, wildfires have increased during the last decades with Pinus halepensis forests being the most affected. Cone differentiation and the early flowering of this species in comparison to other native Spanish species, are traits considered as adaptations to postfire regeneration. The high recurrence of fires promotes a high increase in young and immature pine stands with low capability of regeneration. In this study, silvicultural treatments such as thinning and pruning were carried out 5 years after fire in eleven years old P. halepensis stands located in dry and semi-arid sites in SE Spain. The formation of male and female strobili, production of serotinous grey, mature brown and new green cones were recorded six years after treatments. Seed production and germination percentage were also tested. Results showed acceleration in cone and viable seed production in thinned plots, with some differences between sites being recorded. Serotinous cone production also increased as a result of this treatment.     

Are Phenological Observations Sufficient to Estimate the Quality of Seed Crops from a <Emphasis Type="Italic">Eucalyptus grandis</Emphasis> Open-pollinated Seed Orchard? Consequences for Seed Collections

Reproductive phenology was observed over three years in a Eucalyptus grandis seed orchard in Madagascar to determine the impact of geographic differences of parental selections on the expected genetic composition of seed crops. Pollen flow (observed pollen cloud) was deduced from a paternity assignment based on sampling from one year’s seed production. The two approaches were used to verify whether knowledge of phenology is sufficient to predict the genetic quality of the seeds collected. Despite the high flowering level, with a constant cycle over three years, the results demonstrated wide reproductive phenological differences associated with the parents’ origin, suggesting putative pollination disequilibrium. From both observed and expected pollen clouds, the results showed preferential mating among different provenances, which has consequences for seed crop composition. Phenological observations, which can be made inexpensively, can be used to promote methods of effective seed orchard management in order to improve the genetic quality of seeds.     

Southern pollen sired more seeds than northern pollen in southern seed orchards established with northern clones of Pinus sylvestris

Abstract

Contamination by southern pollen is a considerable problem in Scots pine (Pinus sylvestris L.) seed orchards established with northern clones. This study investigated whether the contamination was due to the competitive superiority of southern pollen by carrying out competition trials using mixtures of pollen from northern and southern populations of Scots pine. Trials were performed in a southerly seed orchard established with clones originating from northern populations. Seed paternity (siring) was determined through the analysis of allozyme variation. Southern genotypes sired significantly more seeds (76%) than their northern competitors and across all mixed-pollen crosses. Maternal genotype had no effect on seed siring success. The mean flower abortion rate was lower in southern pure-pollen crosses and mixed-pollen crosses than in northern pure-pollen crosses. The results show that local pollen may induce high levels of background pollination in southern seed orchards composed of northern genotype grafts. These results must be taken into account when aiming to produce suitable reforestation material for northern areas.     

Evaluation of seed production in a first-generation seed orchard of Chinese pine(Pinus tabuliformis)

We evaluated seed production in a first-generation orchard of Chinese pine(Pinus tabuliformis) during the crucial transition period from first generation to advanced generations.Clones varied significantly in all traits related to seed production.Repeatability of these traits ranged from 0 to 0.96.Seed production per ramet(SPPR),seed producing index(SPI),the number of relative female strobili(RFS),the number of scales,and the number of ineffective scales had comparatively high repeatability at 0.86,0.87,0.89,0.96,and 0.91,respectively.Correlation analysis showed that SPPR was greatly influenced by RFS and by the number of full seeds per cone.Finally,we showed that SPI was the best predictor of the seed-producing ability of clones.Our findings will assist seed orchard managers in effectively predicting and improving seed production of Chinese pine seed orchards.     

Pollen dispersal,mating patterns and pollen contamination in an insect-pollinated seed orchard of <Emphasis Type="Italic">Schima superba</Emphasis> Gardn. et Champ.

Pollen flow and mating patterns are the most important factors influencing the genetic structure of insect-pollinated forest trees and are essential parts of genetic management in seed orchards. We investigated pollen flow, the mating system and the level of pollen contamination in a clonal seed orchard of Schima superba Gardn. et Champ. In total, 328 open-pollination progenies coming from 11 mother trees were identified using 13 polymorphic microsatellite loci. A total of 203 full-sib families were identified and were nested among the 11 studied seed donors. The male reproductive success rate from 0.49 to 7.77% for most male parents, with an average of 2.44%. More than 80% of the crosses were found within a distance of 60 m, and the most frequent pollination distance between female parent and male parents was approximately 20 m. Mating system analysis showed that the outcrossing level was very high (t _m  = 1.000, outcrossing rate = 98.5%) in the seed orchard and that there was an average of 2.3 effective pollen donors (N _ep ) per female parent. In addition, the mating success of individual males within neighborhoods was moderately influenced by their fecundity and the direction of their location relative to mother trees. The pollen contamination from outside the seed orchard was high (7.01%). Our findings are valuable for the assessment of seed orchards, and it may be worthwhile to use pollen management strategies to decrease pollen contamination and increase the genetic quality of the seeds produced.     

Evaluation of seed production in a first-generation seed orchard of Chinese pine (<Emphasis Type="Italic">Pinus tabuliformis</Emphasis>)

We evaluated seed production in a first-generation orchard of Chinese pine (Pinus tabuliformis) during the crucial transition period from first generation to advanced generations. Clones varied significantly in all traits related to seed production. Repeatability of these traits ranged from 0 to 0.96. Seed production per ramet (SPPR), seed producing index (SPI), the number of relative female strobili (RFS), the number of scales, and the number of ineffective scales had comparatively high repeatability at 0.86, 0.87, 0.89, 0.96, and 0.91, respectively. Correlation analysis showed that SPPR was greatly influenced by RFS and by the number of full seeds per cone. Finally, we showed that SPI was the best predictor of the seed-producing ability of clones. Our findings will assist seed orchard managers in effectively predicting and improving seed production of Chinese pine seed orchards.     

Promotion of lateral female strobili production inPinus densiflora by cytokinin application at a specific stage

BAP (N ⁶-benzylaminopurine) was sprayed 5, 10 and 15 times at four different periods to promote flowering of a grafted Japanese red pine (Pinus densiflora) clone. Four stages were I, August 4 to September 1; II, September 6 to October 4; III, October 5 to November 14 and IV, November 30 to December 28, in 1994. Concentration of BAP was 250 mg/l. In May, 1995, no apical female strobili were observed at the top of the new shoots in any treatment. However, BAP application at stage III (floral differentiation period) produced lateral female and bisexual strobili at the lower part of new shoots. The numbers of lateral female and bisexual strobili were 87 out of 147 to 234 out of 162 new shoots. Frequency of BAP application did not affect the numbers. Marked decrease in number of male strobili induced by BAP application at stages I and II was observed. In contrast, the number of new shoots was increased by the BAP application at the above stages. Treatment at stage III and IV did not affect significantly the number of male strobili and new shoots. These results indicate that BAP application at stage III specifically induces the development of lateral female and bisexual strobili in Japanese red pine.     

木荷种子园无性系开花物候及同步性分析

[目的]研究种子园无性系的花期物候特征和花期同步性及影响因子对种子园科学管理具有指导意义。[方法]2015年和2016年连续2 a对木荷无性系种子园中19个无性系进行花期物候调查,分析其开花物候与花期同步指数。[结果]木荷种子园各无性系开花持续时间为10~28 d,2016年较2015年晚2 d进入始花期。相关分析结果表明,开花数多的无性系比开花数少的无性系花期更长、座果数更多,花期持续时间长的无性系比持续时间短的无性系座果数更多。无性系组合间花期同步指数有较大差异,2015年和2016年变化范围分别为0.552~0.857和0.406~0.808,平均分别为0.758和0.713,其中花期同步指数大于0.80的无性系分别在50%和40%以上。年度内花期同步指数变异系数分别为12.016%~46.476%和15.375%~51.202%,无性系之间平均花期同步指数存在极显著变异(P0.01)。年度间花期平均同步指数为0.737,平均花期同步指数间不存在显著变异(P0.05)。年度间花期同步指数相关系数为0.229,表明年度间花期同步指数具有一定的相关性,但不显著。[结论]木荷种子园无性系年度间花期同步指数较为稳定,可根据开花物候和花期同步指数对无性系进行筛选和优化,同时辅以人工授粉等措施弥补花期同步性差异,以达到提高种子园种子产量和质量的目的。