Using SSR markers to study the mechanism of 2n pollen formation in Populus × euramericana (Dode) Guinier and P. × popularis

? Using cytological methods and SSR DNA marker analysis, this study revealed the formation mechanisms and the genetic constitutions of the 2n pollen in Populus × euramericana (Dode) Guinier and P. × popularis.

? In P. × euramericana (Dode) Guinier, four abnormalities in microsporogenesis were observed: parallel spindle, fused spindle, tripolar spindle and premature cytokinesis. The first three can lead to first-division restitution (FDR) 2n pollen formation and the last one can form second-division restitution (SDR) 2n pollen. The SSR marker analysis results of parents and their tetraploidy filial generation confirmed that the genetic constitution of 2n pollen produced by P. × euramericana (Dode) Guinier was FDR.

? In P. × popularis, three of these abnormalities were observed: parallel spindle, fused spindle and premature cytokinesis. The SSR marker analysis results showed the genetic constitution of 2n pollen produced by P. × popularis was SDR. Natural 2n female gametes in P. × euramericana (Dode) Guinier are reported for the first time. SSR analysis indicated that natural 2n female gametes of P. × euramericana (Dode) Guinier did exist and were fertile, which could be FDR genetic constitution.

? The results from this study showed a great potential for using 2n gametes to produce polyploid poplar clones, which can be used effectively for polyploid breeding for poplar species in the section Aigeiros.

     

两种杨树树皮内生真菌多样性及优势种群动态变化

为了解健杨94(转基因抗虫杨94)、三倍体毛白杨2个杨树品种干部树皮内生真菌区系及其优势种群的季节变化情况,本研究利用组织分离法从2个杨树品种996块组织中分离内生真菌1 175株,健杨94和三倍体毛白杨分别分离612、563株.利用形态特征和分子生物学方法鉴定为15个属、35个分类单元,包括担子菌1个分类单元,子囊菌34个分类单元.2个杨树品种内生真菌优势种群包括链格孢、葡萄座腔菌、镰孢属真菌、间座壳属真菌等,其中,仅有链格孢、葡萄座腔菌和桑砖红镰孢是两品种共有的优势种群种类,而且优势种群会随季节变化而变化.在两品种的内生真菌中,链格孢、葡萄座腔菌是最为常见的优势种群.     

黑杨派基因库内无性系生长特性的遗传分析*

黑杨派基因库内无性系生长特性的遗传分析解荷锋，于中奎，陈一山，张绮纹，徐红关键词黑杨派，基因库，生长特性，遗传变异黑杨派杨树在杨属栽培种中速生性能及经济价值最高［１］，其中尤以美洲黑杨为佳。我国从长江流域到黑龙江以南，广大平原地区都适宜生长［２］。为...     

Variability in Populus leaf anatomy and morphology in relation to canopy position, biomass production, and varietal taxon

Twelve poplar (Populus) genotypes, belonging to different taxa and to the sections Aigeiros and Tacamahaca, were studied during the third growing season of the second rotation of a high density coppice culture. With the objective to highlight the relationships between leaf traits, biomass production and taxon as well as the influence of canopy position, anatomical and morphological leaf characteristics (i.e. thickness of epidermis, of palisade and spongy parenchyma layers, density and length of stomata, leaf area, specific leaf area (SLA) and nitrogen concentration) were examined for mature leaves from all genotypes and at two canopy positions (upper and lower canopy). Above ground biomass production, anatomical traits, stomatal and morphological leaf characteristics varied significantly among genotypes and between canopy positions. The spongy parenchyma layer was thicker than the palisade parenchyma layer for all genotypes and irrespective of canopy position, except for genotypes belonging to the P. deltoides × P. nigra taxon (section Aigeiros). Leaves at the upper canopy position had higher stomatal density and thicker anatomical layers than leaves at the lower canopy position. Leaf area and nitrogen concentration increased from the bottom to the top of the canopy, while SLA decreased. Positive correlations between biomass production and abaxial stomatal density, as well as between biomass production and nitrogen concentration were found. A principal component analysis (PCA) showed that genotypes belonging to the same taxon had similar anatomical characteristics, and genotypes of the same section also showed common leaf characteristics. However, Wolterson (P. nigra) differed in anatomical leaf characteristics from other genotypes belonging to the same section (section Aigeiros). Hybrids between the two sections (Aigeiros × Tacamahaca) expressed leaf characteristics intermediate between both sections, while their biomass production was low.     

黑杨三倍体 PtSIP3基因的克隆与功能分析

本研究以黑杨中林46三倍体为材料,依据前期表达谱芯片的分析结果,分离克隆得到三倍体材料中高表达基因PtSIP3,该基因的编码产物为钙调磷酸酶B(Calcineurin B-like Proteins,简称CBL)的靶蛋白。随后构建了双元表达载体pRI101-PtSIP3,并成功的转入拟南芥中。经qRT-PCR表达分析和表型观察,发现PtSIP3 在拟南芥中异源过表达会抑制胚和果荚的发育、促进根的快速伸长。推测该基因在黑杨三倍体中的表达升高,可能会参与黑杨三倍体的营养生长期的调控并促进根的发育。     

Development of Melampsora ciliata rust on nursery‐grown poplars in north‐western Himalayas

Development of leaf rust (Melampsora ciliata) in different species, hybrids and cultivars of poplar (Populus spp.) was studied in nursery‐grown plants. Five different criteria were used to assess the disease development. The mean disease index on 10 August was 9.4% which increased to 70.0% on 10 October. The lowest disease index (5.5%) was recorded in P. yunnanensis whereas the maximum was recorded in P. × euramericana‘Rubra‐Poiret’ (59.5%). The apparent infection rate per unit per day was highest in P. nigra × P. trichocarpa (9.7 × 10^–2) whereas the minimum occurred in P. maximowiczii × P. berolinensis‘Oxford’ (3.3 × 10^–2). The area under the disease progress curve was a maximum in P. × euramericana‘Rubra‐Poiret’ (26.7) and a minimum in P. yunnanensis (1.6). Complete defoliation by the first half of October occurred in P. nigra × P. trichocarpa, which also had the maximum apparent infection rate and area under the disease progress curve. Inoculum production was highest in P. nigra × P. trichocarpa and lowest in P. maximowiczii × P. berolinensis‘Oxford’.     

Induction of unreduced megaspores with high temperature during megasporogenesis in <Emphasis Type="Italic">Populus</Emphasis>

Introduction  

Triploid breeding is one of the most powerful approaches for improvement of the genus Populus L. Pollination with artificial unreduced (2n) pollen was inefficiency, owing to weak competition of 2n pollen. To induce 2n megaspores and improve the efficiency of triploid production, female buds of Populus pseudo-simonii × Populus nigra ‘Zheyin3#’ were exposed to high temperature during megasporogenesis.     

Effect of some chemicals employed in the detached leaf culture of Populus on the infection of Melampsora larici-populina

A study was made on the effect of certain chemicals and their combinations on the longevity of the leaf disks cut from Populus × euramericana‘1-488’and P. alba cv. hickeliana. Benzimidazole and gibberellic acid maintained (up to a maximum of 80 days) the leaf disks better than the others, however, the longevity was dependent on the chemical and the concentration employed. In a factorial experiment of 3 chemicals × 2 concentrations × 2 cultivars × 2 races of the fungus, it was found that. all these factors contributed significantly (P<0.05) to the experimental variance. However, differential effects of the chemicals or the concentrations on the cultivar × race interaction were not observed.     

卡特兰雌配子的发育

采用石蜡切片法观察了卡特兰雌配子的发育过程。结果表明:卡特兰在授粉前,胎座和胚珠并没有开始发育;在成功授粉后,胚珠原基开始发育;卡特兰的胚珠为倒生型,具薄珠心和2层珠被,成熟胚囊为8核;从授粉到胚囊成熟需要112 d左右;外珠被的过早发育可能是造成胚败育的原因之一。     

CONTROL TECHNIQUE OF BACTERIAL CANKER OF POPLAR

The poplar bacterial canker caused by Erwinia sp is a serious disease spreading inmost parts of Northeast China,where poplars are cultivalted.The disease seriously infects thosepoplars belonging to section Aigeiros or the intersection hybrids of section Tacamahaca and section Aigeiros and their clones.Five fine poplar cultivars resistant to the bacterial canker have been ob-tained by screening tests.They are Lz_2(Populus simonii×nigra var.italica),A_(15)(P.×sp.)613(P.×sp.),A_(102)and A_(98)(P.simonii×sp.)T following chemicals are obviously effective,such as:50％DT germicide ,40％ XF-136 and 10％ C.C.M.A.or 12.5％ ramphencol and 50,000 ppm/mlvalidamycin,with which the stems or roots can be watered.     

Factors influencing direct shoot regeneration from leaves,petioles, and plantlet roots of triploid hybrid <Emphasis Type="Italic">Populus</Emphasis> sect. <Emphasis Type="Italic">Tacamahaca</Emphasis>

Since the generation of full-sib artificial triploid families, rapid clone establishment and genetic improvements have been needed. Here, we report an in vitro method of direct shoot regeneration of a triploid hybrid poplar [(Populus simonii × P. nigra ‘Italica’) × (P. × ‘popularis’)]. Using different randomized block designs, we selected one triploid to evaluate the explant type, optimal concentrations of plant growth regulators and agar, and culture time under light or dark conditions over 60 days. The highest rate of shoot induction, 80.0%, was obtained using Murashige and Skoog (MS) medium supplemented with 0.2 mg/L benzyladenine, 0.04 mg/L naphthaleneacetic acid (NAA), and 5.5 g/L agar for the first 30 days in the dark, then 3 g/L agar for the next 30 days in light. This last medium yielded the best rate of shoot induction (6.32 shoots/explant). These three media were also used to evaluate the influence of the genotypes of the parents and hybrid triploids on regeneration. Two parents and three of the four full-sib triploids were regenerated successfully; different genotypes and explant types significantly affected the rate of shoot induction and average number of shoots. Leaves but not petioles were a suitable explant. One genotype produced the highest rate of shoot induction of 96.67%. Half-strength MS medium supplemented with 0.2 mg/L indole butyric acid and 0.04 mg/L NAA was the most effective for rooting; rooting rate was 96.67%, survival rate of transplants was 73.33%, and rooting frequency surpassed 85% for each genotype. Overall, this in vitro regeneration system will be useful for the propagation and genetic modification of triploid poplars.     

Growth analysis of the competition–density effect in non-self-thinning Populus deltoides and Populus?×?euramericana plantations

The competition–density (C–D) effect for non-self-thinning Populus deltoides and Populus × euramericana plantations from 3 to 9 years was analyzed using the reciprocal equation of the C–D effect. The C–D effect was well described by the reciprocal equation, and with the progress of time the C–D curve, on logarithmic coordinates, of the P. × euramericana plantations shifted upward faster than that of the P. deltoides plantations. With increasing physical time t, the biological time τ, i.e. the integral from zero to t of the coefficient of growth λ(t) in the general logistic curve with respect to t, increased rapidly during early growth stages and the increases in τ gradually became slow during later growth stages. This trend was more evident in the P. deltoides plantations than in the P. × euramericana plantations. The coefficients A and B included in the reciprocal equation were calculated at each growth stage. With increasing τ, the coefficient A, the reciprocal of which means the asymptote of yield (=wρ) at a given growth stage, increased abruptly to a maximum value and then tended to decrease gradually to a constant level. On the other hand, the coefficient B, the reciprocal of which means the asymptote of mean stem volume at a given growth stage, decreased exponentially and tended to be close to zero with increasing τ. The λ(t) decreased with increasing stand age, whereas the final yield Y(t) defined as W(t) ρ, where W(t) is the asymptote of w in the general logistic growth curve, increased gradually with increasing stand age. The differences in coefficients A, B, and λ(t) between the two species were reported.     

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.     

川榛柱头可授性及花粉管生长特性的研究

以川榛为实验材料,以平榛和平欧杂种榛为对照,采用荧光显微法观察不同环境条件、不同授粉方式、不同花序形态、不同授粉时间柱头上花粉的附着数量、花粉萌发率和花粉管的生长特性,以期确定川榛最佳可授期和授粉方式,掌握花粉萌发所需的适宜温湿度。结果表明:川榛不同花期柱头可授性大小为:盛花期末花期露红期;川榛疏散状花序的可授性优于直簇状,平刷授粉效率优于点授;在加双层湿滤纸的培养皿高湿度条件下,花粉管萌发和生长速度先于温室和田间条件,说明湿润温和的环境更利于川榛花粉的附着、萌发和花粉管的生长;环境温湿度和种质对花粉管萌发具有重要影响,花粉管的生长速度受环境温湿度的影响较大,与种质无关。     

黑杨DNA甲基转移酶基因片段的分离及表达分析

利用同源序列克隆技术,分离了三倍体黑杨中4类(MET、CMT、DRM、DNMT2)8个特异甲基转移酶片段,其中,5个片段与二倍体同源性达到100%,3个片段发生了剪切变化。通过实时定量PCR技术检测8个甲基转移酶在不同倍性、不同部位、不同生长时期间表达模式的差异,通过对5个不同部位基因表达的检测,表明不同倍性黑杨在相同的部位可能由不同种类的甲基转移酶参与主要的甲基化调控。通过分析植株从分化早期的茎尖到分化晚期的叶和茎整个生长过程中基因表达的情况,发现随着时间的推移,MET家族 (PnD1和PnD2) 基因可能是拮抗调节,CMT (PnD3和PnD4) 家族基因可能是协同调节;而隶属于DNMT2家族的PnD6基因在各部位的表达都比较弱,唯独三倍体茎尖有很高的表达。由于茎尖是生长旺盛的部位,PnD6有可能是影响三倍体速生的重要基因。这些结果可能暗示,DNA甲基转移酶基因可能参与了黑杨发育过程中叶片形态发生的过程。     

青斑蝶雌成虫内生殖器官形态与发育

[目的]青斑蝶是重要的观赏性昆虫,准确掌握和了解其内生殖器官结构和发育状态,能够为其人工培育和利用提供理论依据。[方法]通过对1 12日龄青斑蝶雌成虫内生殖系统解剖,观察内生殖器官结构组成,并测定内生殖器官特征参数,同时根据内生殖器官各组成部分特征对雌性卵巢发育进行分级,从而系统了解青斑蝶雌成虫内生殖器官形态与发育。[结果]1)雌性青斑蝶的内生殖器官由1对卵巢、1对侧输卵管、1根中输卵管、交配囊腺和受精囊组成;2)内生殖器官发育显著受到日龄影响,卵巢管长度随着日龄增长而显著生长,至10日龄最长,卵巢小管和侧输卵管直径随日龄增长而缓慢增大;3)随着日龄的增长,卵巢中成熟卵的数量先增大后减小,未成熟卵数量始终缓慢增加;4)根据卵巢的发育特征,将卵巢发育分为乳白透明期、卵黄沉积期、成熟待产期、产卵盛期、产卵末期。[结论]雌性青斑蝶内生殖器官结构组成与其它蝴蝶类似,其发育均显著受到日龄影响,卵子发育随日龄增长而逐渐成熟。雌性青斑蝶卵巢分级将为人为调控雌蝶产卵量提供理论指导。     

Development and use of new sensitive molecular tools for diagnosis and detection of Melampsora rusts on cultivated poplar

Poplar rusts due to Melampsora larici‐populina (Mlp), M. allii‐populina (Map) and M. medusae f. sp. deltoidae (Mmd) are the most serious disease in Europe on cultivated poplars, that is, Populus × euramericana and P. × interamericana hybrids. These pathogenic species can be identified by the observation of morphological characteristics of urediniospores but this method is not appropriate for high‐throughput analysis and cannot be used on other spore stages, such as aeciospores or teliospores, that are morphologically similar. The aim of this study was to develop a rapid and sensitive molecular method based on PCR amplification that was able to specifically detect these species on various hosts for routine analysis. Three primer pairs ITS‐MLP‐F/ITS‐MLP‐R, ITS‐MAP‐F/ITS‐MAP‐R and ITS‐MMD‐F/ITS‐MMD‐R were designed within the internal transcribed spacer (ITS) sequences of ribosomal DNA to target Mlp, Map and Mmd, respectively, and their specificity were confirmed on a wide range of isolates and species. ITS‐MLP‐F/ITS‐MLP‐R and ITS‐MAP‐F/ITS‐MAP‐R primers proved to be highly specific to Mlp and Map, respectively, whereas ITS‐MMD‐F/ITS‐MMD‐R cross‐reacted with DNA from M. larici‐tremulae and M. pinitorqua. However, these species are not pathogenic on cultivated poplars that all belong to sections Aigeiros and Tacamahaca of the genus Populus. Specific Mmd primers proved to be very sensitive as a positive signal could be obtained with DNA extracts from 6 target urediniospores mixed with 800 000 urediniospores of Mlp. An internal amplification control (IAC) was included to discriminate false negative results due to the potential presence of inhibitory compounds in DNA extracts. ITS‐MMD‐F/ITS‐MMD‐R primers are therefore efficient for the detection of the quarantine pathogen Mmd on samples collected on poplar or larch and are fit for use in official tests. This new PCR assay has been used in routine for ten years, and Mmd has hitherto never been detected in commercial poplar nurseries in France.     

美洲黑杨及其杂种在世界和我国杨树栽培中的地位与作用

本文介绍了美洲黑杨的天然分布, 生长状况。西欧各国对其珍贵基因资源的重视和利用, 我国引种美洲黑杨的现状及其作用等, 并对我国进一步利用美洲黑杨的基因资源, 促进我国杨树栽培的发展提出了一些看法。     

山鸡椒雌花花芽分化形态特征及碳氮营养变化

目的]了解和掌握山鸡椒雌花花芽分化的形态特征及碳氮营养规律,为山鸡椒人工栽培及杂交育种提供参考依据。[方法]采用石蜡切片法观察山鸡椒雌花花芽分化的组织解剖结构,采用生理试剂盒-分光光度法测定雌花不同分化时期的可溶性糖、淀粉、可溶性蛋白、碳氮比等碳氮营养指标。[结果]表明:(1)山鸡椒雌花花芽分化经过未分化期—花序原基分化期—苞片原基分化期—花原基分化期—花器官分化期5个时期。(2)叶片可溶性糖含量随着花芽分化的发展呈不断升高的趋势,最高可达65.07 mg·g~(-1)。叶片淀粉含量随着分化时期的推进呈先升后降的趋势,其最高值出现在苞片原基分化期,达到81.30 mg·g~(-1),最低值出现在花器官分化期,为52.19 mg·g~(-1)。(3)叶片可溶性蛋白含量在花芽前3个分化期呈持续下降趋势,从61.32 mg·g~(-1)下降到52.48 mg·g~(-1),之后基本保持稳定。叶片中的碳氮比在花芽前3个分化期呈持续上升趋势,从1.49上升至2.61,之后基本维持在较高水平。[结论]山鸡椒雌花花芽分化的内部形态特征与雄花基本一致,雌花花芽分化分为5个时期。山鸡椒雌花花芽分化过程中,叶片中可溶性糖不断升高,而可溶性蛋白下降明显,碳氮比升高且保持在较高水平。