Histology of Diplodia pinea in diseased and latently infected Pinus nigra shoots

Diplodia tip blight is a serious disease of >30 conifer species worldwide. Symptoms are particularly severe on non‐native, two‐needled Pinus species, and typically include stunted, necrotic needles and shoots and a general decline of the tree. Latent Diplodia pinea infections occur in current‐year shoots of some symptomless pines, and in some apparently healthy current‐year shoots of diseased pines. Latent infections also occur in symptomless terminal buds. A histological approach was used to investigate the nature of latent infections in shoot stems and terminal buds of landscape Pinus nigra. Fungal colonization was compared in healthy, diseased and latently infected tissues. A nested‐PCR technique that is specific for D. pinea was used to differentiate latently infected tissues from uninfected ones. Latent D. pinea infections were localized in the outer stem cortex, usually in the vicinity of needle scales at leaf axes. In contrast, pathogenic D. pinea infections were characterized by fungal colonization throughout the shoot stem tissues, even very early in symptom development. The presence of necrophylatic periderms in two of the latently infected samples suggests that host defences play a role in the production and maintenance of latent stem infections. Latent infections of terminal buds appeared to originate from the distal bud scales of axillary buds in the terminal bud cluster, and not from the subtending shoot. Fungal tissues were never observed inside asymptomatic, PCR‐negative shoots.     

11条白桦BpSPL家族基因的生物信息学和表达分析

[目的]研究白桦中SPL转录因子的基因序列特征及其在不同时期不同组织中的表达规律。[方法]依据白桦45个转录组测序结果,共获得12条全长SPL基因,依次命名为白桦BpSPL1-BpSPL12,对其中11条BpSPL进行了生物信息学及基因表达特征分析。[结果]生物信息学分析发现,11条BpSPL均含有1个高度保守的SBP结构域,且基因长度差异较大,含有2 10个不等数目的外显子,系统进化分析发现11条白桦SPL分属于六大类SPL蛋白;qRT-PCR分析结果显示,白桦BpSPL基因在不同时期的叶、顶芽、茎、雄花序中表达变化显著,多数SPL基因在顶芽中7月5日和8月20日至9月20日时期表达较高,除BpSPL1基因外在雄花序从6月份到9月份的生长阶段中,SPL的表达水平呈现逐渐升高的总体趋势。[结论]BpSPL基因可能参与到了白桦顶芽和雄花序的生长发育过程。     

Adventitious micropropagation of mature Larix decidua using dormant versus flushed axillary vegetative buds

Tissues of winter-dormant buds were collected from mature Larix decidua, and used for adventitious shoot organogenesis. Tissues of dormant buds merely died on the cytokinin-supplemented, bud-initiation medium. In contrast, buds allowed to flush under greenhouse conditions subsequently yielded approximately two adventitious shoots per bud explant tissue. These shoots elongated normally and several rooted spontaneously. Microbial contamination of in vitro cultures was negligible using explants from buds orgininally collected in a dormant state.     

Beech leaf disease symptoms caused by newly recognized nematode subspecies Litylenchus crenatae mccannii (Anguinata) described from Fagus grandifolia in North America

Symptoms of beech leaf disease (BLD), first reported in Ohio in 2012, include interveinal greening, thickening and often chlorosis in leaves, canopy thinning and mortality. Nematodes from diseased leaves of American beech (Fagus grandifolia) sent by the Ohio Department of Agriculture to the USDA, Beltsville, MD in autumn 2017 were identified as the first recorded North American population of Litylenchus crenatae (Nematology, 21, 2019, 5), originally described from Japan. This and other populations from Ohio, Pennsylvania and the neighbouring province of Ontario, Canada showed some differences in morphometric averages among females compared to the Japanese population. Ribosomal DNA marker sequences were nearly identical to the population from Japan. A sequence for the COI marker was also generated, although it was not available from the Japanese population. The nematode was not encountered in Fagus crenata (its host in Japan) living among nematode‐infested Fagus grandifolia in the Holden Arboretum, nor has L. crenatae been reported in American beech in Japan. The morphological and host range differences in North American populations are nomenclaturally distinguished as L. crenatae mccannii ssp. n. from the population in Japan. Low‐temperature scanning electron microscopy (LT‐SEM) demonstrated five lip annules and a highly flexible cuticle. Females, juveniles and eggs were imaged within buds with a Hirox Digital microscope and an LT‐SEM. Nematodes swarmed to the tips of freshly cut beech buds, but explants could not be maintained. Inoculation of fresh nematodes from infested leaves or buds to buds or leaves of F. grandifolia seedlings resulted in BLD leaf symptoms. Injuring dormant buds prior to nematode application, in fall or spring, promoted the most reliable symptom expression. The biogeography and physiology of anguinid nematode leaf galling, and potential co‐factors and transmission are discussed.     

Field performance of Pinus contorta trees propagated vegetatively via adventitious buds

Abstract

Pinus contorta plants regenerated from adventitious buds induced on zygotic embryos were planted in a field trial where growth, straightness and flowering were followed over 7 years. Seedlings were taller than the plants derived from adventitious buds at the start of the field trial. Adventitious plants did not catch up with seedlings in height after 7 years in the field, but the relative height growth was equal. The higher frequency of plagiotropic growth that was displayed by adventitious plants the first year in the field was not reflected by differences in the frequency of basal sweeps 6 years later. However, the frequency of crooked stems was comparatively high but similar among the plant types. Flowering was more abundant for the seedlings than for adventitious plants. Taken together, the results show that P. contorta can be cloned via adventitious buds from zygotic embryos, and that the resulting plants show satisfactory growth for inclusion in breeding.     

The emergence of beech leaf disease in Ohio: Probing the plant microbiome in search of the cause

American beech (Fagus grandifolia) is the target of a newly emerging disease in North America called beech leaf disease (BLD) that affects and disfigures leaves and which can lead to tree mortality. Beech leaf disease may be caused by a newly recognized subspecies of the anguinid nematode Litylenchus crenatae subsp. mccannii, but the associations of this nematode with bacterial and fungal taxa are unknown. We examined microbial communities associated with beech leaves affected by BLD in a 16‐year‐old American beech plantation using molecular methods. We detected L. crenatae subsp. mccannii in anywhere from 45% to 90% of leaves depending on the degree of visual BLD symptoms. Approximately 37% of asymptomatic leaves contained L. crenatae subsp. mccannii, whereas 90% of buds associated with symptomatic leaves contained L. crenatae subsp. mccannii. We found that fungal communities on leaves and buds were unaffected by BLD, but bud and leaves had significantly different fungal communities. Bacterial communities on buds also were unaffected by BLD, but bacterial communities were significantly different between symptomatic and asymptomatic leaves suggesting that the nematode could be altering the community of bacteria on the leaves. Clone libraries indicate that Wolbachia, an intracellular endosymbiont of arthropods, was found only on symptomatic leaves and buds associated with either symptomatic or asymptomatic leaves. In addition, only symptomatic leaves contained taxa in the genus Mucilaginibacter, which previous studies suggest could produce exopolysaccharides. These bacterial taxa could represent a marker for the vector of L. crenatae subsp. mccannii that enables spread between trees and a possible endosymbiont that could facilitate nematode feeding and establishment on nematode infested leaves. Our results are the first to examine changes to the leaf microbiome of this newly emerging pest and may aid identification of mechanisms associated with the spread and success of L. crenatae subsp. mccanni.     

Foliar nematode,Litylenchus crenatae ssp. mccannii,population dynamics in leaves and buds of beech leaf disease‐affected trees in Canada and the US

A foliar nematode, Litylenchus crenatae ssp. mccannii, is associated with beech leaf disease (BLD) symptoms. Information about the types of tissues parasitized and how nematode populations fluctuate in these tissues over time is needed to improve surveys as well as understand the nematodes role in BLD. During this study, the nematode was detected throughout the known range of BLD by researchers at both Canadian and US institutions using a modified pan method to extract nematodes. Monthly collections of symptomatic and asymptomatic leaves during the growing season (May–October), and leaves and buds between growing seasons (November–March), revealed that nematodes were present in all tissue types. Progressively larger numbers of nematodes were detected in symptomatic leaves from Ohio and Ontario, with the greatest detections at the end of the growing season. Smaller numbers of nematodes were detected in asymptomatic leaves from BLD‐infected trees, typically at the end of the growing season. The nematode was detected overwintering in buds and detached leaves. The discovery of small numbers of nematodes in detached leaves at one location before BLD was detected indicates that nematodes may have been present before disease symptoms were expressed. Other nematodes, Plectus and Aphelenchoides spp., were infrequently detected in small numbers. Our findings support the involvement of the nematode in BLD and indicate that symptoms develop only when certain requirements, such as infection of buds, are met. We also found that the nematode can be reliably detected in buds and leaves using the modified pan extraction method.     

The effect of bagging branches on levels of endophytic fungal infection in Japanese beech leaves

The species composition of the endophytic mycobiota in leaves of Japanese beech trees (Fagus crenata) and the sources for leaf infections were studied in a forest reserve situated in central eastern Honshu, Japan. To clarify the mechanism of infection of leaves, half of the branches were covered with polyethylene bags and species composition and levels of endophytic fungal infection were then compared with those of unbagged controls. Isolations were carried out from the leaves, petioles, and current‐year twigs of both, bagged and unbagged branches. Additionally, species composition was detected in overwintered terminal buds of beech trees and in the leaves of potted seedlings that had been placed in the field in different seasons. The species assemblage of the unbagged leaves, petioles, and current‐year twigs was dominated by Mycosphaerella buna, Ascochyta fagi, Periconiella sp., and Tritirachium sp. Other frequently recovered species were Xylaria sp., Phomopsis sp., and Tubakia dryina. Mycosphaerella buna and A. fagi were never isolated from leaves on bagged branches. A. fagi was, however, detected on both bagged and unbagged petioles and current‐year twigs at comparatively low isolation frequencies. The detection of Periconiella sp. on all occasions in both bagged and unbagged leaves was a characteristic feature that differs from those of the other three dominant endophytic fungi. The fungus was also detected without significant differences in bagged and unbagged petioles and current‐year twigs on most sampling dates. Furthermore, Periconiella sp. was isolated from immature twigs inside the bud scales. Tritirachium sp. was frequently detected in unbagged leaves and petioles and in both bagged and unbagged current‐year twigs, and rarely in bagged leaves and petioles, but was never recovered from terminal buds. The results of the potted seedling experiments revealed that all four dominant species had airborne inocula. The infection of leaves by M. buna occurs exclusively by airborne propagules, i.e. ascospores in spring and conidia in autumn. In Periconiella sp. hyphal growth of the fungus from immature twigs inside the buds into the leaf tissues was suggested in addition to infection by airborne inocula. Tritirachium sp. hyphae were suggested to grow from previous‐ to current‐year twigs. Ascochyta fagi was present in the outermost scales of overwintered terminal buds, but no systemic growth of the fungus into the petioles and current‐year twigs was observed. Our technique of covering the branches before new leaves unfolded was effective in preventing infection by airborne inocula of endophytic fungi.     

Strawberry blossom weevil Anthonomus rubi Herbst (Col.: Curculionidae): relationships between bud damage, weevil density, insecticide use, and yield

The activity of overwintered strawberry blossom weevils, Anthonomus rubi Herbst, measured as cut buds and adult weevils, was recorded weekly in sprayed and unsprayed quadrats in young and old strawberry plantings. The study was done in nine ‘Korona’ fields in a district with high A. rubi density, in the southeast of Norway, in 2000. During the period of study, the cumulative number of cut buds and adult weevils found by tapping whole plants above a bowl were highly correlated. An increase in sampled weevils was followed by an increase in cut buds 1–2 weeks later. Thus, sampling adults has potential for predicting the bud damage. In five of the six fields where yield was measured, a negative relationship between cut buds and the saleable yield was found. The effect of spraying was relatively low: A. rubi activity was reduced by 21% and the saleable yield increased by 16%. We conclude that present spraying practices are not very efficient in preventing A. rubi damage in the area studied.     

白刺幼苗芽库及枝系构型对不同氮添加水平的响应

[目的]研究不同氮添加梯度下白刺幼苗芽库组成及空间结构的差异,探讨了幼苗的养分限制、氮素利用以及根系和插穗的品质与芽库的关联,阐明了白刺芽库响应氮素有效性所采取的适应性策略。[方法]本研究通过盆栽控制实验,对唐古特白刺幼苗的芽库特征进行了测量分析。[结果]氮添加显著增加了白刺幼苗芽和营养枝数量,显著降低了休眠芽和休眠枝数量,并对二级枝和三级枝出芽率具有明显促进作用;随着氮添加量的递增,营养枝有从基部向顶端移动趋势;氮平衡指数与白刺幼苗出芽强度、分枝强度、二级枝和三级枝出芽率均呈二次非线性正相关关系;植株氮含量、氮积累量和地下部分形态特征分别与芽和营养枝数量呈正相关关系,与休眠芽和休眠枝数量呈负相关关系。[结论]氮添加影响幼苗芽库容量。随着氮添加水平的提高,各指标多数在N3和N4处理达到最大值后逐渐降低,因此6个氮添加水平中36 mmol·L-1和48 mmol·L-1是白刺幼苗最适氮添加量。氮添加对幼苗芽库内组分的相对位置产生了不同影响趋势,所以白刺幼苗可以通过改变芽库容量和空间分布来响应养分有效性变化。     

Detection of the endophyte Discula umbrinella in buds and twigs of Fagus sylvatica

Endophytic fungi have been isolated from buds and twigs of beech trees collected at four different sites in Switzerland. Discula umbrinella. was recovered at high frequencies from the bud scales and the twig pieces contiguous to the buds, but was virtually absent from the rolled up leaves enclosed by the scales. In addition to infection by air-borne inoculum, thalli of D. umbrinella may grow from the twigs into the leaf tissues.     

Plant regeneration in <Emphasis Type="Italic">Actinidia polygama</Emphasis> Miq. by leaf,stem, and petiole culture with zeatin,and from stem-derived calli on low-sucrose medium

We examined the effect of zeatin on the formation of shoot buds from explants and callus tissues derived from stem segments of Actinidia polygama Miq. (matatabi or silver vine). Stem and petiole segments cultured on combined broad-leaved tree medium and woody plant medium (BW medium) supplemented with zeatin for 40 days formed many shoot buds. Callus tissues derived from the stem segments and subcultured on BW medium supplemented with 6-benzyladenine and 1-naphthaleneacetic acid formed shoot buds when the medium contained 13.7µM zeatin. BW medium containing low concentrations of sucrose strongly induced the formation of shoot buds from the callus.     

Studies on the damage potential of Closterotomus (=Calocoris) trivialis and Aphanosoma italicum (Hemiptera: Miridae) on citrus

This study aimed to investigate the damage potential of two hemipteran species that are commonly recorded in citrus orchards of the eastern Mediterranean region. These were Closterotomus trivialis Costa and Aphanosoma italicum Costa (Hemiptera: Miridae). Shoots on sweet orange trees were enclosed in muslin cages into each of which eight nymphs or eight adults were introduced not only in the early development of the buds but also in the different stages of the flowering period in 2008 (i.e., 6 and 19 March, 10 April and 2 May). In the experiments, adults of C. trivialis, and nymphs and adults of A. italicum were used. According to the results, in a few cases these insects caused a significant reduction in the number of buds or flowers and fruits dropped in comparison to the controls. However, the number of fruits that finally remained on the trees was not significantly affected. Therefore, the damage potential of these mirids on the fruit production of citrus was not shown to be significant under our experimental conditions.     

Clonal micropropagation of mature Larix

Adventitious buds were developed in vitro from stems of elongating short shoots of mature European larch (Larix decidua Mill.). Tissue culture propagule populations thus generated, were similarly remultiplied in vitro. Elongation and rooting were routine. No vitreous or otherwise abnormal shoots were noted.     

The effect of late season damage on the apical bud development of Scots pine seedlings

The effect of Lygus rugulipennis Popp. feeding and artificial damage on the apical bud of Scots pine (Pinus sylvestris L.) late in the growing season was studied on two-year-old seedlings. Lygus feeding in late July and August caused inhibition of bud formation in the subsequent year's shoots, which led to loss of apical dominance and formation of interfascicular buds. Mechanical damage caused by piercing the apical bud with a needle in July, August and October produced scars and malformations on subsequent year's shoots and buds, but did not inhibit bud formation. Damage to the apical meristem could not be detected visually on the dormant bud before shoot elongation.     

Histology of witches' brooms on Betula pubescens

Witches' brooms caused by Taphrina betulina on Betula pubescens formed from the development of large numbers of axillary shoots on infected buds. Broom shoots showed vigorous growth during the first season, but died in the second or third year after initiation. Occasional adventitious root production was noted at the base of broom shoots. Cortex and secondary phloem tissues of the central gall-like structure of the brooms expanded. The bark of infected material lacked brachysclereids, and tangential ray parenchyma of the secondary phloem were also absent. Numerous shoot bases and aborted axillary buds were present within the bark. Abundant starch granules were present, especially in the phellogen and phelloderm layers. Distortion of the internal lignified tissues was due to the presence of multiple vascular connections from the proliferation of shoots on the broom.     

Analysis on age structure of Zoysia japonica（Poaceae） population

The age-structure of natural population of Zoysia japonica in Xiuyan County of Liaoning Province was studied by generational method.The results showed that the highest tiller age class was three,but 1st age class tillers held dominant position with proportions over 95% in each month during the growing seasons.The 2nd age class and 2rd age class tillers were minority in the population.So Z.japonica population was an expanding population.The zero age class buds on the rhizomes were dominantin buds age structures.The proportion of buds to tillers on quantity in each month was about 30% to 40% and reached the highest at the end of September.The increasing of buds proportion before dormancy guaranteed the quantity of tillers in the next spring.The biomass of 1st age class tillers changed with time.The biomass kept increasing from April to July and reached the highest at the end of July and then decreased.     

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.     

Caulogenic callus induction and adventitious bud formation from embryos of long-term stored seeds ofPicea jezoensis

Caulogenic calli with a high differentiation potency were induced from mature embryos ofPicea jezoensis seeds stored over a long time, for 29 years, resulting in the active formation of adventitious buds. Embryos began to induce calli within 3 weeks of cultivating on LP medium containing 3 μM BAP and 1 μM 2,4-D. Then, the calli proliferated and transformed into caulogenic calli with bud primordia in 8 weeks. The caulogenic calli increased actively with the addition of 500 mg/l ofl-glutamine in the medium. Furthermore, caulogenic calli, induced on LP medium containingl-glutamine, resulted in the formation of adventitious buds, which elongated after transferring the calli into LP medium with 0.1 μM BAP, but withoutl-glutamine. It appears that the number of adventitious buds and the process of shoot elongation are influenced by the kind of nitrogen contained in the medium for callus induction. A part of this study was presented at the 107th Annual Meeting of the Japanese Forestry Society (1996).     

Overexpression of xyloglucanase (AaXEG2) accelerates heteroblastic development in mangium leaves

Transgenic mangium trees (Acacia mangium) overexpressing xyloglucanase (AaXEG2) were generated by spraying flower buds with Agrobacterium solution and allowing seeds to develop. The overexpression of xyloglucanase decreased xyloglucan content in the cell walls and increased stem length and diameter. The leaves of the transgenic seedlings exhibited accelerated heteroblastic development, proceeding from the stage of three bipinnate leaves to that of enlarging petiole 2 weeks earlier than wild type seedlings did.