内生真菌对羽茅抗病性的影响

为探讨不同种类内生真菌对宿主植物羽茅(Achnatherum sibiricum)抗病性的影响, 以感染不同内生真菌的天然禾草羽茅为实验材料, 进行了体外纯培养的内生真菌、感染内生真菌的离体叶片和在体叶片对3种植物病原真菌的抑菌实验。结果表明: 体外纯培养条件下, 分离自羽茅的内生真菌Neotyphodium sibiricum、Neotyphodium gansuensis和Epichloë gansuensis对新月弯孢霉(Curvularia lunata)、根腐离蠕孢(Bipolaris sorokiniana)和枝孢霉(Cladosporium sp.)等3种病原真菌都具有抑制作用, 其中N. sibiricum的抑制作用最强, 对新月弯孢霉、根腐离蠕孢和枝孢霉的抑菌率分别为47.8%、40.1%、39.4%; 内生真菌培养滤液也可以有效抑制这3种病原真菌的孢子萌发, 其中N. gansuensis的抑制作用最强, 新月弯孢、根腐离蠕孢和枝孢霉的孢子萌发率分别为9.8%、8.7%、8.5%。对于离体叶片, N. sibiricum和N. gansuensis感染可以有效降低叶片受3种病原真菌侵染后的病斑数和孢子浓度, 其中N. sibiricum对根腐离蠕孢的抑制作用显著高于N. gansuensis, 而E. gansuensis只降低新月弯孢和枝孢霉侵染的病斑数以及枝孢霉侵染的孢子浓度。在体条件下, 内生真菌均可以显著降低病原真菌侵染羽茅后的病斑数、病斑长度和孢子浓度, 其中E. gansuensis的抑菌作用趋于最弱, 而N. sibiricum的抑菌作用趋于最强。     

中国光萼苔科植物增补

本文描述了中国光萼苔科1新种, 1新变种, 1新变型, 分别是Porella sichuanensis.P, densifolia var.pilosa和P.perrotetiana fo.flaccida。另外有3个新组合(1种, 2变种)和5个中国分布新记录。     

东北罂粟属植物的分类

本文对东北罂粟属植物的分类及其中的问题进行了讨论,并提出了一些新的见解,其中主要的是扩大了野罂粟(Papaver nudicaule.L.)(sensu ampl.),基本型分类性状的含义,缩小了应看做是其亚种的黑水罂粟(P.nudicaule L.subsp.amurense N.Busch.的分类性状含义,发现并提出了两个新的变种,即光果野罂粟P.undicaule L.var.glabricarpum P.Y.Fu及毛果黑水罂粟P.nudicaule L.subsp.amurenseN.Busch.var.seticarpum P.Y.Fu,同时提出了包括种下分类群的便于鉴别的东北罂粟属植物分类检索表。     

豌豆茎基腐病病原菌分离鉴定及其对杀菌剂的敏感性测定

豌豆(Pisum sativum)是我国重要的豆类经济作物, 病害对豌豆生产造成重大经济损失。通过形态学观察、分子鉴定以及致病性测定, 最终确定引起豌豆茎基腐病的3种病原菌分别为尖孢镰刀菌(Fusarium oxysporum)、芸苔链格孢菌(Alternaria brassicae)和格氏镰刀菌(F. grosmichelii), 优势菌株为尖孢镰刀菌, 分离率为53.6%。室内毒力测定结果表明, 5种供试杀菌剂对3种病原菌的菌丝生长均有抑制作用, 其中咯菌腈和戊唑醇的抑菌效果最好。研究结果为豌豆茎基腐病的防治提供了科学依据。     

基于DNA条形码技术对浙江省外来入侵福寿螺进行分子鉴定

外来入侵福寿螺对我国农业生产和水生生态系统平衡等造成严重危害。2010年, 种类鉴定研究首次揭示我国外来入侵福寿螺包括Pomacea canaliculata和P. maculata两个种, 而浙江省仅见P. canaliculata一种报道。P. canaliculata和P. maculata种间形态近似, 且受环境、食物源等因素影响, 同种内外壳形态特征多样, 因而基于形态特征进行种类的准确鉴定极为困难。本研究在采集浙江省7个区县的福寿螺样本的基础上, 利用DNA条形码技术扩增了101个不同个体的COI序列, 并从BOLD数据库下载了“P. canaliculata种团”的5个近缘种的55条COI序列用于分析, 其中包括P. lineata, P. dolioides和P. paludosa所有已发表序列, 以及P. canaliculata和P. maculata的南美洲样品的序列等。序列相似度比对、DNA条形码间隙和系统发育树等分析表明, COI序列可以实现近缘福寿螺的有效鉴别。待测的浙江省福寿螺样品中, 杭州江干区检测到P. canaliculata和P. maculata两种, 而舟山普陀区、绍兴上虞区和新昌县、温州瓯海区及杭州西湖区仅检测到P. canaliculata, 表明P. canaliculata在浙江省具有更广的分布范围。P. canaliculata和P. maculata分别形成4种和2种单倍型, 各区县样点分别包含1-3种单倍型, 浙江省各发生地呈现较低的遗传多样性。依据系统发育关系推测, 浙江省分布的P. canaliculata和P. maculata分别可能来源于阿根廷和巴西。     

穆坪耳蕨及其近缘种研究

作者对耳蕨属的穆坪耳蕨群作了分类学的研究, 共记录了国产种类18种。其中P.habaense, P.obtusipinnum, P.salminense, P.nigrum, P.melano-stipes等为新种;P.lichiangense(Wright) Ching为新组合。对P.tsuchuense等4种及P.qamdoense var.elongatum作了归并。本文还附有分类检索表。     

三峡库区消落带常见野生植物AM真菌

从三峡库区消落带3种常见野生植物的15个根际土壤样本中共分离鉴定出24种丛枝菌根真菌（Arbuscular mycorrhizal fungi,AMF）,包括球囊霉属Glomus 15种、无梗囊霉属Acaulospora 3种、近明球囊霉属Claroideoglomus 2种和多样孢霉属Diversispora、内养囊霉属Entro phospora、巨孢囊霉属Gigaspora及类球囊霉属Paraglomus各1种。其中地表多样孢囊霉D. epigaea 和伯氏类球囊霉P. pernambucanum是我国的新记录种。研究发现这3种植物根际土壤中丛枝菌根真菌的优势属为球囊霉属,优势种为地球囊霉G. geosporum、单孢球囊霉G. monosporum、地表球囊霉G. versiforme、副冠球囊霉G. coronatum 和黄孢球囊霉G. flavisporum。研究还发现在这3种植物根际土壤中丛枝菌根真菌的孢子密度平均为839±170个/100g土,物种丰富度在14-22种之间,Shannon-Wiener多样性指数和均匀度分别在1.97-2.21、0.409-0.479之间波动。结果表明三峡库区消落带是一个AMF多样性研究的资源库。     

生姜共生真菌的分离及其体外抑菌效应初探

为筛选得到优良植物病害生防菌,对广西生姜Zingiber officinale种植区健康生姜根系和叶片中的共生真菌进行了组织分离,以生姜茎腐病菌群结腐霉Pythium myriotylum和香蕉枯萎病菌尖孢镰刀菌古巴专化型4号生理小种Fusarium oxysporum f. sp. cubense race 4为指示菌,通过平板对峙培养法和发酵液菌落直径法试验进行筛选评价,并结合形态学观察及ITS序列分析对筛选出的生防效果最好的共生真菌进行了鉴定。结果表明,从生姜植株共分离得到34株共生真菌,其中根系分离22株,叶片分离12株;对峙培养发现有6株共生真菌对生姜茎腐病菌和香蕉枯萎病菌均有抑制作用;其中菌株SBM-11拮抗作用最强,对生姜茎腐病菌抑制率达到93%,对香蕉枯萎病菌抑制率达到82%;SBM-11的发酵液对生姜茎腐病菌和香蕉枯萎病菌抑制率分别为82%、73%,与其他菌株发酵液抑制效果相比差异明显;结合形态和分子鉴定结果表明SBM-11菌株为绿色木霉Trichoderma viride,极具生防潜力。     

板栗褐缘叶枯病协同致病菌Ophiognomonia castaneae的生活史

采用室外定点观察,子实体诱导及rDNA ITS、MS204、tef1-α 3种分子标记进行系统发育分析等方法,对板栗褐缘叶枯病Phomopsis castaneae-mollissimae的协同致病菌板栗蛇孢日规壳Ophiognomonia castaneae的生活史进行了研究。结果表明,每年7月下旬至8月初叶片发病初期很少分离到O. castaneae,随着病斑扩大该菌的分离频率逐渐增大,至发病后期其分离频率可高达78.5%,甚至可超过致病菌P. castaneae-mollissimae,10月下旬板栗落叶背面的病斑上开始形成O. castaneae的分生孢子盘,11月下旬开始形成O. castaneae的子囊壳原基,次年5、6月越冬落叶背面的病斑上长出子囊壳;带病斑的叶片经室内外诱导,0-25℃范围均可产生成熟子囊壳;湿度是决定子囊壳能否形成的关键因素,强光照不利于子囊壳的产生;分离物的菌丝体在PDA培养基上培养,易产生分生孢子;将分离物分为两种交配型,相互交配后6个月所有处理均未长出该菌的有性型子实体。室外定点观察及rDNA ITS、MS204、tef1-α 3种分子标记表明分离物和病斑上的子囊孢子及其萌发菌丝为O. castaneae的不同生长发育阶段。     

青海豌豆根腐病病原菌种类及致病性的研究

豌豆极腐病是青海东部干旱地区豌豆生产上的一种新病害,近年来危害逐年加重,致使豌豆产量遭受严重损失。根据分离鉴定和致病性测定结果,青海豌豆根腐病病原真菌是由茄镰刀菌、尖孢镰刀菌、豌豆丝囊霉、根串珠霉、立枯丝核菌、腐霉、链孢粘帚霉等复合反染所引起的。经回接试验：镰刀菌和豌豆丝囊霉对豌豆具有较强的致病力;腐霉及其他病原菌则有加强腐烂作用。     

Augmentation of Soil with Sporangia of Actinoplanes spp. for Biological Control of Pythium Damping-off

A method was developed for applying strains of Actinoplanes spp. that are hyper-parasites of oospores of Pythium ultimum to soil for reducing Pythium damping-off of plants. The method is based on the augmentation of soil with sporangia of a strain of Actinoplanes spp. borne on clay granules. In vitro sporulation of strains K30, W57, W257 and 25844 was: (1) greater for most strains on dilute Czapek-Dox agar than on four other agar media; (2) inhibited by continuous exposure to fluorescent light of intensity 4-150 μEm^-2s^-1, but not by exposure to 1 μEm^-2s^-1 or darkness; (3) greater at 20-307deg;C than at 10°C;and (4) greater at pH 6-7 than at pH 5 or 8. On solid carriers treated with dilute Czapek-Dox broth (pH 7) and incubated in the dark at 30°C for 3 weeks, strains sporulated poorly or not at all on vermiculite, perlite and rice hulls, but sporulated abundantly (10⁷-10⁹ colony-forming units (CFU) g^-1 of granules) on montmorillonite clay granules. When strains 25844, W57 and W257 were applied as granules (4 10⁷ - 4 × 10⁸ CFU g^-1) at 5% (w/w) to field plots infested with 750-1000 oospores of P. ultimum g^-1 of soil, only strain 25844 consistently increased emergence and reduced root rot of table beets 8- 1 at 24-28 days after planting compared with controls. Strain 25844 (10⁸ CFU g^-1 of granules) at 1% (w/w) also increased the emergence of bush beans at 28 days after planting in P. ultimum-infested plots, but lower rates were ineffective. The inoculum viability of strain 25844 on clay granules declined 100-fold during 2 months of storage at 5-35°C, but thereafter remained stable for another 4 months. Strain 25844 on 6-month-old granules retained a high degree of hyper-parasitic activity toward oospores of P. ultimum. Augmentation of field soil with sporangia of Actinoplanes spp. is a valid approach to the biological control of pythium damping-off.     

Effects of Pseudomonas chlororaphis on Pythium aphanidermatum and Root Rot in Peppers Grown in Small-scale Hydroponic Troughs

The ability of Pseudomonas (Ps.) chlororaphis isolate Tx-1 to suppress Pythium aphanidermatum and control root rot was investigated in sweet peppers grown in small-scale hydroponic trough units with recirculating nutrient solution. The agent was introduced to the nutrient solution 3 days after the peppers were inoculated with P. aphanidermatum, or 4 days before and 3 days after inoculation, or 4 days before and 3 and 10 days after. Controls either received no agent or pathogen, or the agent only (applied once, 4 days before the time of pathogen inoculations), or the pathogen only. The experiment was conducted in a greenhouse with repetitions beginning in January, March, and May. Severity of root browning associated with P. aphanidermatum, in the absence of Ps. chlororaphis, increased in the first, second and third repetitions, respectively, to 74% at 21 days, 28% at 21 days, and 68% at 11 days. In treatments with one, two, or three applications of Ps. chlororaphis, respectively, areas under root browning progress curves were reduced by 18-48%, 50-73%, and 62-79% compared to the pathogen controls, and recovery incidence of the pathogen from roots at 21 days after inoculation was reduced by 14-47%, 62-82%, and 60-89%. Roots not inoculated with P. aphanidermatum were whitish and not colonized by the pathogen. Density of Ps. chlororaphis in treated plants was usually 2×10³-1.2×10⁵ cfu g^-1 fresh roots. The Ps. chlororaphis treatments almost invariably prevented reductions in plant height, leaf area, fresh mass and dry mass of the shoots and of the roots, and root volume associated with P. aphanidermatum and root browning. It is concluded that Ps. chlororaphis Tx-1 strongly antagonized P. aphanidermatum in the pepper roots and that the agent has considerable potential for controlling root rot and maintaining productivity in commercial hydroponic peppers.     

Interactions of Gliocladium virens with Rhizoctonia solani and Pythium ultimum in Non-sterile Potting Medium

Interactions of Gliocladium virens with Pythium ultimum and Rhizoctonia solani under simulated in vivo conditions were observed microscopically. Different types of propagules of the three fungi were paired on nitrocellulose membranes and incubated at 25°C in non-sterile potting medium in Petri dishes for 1-5 days. Alginate-wheat bran prill were used as carriers for G. virens. Prill inoculated with G. virens and pre-incubated in potting medium for 3-5 days before placement on membranes did not inhibit the germination of Pythium sporangia, but subsequent Pythium growth was markedly stunted and distorted, with some hyphal collapse and cytoplasmic leakage. G. virens had no visible effect on older Pythium mycelium. Two to 5 days' growth of G. virens caused cytoplasmic leakage of Rhizoctonia mycelium, prevented secondary branching of hyphae and occasionally coiled around Rhizoctonia hyphae. Prill that were newly colonized by G. virens, but not prill pre-incubated for 3 or 5 days, stimulated the growth of Pythium mycelium and sporangia, Rhizoctonia mycelium and unprimed monilioid cells, probably by supplying nutrients. The timing of the interactions and their specificity for the different pathogen propagules were consistent with the production of gliotoxin by G. virens. This view was supported by in vitro experiments, in which pathogen propagules were incubated in a range of concentrations of gliotoxin in potato dextrose broth. Pythium sporangia and mycelium were inhibited by 1 or 2 μmg ml^-1, but Rhizoctonia monilioid cells and mycelium required 3-5 μmg ml^-1 for inhibition. At the lowest effective concentrations the inhibition was sometimes reversible, but propagules were killed at high concentrations of gliotoxin.     

Induction of plant defence compounds by Pseudomonas chlororaphis PA23 and Bacillus subtilis BSCBE4 in controlling damping-off of hot pepper caused by Pythium aphanidermatum

Bacillus subtilis strain BSCBE4 and Pseudomonas chlororaphis strain PA23 (=P. aureofaciens) were effective biocontrol agents against Pythium aphanidermatum, the causal agent of damping-off of hot pepper (Capsicum annum L.) in greenhouse vegetable production systems. Application of strains BSCBE4 and PA23 at the rate of 20 g kg^-1 of seed significantly increased the growth of hot pepper seedlings. The efficacies of various carriers in sustaining the population of these strains in storage were assessed. Both the antagonists survived up to 180 days of storage in peat and talc-based formulations. The two bacterial strains induced development of plant defence-related enzymes including phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, phenol content, suppressed incidence of damping-off and increased growth of hot pepper seedlings.     

Biocontrol of Pythium tracheiphilum in Chinese Cabbage by Clonostachys rosea under Field Conditions

Control of leaf and head rot of Chinese cabbage (Brassica campestris L. ssp. pekinensis), caused by Pythium tracheiphilum, was obtained by Clonostachys rosea (isolate IK726) in field trials conducted in 1995 and 1999 on naturally infested land in a commercial crop in Denmark. A significant 2-3-fold disease reduction was obtained at an application rate of 10⁸-10⁹ conidia m^-2 (high application rate) in both years, but not at a 10-fold reduced rate in 1999. Disease reduction by Trichoderma harzianum (Supresivit) was almost significant at the high application rate (1 g product m^-2 corresponding to 7×10⁹ colony forming units (CFUs) m^-2) in both years, but not at a 10-fold reduced rate applied in 1999. In both 1995 and 1999 trials, the percentage of marketable heads increased significantly by 10% following a full application rate of C. rosea. Supresivit applied at the full application rate gave a significant 13% yield improvement in 1995 but not in 1999. No yield improvement was found when the two agents were applied at 10-fold reduced rates. A Danish T. harzianum isolate significantly increased yield by 13% in 1995, but gave no disease control. Plant growth promotion may have been responsible for yield improvements obtained by Supresivit and the Danish isolate of T. harzianum. The 1995 trial also evaluated the products Binab T (T. harzianum+T. polysporum), Mycostop (Streptomyces griseoviridis), Polyversum (P. oligandrum) and Aliette (fosetyl-Al) and Danish isolates of P. oligandrum (2) and T. virens (1), none of which gave disease control or yield effects.     

长白松的木材解剖及其分类问题

关于长白松的分类问题一直有争议^[6-11],根据作者对吉林省安图县二道白河产的长白松的木材解剖研究,发现其木材的射线管胞内壁具深锯齿,颇与欧洲赤松(Pinus sylvestris Linn.)相近^{[14] [15]},但以其木射线薄壁组织细胞水平壁无纹孔和晚材最后数列管胞弦壁上存在着排列成轴向单行小形具缘纹孔来看,这又显属赤松(Pinus densiflora Sieb.et Zucc.)的特征。因此,作者认为长白松的木材结构是处于欧洲赤松与赤松之间而更偏于赤松^[10],故确认其当归属于赤松的种群范围。     

Field application of an insect virus in the Mekong Delta: Effects of a Vietnamese nucleopolyhedrovirus on Spodoptera litura (Lepidoptera: Noctuidae) and its parasitic natural enemies

A new Vietnamese isolate of Spodoptera litura nucleopolyhedrovirus (NPV) was applied in local soybean fields, and the effect of its application on S. litura larvae was examined. The virus was propagated in vivo and a crude extract was prepared for spraying at high and low doses (1.7×10⁸ and 3.3×10⁷ occlusion bodies/m², respectively). The percentage of larvae infected with NPV increased from 22.2% on the day before NPV application (Day 0) to 50.8% (Day 6) in the high dose treatment plot, and from 7.9% (Day 0) to 35.7% (Day 6) in the low-dose plot. Microsporidium sp. was observed as another major pathogen of S. litura larvae. Three dominant parasitic natural enemies were found in S. litura larvae: Microplitis manilae (Braconidae), Chelonus sp. (Braconidae), and Peribaea orbata (Tachinidae). The fate of parasitoids developing within virus- and Microsporidium- infected hosts differed between these three parasitoids; more Chelonus sp. emerging from infected hosts died during their larval stage before spinning cocoons, or failed to reach adult eclosion, than did P. orbata. This suggests that the impact of virus application on the survival of parasitoids varies from species to species.     

Laboratory investigations on the potential of entomopathogenic fungi for biocontrol of Helicoverpa armigera (Lepidoptera: Noctuidae) larvae and pupae

The susceptibility of third instar Helicoverpa armigera to seven strains of three entomopathogenic fungal species, i.e. Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus, was tested under laboratory conditions using the larval immersion method. High efficacies ranging from 68 to 100% corrected mortality were recorded with more profound effects in treatments with B. bassiana and P. fumosoroseus strains. The median lethal concentration (LC₅₀) for L3 was 6.0×10⁵ in M. anisopliae 79, 1.5×10⁵ in B. bassiana 124 and 4.2×10⁴ in P. fumosoroseus 14. These three strains were further used to characterize the age-dependent mortality of different larval stages (L2-L5) and the effect against pupae of H. armigera. Larval stages did not differ in their mortality but differed i in median lethal time, with shorter values recorded in the second instar. Tested fungi also caused a high reduction between 74.4 and 100% in the emergence of pupae using the soil inoculation method and the pupal immersion technique. All three fungal species, especially P. fumosoroseus, have a high potential for biocontrol of H. armigera larvae and also as a soil treatment targeting the pupae.     

Chitinolytic and cellulolytic Pseudomonas sp. antagonistic to fungal pathogens enhances nodulation by Mesorhizobium sp. Cicer in chickpea.

Pseudomonas strains isolated from the rhizosphere of chickpea (Cicer arietinum L.) and green gram (Vigna radiata L.) were screened for the production of chitinases and cellulases. Five Pseudomonas strains were found to produce appreciable amounts of both enzymes in culture-free supernatants and showed growth inhibition of the two fungi Pythium aphanidermatum (Oomycete) and Rhizoctonia solani (Basidiomycete) in plates on potato dextrose agar medium. The fungal growth inhibition was not correlated with cell wall-degrading enzyme activity, which suggested that other antifungal compounds produced by these rhizobacteria were also involved in antagonism. Coinoculation of the Pseudomonas strains with the Mesorhizobium sp. Cicer strain Ca181 resulted in a significant increase in nodule biomass when grown under sterilized chillum jar conditions. The results suggest that hydrolytic enzymes produced by Pseudomonas sp. contribute to suppression of plant diseases by inhibiting growth of phytopathogenic fungi and also promote nodulation of legumes by rhizobia.     

Inhibition of soil-borne plant pathogens by the treatment of sinigrin and myrosinases released from reconstructed Escherichia coli and Pichia pastoris

Myrosinases (thioglucoside glucohydrolase, EC 3.2.3.1) are able to hydrolyse glucosinolates in natural plant products. In Arabidopsis thaliana three different genes with different tissue-specific expressions and distribution patterns encode myrosinases. cDNAs of myrosinase genes (TGG1 and TGG2) were isolated from A. thaliana and expressed in Escherichia coli and Pichia pastoris. The enzyme activities of myrosinase TGG1 and TGG2 genes expressed in P. pastoris were higher than those expressed in E. coli. Among six glucosinolates tested for specificity to myrosinases TGG1 and TGG2, the suitable substrates for these two genes expressed in P. pastoris and E. coli were sinigrin, gluconapin, glucobrassicanapin and glucoraphanin. Treatment of sinigrin with myrosinases excreted from reconstructed E. coli and P. pastoris with TGG1 and TGG2 genes showed strong fungicidal effects on mycelial growth of Rhizoctonia solani AG-4, Sclerotium rolfsii, and Pythium aphanidermatum. This study suggests that the combination of glucosinolate with myrosinases excreted from the reconstructed microbes may be of potential for control of soil-borne diseases.