Detecting mislabeling and identifying unique progeny in Acacia mapping population using SNP markers

Acacia hybrids offer a great potential for paper industry in Southeast Asia due to their fast growth and ability to grow on abandoned or marginal lands. Breeding Acacia hybrids with desirable traits can be achieved through marker assisted selection(MAS) breeding. To develop a MAS program requires development of linkage maps and QTL analysis. Two mapping populations were developed through interspecific hybridization for linkage mapping and QTL analysis. All seeds per pod were cultured initially to improve hybrid yield as quality and density of linkage mapping is affected by the size of the mapping population. Progenies from two mapping populations were field planted for phenotypic and genotypic evaluation at three locations in Malaysia,(1) Forest Research Institute Malaysia field station at Segamat, Johor,(2) Borneo Tree Seeds and Seedlings Supplies Sdn, Bhd.(BTS) field trial site at Bintulu, Sarawak, and(3) Asiaprima RCF field trial site at Lancang, Pahang. During field planting, mislabeling was reported at Segamat, Johor, and a similar problem was suspected for Bintulu, Sarawak. Early screening with two isozymes effectively selected hybrid progenies, and these hybrids were subsequently further confirmed by using species-specific SNPs. During field planting, clonal mislabeling was reported and later confirmed by using a small set of STMS markers. A large set of SNPs were also used to screen all ramets in both populations. A total of 65.36% mislabeled ramets were encountered in the wood density population and 60.34% in the fibre length mapping population. No interpopulation pollen contamination was detected because all ramets found their match within the same population in question.However, mislabeling was detected among ramets of the same population. Mislabeled individuals were identified and grouped as they originated from 93 pods for wood density and 53 pods for fibre length mapping populations.On average 2 meiotically unique seeds per pod(179 seeds/93 pods) for wood density and 3 meiotically unique seeds per pod(174 seeds/53 pods) for fibre length mapping population were found. A single step statistical method was used to evaluate the most informative set of SNPs that could subsequently be used for routine checks for mislabeling in multi-location field trials and for labelling superior clones to protect breeder's rights. A preliminary set of SNPs with a high degree of informativeness was selected for the mislabeling analysis in conjunction with an assignment test. Two subsets were successfully identified,i.e., 51 SNPs for wood density and 64 SNPs for fibre length mapping populations to identify all mislabeled ramets which had been previously identified. Mislabeling seems to be a common problem due to the complexity involved in the production of mapping populations. Therefore, checking for mislabeling is imperative for breeding activities and for analyses such as linkage mapping in which a correlation between genotypic and phenotypic data is determined.     

濒危植物格木天然种群的表型多样性及变异

【目的】通过对濒危植物格木在我国中心分布区种群的果荚和种子表型性状测量分析,研究其表型多样性及变异规律,结合地理气候信息探讨影响表型变异的关键因子,为格木天然种群的遗传多样性保护恢复和引种栽培提供科学依据。【方法】对8个格木天然种群114个单株的13个果荚及种子表型性状进行测量,利用巢式方差分析、多重比较、相关性分析、主成分分析、聚类分析等分析方法,探讨格木天然种群的表型多样性水平、进化和适应性潜力、表型变异规律、表型分化水平、种群聚类特征及其与地理气候因子的相关性。【结果】格木表型多样性水平中等,13个性状的Shannon-Wiener指数在1. 911 1(种形指数)～2. 103 9(果荚周长)之间,平均2. 027 8;8个种群的Shannon-Wiener指数在0. 920 9(P6)～1. 885 6(P8)之间,均值为1. 474 7。格木表型性状离散程度较轻,13个性状的变异系数在7. 544 6%(种子长)～18. 868 5%(果荚面积)之间,均值为12. 410 9%; 8个种群的变异系数在8. 852 9%(P1)～13. 984 8%(P5)之间,均值为12. 410 9%。13个表型性状的极大值为极小值的1. 395 2(种形指数)～2. 805 6(每荚种子数)倍,均值为1. 791 7。巢式方差分析表明,格木13个表型性状在种群间和种群内均存在极显著差异(P<0. 01),说明这些性状在种群间和种群内均存在丰富的变异;种群内的变异(40. 387 1%)远大于种群间(11. 948 9%)。种群平均表型分化系数为21. 857 9%,各性状的分化系数变化幅度在7. 476 5%(种形指数)～38. 674 0%(果荚宽)之间。相关性分析表明,13个表型性状之间大多存在显著或极显著相关,果荚宽度、种子周长等性状和温度指标呈负相关,与纬度呈正相关,格木在气温高、降雨量大的地区往往具有小果荚、小种子的特征,随着温度指标降低和纬度增加,格木果荚及种子有变大的趋势。格木13个表型性状可提炼为4个主成分,累积贡献率为87. 382%。8个天然群体基于欧氏距离可分为3类:第1类2个种群呈现果荚小、种子小的特征,该类表型主要出现在格木分布区的南端;第2类5个种群呈现果荚大、种子大的特征,该类表型主要出现在格木分布区的北部,仅1个种群位于分布区的南部;第3类1个种群呈现果荚小、种子少、种子大而薄的特征,该类表型出现在格木南北分布区的中间衔接部。【结论】格木的表型多样性处于中等水平,在个体上存在较为丰富的变异。在群体及物种水平上,格木表型性状离散程度较轻,各个性状分布相对集中,较为稳定,不易发生群体分化,拥有特殊表型的格木单株具有较高的保存价值。果荚及种子大小进化及适应性潜力相对较大,种形指数最不易产生变异。种群内的变异是格木表型变异的主要来源,格木的种质资源收集保存可减少种群数量,增加种群内个体数量,并优先收集有特殊表型的单株。广西省容县和龙圩区的格木种群是格木遗传多样性保护及种质资源收集保存的重点区域。年均温等温度因子是影响格木表型及分布的最主要因子。果荚及其中种子的大小是格木种群间聚类的主要依据。     

印度,孟加拉西部的Terai地区的大叶合欢预处理及标准化豆荚采集时间和豆荚长度

在孟加拉西部的Pundibari地区,研究在4个采集（豆荚）时间,3个采集豆荚长度（〈10cm,10-20cm和〉20cm）,以及进行的13种预处理对大叶合欢种苗生长和萌发率的影响。豆荚采集时间为2月19日,且没有任何预处理,大叶合欢种子萌发率为92.5%。这明确表明,在2月19日采集的种子具有非休眠,有活力和生理成熟的特性。从长度为10-20 cm豆荚剥离出的种子,2月19日播种,种苗生长良好。利用硫脲预处理种子后,种子的萌发率并没有表现出明显提高,但是当利用低浓度硫脲（100 μL&#183;L-1）预处理种子后,有利于种苗生长。     

Characterization of mean stem density,fibre length and lignin from two Acacia species and their hybrid

The objective of this study was to compare the wood properties related to wood pulp quality of two widely planted Acacia species viz.Acacia mangium Willd.and Acacia auriculiformis A.Cunn.Ex Benth.and their hybrid.Acid insoluble lignin content(Klason),mean stem density and fibre length differed considerably among the species and hybrids.A.mangium possessed a high percent of lignin content compared to A.auriculiformis and the Acacia hybrid.However,mean stem density of A.auriculiformis was higher than A.mangium and the hybrid.Fibre length of heartwood tissues was generally shorter than that of sapwood tissues.The hybrid had longer fibres than the parent species.Lignin was negatively correlated with mean stem density.Generally,the wood properties of the hybrid were superior to its parent species.The significant intraspecific variation observed for wood properties of Acacia species could be used in breeding superior hybrids combining desirable traits of the two species.Considering thedifficulty involved in accurately measuring the lignin content compared to mean stem density,selection for plants with low lignin content can be achieved by indirect selection of high mean stem density.     

Fruit infestation of Acacia tortilis (Forsk) hyne by Bruchidius andrewesi Pic. (Coleoptera: Bruchidae) in the Thar desert

Bruchidius andrewesi Pic. has been recorded as a serious pest of pods and seeds of Acacia tortilis in the Thar desert of India. Pest infestation on developing pods and its relationship with morphological traits is reported. Pod infestation varied from 5 to 19% with 5–29% infestation of seeds. Infestation of pods is directly related to infestation of seed (r = 0.72^**), and both pod and seed infestation are also directly correlated with loss in seed biomass (r = 0.79^** and R = 0.88^**). The infestation of pods starts in November and increases steady until harvest. Seeds kept in the laboratory for further studies were found to be 100% infested with B. andrewesi, as the insect multiplied faster under these conditions. The heavy infestation is damaging not only to A. tortilis but also to other leguminous trees of the desert. Bruchidius andrewesi has also been found on pods and seeds of Prosopis cineraria, an important indigenous tree of the region.     

尾叶桉与赤桉正反析因交配杂种F_1材性遗传参数的估算

[目的]通过尾叶桉与赤桉杂种F1材质性状的遗传参数估算及其分析,为桉树杂交育种材性改良亲本选配和交配设计提供理论依据。[方法]以尾叶桉与赤桉6×6正反析因交配杂种F1测定林为材料,并以相应亲本自由授粉的半同胞子代作对照。9年生时,对各小区的杂种和家系(5株小区,6次重复)选取2株平均木,在胸高1. 3 m处沿南北向钻取木芯,共984个,测定木材基本密度、纤维长、纤维宽和纤维长宽比,利用ASReml-R估算4个材质指标的特殊配合力、杂种优势、父母本单株狭义遗传力、遗传相关和表型相关系数。[结果]表明:尾叶桉与赤桉正反交的杂种木材基本密度优于对照,呈明显的杂种优势;反交组合杂种其木材基本密度优于正交组合杂种,正交组合杂种的纤维特性优于反交组合的;在正交组合杂种中,木材基本密度、纤维长和纤维长宽比的母本效应低于父本效应;而反交组合杂种中,4个材质指标的母本效应均高于父本效应。木材基本密度和纤维特性指标受中至低遗传控制。正反交组合杂种中,除纤维长宽比外,其余3个材质指标的显性效应大于加性效应。对于遗传相关,杂种的木材基本密度与纤维宽呈极显著的负相关,与纤维长宽比呈极显著的正相关,与纤维长呈不显著负相关,纤维长与纤维宽呈显著正相关。对于表型相关,除纤维宽与纤维长宽比呈不显著正相关外,其余性状间均呈极显著正相关。[结论]尾叶桉与赤桉杂种的材质性状遗传差异因亲本和交配方式而异,表明通过种间杂交和正向选择进行材质性状的遗传改良具有潜力。     

尾叶桉×细叶桉木材密度与生长的联合选择

[目的]评估Resistograph钻刺法间接测定尾叶桉×细叶桉木材密度的可靠性,检测杂交亲本对子代表型的效应以及生长与木材密度的相关,评选速生、优质的尾细桉杂种。[方法]基于10株尾叶桉与10株细叶桉不完全析因交配产生的56个杂交组合的7.5年生试验林,利用79株分析容积法与Resistograph钻刺法测定的木材密度的相关,通过方差分析检测亲本对杂种生长和木材密度的效应,结合多重比较和独立淘汰法进行材积和木材密度的联合选择。[结果]容积法与Resistograph钻刺法测定的木材密度的表型相关系数为0.52(P0.001),遗传相关系数为0.55(P0.05);树高、胸径及材积的母本间和父本间均呈极显著差异(P0.001或0.01),但母本×父本互作的效应不显著;对钻刺木材密度,父本间呈极显著差异(P0.001),母本×父本互作显著(P0.05),但母本间差异不显著;树高、胸径和材积间的表型相关和遗传相关均极显著(P0.001),其与钻刺木材密度的表型相关极显著(P0.001),但遗传相关不显著;评选出速生、木材密度较高的杂交组合14个、单株17株。[结论]Resistograph钻刺法是一种间接测定尾细桉木材密度的简便、经济和可靠的方法;母本和父本选择以及母本与父本的组配对培育速生、材质优良的尾细桉杂种均较重要;尾细桉生长与木材密度的遗传相关不显著,需要对这两类性状分别进行选择;评选的尾细桉杂交组合和单株为培育速生、优质的桉树良种提供了有用的材料。     

Correlations and clines in tree growth and wood density of <Emphasis Type="Italic">Balanites aegyptiaca</Emphasis> (L.) Delile provenances in Niger

Balanites aegyptiaca (L.) Delile provides wood and other essential products for farming and pastoralist communities in the West African Sahel. In this region, mean annual rainfall decreases in general from south to north and from west to east, so one might expect clinal variation in fitness-related traits in relation to the rainfall gradients. This paper presents results from a provenance/progeny test of 11 provenances sampled in Niger and established at one relatively dry site in Niger. The objectives were to evaluate phenotypic correlations (Pearson r) between tree growth (height, stem diameter) and basic density of the wood, and clines in growth and density at 13 years. Tree growth and wood density were positively correlated, and the correlations differed among provenances. Mean tree height and stem diameter of provenances increased, in general, from the more humid to the drier parts of the sample region. Results suggest that selection of faster-growing trees could lead to an increase in wood volume and density in the subsequent generation, especially in certain provenances; that provenances from drier areas are better adapted to drought; and that tree improvement and conservation programs should collect seeds from populations in the drier areas for planting and conservation.     

毛乌素沙地中国沙棘种群数量动态研究

沙棘不仅是优良的多用途树种 (multiplepurposetree) ,而且无性繁殖能力极强 ,能通过串根萌蘖而“独木成林”并赖以维持群落的持久性 ,在黄土高原生态环境建设和区域经济发展中起着积极的作用。我国拥有沙棘属植物资源 92万km2 ,中国沙棘 (HippophaerhamnoidesL .subsp .sinens     

Genetic evaluation of Prosopis alba (algarrobo) in Argentina for cloning elite trees

Algarrobo (Prosopis alba) is an important nitrogen-fixing tree adapted to the semiarid regions of northwestern Argentina. The tree provides fuelwood and dimensionally stable lumber for flooring and furniture; its pods that have a high sugar content are consumed by livestock and humans. P. alba has been one of the most heavily harvested species. This paper reports on the evaluation of a nine-year-old Prosopis alba progeny trial containing 57 half-sibling families of eight provenances in northwestern Argentina (of the 1,596 trees planted, 1,289 survived in 1999). Considering the multiple uses of P. alba, the selection criteria included: total biomass production (from basal diameter using regression equations), height, rate of pod production, and pod sensory characteristics. The family-narrow-sense-heritability was 0.487 for height. 0.548 for biomass production, and 0.244 for pod production. In 1998, 12 of the 1,289 trees were more than 4 m tall and had more than 1.75 kg pods per tree; and their pods had a sweet or very sweet non-astringent taste. Scions from these 12 trees were successfully grafted onto unselected P. alba rootstock. These are currently being used to produce rooted cuttings. The pod production, biomass, and height of the 12 clones ranged from 6.55 to 14.4, 1.57 to 13.5, and 1.29 to 1.70 times the respective population means. The genetic gains of 13.5%, 77%, and 147% for height, biomass, and pod production respectively are greater than genetic gains for other tree species and are probably due to the great genetic variability in the local population and the lack of any prior genetic improvement program.This revised version was published online in November 2005 with corrections to the Cover Date.     

Influence of prior land use on wood quality of Pinus radiata in New South Wales,Australia

Wood quality data from 33 sites aged between 15 and 18 years old were analysed to determine the effect of prior land use (pasture, cleared grazing land or timber) on wood density, wood stiffness, fibre length and kraft pulp yield. Sampling sites covered 6 different forest areas within the estate. Prior land use (PLU) significantly affected wood stiffness, density and fibre length but not the kraft pulping traits. In contrast, differences were found between different forest areas for the kraft pulping traits plus fibre length but not for stiffness or density.     

Genetic control of wood density and bark thickness,and their correlations with diameter,in pure and hybrid populations of Eucalyptus grandis and E. urophylla in South Africa

Tree diameter under and over bark at breast height (dbh), wood density and bark thickness were assessed on samples from control-pollinated families of Eucalyptus grandis, E. urophylla, E. grandis × E. urophylla and E. urophylla × E. grandis. The material was planted in field trials in the coastal Zululand region of South Africa. At 75 months, between three and seven of the best trees per family were felled and wood samples collected. Genetic parameters for wood density, bark thickness and bark percentage (ratio of double bark thickness to overbark diameter) and the inter-trait correlations for the different species and hybrids were calculated. Genetic parameter estimates for wood density, bark thickness and bark percentage in the E. urophylla × E. grandis hybrids showed these traits to be under total additive genetic control. This was confirmed by the intermediate hybrid means for these traits relative to those of the parental species. There was a very low correlation between dbh and wood density for the E. urophylla × E. grandis hybrids (r_G = –0.07 and r_P = 0.064). Amongst the E. urophylla families there was a moderate positive and significant phenotypic correlation between wood density and bark thickness (r_P = 0.391), and between wood density and bark percentage (r_P = 0.442).     

四川桤木木材密度和纤维特性研究

通过对四川桤木6个家系,2～9a生样本连续进行生长、材性测定,得出如下主要结论:不同树龄之间、不同家系之间木材基本密度、纤维长度、纤维长宽比、纤维含量都达到极显著差异水平。纤维长度、纤维含量、基本密度第8年、第7年、第5年以后基本稳定。4个指标两两之间均达到极显著相关,生长速度与其材性没有必然的联系。     

大青杨等天然群体幼苗基本材性变异研究

本文对８个产地大青杨天然群体，１个产地大青杨群体内不同个体，２个产地香杨群体幼苗基本材性进行了研究，测定了纤维长度、纤维宽度和基本密度。结果表明：８个大青杨群体间，１个大青杨群体内１０个个体幼苗３个材性主要指标，纤维长度、宽度和基本密度差异显著；纤维长度和基本密度的广义遗传力，大青杨天然群体间为０．２２５和０．２１７，群体内为０．４７５和０．３４５。本研究同时对大青杨群体内、群体内的个体进行了优选     

Influence of initial planting spacing and genotype on microfibril angle,wood density,fibre properties and modulus of elasticity in Pinus radiata D. Don corewood

Pinus radiata D. Don trees from six clones, grown at initial spacings of 2500 stems ha⁻¹ and 833 stems ha⁻¹ were destructively harvested. For these trees wood properties were measured on radial slices sampled at a height of 1.4 m above the ground. Relative to wide spacing, close initial stand spacing significantly reduced microfibril angle (MFA) and ring width and significantly increased dynamic modulus of elasticity (MOE), fibre length, latewood percentage and cell wall thickness. Density and fibre width were not significantly different between spacing treatments. Examination of the influence of genetic population on wood properties indicated that genotype significantly influenced MFA, MOE and ring width. The key wood properties MFA, MOE and fibre length were regressed against tree diameter, height and stem slenderness. All three wood properties were most strongly correlated with stem slenderness. Multiple regression models developed for MFA, MOE and ring width accounted for respectively 62%, 81% and 58% of the variation in these variables. The following changes occurred in sampled properties with increasing ring number: MFA and ring width declined markedly; MOE and fibre length increased markedly; latewood percentage and cell wall thickness increased slightly; and density and fibre width did not show any radial trend.     

Methods for the determination of wood properties,Kraft pulp yield and wood fibre dimensions on small wood samples.

A trial set-up with methods for sampling, treatment and analysis of small wood chips are presented in this paper, to determine important wood and fibre properties, like basic density, dry density, volume swelling of wood, Kraft pulp yield, fibre length, fibre coarseness, fibre width, lumen width and fibre wall thickness. The required time for one sample is about 1.5 man-hour, but this requires relatively larger series and trained personnel. Acceptable measurement accuracy is achieved when the volume of the wood sample is at least 1.5 ml, except that of wood volume swelling. To gain acceptable measurement accuracy for volume swelling, the wood volume should be increased to at least 3 ml, and preferably more than 5–6 ml per sample. The level of pulp yield and wood density do not show a significant effect on the measurement accuracy for fibre cross-section dimensions. Fibre coarseness, on the other hand, has a significant influence on these accuracies. A double measurement of fibre coarseness will improve the accuracy to an acceptable level. The method presented here may, together with information about trees and growth locations, form the basis for greater insight into the mechanisms involved in development of wood and fibre properties in trees, which in turn may provide better control and utilisation of wood for pulp and paper production.Abbreviations CWD cell wall density in dry wood=1500 kg/m³ - Ww dry weight of wood (kg) - Vmax green (wet) volume of wood (m³) - Vmin dry volume of wood (m³) - BD basic wood density (kg/m³) - DD dry wood density (kg/m³) - VS maximum volume swelling of wood (%) - Wp dry weight of pulp (kg) - PY pulp yield (%) - C fibre coarseness, the average weight of a unit length of fibre (g/m) - CL average chip length (mm) - CWT average cell wall thickness (m) - FW average fibre width (m) - l average native fibre length in solid wood - L chip length - lc average fibre length in wood chip (mm) - Lc length-weighted fibre length in wood chip (mm) - lw native average fibre length in wood (mm) - Lw native length-weighted fibre length in wood (mm) - LW average lumen width (m) - llw average native fibre length, length weighted, in wood - X average fibre length in chip - Xlw average fibre length, length weighted, in chip     

Determination of the cellulose and lignin content on wood fibre surfaces of eucalypts as a function of genotype and site

We compared the chemical composition of wood fibres and fibre surfaces of several eucalypt species and hybrids originating from various growth sites in South Africa. The objective was to test for differences in chemical surface composition due to genetics or site with the ultimate aim to facilitate a tailor-made supply of wood for pulping that results in an optimal blend of fibres that can be pulped together with similar yields. This, however, requires a sound knowledge of the fibre properties. The surface functionality on the single fibre level is a key property, because it determines how good inter-fibre bonding will be when paper is formed, which depends amongst other fibre properties on the amount of free hydroxyl groups that are available and therefore on the cellulose content on the fibre surface. The cellulose and lignin content on the fibre surface were determined with chemical force microscopy, a variation of atomic force microscopy. Since the general bulk composition of the fibre and the surface composition might differ, both parameters were determined. We found significant differences in the cellulose and lignin content on fibre surfaces, with regard to genotype and site, respectively. In some, but not all, cases, the surface composition of wood fibres followed the bulk composition, and differences were generally more pronounced. Differences due to genotype were significant, especially with regard to the surface lignin content—but variation due to site was also distinctly recognisable. This variation in surface functionality could be the reason why some pulpwood blends result in a lower pulp yield and different quality.     

林木遗传图谱构建和数量性状基因定位

本文简要介绍了９０年代以来利用ＲＦＬＰ和ＲＡＰＤ分子标记的构建林木的遗传连锁图谱，并应用高密度的遣传连锁图谱上的分子标记进行林木重要数量性状的ＱＴＬ作图，进而阐明了控制多基因的数目，确定ＱＴＬ在染色体上的位置，测定单个基因的作用效应，遗传连锁图谱构建和数量性状基因定位预示林木遗传育种研究将产生深刻的变革。     

Reproduction of a Robinia pseudoacacia population in a coastal Pinus thunbergii windbreak along the Kujukurihama Coast, Japan

We investigated reproduction of the clonal tree Robinia pseudoacacia in a Pinus thunbergii windbreak. Microsatellite polymorphism analysis revealed that asexual reproduction through horizontal root elongation and ramet formation was the primary mode of reproduction in this population. Germination experiments indicated that the inability of established genets to produce viable seeds inhibited sexual reproduction. The boundaries between genet areas were relatively distinct, indicating that ramets within genets spatially excluded one another. Analyses of the number of annual rings and thickness of excavated horizontal roots provided new insight regarding photosynthate transfer through horizontal roots. The number of annual rings and thickness were highest for horizontal roots near the base of ramets and decreased along portions far from ramets. This result indicates that the new horizontal root is supplied with photosynthate from the mother ramet for a few years only, and that resource translocation between neighboring ramets within the same clone ceases after several years. Hypotheses about horizontal root expansion and cessation were considered to explain the exclusive distribution pattern of the horizontal root between genets.