Effects of accelerated pollen-cone development on pollen cytology and fertilizing potential in western hemlock (Tsuga heterophylla)

Western hemlock (Tsuga heterophylla (Raf). Sarg.) pollen cones on cut branches were collected at different stages of development and maintained at room temperature in flasks of water to accelerate pollen-cone development and produce forced pollen. Accelerating pollen-cone development at early developmental stages reduced the number of cones that matured, decreased the quantity of pollen produced, increased the proportion of abnormally developed pollen and reduced the fertilizing potential of the pollen, as determined by seed efficiency. If the branches were collected after the pollen cones were at least 50% emerged beyond the bud scales, pollen-cone development could be successfully accelerated without decreasing pollen yield or fertilizing potential. Pollen collected from cones which were accelerated at early stages and stored for two years had a lower fertilizing potential and produced fewer seeds per cone than either unstored pollen or stored pollen collected from cones accelerated at later stages of development. The feasibility of accelerating pollen-cone development to ensure adequate supplies of pollen for controlled crosses or supplemental mass pollination is discussed.     

Evaluating Supplemental Mass Pollination Efficacy in a Lodgepole Pine Orchard in British Columbia using Chloroplast DNA Markers

Supplemental mass pollination (SMP) success in a grafted lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm.) seed orchard in southern British Columbia was studied by employing four chloroplast DNA (cpDNA) markers to determine fertilization success of 10 pollen parents. SMP was conducted operationally with a bulked pollen mix twice during peak receptivity in the seed orchard. Fertilization success of the 10 SMP parents averaged 16% greater than for wind-pollinated controls in a different section of the orchard. SMP also increased the uniformity of the male contribution in treated seeds.     

Flowering and Airborne Pollen Occurrence in a Pinus sylvestris Seed Orchard Consisting of Northern Clones

To explain the high background pollination rate detected in earlier studies, this study investigated the flowering phenology of a Scots pine seed orchard with respect to airborne pollen occurrence during 1992 and 1993. The receptive period of the female strobili and the shedding period of the male strobili were observed visually. Airborne pollen was monitored with Rotorod traps located inside and outside the seed orchard. There was no time gap in the onset of flowering between the earliest female and male strobili in 1992. Female flowering began 3 days earlier than male flowering in 1993, and the earliest strobili were probably pollinated by pollen transported long distance from more southerly areas. The almost simultaneous peak in pollen occurrence inside the seed orchard and in surrounding pine stands indicates that the seed orchard clones are not temporally isolated from sources of contamination. However, pollen concentrations inside the seed orchard were higher than those in the surrounding stands. Therefore, flowering phenology cannot sufficiently explain the observed pollen contamination rates in this seed orchard.     

Estimates of pollen contamination and selfing in a coastal Douglas-fir seed orchard

Seed orchards are the link from tree breeding to reforestation programs and are theoretically expected to function as closed, perfect populations, ensuring gain and diversity are consistently and predictively delivered as improved seed and seedlings. Seed orchard populations often deviate from panmixia due to fertility variation, reproductive asynchrony, and gene flow, leading to reduced seed crops genetic quality. Here, as a part of multiyear monitoring study, we used DNA fingerprinting (simple sequence repeat markers) to assess a Douglas-fir (Pseudotsuga menziesii) seed orchard's seed crop genetic quality (2009 seed crop). The studied seed crop was produced under ambient temperature (i.e. no reproductive phenology manipulation) and pollination was augmented by pollen from within orchard's pollen donors. DNA fingerprinting of the parental population (66 parents) along with 207 gametophyte (1n) – embryo (2n) pairs of random bulk sample of seed allowed parentage (maternal and paternal) assignment and the direct assessment of pollen contamination (0.18 ± 0.027) and selfing (0.17 ± 0.025) rates as well as parental (pollen, ovule and individual parent) gametic contribution was compared to a previous year's crop (2005). The extended reproductive phenology coupled with variable within-orchard pollen availability has created opportunities for both self and foreign pollen to be successful at various times resulting in the seemingly paradoxical scenario of high selfing and gene flow. These results (2005 and 2009) allowed comparison of seed orchard's crop management practices and are expected to provide scientific foundations to effective seed crops genetic quality improvement.     

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.     

Pollination dynamics in a Douglas-fir seed orchard as revealed by pedigree reconstruction

• ? Pollination dynamics was studied in a Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seed orchard using 8 nuclear microsatellite markers and pedigree reconstruction.
• ? The seed orchard consisted of 49 parents (clones). Cone-crop management included bloom delay and supplemental mass pollination (SMP) using 12 internal and 4 external pollen donors.
• ? A random sample of 801 bulk seeds was genotyped for both haploid megagametophyte and corresponding diploid embryo.
• ? Using the parental population’s multilocus genotypes, full pedigree reconstruction generated all the information needed to estimate the maternal, paternal, and parental reproductive success, selfing, pollen contamination, and pollination success of the 4 external pollen donors.
• ? Maternal, paternal, and parental reproductive success varied with 80% of gametes being produced by 23, 45, and 37% of the orchard’s parents, respectively, resulting in a drastically reduced effective population size as compared to the census number (14 vs. 53).
• ? Selfing, pollen contamination, and aggregate SMP success (internal and external) were estimated to be 15.2, 10.4, and 15.0%, respectively.
• ? Full pedigree reconstruction was effective in unraveling the orchard’s pollination dynamics and both female and male reproductive success.

     

取食不同受害程度的马尾松对马尾松毛虫种群数量的影响

利用不同受害程度的马尾松Pinus massoniana针叶饲养马尾松毛虫Dendrolimus punctatus，研究其对马尾松毛虫生长发育的影响.结果表明：马尾松毛虫取食不同受害程度松针后，幼虫的取食量、排粪量、蛹重、对食物的消耗和利用指数、化蛹率、生殖力、发育历期和死亡率均有所不同，其中幼虫的取食量、排粪量、蛹重、对食物的消耗和利用指数、化蛹率和生殖力以取食未受害松树（针叶损失0）的最高，重度受害（针叶损失75%）次之，轻度受害（针叶损失25%）再次，中度危害（针叶损失50%）最低，而发育历期和死亡率则相反，由此可见马尾松毛虫取食不同受害程度的松针对其种群数量有明显的影响。     

普通油茶及其优树生殖生态研究

本文着重研究了普通油茶 (Camellia oleifera Abel)霜降类型及其优良单株生殖器官的形态结构及其生殖过程 ,并对油茶的生殖生态开展了一个周期 (5 78d)的研究。结果表明 ,普通油茶由花芽分化到种子成熟共需 5 0 0 d;花果相会和花粉管萌发快是油茶生殖的两个主要特点 ,油茶的生殖过程包括五个阶段 ;花期或花时不遇是影响油茶授粉乃至落花的重要原因 ,花期阴雨、低温是影响油茶授粉和落花的主要气候因素 ,水分是油脂形成期的主导因子。油茶优良单株的生殖结构比一般油茶单株表现四方面的优势。用油茶优良单株为繁殖材料 ,建立花期、花时、果熟期一致的油茶林分 ,是提高油茶单位面积产量的关键途径 ;提高集约经营度 ,提供良好的生态环境 ,是改造油茶低产林分的有效措施。     

Success rate of supplemental mass pollination in a seed orchard of Pinus sylvestris L.

Two experiments (Expt 1, Expt 2) with supplemental mass pollination (SMP) were carried out in a seed orchard of Pinus sylvestris L. in order to assess the biological and operational prospects for SMP. The success of the SMP was studied by means of allozyme markers. In Expt 1 the pollen mix was applied very close to each female strobilus. The estimated success rates for treatments varied between 66% and 84%. When supplemental pollination was done at peak receptivity, one single pollination was enough to obtain high success rates. In Expt 2 the pollen mix was dusted over clusters of receptive strobili on whole‐trees. The estimated success rates for treatments varied between 10% and 23%. The success rates improved when the trees were pollinated on more than one occasion per day during the receptive period. The overall conclusion is that SMP is practical for P. sylvestris seed orchards.     

马尾松雄球花生长发育特性及花粉采收特点

马尾松(Pinus massoniana Lamb.)在我国松属(Pinus)树种中分布最为广泛,其适应性强、生长快,是一种多功能、多用途、高效益的树种.除生产木材、采割松脂、培养茯苓等外,还可生产大宗的松花粉.     

油杉花粉个体发育与传粉过程

应用常规压片法和整体染色透明法对油杉小孢子的发生和雄配子体的发育过程进行观察,研究其传粉生物学特征.结果表明:油杉小孢子母细胞在1月26日-2月5日进行减数分裂,在减数分裂过程中胞质分裂为同时型,四分体排列方式多为左右对称型;小孢子经过连续4次有丝分裂后,于2月底形成5-细胞型的成熟花粉粒,成熟花粉由两个原叶细胞、一个不育细胞、一个精原细胞和一个管细胞组成,其中两个原叶细胞在花粉成熟时已退化;经TCC法检验,花粉生活力为92.6%;3月1日-8日为油杉传粉期,3日-5日为传粉高峰;在传粉期间,胚珠珠孔端无传粉滴产生,珠孔端耳状结构边缘粘附有花粉.虽然花粉母细胞发育表现出不同步现象,产生少数具3个或4个气囊的异形花粉,但从小孢子的发生及雄配子体发育的整个过程看,未见败育或其他异常现象,能形成大量正常的成熟花粉,因此,花粉个体发育不是影响油杉结籽率低的主要因素,而造成油杉球果结籽率低的主要原因可能在于其传粉过程.     

In vivo interspecific pollination success between Pinus radiata,P. maximinoi,P. oocarpa and P. tecunumanii

The objective of the study was to investigate in vivo interspecific pollination success between Pinus radiata, P. maximinoi, P. oocarpa and P. tecunumanii. Pinus radiata was control pollinated with pollen lots of P. maximinoi, P. oocarpa and P. tecunumanii in a P. radiata seed orchard at Karatara (Southern Cape, South Africa). Pollination success was determined by counting the number of visible ovules, pollen grains inside and outside P. radiata ovules, as well as pollen tubes visible inside P. radiata ovules. Conelets were harvested and studied at eight time intervals, including 24 h after pollination, and weekly for 7 weeks after pollination. Histology studies with a standard fixation-dehydration-embedding sequence and paraffin wax method were used to determine the number of visible pollen grains inside versus outside the ovules and number of pollen tubes. Results indicated that pollen grains did sift through the cone scales within 24 h after pollination. However, P. radiata differed significantly (time by type of cross interaction) from the other three hybrid combinations in terms of number of visible ovules, visible pollen grains inside and outside of the ovules as well as pollen tubes, confirming limited interspecific hybridisation success. Future studies need to determine the percentage of fertile ovules in cross combination as a tool in predicting pollination success.     

华北落叶松传粉生物学的研究

本文研究了华北落叶松雌雄球花在树冠上的分布和树冠周围花粉密度变化规律、传粉机制、胚珠接收花粉量及结实状况等。雌球花主要分布在树冠中上部2－4龄枝上，雄球花分布范围较广，且常与雌球花混生；树冠周围花粉密度与雄球花的分布密切相关，中部最大，依次向上下两端减少。依据雌球花形态特征的变化，可将传粉期为5个阶段，整个期间无传粉滴出现，由球被顶端大裂片的珠被毛接收花粉。最佳传粉期约为2天。胚珠平均接收4粒花粉，虽然胚珠接收的花粉数与饱满种子的产量有一定的关系，但并不是造成空粒的主要原因。此外，讨论了不同授粉方式对种子产量的重要意义。     

Low rates of pollen contamination in a Scots pine seed orchard in Sweden: the exception or the norm?

We investigated mating structure and gene flow in a clonal seed orchard of Scots pine (Pinus sylvestris L.) over three consecutive pollination seasons (2010–2012) with nine nuclear microsatellite markers. The paternity of 1991 offspring from four maternal parents was assigned to 28 candidate fathers using an exclusion procedure and a likelihood-based method implemented in the program CERVUS. Relative reproductive success was highly variable among pollen parents but consistent across years and ranged from 0.1% to 18.3%. Consequently, the seed crops’ effective number of fathers was reduced to 52.9%, 48.8%, and 45.7% of the census in the three seasons, respectively. Self-fertilization fluctuated around the orchard's expected value of 5.1%, reaching 4.05%, 7.71%, and 6.61%, respectively. Pollen contamination was estimated to be 5.64%, 7.29%, and 4.89%, respectively, after correction for cryptic gene flow. CERVUS provided similar results as the exclusion method, but estimates greatly varied depending on the input parameters, mainly the proportion of fathers sampled. These results indicate the studied seed orchard is a well-functioning production population with only minor negative effects of self-fertilization and pollen contamination on the quality of seed crops. Genotyping issues associated with microsatellites as a potential source of false paternity assignment and exclusion are discussed.     

鹅掌楸属两种植物花粉品质和花粉管生长的研究

应用ＦＤＡ荧光染色法、离体和活体培养及看护培养等方法检验了鹅掌楸属两种木本植物花粉的品质，观察了开花期间花粉品质的变化，同时也观察统计了自然和人工授粉情况下，花粉在柱头上萌发及花粉管在花柱中生长的情况。研究结果表明：在离体条件下，柱头和子房对花粉的萌发和花粉管的生长有明显的促进作用；花粉在自己或异己的柱头和胚珠上萌发均良好，是亲和的，但花粉管通过花柱的比率较低，仅２４％。与本课题前期工作的研究结果相一致。并从数量调查和活体观察两个方面，再一次证实雄配子体的发育水平是影响结籽率的重要因子之一，花柱是不亲合性反应的发生区域。     

How meso-scale pollen dispersal and its gene flow shape gene conservation decisions

Pine pollen dispersal has been well-studied over a century due to its ubiquity, robust shape and unusual longevity; this knowledge can be brought to bear on forest population genetics applications and gene conservation programs for pines and other high-latitude wind-pollinated species. Dispersal models are shifting towards meso-scale transport processes so I assert here that this shift in transport scale is important to population genetics assumptions inherent to gene conservation decision-making. Support comes from the following: (1) aerodynamic properties for pollen is more akin to spores than seeds. (2) Gradient-free dispersal is typical of pollen transported at meso-scale distances. (3) Importance of vertically uplifted pollen on meso-scale transport has been overlooked and its interaction with atmospheric processes is not yet understood. (4) A fraction of pine pollen retains its capacity for germination and seed fertilization after meso-scale transport. These findings raise the question of whether forest fragmentation aligns with genetics theory of small populations; this question shapes ex situ and in situ collections. The shift to meso-scale transport of pine pollen can re-shape forest gene conservation decision-making about ex situ and in situ collection strategies.     

Mating patterns and pollen dispersal in four contrasting wild cherry populations (Prunus avium L.)

Although pollen dispersal has been extensively studied in trees, parameters influencing between-population variation are still poorly understood. In this study, we conducted paternity analyses on open-pollinated seeds in four natural populations of wild cherry (Prunus avium) with contrasting density and clonal propagation, using eight microsatellite loci and one self-incompatibility system locus. We also measured four quantitative traits and spatial positions as potential correlates of reproductive success. Levels of polyandry differed among populations and 30% of the seed families exhibited unequal paternal contributions, suggesting variation in reproductive success rather than variation in mate availability. Mating occurred preferentially among neighbours in all populations, suggesting that it is a common pattern in wild cherry and probably results from pollinator behaviour. Paternal success was positively correlated with diameter at breast height, as indicated in previous studies and tree dominance only resulted in higher paternal success in low density plots. Mating patterns were thus also affected by both density and tree size. Large-scale studies are needed to disentangle relative influences of these factors on the mating system and pollination success.     

宝兴百合花柱偏转促进延迟自交试验研究

2011年7~8月对云南香格里拉高山植物园的宝兴百合(Lilium duchartrei Franch.)种群进行了传粉生物学研究实验,以期找到这种花朵开放初期雌雄异位的植物传粉生物学特点。结果表明,记录到的178次昆虫对宝兴百合的访问中能触碰到柱头的仅有蝴蝶类1次,有效传粉者传粉频率严重不足;单花期5.5~6.5 d,花开放的前3天,柱头远离花药,从花开放的第二天起花柱缓慢向上偏转,第四天将柱头从远离花药位置送到刚好可以接受自花花粉的位置,利于早期没有昆虫传粉的花朵延迟自交,自花授粉发生在花期末期,自交之前仍然保持异交传粉机制;自然状态(92.3%)及完全套袋处理(90%)的结实率接近,说明宝兴百合传粉存在花柱偏转促进的延迟自交,但2种处理的每果实种子数自然状态(121)显著高于完全套袋处理(78),又说明由昆虫传粉的异交更利于宝兴百合结实。延迟自交避免了自交与异交竞争造成的花粉或者种子的折损,为宝兴百合在有效访花昆虫不足导致异交失败时提供繁殖保障。     

The natural regeneration pattern and process of Korean pine population under natural Poplar-birch forest in Xiaoxing’an Mountains

According to a great deal of field investigation and detailed indoor analysis, the natural regeneration pattern and process of Korean pine population under natural Poplar-birch forest has been uncovered. The results show that the regeneration quantity and quality of Korean pine population under mountainous Poplar-birch forest and mountainous white birch forest are better than those under valley moss-grass White Birch forest and virgin broad-leaved Korean pine forest. Korean pine population shows aggregated distribution, the mass-occurrence period of Korean pine population is behind the mass-occurrence period of Poplar-birch population. Various aged Korean pines growth is affected by different stand structure factors. Man-made lighting tending can accelerate the regeneration process and increase the regeneration quality of Korean pine population. It is feasible to set up regeneration recombination of upper layer White Birch and lower layer Korean pines.     

Belowground interactions for water between trees and grasses in a temperate semiarid agroforestry system

A fundamental hypothesis of agroforestry is the complementary use of soil resources. However, productivity of many agroforestry systems has been lower than expected due to net competition for water, highlighting the need for a mechanistic understanding of belowground interactions. The goal of this study was to examine root–root interactions for water in a temperate semiarid agroforestry system, based on ponderosa pines and a Patagonian grass. The hypotheses were: (a) A greater proportion of water uptake by pines is from deeper soil layers when they are growing with grasses than when they are growing alone; (b) Growth of grasses is improved by the use of water hydraulically lifted by pines. We used stable isotopes of O to analyze water sources of plants, and we measured sapflow direction in pine roots and continuous soil water content with a very sensitive system. We also installed barriers to isolate the roots of a set of grasses from pine roots, in which we measured water status, relative growth and water sources, comparing to control plants. The results indicated that pines and grasses show some complementary in the use of soil water, and that pines in agroforestry systems use less shallow water than pines in monoculture. We found evidence of hydraulic lift, but contradicting results were obtained comparing growth and isotope results of the root isolation experiment. Therefore, we could not reject nor accept that grasses use water that is hydraulically lifted by the pines, or that this results in a positive effect on grass growth. This information may contribute to understand the complex and variable belowground interactions in temperate agroforestry.