Open-pollinated, interspecific F1 hybrid seed orchards of Leucaena: I. Rootstock effects

Seed orchards of Leucaena were established at two experimental sites in Hawaii to produce either triploid or tetraploid interspecific hybrid seed by interplanting clonal self incompatible diploid (L. diversifolia, L. esculenta) or tetraploid (L. pallida) species with self- compatible L. leucocephala lines and the F1 hybrid of L. diversifolia (2n = 4x) × L. leucocephala. The seed parents were grafted onto three seedling rootstocks (L. diversifolia (2n = 4x), shrubby L. leucocephala, and giant L. leucocephala) to compare rootstock effects on scion growth. This paper reports on the effects of rootstock on scion growth during the first year based on total truck cross sectional area, height growth, and crown habit. In the triploid seed orchard, rootstock significantly affected the growth of four of the six clones, while in the tetraploid orchard the growth of two of the five clones was significantly affected by rootstock. Crown habit was independent of rootstock. Our study suggests that rootstock effects depend on the specific scion and rootstock combination, and site. This revised version was published online in June 2006 with corrections to the Cover Date.     

Production of hybrid seed in a seed orchard of two larch species,Larix sibirica and Larix decidua

The larch seed orchard studied is intended to produce interspecific hybrid seed between Larix sibirica and Larix decidua. Only the L. sibirica clone (half of the grafts) in the orchard was intended to act as the seed parent, and the six L. decidua clones are pollinators. The proportion of hybrid seed produced was determined using Mdh3 locus as a genetic marker. The mean percentage of hybrid seed produced by the grafts of L. decidua was 93, but that of the hybrid seed produced by L. sibirica was only 18. The results can be explained by differences in flowering abundance. The grafts of L. sibirica produced several times more pollen than those of L. decidua. Contrary to the original plans, L. decidua should be used as the seed parent and L. sibirica only as the pollen parent in the studied seed orchard.     

Forage yield and compositional analysis of Leucaena species and hybrids adapted to cool sites

Four species of Leucaena (L. leucocephala, L. diversifolia 2n and 4n, andL. pallida) and three interspecific hybrids KX1 (L. diversifolia × L. pallida), KX2 (L. leucocephala × L. pallida), and KX3 (L. leucocephala × L. pallida) were evaluated for forage yield on a cool upland site at the Mealani Research Station on the island of Hawaii (900 m elevation). Two-month-old seedlings were planted at a density equivalent to 40,000 trees/ha and coppiced (harvested) every six months for a two-year period. Leaf material collected from the plots was oven-dried, finely ground, and evaluated for forage components and digestibility.Two Holstein steers fitted with cannulae in the rumen and proximal duodenum were used to conduct a replicated trial to examine rumen and post-rumen dry matter and crude protein digestibility of leaf material. Nylon bags containing leaf material from K636 (L. leucocephala), KX2 (L. leucocephala × L. pallida), or K376 (L. pallida) were suspended in the rumen for periods of 0, 4, 8, 12, 24, 36, and 48 hours. Weight loss was measured and nitrogen content of each sample was determined using a macro-Kjeldahl procedure. Subsequently, rumen-incubated material was placed in nylon bags, inserted into the duodenal cannulae of the two steers and collected in the feces for periods of 24 h after insertion.Results indicate that varieties and hybrids of Leucaena are adapted to cool sites and produce good yields of high quality forage. No significant difference was found in either ruminal or intestinal dry matter or crude protein disappearance between K636 and KX2 whereas K376 was significantly lower for both values. Nearly 40% of dietary crude protein in the K636 and KX2 leaf material escaped ruminal digestion whereas nearly 70% was non-digested for K376. For K636 and KX2, intestinal availability of that protein which escaped ruminal digestion was nearly 50% whereas only 20% of ruminal escape protein of K376 was available in the intestines. Because KX2 produced higher forage yields, plantings on cool sites would favor the use of KX2 over K636 if animal acceptability and rumen retention of dry matter are not problems.     

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.     

Pitch canker ratings of longleaf pine clones correlate with Fusarium circinatum infestation of seeds and seedling mortality in containers

The incidence and severity of pitch canker was rated among 16 clones of mature longleaf pine (Pinus palustris) in a seed orchard and cones were collected from five of those clones across the range of pitch canker disease ratings. Seed were extracted and commercially processed by clone and the percentages contaminated with Fusarium circinatum determined. Seed from each clone were sown either without treatment or with one of three fungicide treatments in a soil‐less mix at a commercial container nursery to evaluate the effects of F. circinatum (syn. F. subglutinans) on seed and seedling survival. The percent of seed with F. circinatum correlated with the pitch canker ratings in the orchard for the year of collection (r = 0.88, p = 0.05) and, when sown without fungicide treatment, with the number of seedlings produced (r = ?0.94, p = 0.01) and with seedling mortality after germination (r = 0.92, p = 0.02). The same orchard clones were more symptomatic of pitch canker through three annual surveys. Fusarium circinatum was isolated from a higher percent of seed from more symptomatic clones and a lower percent of their seed produced plantable seedlings. We propose that removing such clones from seed collections should reduce seedling mortality at the nursery.     

火炬松种子园无性系花粉产量变异研究

为探讨火炬松(Pinus taeda)无性系花粉产量的变异,在火炬松1.5代种子园选出20个无性系,每无性系5株,共100株火炬松作为研究材料,调查胸径、树高、冠幅和花粉量,通过对比单株花粉量、单位面积花粉量和单位面积有效花粉量来筛选高花粉产量的无性系.结果表明,单株花粉量与冠幅呈显著正相关(P<0.05),单株花粉量最高的是243无性系,为1302.55 g;单位面积花粉产量最高的是201无性系,为36.70 g/m2;单位面积有效花粉量最高的是243无性系,为34.14 g/m2;花粉活力最大的是298无性系,为100%.火炬松种子园的花粉产量变异主要来源于无性系间,最终筛选出243、201和298无性系3个优良无性系.     

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.     

Balancing seed yield and breeding value in clonal seed orchards

Seed orchards should produce seeds that are both abundant and of high genetic value. This study suggests methods to achieve such a compromise and study their efficiency. The methods were applied on data obtained from 41 seed orchard clones of Scots pine from mid-Sweden. The value of the seed orchard crop was set as a function of its breeding value, the amount of seeds produced and their gene diversity, measured as the effective number of clones. The proportion of ramets of different clones that maximized this value was regarded as the optimum for deployment of the clones in a seed orchard. The results were compared with truncation selection for breeding value, truncation selection for clone benefit (the product of seed production and breeding value) and linear deployment (where ramets are deployed linearly in relation to breeding value). The influence of two parameters was studied: the relative importance of breeding value for seed value and the size of the penalty for reducing the value of the seed crop with respect to lost gene diversity. The conventional wisdom is to select the clones with the highest breeding values, but that turned out to be the most inferior alternative studied. Clone benefit truncation provided a good approximation to optimal benefit for cases, where the effective number was low and dependence of breeding value limited.     

Growth, cold-hardiness, protein content, and digestibility of 70 Leucaena seedlots on three sites in Texas, USA

Seventy seedlots of Leucaena leucocephala, L. pallida, L. diversifolia, L. retusa, L. esculenta, L. confertifolia, L. greggii and L. pulverulenta and various hybrid combinations were examined for survival to freezing weather, biomass production, leaf protein content and in vitro dry matter digestibility in Texas. Three sites along a 600 km north-south transect were chosen to provide a wide range in exposure to freezing weather. The minimum temperature on the warmer Three Rivers site was −7 °C while the minimum temperatures on the colder Vernon site was −14 °C. At the Three Rivers site where −7 °C temperatures occurred and where forage production was a priority and some winter frost damage was not a problem, L. leucocephala K636 and hybrids containing L. diversifolia and L. pallida have the most promise. If total lack of damage to −7 °C is essential, species such as L. retusa, L. pulverulenta (seedlot 61, 62) and hybrids such as L. retusa × L. greggii (53) and L. retusa × L. pulverulenta (55) have the greatest potential. This study examined only one family from each of the cold-hardy species as a parent in the promising hybrids. As extensive intraspecific genetic variation is present within these leucaena species there is great potential to find hybrids with much better combinations of cold hardiness, forage production, leaf protein and dry matter digestibility. This revised version was published online in June 2006 with corrections to the Cover Date.     

Mating patterns in an indoor miniature <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> seed orchard as revealed by microsatellite markers

To evaluate the practical application of an indoor seed orchard, we compared the seed production, final germination rates and mating patterns of potted Cryptomeria japonica D. Don clones in two miniature seed orchards: one in a glasshouse and the other outdoors. There was no statistically significant difference in total seed production between the orchards (P = 0.275, ANOVA). However, the final germination rate of seeds produced in the indoor orchard was significantly lower than that of seeds produced in the outdoor orchard (P < 0.01, Wilcoxon test). The average self-fertilization rate was higher in the indoor orchard (27.2%) than in the outdoor orchard (5.6%), and this parameter was strongly associated with the reduced final germination rates of the seeds harvested in the glasshouse. Pollen contamination was considerably lower in the indoor seed orchard (48.9% in the outdoor seed orchard and 4.4% in the indoor seed orchard). χ² tests indicated that the paternal contributions of each constituent clone differed significantly in both orchards (P < 0.001 in both cases). The strong variations in paternal contributions among indoor orchard clones was significantly correlated with their respective pollen production capacities (Spearman ρ = 0.693, P < 0.001). To reduce the high self-fertilization rates, we recommend the use of orchard clones with similar pollen production capacity in indoor seed orchards.     

Elevated temperature during reproductive development affects cone traits and progeny performance in Picea glauca x engelmannii complex

Two temperature regimes were applied during reproductive development of seed and pollen cones of interior spruce (Picea glauca (Moench) Voss and Picea engelmannii (Parry) complex) to determine temperature effects on the adaptive traits of progeny. In Experiment 1, identical crosses were made on potted interior spruce using untreated pollen followed by exposure to a day/night temperature of 22/8 or 14/8 degrees C with a 12-h photoperiod during the stages of reproductive development from post-pollination to early embryo development. Frost hardiness and growth of progeny from seed produced in the two temperature treatments were measured over a 4-year period. Elevated temperature significantly affected both seed-cone development and the adaptive properties of the progeny. Seed cones exposed to the 22/8 degrees C treatment reached the early embryo stage in 53 days versus 92 days in the 14/8 degrees C treatment. Seed yields, cotyledon emergence and percent germination were also significantly enhanced by the 22/8 degrees C treatment. Progeny from seed produced in the higher temperature treatment showed significantly reduced spring and fall frost hardiness, but the elevated temperature treatment had no significant effects on time of bud burst, growth patterns or final heights. In Experiment 2, single ramets of the same clone were subjected to a day/night temperature of 20/8 or 10/8 degrees C during pollen cone development, starting from meiosis and ending at pollen shedding. The two populations of pollen were then crossed with untreated seed cones. Compared with pollen cones exposed to the 10/8 degrees C treatment, pollen cones exposed to the 20/8 degrees C treatment during development reached the shedding stage 2-4 weeks earlier, whereas pollen yields, in vitro viability and fertility (seed set) were significantly lower; however, the resulting progeny displayed no treatment differences in frost hardiness or growth after 1 year. Results suggest that seed orchard after-effects could be caused by temperature differences between orchard site and parent tree origin and that this effect acts on maternal development. Gametophytic (pollen or megagametophyte or both) and early embryo (sporophytic) selection are possible mechanisms that may explain the observed results. Although the effects are biologically significant, they are relatively small and do not justify changes in current deployment strategies for seed orchard seed.     

Condensed tannins in Leucaena and their relation to psyllid resistance

Leucaena species and hybrids were examined to determine the relative content of condensed tannins and its relationship to susceptibility to defoliation by the Leucaena psyllid (Heteropsylla cubana). The interspecific Leucaena hybrid KX2 (L. leucocephala × L. pallida) was examined for variability of condensed tannin levels. A broad range of condensed tannin values were found which indicate potential for improved forage value through selection for KX2 trees with lower condensed tannin while maintaining psyllid resistance. A significant correlation was not found linking condensed tannins and psyllid resistance.     

Allozyme frequencies,outcrossing rate and pollen contamination in Picea abies seed orchards

Allozymes of Swedish seed orchard clones of Norway spruce (Picea abies Karst.) were studied. There were differences between gene frequencies of origins from different regions. The gene pool of the fertilizating pollen of two different orchards was different. Pollen contamination and outcrossing rates were estimated in two seed orchards. The fraction of paternal gametes which did not originate from any seed orchard clone was 10% and 17%, respectively. Based on simulation it was estimated that only a quarter of the gametes with origin outside the orchard would be detected. Estimation of multilocus outcrossing rates were 0.95 and 0.98.     

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.     

马尾松第一代改良无性系种子园建园材料选择

以福建省漳平五一国有林场马尾松第一代无性系种子园为研究群体,在对园内无性系及其子代家系测定数据分析基础上进行无性系再选择,重建改良代种子园。结果表明,子代生长量与当代无性系产籽量和雌雄球花量,在家系与无性系水平上差异极其显著,且不同程度受遗传控制,具有一定选育潜力。采用综合评定和独立淘汰相结合的方法,能实现性状兼优的无性系再选择。以一代(CK1)为对照,从160个无性系中再选择30个无性系建立的改良代种子园育种效果显著,其遗传增益估值,8年生子代材积提高10.9%,产籽量和雌、雄球花量分别提高49.1%、18.9%、7.6%;以母树林混合种(CK)为对照,2个选择阶段,8年生子代材积累计遗传增益估值18.1%。     

落叶松种间交配结实力变异和自交衰退的研究

落叶松种间杂交的结实率普遍较高，但不同种之间有一定差异，其中日本落叶松与兴安落叶松交配结实率最低。种内自由授粉和自交的对比试验显示，示粒种子产生的主要原因是自交。本文讨论了自交不孕无性系的可能利用途径。在不同交配组合下研究了华北落叶松捕捉花粉的过程，确定在传粉期花粉和雌性生殖器官存在一定的相互作用，但无特异性。传粉后约4周，珠心顶端破裂，分泌物进入珠孔道，并与珠被分泌物共同作用于花粉，使花粉向珠心移动。组织化学分析表明，从传粉到受精期间，蛋白质的分布发生区域性变化，传粉期蛋白质几乎分布于整个胚珠，而在受精期则主要集中在颈卵器之上的珠心组织中。初步确定在花粉萌发和受精期，华北落叶松中存在花粉识别与选择系统。     

Variability in seed traits and genetic divergence in a clonal seed orchard of Dalbergia sissoo Roxb.

The variations in seed and pod traits, genetic superiority and genetic divergence were evaluated for a Clonal Seed Orchard (CSO) of Dalbergia sissoo Roxb. at Bithmera, India consisting of 20 clones from different agro-climatic conditions of four northern states (Uttar Pradesh, Uttarakhand, Haryana and Rajasthan). The seeds and pods of various clones in the orchard exhibited significant variability in size, weight and other characters. Significant positive correlations were observed between seed length and seed width (p<0.05), seed length and seed thickness (p<0.01), seed length and seed weight (p<0.01), seed thickness and seed weight (p<0.01), seed length and germination value (p<0.05). The genetic parameters for seed and pod traits also showed a wide range of variations in the orchard. Heritability values were found to be over 50 percent for most of the seed and pod traits. Seed weight, seed length and seed thickness showed high heritability values coupled with maximum genetic gain for these characters. Ward’s minimum variance dendrogram of clones of D. sissoo showed three distinct clusters; cluster 1 was the largest with 12 better clones whereas cluster 2 and 3 consisting of seven moderate clones and one poor clone, respectively. Mean cluster values showed sufficient variation among the clusters for seed weight, germination value and seed length. The possible hybridization between best clones of cluster 1 to the disease resistant clone of cluster 2 (resistant against deadly Gandoderma lucidum root rot disease of D. sissoo) is also suggested for further breeding programmes of the species. The deployment of clone 194 (better performed and disease resistant) is also recommended in future plantation programmes of D. sissoo in northern India.     

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。     

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.     

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.